>> GOOD MORNING, EVERYBODY. WELCOME TO THE SECOND DAY OF THE SUMMIT. THIS IS THE FIRST SESSION OF METABOLISM. MY NAME IS AARON POLLACK AT THE NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE DISEASES. MY CO--- ENVIRONMENTAL HEALTH SCIENCES AND RON KOHANSKI FROM NIA. WE'RE ALL MEMBERS OF THE JR. ROW SCIENCE INTEREST GROUP RESPONSIBLE FOR PUTTING THIS TOGETHER. THE EXTERM CO-CHAIRS FOR THE METABOLISM SESSION ARE CHRISTOPHER NEWGARD, HE'S PROFESSOR DEPARTMENT OF PHARMACOLOGY AND CANCER BIOLOGY AND STAIR RA STUDMAN METABOLISM CENTER DUKE UNIVERSITY. HE WILL GIVE AN INTRODUCTORY OVERVIEW OF THE TOPIC AS WELL AS INTRODUCE SPEAKERS THROUGHOUT THE SESSION. PRESTON IS DIRECT TO OF DIABETES RESEARCH CENTER ALBERT EINSTEIN COLLEGE OF MEDICINE WHO WILL HANDLE THE DISCUSSION AFTER THE PANELISTS GIVE THEIR PRESENTATION AS WELL AS A SUMMARY OF THE SESSION. SO WITHOUT FURTHER ADIEU. CHRIS WILL TAKE OVER. Q. THANKS VERY MUCH, AARON. PLEASURE TO BE HERE TODAY. THIS HAS BEEN AN INTERESTING SYMPOSIUM, I THINK SO FAR, I THINK WE CAN AGREE TO THAT. MARY LAUGHLIN SENIOR PROGRAM MANAGER FOR NITDK, I SAW HIM YESTERDAY P AND ASKED HOW MANY OF THESE NIH WORKSHOPS HAVE YOU BEEN TO OVER YEARS, CHRIS? AND I COULDN'T FRANKLY ANSWER. I KNOW IT'S BEEN A COUPLE AT LEAST. A NUMBER OF, A NUMBER OF THEM. BUT I HAVE TO SAY THIS ONE IS UNIQUE. THIS -- THERE IS A DIVERSITY OF PRESENTATIONS ABREADTH OF SCIENCE AND ENGAGEMENT IN TERMS OF THE QUESTIONS THAT OCCUR AFTER THE SESSIONS THAT HAS BEEN REALLY REMARKABLE. IT'S PARTLY BECAUSE AGING REMAINS A TOPIC DIFFICULT TO GET OUR ARMS AROUND ABOUT DEFINING THE MOST CONCISE CAUSE AND EFFECT EXPERIMENTS AND PICKING OUT CAUSAL DRIVERS OF AGING PHENOTYPES. IT SEEMS THAT FOR ALMOST EVERY TARGET AND EVERY IDEA WE HAVE THERE IS P PRO AND CON INFORMATION. EVERYTHING IS GOING TO CHANGE AND BE CRYSTAL CLEAR BUT IN FACT, WE ARE BEDEVILED BY SAME ISSUES HERE IN SPEAKING OF METABOLISM IN TERMS OF CAUSE AND EFFECT. TO MAKE IT MORE COMPLEX OUR PANELISTS ARE NOT NECESSARILY CARD CARRYING AGING RESEARCHERS. SOME LIKE BILL EVANS THAT IS KNOWN TO MANY OF YOU, CERTAIN HAS WORKED IN THE AGING FIELD FOR QUITE A NUMBER OF YEARS BUT OTHERS SUCH AS MYSELF JEFF AND MARY BIRNBAUM WE CHARACTERIZE OURSELVES METABOLIC DISEASE RESEARCHERS BUT NOT NECESSARILY FOCUSED ON AGING. I HOPE BRINGING PEOPLE TOGETHER OF DIFFERENT STRIPES GIVES A DIVERSITY OF OPINION, A FRESHNESS OF APPROACH THAT WILL SOMEHOW BE USEFUL TO THE GATHER ED GROUP HERE. SO I'M GOING TO BRIEFLY MAKE A FEW REMARKS. THIS IS A AN ATTEMPT TO SUMMARIZE MOLECULAR PATHWAYS OF AGING AND THIS WAS A REALLY PAINFUL EXPERIENCE IN WRITING A LITTLE REVIEW ARTICLE FOR JCI WITH NED SHARPLUS. AS A NON-AGING RESEARCHER TO COME UP WITH ONE DIAGRAM THAT SUMMARIZES THE MOLECULAR PATHWAYS OF AGING WAS A TORTURE EXPERIENCE I HAVE TO SAY. WITH REGARD TO THIS MORNING'S TOPIC, ALREADY THERE'S SOMETHING PRESUMPTION AND PROVOCATIVE ABOUT THE SLIDE IN THAT IT PUTS THE METABOLIC EVENTS VERY PROXIMAL IN THE SCHEME. SO WE THINK OVERNUTRITION PANDEMIC OF WORLD OBESITY, ANABOLIC HORMONE SIGNALING IS TALKED HERE, INSULIN AND IGF 1 AS DRIVERS OF AGING PHENOTYPES, CHRONIC MTOR ACTIVATION, DECREASED SIRTUANS. REACTIVE OXYGEN SPECIES. MITOCHONDRIAL DYSFUNCTION FUELED BY A SLOW RATE OF MITOCHONDRIAL DNA MUTATION. WE HAVE PUT IN THIS PARTICULAR DIAGRAM THESE METABOLIC EVENTS AS PROXIMAL IN THE SCHEME LEADING TO THE MORE CORE CELL BIOLOGICAL CHANGES THAT WE HAVE BEEN DISCUSSING AT THIS MEETING SUCH AS SENESCENCE, THE SAS RESPONSE, INCREASED CYTOKINE PRODUCTION, IMPAIRED STEM CELL RENEWAL. THESE CELL BIOLOGICAL EVENTS LEADING TO AGING PHENOTYPES THAT HAVE ALSO BEEN UNDER DISCUSSION HERE, REDUCED IMMUNE RESPONSE. CHRONIC INFLAMMATION, FRAILTY. IMPAIRED CAPACITY TO RESPOND TO REGENERATION, THERAPIES. BUT RIGHT OFF THE BAT, PERHAPS I'M INTRODUCING SOMETHING THAT WE CAN DEBATE AND CONSIDER TO BE CONTROVERSIAL, IS METABOLISM EARLY L AND CAUSAL OR IS IT MORE A REFLECTION OF THE PROCESS OF AGING, DOES METABOLISM CHANGE AS CONSEQUENCE OF AGING, DOES METABOLISM CHANGE EARLY AND DRIVE AGING, IS ONE OF THE FUNDAMENTAL QUESTIONS THAT WE CAN ADDRESS FOR WHICH I DON'T THINK WE HAVE AN ABSOLUTE CLEAR ANSWER BUT MAYBE SOME OF THE WAYS TO ATTACK IT COURTROOM OUT IN THIS SESSION. SO ANOTHER WAY THINKING ABOUT THIS IS TO SAY WHAT DO WE THINK WE KNOW ABOUT AGING AND METABOLISM. THESE ARE SOME SIGN POTSES YOU CAN HANG YOUR HAT ON FROM THE LITERATURE. AGENTS THAT SUPPRESS ANABOLIC PROCESSES. RAPAMYCIN HAS BEEN TALKING ABOUT A LOT HERE INHIBITOR OF MTOR. INCREASED LIFE SPAN. BUT INTERESTINGLY, OTHER AGENTS THAT STIMULATE OXIDATIVE FUEL METABOLISM, METAPHORMAN, WE HAVE NOW LEARNED IS AN INHIBITOR OF MITOCHONDRIAL ELECTRON TRANSPORT CHANGE AT SITE 1 OF THE ELECTRON TRANSPORT CHAIN RESULTING IN A RISE IN AMT ATP RATIO. ACTIVATION OF THE AMP RESPONSE FIVE PRIME AMP KINASE LEADING TO ACTIVATION OF FATTY ACID OXIDATION, METAPHORMAN HAS BEEN ASCRIBED LIFE SPAN PROMOTING ACTIVITIES T. ANOTHER METABOLICALLY RELEVANT INTERVENTION WE DISCUSSED HERE IS CALORIC RESTRICTION INCREASES LIFE SPAN. THERE'S A LOT OF LITERATURE AND DEBATE AND CONTROVERSY AROUND THE IDEA THAT AGENTS THAT LIMIT MITOCHONDRIAL STRESS AND PRODUCTION OF REACTIVE OXYGEN SPECIES, ANTIOXIDANTS WILL ENHANCE LIFE SPAN. AND WE TO CUTSD ON SER T 1 HERE, SER T-3 MAYBE INVOLVED IN PROCESSES. AND AND METABOLISM WHICH IS AN INTERESTING AREA. BULLETS WE THINK WE KNOW, BUT WHEN WE LOOK AT THIS A LITTLE HARDER, FUNDAMENTAL ISSUES ARISE. DOES RAPAMYCIN SLOW APPEARANCE OF AGING PHENOTYPES OR DOES IT INCREASE LIFE SPAN INDEPENDENT OF PHENOTYPES. THERE'S LITERATURE OUT NOW THAT SAYS THESE ARE POTENTIALLY SEPARABLE. IF REACTIVE OXYGEN SPECIES GENERATION IS TOXIC, WHY DOES AN AGENT THAT INCREASES METABOLIC RATE METAPHORMAN AND PRESUMABLY ROSS GENERATION HAVE POSITIVE EFFECTS ON LONGEVITY AND CONVERSELY, WHY DOES CALORIC RESTRICTION AND INTERVENTION THAT SLOWS ENERGY EXPENDITURE ALSO ENHANCE LONGEVITY? ONE INTERESTING THING FROM A FUEL SELECTION POINT OF VIEW, A COMMON FEATURE OF TWO MANEUVERS METAPHORMAN AND CALORIC RESTRICTION IS THEY FAVOR A SHIFT TOWARDS FATTY ACID OXIDATION AN WAY FROM OXIDATION OF GLUCOSE AND OTHER FUELS. IS THERE SOMETHING RELEVANT TO PRECEDENCE FOR FATTY ACID OXIDATION THAT HAS A LINK TO LONGEVITY AND HOW DOES NAD REGULATE AGING AN ARE SIR TUANS INVOLVED. SO THESE ARE THE BEDEVILING QUESTIONS OF THE METABOLISM AGING FIELD AND THREE OTHER TOPICS COVERED BY OUR SPEAKERS, HOW ARE OBESITY RELATED AND AGING RELATED METTLE BOLLIC DYSFUNCTION -- METABOLIC DYSFUNCTION SYNDROMES SIMILAR BUT ALSO DISTINCT? DO METABOLIC FUEL CYCLES HAVE A ROLE IN REGULATION OF THE AGING PROCESS, THAT WILL BE COVERED BY DEBBIE. AND WHAT'S THE ROLE OF CENTRAL REGULATION METABOLISM INCLUDING CONTROL OF CIRCADIAN RHYTHM IN AGING BIOLOGY. SO I HAVEN'T HELPED YOU GET CLARITY. I HAVE JUST THROWN A BUNCH OF THINGS TO CONFUSE YOU T. I'M GOING TO RELY NOW ON OUR SPEAKER PANEL AND DISCUSSION SECTION TO PROVIDE THAT CLARITY. WE HAVE A WONDERFUL GROUP OF SPEAKERS THAT WILL COVER TOPIC AREAS THAT I JUST DESCRIBED. WE'LL START WITH MARY BIRNBAUM OVER MANY YEARS A LEADER IN MOLECULAR SIGNALING PATHWAYS THAT CONTROL METABOLISM. MARY. -- MORRY. >> THANK YOU VERY MUCH, CHRIS AND JEFF. THANKS FOR THE OPPORTUNITY TO TALK TO YOU BUT MORE THAN THAT THE OPPORTUNITY TO LISTEN TO THE INTERESTING AND PROVOCATIVE PRESENTATIONS THAT WHEN ON YESTERDAY AND SURE WILL HAPPEN TODAY. SO MY TASK IF IT SHOWS UP, MY TASK OUR TASK WITH TALKING TO YOU ABOUT SIGNALING PATHWAYS INVOLVED IN ANABOLIC METABOLISM AND IN HOW THEY RELATE TO AGING AND OBVIOUSLY THAT'S A CHALLENGE WHICH IS MUCH TOO COMPLICATED AND EXTREME TO COVER IN TEN MINUTES EVEN THE IF I WERE ABLE TO SHOW REAMES OF DATA. SO INSTEAD WHAT I'M GOING TO DO IS TALK ABOUT THIS RELATIONSHIP BETWEEN ANABOLIC METABOLISM BETWEEN MORE CONVENTIONAL METABOLIC REGULATION RELATING TO DIABETES, GLUCOSE METABOLISM AND POSE SOME QUESTIONS AND CONTEXT TO THINK ABOUT IT AND IF THERE ARE MORE SPECIFIC ISSUES TO COVER WE CAN DO THAT IN THE PANEL DISCUSSION WITH THE OPTION OF DEFERRING TO COLLEAGUES. THERE WE GO. YESTERDAY A NUMBER OF SPEAKERS SHOWED SLIDES OF VARIOUS EXPERIMENTS REAFFIRMING THE LONG HELD OBSERVE RATION THAT CALORIC RESTRICTION EXTENDS LIFE SPAN. ONCE ONE ACCEPTS THAT IT'S LOGICAL EXTENSION TO BELIEVE THAT CALORIC RESTRICTION IS WORKING BY SUPPRESSING AN ANBOTIC RESPONSE. THE KEY TO ANABOLIC RESPONSE OR AT LEAST WHAT'S BELIEVED CENTRAL INTEGRATING NODE FOR STIMULUS CATABOLISM IS THE TOR COMPLEX WHICH I IS WHAT I'LL TALK ABOUT TODAY. SUPPORT FOR THE CRITICAL ROLE OF COMPLEX IN ANABOLIC ME TAB HIM IN AGING PROCESS COMES FROM PARALLEL WORK IN THREE CLASSES OF ORGANISMS WHICH ARE SHOWN IN THIS SLIDE. SO I JUST WANT TO BEGIN BY EMPHASIZING THE DIFFERENCES IN THE WAY THE SIGNALING PATHWAYS LINK UP IN THESE THREE ORGANISMS. YEAST IS UNIQUE IN THE SENSE THAT IT'S PRIMARILY A CELL AUTONOMOUS ORGANISM RELYING ON NUTRITIONAL SIGNALS TO REGULATE THE GROWTH RESPONSE. FLIES AND WORMS ARE MOR COMPLICATED THAT THEY HAVE THE NON-AUTONOMOUS HORMONAL INPUT REGULATED BY INSULIN IGF-1 SIGNALING PATHWAY BUT ALSO WORTH REMEMBERRENING THESE TRACTABLE INVERTEBRATES THERE IS A SINGLE SYSTEM THAT REGULATES THIS PATHWAY. THERE ARE MULTIPLE LIGANDS, FIVE OR SO IN FLIES, MORE IN WORMS BUT THERE'S A SINGLE RECEPTOR THAT CONVEYS THE SIGNAL BOTH FOR METABOLISM AND GROWTH AND CLEARLY ORGANISMS THE GROWTH RESPONSE IS DOMINANT, MUCH DOMINANT, METABOLISM THOUGH THERE'S METABOLIC PATHWAY. THIS VERY SKETCHY DIAGRAM IS NOT TO IMPLY THIS IS THE ONLY THING INSULIN DOES, IT REGULATES ANOTHER PATHWAY, THE FOX ANTI-STRESS PATHWAY, BUT THIS IS REALLY TO EMPHASIZE THERE'S A SIGNAL RECEPTOR THAT TRANSMITS THIS. THEN THE ORGANISMS WE CARE MOST ABOUT MAMMAL, JUST REALLY EMPHASIZES THIS SYSTEM IS DUPLICATED TO TWO PARALLEL PATHWAYS ONE WHICH PRIMARILY NOT EXCLUSIVELY DEVOTED TO REGULATION OF METABOLISM, INSULIN, THE OTHER ONE, THE IGF SUGGESTIONNAL REGULATED PRIMARILY THOUGH NOT EXCLUSIVELY THE REGULATION OF GROWTH. THIS COMPLICATES THINGS ENORMOUSLY BECAUSE THESE TWO PATHWAYS TALK TO EACH OTHER, THEY INFLUENCE OPEN OTHER AND WHEN -- EACH OTHER AND WHEN CONSIDERING THE PATHWAY, THE ANABOLIC PATHWAY IN ETIOLOGY OF AGING OR EVEN MORE PROFOUNDLY AS A THERAPEUTIC TARGET, IT IS IMPOSSIBLE NOT TO CONSIDER THE IMPLICATIONS FOR THE PARALLEL METABOLIC PATHWAY. THAT'S ONE THING I'M GOING TO TALK ABOUT TODAY. THE FACT BLOCKING THIS TOR PATHWAY AND SUPPRESSING CANONCAL GROWTH RESPONSE IN THESE ORGANISMS HAS BEEN GIVEN RIGHTLY AS ONE OF THE MOST COMPELLING PIECES OF EVIDENCE THAT IT IS TOR ITSELF REGULATING THE AGING PROCESS AND NOT SECONDARY CONSEQUENCE OF ALTERING THAT SIGNALING NODE. THAT OBVIOUSLY IS A COMPELLING ARGUMENT BUT I WANT TO EMPHASIZE THAT IT STILL REMAINS AN ARGUMENT. BECAUSE ALTERING ONE PARTICULAR SIGNALING MOLECULE HAS PARALLEL EFFECTS IN TWO CLASSES OF ORGANISMS, IT DOES NOT IN ITSELF PROVE IT'S ACCOMPLISHING THAT BY THE SAME MECHANISM. HUMANS ARE MORE COMPLICATED. THAT'S BECAUSE THIS ELEMENT IS CHOICE. MOUSE CAN CHOOSE HOW MUCH IT CAN EAT IN A CAGE BUT CAN'T CHOOSE SUBSTRATE SELECTIVITY THE WAY PEOPLE CAN. I DON'T KNOW IF THIS IS THE REASON BUT IT'S IMPORTANT TO REALIZE THAT IN TRYING TO APPLY WHAT WE HAVE LEARNED FROM MOUSE ENERGY METABOLISM TO HUMANS, IT'S BEEN REMARKABLY UNSUCCESSFUL, THAT IS TO SAY, IT IS VERY EASY AT THIS POINT TO INFLUENCE OBESITY GENETICALLY IN A MOUSE, UNCOUPLE THE LINK BETWEEN OBESITY UNDINES LINS RESISTANCE AND DIABETES IN A MOUSE BUT SO FAR THESE OBSERVATIONS HAVE NOT BEEN TRANSFERABLE TO HUMANS. ONE IS THE FACT HUMANS HAVE MORE CONTROL OVER THE ENVIRONMENT, DIET, EXERCISE THAN MICE DO. WE'RE TALKING THIS MOLECULE HERE, THIS IS A COMPLICATED SLIDE WITH DIFFERENT COMPONENTS. I WON'T TALK ABOUT IT EXCEPT TO SAY THERE ARE MANY THINGS THAT INFLUENCE IT. AND FOR THE REST OF THE TALK I'LL BREAK THEM TO ENERGY STRESS WHICH SUPPRESSES IT AND ANABOLIC INPUTS FROM NUTRITIONAL QUEUES OR HORMONES WHICH STIMULATE IT. ALSO EMPHASIZES THAT TOR OUTPUTS AN THOUGH THIS REVIEW WAS PUBLISHED IN 2012, THIS LIST IS ALREADY INCOMPLETE. THE OTHER QUESTION OF COURSE IS, WHICH OF THESE DOWNSTREAM PATHWAYS ARE RESPONSIBLE FOR THE AGING EFFECTS OF TOR AND ANTI-AGING BLOCKING IT, I'M NOT GOING TO TREAT THAT BUT WE CAN TALK LATER IN DISCUSSION IF PEOPLE ARE INTERESTED. SO THE TOR PATHWAY IS COMPLICATED BY TWO TOR SIGNALING PATHWAYS. ONE SIGNALS TO GROWTH, THOUGH I HAVEN'T PUT IT UP HERE REGULATES F O SO IN LOOKING AT VARIOUS STUDIES DONE LOOKING AT THIS PATHWAY, WE HAVE TO REALIZE SOME INFLUENCE BOTH OR ONE. WHEN USING HYPOMORPHIC APPROACHES TO INFLUENCING THE PATHWAY WE DECREASE THE TOR 1 AND TOR 2 RESPONSE AND THAT CHEERILY EXTENDS LIFE SPAN GENETICALLY. THE ONE THING I SHOULD EMPHASIZE IS THOUGH MTOR 2 REGULATES AND IS NECESSARY FOR THIS PATHWAY, THERE'S SPARE SIGNALING IN THIS PATHWAY SO WHEN YOU TURN DOWN THE TOR 2 ARM YOU CAN DO IT WITHOUT FUNCTIONAL CONSEQUENCES AT LEAST UNDER STEADY STATE CONDITIONS, THAT'S WHY DROPPING TOR TO BOTH HAS PROFOUND ON GROWTH HAS LESS EFFECT ON METABOLISM. THE OTHER STRATEGY IS TO SELECTIVELY DECREASE TOR 1 GENETICALLY OR THERAPEUTICALLY, IN A WAY THAT DOESN'T AFFECT TOR 2, ONE IS TO KNOCK OUT A MAJOR TARGET OF TORK 1, THAT'S KINASE, THAT EXPERIMENT IS DONE AND SHOWN TO EXTEND LIFE SPAN. INTERPRETATION OF THAT EXPERIMENT TEACH AS VERY IMPORTANT LESSON IN TERMS OF INTERPRETING METABOLIC PHENOTYPES. HERE IS THE KNOCK OUT WHICH EXTENDS LIFE SPAN BUT THIS IS IS KEY FIE GUR IN THAT PAPER WHEN BODY GERM LIKE KNOCK OUT KINASE IS STUDIED, ADIPOSITY IS DECREASED. WHEN YOU KNOCK OUT A KINASE A PRODUCT OF TORK 1 SIGNALING ONE LIKES TO BELIEVE THAT YOU'RE MIMICKING CALORIC RESTRICTION, THAT'S THE PATHWAY GOES CALORIC RESTRICTION TO T OBJECTIONR KINASE TO LIFE EXPANSION. IN THIS MODEL THAT PATHWAY IS REVERSED. WHAT'S HAPPENING IS THE EFFECTIVE LOSS OF S-6 KINASE IN THE BRAIN IS CAUSING A CHANGE IN ENERGY METABOLISM SO THE WAY THE PATHWAY GOES IS DECREASE SIGNALLENING THE BRAIN, DECREASED CALORIC INTAKE, THERE'S A CHANGE IN FONTS THERE AND INCREASED LIFE SPAN. THAT IS TO SAY IS THIS IS UPSTREAM OF CALORIC RESTRICTION AND CLASSICAL RESPONSE BECAUSE OF THE CELL AUTONOMY OF THE RESPONSE. THERE IS A MORE COMPLICATED PATHWAY WITH RAPAMYCIN. RAPAMYCIN THROUGH A SIGNALING ROUTE HERE, NOT ONLY BLOCKS TORK 1 SIGNALING BUT ALSO TORK 2 SIGNALING. THAT HAS BEEN REALLY NICELY SHOWN IN A RECENT STUDY. FROM JOE BAUER COLLEAGUE OF MINE UNIVERSITY OF PENNSYLVANIA IN COLLABORATION WITH DAVID SABATINI, WHEN YOU TREAT ANIMAL WITH RAPAMYCIN YOU EXTEND LIFE SPAN BUT YOU MAKE THAT ANIMAL INSULIN RESISTANT. WHICH CREATES THAT INTERESTING PROBLEM. HOW IS IT THAT YOU CAN HAVE AN ANIMAL WHICH IS INCREASED INSULIN RESISTANT BUT ALSO HE CAN PENDS LIFE SPAN. WE KNOW INSULIN RESISTANCE. THERE'S TWO POSSIBLE EXPLANATIONS. ONE IS THAT RAPAMYCIN ACTUALLY CAUSES INSULIN RESISTANCE IN A WAY FUNDAMENTALLY DIFFERENT THAN OBESITY. THAT IS CERTAINLY POSSIBLE. THE OTHER IS, THE ONE WE WORRY ABOUT IS MOUSE IS NOT A SENSITIVE ORGANISM FOR LOOKING AT THE ADVERSE CONSEQUENCES OF EXTENDED LIFE SPAN. I THINK THERE'S REALLY DATA TO SUPPORT THAT. DIABETES AND MICE DOESN'T CAUSE PROFOUND CARDIOVASCULAR DISEASE IN HUMANS TWO-THIRDS OF MORBIDITY MORTALITY AND EXPENSE IS CARDIOVASCULAR DISEASE AN SECONDARILY DIABETES AND OBESITY DON'T CAUSE SEVERE LIVER DISEASE SO WE HAVE TO BE CAUTIOUS THAT THE ADVERSE SIDE OF RAPAMYCIN MIGHT NOT BE HAVING THE CONSEQUENCES THAT IT MIGHT WELL HAVE IF GIVEN TO HUMAN BEINGS. THANK YOU. [APPLAUSE] >> THANK YOU, MORRY. OUR NEXT SPEAKER IS BILL EVANS FROM GLAXO SMITHKLINE WHO WILL SPEAK TO US ABOUT IMPLICATIONS OF AGE ASSOCIATED INCREASED NUTRIENT REQUIREMENTS AND DECREASED ENERGY NEEDS. BILL. >> THANK YOU VERY MUCH. IT IS INDEED A PLEASURE AND HONOR THANKS TO THE ORGANIZERS AND TO THE PREVIOUS SPEAKER, SETS US UP A BEAUTIFUL TALK ON THE IMPLICATIONS OF WHAT WE CALL ANABOLIC RESISTANCE. I WILL START BY SAYING SOME YEARS AGO WE CONDUCT AD STUDY TO LOOK AT EFFECTS OF DIETARY PROTEIN AND EXERCISE ON CHANGES IN MUSCLE MASS. ONE THING WE OBSERVED WHEN WE PUT HEALTHY OLDER PEOPLE ON THE RECOMMENDED DIETARY ALLOWANCE FOR PROTEIN IS THAT THEY WERE ALL IN NEGATIVE NITROGEN BALANCE. IF THE RDA FOR PROTEIN IS ADEQUATE, WE SHOULDN'T BE SEEING THAT. WE SPECULATED AT THE TIME THERE WAS INADEQUATE DATA TO SAY ANYTHING ABOUT DIETARY PROTEIN NEEDS OF OLDER PEOPLE EXCEPT TO SAY THIS WAS AN INTERESTING RESULT. IF YOU GO BACK AND LOOK AT THE DATA HOW THE RDA WAS ARRIVED AT, FUNDAMENTALLY, IT IS IN YOUNG MIT AND BERKELEY STUDENTS 90% WERE MALE, ALMOST NO OLDER PEOPLE. SO THAT I THINK IS AN EARLY INDICATION THERE IS ANABOLIC RESISTANCE TO EFFECTS OF DIETARY PROTEIN. WE TEST THE HYPOTHESIS IF THE RECOMMENDED DIETARY ALLOWANCE IS INADEQUATE TO MEET THE NEED OF OLDER PEOPLE WHAT HAPPENS IF P YOU GIVE THAT AMOUNT OVER LONG PERIOD OF TIME? WHAT I TELL YOU IS WE GIVE HEALTHY OLDER PEEP IT WILL RDA FOR PROTEIN THEY MOVE INTO POSITIVE NITROGEN BALANCE OVER THAT THREE MONTH PERIOD BUT DO IT AT THE EXPENSE OF MUSCLE. WE HAVE A STRONG INDICATION DIETARY PROTEIN RECOMMENDATION IS INCH ADEQUATE. THAT IS IMPORTANT INFORMATION BECAUSE AT THE SAME TIME AGING IS ASSOCIATED WITH LOSS OF ENERGY REQUIREMENTS. THAT ALMOST CERTAINLY IS DUE TO THE RESULT OF A DECREASE IN THE AMOUNT OF MUSCLE. DATA FROM THE BALTIMORE INSTITUTE ON AGING SHOW FUNDAMENTALLY ALL REDUCTION IN METABOLIC RATE IS SIMPLY A CONSEQUENCE OF REDUCTION MUSCLE MASS. I'LL SHOW YOU THIS SLIDE YOU SAW IT YESTERDAY FROM LINDA, THIS IS HER KIND OF OPERATIONAL VIEW OF FRAILTY. BUT I WILL POINT OUT THIS ONE HERE. CHRONIC UNDERNUTRITION OF PROTEIN IN PARTICULAR, BUT A NUMBER OF OTHER NUTRIENTS IS FUNDAMENTAL TO THE LOSS O OF HOMEOSTASIS AND CERTAINLY LOSS OF SKELETAL MUSCLE. AT THE SAME TIME DIETARY PROTEIN INTAKE OR REQUIREMENTS GOING UP AND ENERGY REQUIREMENTS GOING DOWN, THAT SETS UP AN EXTRAORDINARILY DIFFICULT SITUATION. TRY TO COME UP WITH STRATEGIES TO FEED OLDER PEOPLE AN ADEQUATE DIET. ONCE OF THE CONSEQUENCES IS INCREASE IN BODY FATNESS. THAT IS ALSO ONE OF THE MOST FREQUENTLY MEASURED CONSEQUENCES OF AGING. AND IT'S INTERESTING IF YOU LOOK AT THIS STATISTICS, WE KNOW THERE IS THIS OBESITY EPIDEMIC IN THE UNITED STATES AMONG WOMEN 20 TO 39 THE OBESITY PERCENTAGE IS 32%, AMONG WOMEN 36% AND AMONG WOMEN OVER 60 IT'S 43% IN THE US. SO 43% OF ALL WOMEN IN THE U.S. ARE OBESE. WHICH WE DO KNOW IN STUDIES DONE AT WASU, 96% OF OBESE ELDERLY ARE FRAIL. BY LINDA'S CRITERIA. OBVIOUSLY OBESITY ISN'T ASSOCIATED WITH SHRINKAGE OR LOSS BUT IT IS ASSOCIATED WITH POOR FUNCTION. BODY FATNESS IS ONE OF THE MOST POWERFUL PREDICTORS OF LATE LIFE DISABILITY. THESE ARE DATA FROM THE HEALTH ABC STUDY, I WANT TO SHOW YOU, VERY INTERESTING INFORMATION IN POST MENOPAUSAL WOMEN. IN YOUNG PEOPLE, GAIN WEIGHT ABOUT A THIRD OF THE WEIGHT GAIN IS LEAN MASS. BUT LOOK AT THIS. AS OLDER WOMEN INCREASE BMI, BMI ABOUT OVER 26 THERE IS NO FURTHER INCREASE IN LEAN BODY MASS. 100% WEIGHT GAIN IS FAT. WHEN THEY ATTEMPT TO LOSE WEIGHT VIA ENERGY RESTRICTION, 50% OF THE WEIGHT LOSS IS LEAN. IN FACT, WEIGHT LOSS AMONG POST MENOPAUSAL WOMEN IS ASSOCIATED WITH A TWOFOLD INCREASE IN RISK FOR HIP FRACTURE IRRESPECTIVE WHEN THEY START LOSING WEIGHT OR INTENT TO LOSE WAIT. -- WEIGHT. WE TALKED ABOUT THE EFFECTS OF EXERCISE. AND THE LAST THING I WANTED TO SHOW YOU ARE SOME DATA WE GENERATED SHOWING THE OPPOSITE EFFECT. A SITUATION ELDERLY PEOPLE ARE FAR MORE LIKELY TO ENCOUNTER, THAT IS BEDREST. CHANGES ASSOCIATED WITH AGING INCLUDE DECREASED FATTY ACID OXIDATION, DECREASE MUSCLE MASS AND ALL OF THESE THINGS THAT WE HAVE BEEN TALKING ABOUT BUT WHEN YOU PUT SOMEONE TO BED, MANY OF THESE THINGS ARE ACCELERATED AND ACCENTUATED. THERE IS A REAL DECREASE IN THE RATE OF PROTEIN SYNTHESIS, INCREASE IN SENSITIVITY TO CORTISOL, WHICH FURTHER DOWN REGULATES PROTEIN SYNTHESIS. INSULIN REGULATION WHICH -- DECREASE IN FATTY ACID OXIDATION WHICH CAUSES INCREASE IN INTRACELLULAR TRIGLYCERIDE CONTENT WHICH CAUSES INSULIN RESISTANCE WHICH FURTHER REGULATES MUSCLE PROTEIN SYNTHESIS RESULTING IN PROFOUND LOSSES OF MUSCLE LEADING TO DECREASE STRENGTH AND FRAILTY WHICH HAS CYCLICAL EFFECT. I'LL SHOW YOU WHEN WE PUT HEALTHY OLD PEOPLE TO BED FOR TEN DAYS THIS IS WHAT WE SEE IN TERMS OF NITROGEN BALANCE. FIRST THING, THIS IS THE PRE-BEDREST PERIOD SO WE FEED ELDERLY PEEP IT WILL RDA FOR PROTEIN ADEQUATE AMOUNTS OF ENERGY. THE FACT THAT BEFORE THEY START BEDREST THEY'RE IN NEGATIVE NITROGEN BALANCE RDA FOR PROTEIN IS INADEQUATE AND A RESULT OF ANABOLIC RESISTANCE ACTIVATE MTOR FAILS TO INCREASE BLOOD FLOW BUT WHAT YOU CAN SEE IS INACTIVITY RESULTS IN THESE SUBJECTS MOVING TO PROFOUNDLY NEGATIVE NITROGEN BALANCE. CONS P WEDNESDAY OF THAT IS INTERESTING. YOUNG PEOPLE BED 28 DAYS THEY LOSE 400-GRAMS OF MUSCLE FROM THEIR LEGS. , ELDERLY PEOPLE TO BED FOR TEN DAYS THEY LOSE 1.2-KILOGRAMS OF MUSCLE. IT'S AN ASTONISHING CHANGE AND VERY, VERY SHORT PERIOD OF TIME. WHAT WE PUBLISHED IS THERE IS A REALLY STRIKING INCREASE IN HEPATIC INSULIN RESISTANCE WE DIDN'T RECOGNIZE STARTING OUT, ALSO INCREASE IN PERIPHERAL INSULIN RESISTANCE BUT HEPATIC INSULIN RESISTANCE MAYBE DRIVING CONTINUED INCREASE GLUCOSE PRODUCTION THAT MAYBE DRIVING DOWN FATTY ACID OXIDATION IN MUSCLE WHICH MAY CAUSE ELEVATION IN TRIGLYCERIDE LEVELS BUT THIS LOSS OF MUSCLE MASS IS STRIKING. IN OLDER PEOPLE, 40% REDUCTION IN MUSCLE PROTEIN SYNTHESIS. THIS IS AN AREA RODENTS AND HUMANS DIFFER BECAUSE INACTIVITY IN RHODENS IS ASSOCIATED WITH INCREASE IN PROTEIN DEGRADATION. WE DON'T SEE ANY EVIDENCE OF THAT. WE SEE CHANGE IN RATE OF SYNTHESIS. SO IN CONCLUSION, THE EMERGING BABY BOOM GENERATION ENTERS OLD AGE WITH SARCOMEANIC OBESITY, OBESITY ASSOCIATED WITH LOW AMOUNTS OF MUSCLE MASS. IT WILL BE AN EXTRAORDINARY CHALLENGE TO EVERYBODY HERE ON HOW TO DEAL WITH THAT. BECAUSE TRADITIONAL ENERGY RESTRICTION TO LOSE BODY WEIGHT IS NOT THE APPROPRIATE STRATEGY FOR OLDER PEOPLE. WE NEED THE BEST AND SAFEST WAY TO MAINTAIN MUSCLE MASS AND BONE, AT THE SAME TIME DECREASE BODY FATNESS. THIS ANABOLIC RESISTANCE TO PROTEIN IS AMPLIFIED BY INACTIVITY INDITE CREASED ENERGY INTAKE. THANK YOU. [APPLAUSE] >> THANK YOU, BILL FOR THOSE VERY PROVOCATIVE AND INTERESTING COMMENTS. NOW MY PLEASURE TO INTRODUCE MY FRIEND AND COLLEAGUE FROM THE STEADMAN NUTRITION AND METABOLISM AND DUKE INSTITUTE OF MOLECULAR PHYSIOLOGY, DEBBIE MUOIO, SHE'S TAKEN ON BRAVELY A REALLY DIFFICULT TOPIC, FUTILE CYCLES AS TARGETS OF METABOLIC CONTROL AND AGING DISEASE. GLAD SHE'S DOING IT AND NOT ME. >> FIRST THANK CHRIS AN JEFF AND ORGANIZERS FORGIVING ME THE OPPORTUNITY TO BE HERE AND A SPECIAL THANK YOU FOR SIGNING ME A CHALLENGING TOPIC, FUTILE CYCLES AND KEEP IT SIMPLE. I ALSO WANT TO THANK MOR,RI AND RON FOR THE CONTEXT OF MY TALK AND I JUST NEED THE FORWARD BUTTON. THAT WILL WORK. SO THE CONCEPT THAT I'M GOING TO COVER TODAY IS REALLY THE POTENTIAL APPROACH TO EXTENDING HEALTH SPAN BY TARGETING FETAL METABOLIC CYCLES. THE SIMPLEST DEFINITION OF FETAL CYCLE HAS THE POTENTIAL TO WASTE ENERGY. SOILY START WITH A QUESTION OF WHY THIS MIGHT BE BENEFICIAL PARTICULARLY IN THE CONTEXT OF AGING. THE RATIONALE STEMS FROM WHAT I REFER TO AS THE ENERGY BALANCE MODEL OF AGING WHICH SUGGESTS THAT CHRONIC POSITIVE ENERGY BALANCE RESULTS IN EXCESSIVE STORAGE OF ENERGY OR EXCESSIVE POTENTIAL ENERGY AND THIS ENERGY FUELS DAMAGE TO CELLULAR CONSTITUENTS SUCH AS DNA AND PROTEIN AN LIPID THROUGH A BROAD RANGE OF MECHANISMS THAT WE DISCUSSED AT THIS MEETING SO THE IDEA IS EXCESS ENERGY FUELS DAMAGE AND THERE BY ACCELERATES IS AGING PROCESS. BY CONTRAST, IF ENERGY INTAKE CAN BE MATCHED MORE PRECISELY TO MEET ONLY WHAT'S REQUIRED FOR OPTIMAL CELLULAR FUNCTION AND ENERGY STORAGE AND DAMAGE ARE MINIMIZED IN DELAYING OR AGING IS DELAYED. SO WHAT IS THE EVIDENCE OF THIS MODEL? THE EVIDENCE STEMS FROM THREE BASIC OBSERVATION OBSERVATIONS. THE FIRST IS OBESITY LOWERS HEALTHY LIFE EXPECTANCY. OBVIOUSLY OBESITY EXCESSIVE ENERGY STORAGE. THE SECOND OBSERVATION IS HABITUAL PHYSICAL ACTIVITY WHICH INCREASES FUEL BURNING AN MINIMIZES FUEL STORAGE IS ASSOCIATED WITH ENHANCED LIFE EXPECTANCY. THIRD, HOT TOIC OF DISCUSSION HERE, STUDIES APPLYING MODELS RANGING FROM WORMS TO NON-HUMAN PRIMATES CALORIC RESTRICTION EXTENDS LIFE SPAN AND LOWERS DISEASE INCIDENCE. SO CONCLUSION WE COME TO BASED ON OBSERVATIONS IS THAT ENERGY BALANCE MATTERS PERHAPS THE SIMPLEST PRESCRIPTION FOR LONG HEALTHY LIFE IS EAT LESS AND MOVE MORE. THE PROBLEM WITH THAT AS MANY ARE WELL AWARE THAT LIFESTYLE MODIFICATION IS EASIER SAID THAN DONE. SO THAT BRINGS US TO THE NEXT QUESTION WHETHER OR NOT WE CAN DEVELOP ALTERNATIVE STRATEGIES TO MINIMIZE ENERGY SURPLUS. THAT IS WHERE WE COME TO THE POSSIBILITY OF TWEAKING METABOLIC CYCLES. MORE ELABORATE DEFINITION OF FETAL CYCLE IS NON-EQUILIBRIUM REACTION CATALYZED BY DIFFERENT ENZYMES THAT ACT SIMULTANEOUSLY. SO I'M GOING TO HIGHLIGHT TWO IMPORTANT ELEMENTS OF THE CYCLES. FIRST IS THE FORWARD REACTION SHOWN IN GREEN, REVERSE REACTION IN BLUE IS CATALYZED BY DIFFERENT ENZYMES. THIS IS IMPORTANT BECAUSE IT ALLOWS REACTIONS TO OCCUR SIMULTANEOUSLY AND ALSO PRESENTS THE OPPORTUNITY TO REGULATE FLUX AT TWO INDEPENDENT SITES. THIS REACTION, THE FORWARDS -- THE FORWARD REACTION IS CATALYZED BY A, REVERSE REACTION BY B AND THE NET PHLOX IS RATE OF FORWARD REACTION MINUS RATE OF REVERSE REACTION. ANOTHER CRITICAL ELEMENT TO THE CYCLES IS AT LEAST ONE REACTION MUST DEPEND ON ATP HYDROLYSIS. THESE CYCLES CONSUME ATP AND GENERATE HEAT WITHOUT CHANGE IN RATIO OF SUBSTRATE TO PRODUCT. ANOTHER QUESTION FORTED BAIT IS WHY THE CYCLES EXIST. SO I THINK THE CONVENTIONAL THINKING IS THAT EVOLUTION FAVORED PROCESSES THAT ARE MORE ENERGY THRIFTY SO MIGHT SEEM COUNTER INTUITIVE PRESERVED A CYCLE FROM ENERGY. THE THEORETICAL ANSWER ORIGINALLY PROPOSED BY ERIC NEWSHOLD IS THESE CYCLES INTRODUCE ARE IN PART SENSITIVITY SO A CELL OR SYSTEM THAT OPERATES WILL BE MORE RESPONSIVE TO REGULATORY QUEUES THAT TARGET ONE OR BOTH OF THESE ARMS OF THE PATHWAY. THE ADVANTAGE OF THE CYCLE SENSITIVITY AND FLEXIBILITY AND THE COST IS ATP CONSUMPTION. THE NEXT QUESTION, WHY WOULD SENSE ACTIVITY BE IMPORTANT? ONE REASON IS BECAUSE IN MANY CASES METABOLIC INTERMEDIATES GENERATED BY CYCLES ARE USE ADS SUBSTRATES FOR ENERGY PROVISION UNDER CIRCUMSTANCES WHEN ENERGY DEMANDS INCREASES. THE ANALOGY THAT'S OFTEN DRAWN IS ONE OF AN IDOLLING ENGINE. AN ENGINE THAT IS IDOLLING AT HIGH RPM IS WELL POISED TO ACCELERATE FROM 0 TO 60 IN SHORT NOTICE. SO YOU CAN THINK THE SAME MIGHT BE TRUE OF RESTING SKELETAL MUSCLE IN AN ANIMAL THAT IS ANTICIPATING THE NEED TO MOVE QUICKLY. SO THEN THE ANTICIPATION -- OKAY. SO THE ANTICIPATION MIGHT INCREASE THE CYCLING RATE AND THE RIGHT REGULATORY QUEUE THAT THEN INHIBITS THE REVERSE REACTION WOULD CAUSE A VERY DRAMATIC INCREASE IN SUBSTRATE FEE THAT CAN PROVIDE ENERGY AND ATP PRODUCTION, THERE BY PROVIDING A VERY ROBUST FIGHT OR FLIGHT RESPONSE. IN ADDITION TO PROVIDING ENERGY, SOME OF THESE INTERMEDIATE CYCLES CAN BE USED IN A SIGNALING CAPACITY OR CAN BE USED FOR BIOSYNTHESIS OF MACRO MOLECULES SUCH AS DNA OR PROTEIN AND LIPIDS SO THEN ANOTHER ADVANTAGE OF THESE CYCLES ENMANS SIGNALING CAPACITY OR PROVIDE SUBSTRATES FOR METABOLIC BIOSYNTHESIS WHICH COULD BE IMPORTANT IN CIRCUMSTANCES SUCH AS GROWTH AND REPAIR. SO THIS BRINGS ME TO THE TRIGLYCERIDE FATTY ACID SIGNAL WHICH IS ONE EXAMPLE OF SUBSTRATE CYCLE THAT I THINK CAN BE RELEVANT IN CONTEXT OF AGE RELATED OBESITY. TRIGLYCERIDE MOLECULE IS MAJOR STORAGE FORM OF ENERGY IN MOST CELLS T THREE FATTY ACIDS. MOST IF NOT ALL CELLS ENERGY RESERVOIRS ARE UNDERGOING CONSTANT REMODELING. SO THE PROCESS OF GLYCOLYSIS WILL CONVERT THE TRIGLYCERIDE MOLECULE TO FATTY ACID AND GLYCEROL AN THESE PRODUCTS FROM NOT NEEDED FOR ENERGY PRODUCTION OR SOME OTHER PURPOSE THEN THEY WILL BE QUICKLY REVERIFIED THROUGH REACTIONS THAT REQUIRE ATP. ANOTHER ASPECT OF THIS PARTICULAR CYCLE IS THE GLYCEROL GENERATED THROUGH GLYCOLYSIS CAN ALSO ENGAGE IN A SECOND CYCLE WITH POTENTIAL TO PRODUCE NAD. THAT'S A REQUIRED CO-FACTOR FOR THE SIRTUIN AND THERE BY IS WIDELY CONSIDERED A POTENTIAL LONGEVITY SIGNAL. SO THEN IF WE PULL THIS INFORMATION TOGETHER WE MIGHT SUGGEST THAT WE MIMIC CALORIC RESTRICTION BY ACTIVATING CYCLES WHICH IN THEORY CONSUME ARCTP, REDUCE ENERGY STORAGE AND POTENTIALLY GENERATE LONGEVITY SIGNAL. ALSO INTERESTING IS MANEUVERS THAT ARE CONSIDERED TO BE MORE ANTI-AGING SUCH AS CALORIC RESTRICTION AND EXERCISE ACTUALLY ACTIVATE THE CYCLE AND SIGNALS MORRI TALKED ABOUT, INSULIN SIGNALING AND HFO SIGNALING INHIBIT THE CYCLE. SO BRINGS US TO THE PROVOCATIVE IDEA WE MIGHT BE ABLE TO MIMIC CALORIC RESTRICTION BY ACTIVATING THE CYCLE. SO TRIGLYCERIDE FATTY ACID CYCLE IS ONE OF MANY POTENTIAL THERMO GENIC CYCLES, I HAVE LISTED SEVERAL HERE. THESE INCLUDE PROTEIN AND GLYCO GENERAL TURN OVER. I WILL END BY RAISING IMPORTANT KNOWLEDGE GAPS IN THE FIELD, FIRST WE NEED TO UNDERSTAND WHERE AND WHEN CYCLINGS ARE ACTIVATED IN VIVO AND ARE THEY AFFECTED BY AGING. WHETHER OR NOT WE CAN MIMIC CALORIC RESTRICTION BY INCREASING SUBSTRATE CYCLING, WHETHER THEY'RE INVOLVED IN METABOLIC FINE TUNING AND GENERATING LONGEVITY SIGNALS AND ARE THEY MODIFIABLE. THANK YOU VERY MUCH. [APPLAUSE] WE'LL MOVE TO THE PEN ULTIMATE TALK BY DON DONGSHENG CAI, HE DID WORK ON CENTRAL INFLAMMATION AND TALK ABOUT HYPOTHALAMIC NEURODEGENERATION, A COMMON LINK BETWEEN METABOLIC DISEASE AND AGING AND I'LL FOLLOW YOU. THANK YOU VERY MUCH. THANK CHRIS AND JEFF FOR THE INVITATION AND ORGANIZERS FOR PUTTING TOGETHER THIS TERRIFIC PROGRAM, IT'S EXCITING AND STIMULATING. SO I HAVE HEARD INTERESTING TOPICS FROM MORRIS TO DIFFERENT METABOLIC CHANGES RELEVANT TO AGING. YOU ALSO HEARD SOME JUST IN CONCEPTS ABOUT INFLAMMATION YESTERDAY. THERE WAS A WHOLE SECTION ABOUT THAT FOLLOWED BY STRESS INFLAMMATION. I ALSO NOTICED SOME SPEAKERS MENTION ABOUT THE BRAIN DYSFUNCTION FUNCTIONS HAS BEEN DESCRIBED WHETHER BRAIN FUNCTION COULD HAVE CAUSATIVE -- IN DEVELOPMENT AGENT OR AGENT BIOLOGY SO I'M TRYING TO PUT ALL THESE FEW POINTS TOGETHER, CONSIDER PERHAPS THE CONNECTIONS OF THOSE MAJOR POINTS MAY HAVE PARTICULARLY IMPORTANT ROLE FOR OUR UNDERSTANDING OF AGING BIOLOGY AN AGING RELATED DISEASES, SUGGESTING POTENTIAL PATHWAY NEW INFLAMMATION, HYPOTHALAMIC INFORMATION MAY HAVE FORM BARRIERS BETWEEN THE METABOLIC DISEASE AND AGING AND PARTIALLY THROUGH NEURODEGENERATION. SO TO DISECT THIS COMPREHENSIVE NETWORK, TWO LABELS WE CAN APPROACH. SO WHY IS ON THE LEFT HAND IN THE CONTEXT OF CALORIC OVERNUTRITION WE KNOW THIS IS -- PROVIDE IMPORTANT BACKGROUND FOR DEVELOPMENT OF METABOLIC SYNDROME, A COLLECTIVE TERM OF NUMBER OR INTERCONNECTED DISEASE OR DISORDERS SUCH AS OVERWEIGHT, HYPERLIPIDEMIA, INSULIN INTOLERANCE, HYPERTENSION, ALL PARTS OF METABOLIC SYNDROME HAS BEEN ADDED AS PART OF METABOLIC SYNDROME. SO WE KNOW UNCONTROLLED CONDITION OF METABOLIC SYNDROMES LEAD TO DEVELOPMENT OF TYPE 2 DIABETES, NEURODEGENERATIVE DISEASE ET CETERA. ALL THESE PROBLEMS CAN BE FURTHER PROMOTED UNDER AGING CONDITION SO OVER SOME YEARS, 6, 7 YEARS, OUR LAB AS WELL AS OTHER LABS OBTAIN SIGNIFICANT EVIDENCE SUGGESTING THAT THE NEW INFORMATION, THE HYPERTHALAMIC INFORMATION MAY REPRESENT A LINK BETWEEN METABOLIC ENVIRONMENTAL INDUCED METABOLIC CHALLENGES AND METABOLIC CONSEQUENCES. THIS IS THE ONE ARM, MEANWHILE ON THE RIGHT HAND, THIS NEW INFORMATION OR HYPOTHALAMIC INFORMATION SYSTEM CAN BE ALSO EMPLOYED OR UTILIZED UNDER CONDITION OF AGING INCREASE, POST DEVELOPMENTALLY AND AGING DEVELOPMENT INCLUDING AGING RELATED DISEASES. IN REALITY, LEFT AND RIGHT QUITE OFTEN P SIMULTANEOUSLY TAKE PLACE. BUT AT THE LEVEL TRY TO DISSECT FOR EXAMPLE CAN USE ANIMAL MODELS WITH OVERNUTRITION AT YOUNG AGE TO UNDERSTAND THIS, OR WE CAN STUDY ANIMALS UNDER NORMAL NUTRITION TO START THIS AND THEN PUT TOGETHER TO GET MORE VIEW. I THINK THIS TYPE OF INFORMATION PROBABLY A LITTLE BUILT DIFFERENT THOUGH A LOT OF SIMILAR CHANGES COMPARED TO THE CLASSICAL INFORMATION AND ONE SIMILARITY IS VERY IMPORTANT CONTROL COST INFORMATION NF KAPPA B TRANSCRIPTION FACTOR AND UPSTREAM KINASE IKK PATHWAY. SO I JUST A LITTLE BIT BACK GROWN OF THIS PATHWAY, SO WE HAVE SIGNIFICANTLY APPRECIATE THAT ACTIVATION OF THIS PATHWAY AND THE IMMUNITY INFLAMMATION REACTION IN RESPONSE TO PATHOGENS AND ENVIRONMENTAL INNOVATIONS LOAD TO ROBUST QUICK OR CHRONICALLY ACTIVATION OF THIS PATHWAY AND THE HIGH MAGNITUDE, ASSOCIATED WITH MORPHOLOGY CHANGES AND THE PATHWAY HUNDREDS O GENES THAT I INVOLVED FOR THE INFLAMMATION, MUTAGENESIS APOPTOSIS PATHWAY. THIS PATHWAY TYPICALLY IN THE IMMUNE SYSTEMS IMMUNE CELLS, HOWEVER, IN THE CONTEXT OF AGING OR METABOLIC SYNDROME THIS SYSTEM CAN BE ACTIVATED IN NON-IMMUNE SYSTEM -- IMMUNE CELLS, VERY SENSITIVE IF YOU HAVE FOR EXAMPLE A (INAUDIBLE) OR VERY EARLY STAGE AGING SO THIS PATHWAY CAN BE IN THE HYPOTHALAMUS. AND WE HAVE PARTICULARLY APPRECIATE THE ACTIVATION OF THIS SYSTEM INVOLVED A NUMBER OF INTRACELLULAR ORGANELLE OR DYSFUNCTIONS. FOR EXAMPLE IN THE FORM OF (INAUDIBLE) MITOCHONDRIAL CHANGES DEFECT LYSESOME CHANGES, ALSO NUCLEAR ABNORMALITIES SUCH AS DNA DAMAGE HISTONE DYSFUNCTIONS. SO ALL THIS ORGANELLE CHANGES, THEY HAVE THEIR OWN WAYS TO CROSS TALK TO THIS INTRACELLULAR INFLAMMATORY MACHINERY. TO INDUCE ACTIVATION. INITIALLY TO TRY TO ADAPT, TRY TO RESOLVE SOME OF BAD EVENTS BUT THIS IS -- WHEN THIS IS NOT RESOLVED, IT'S ALWAYS ON AND HAS BEEN KNOWN BE P PROBLEMATIC. AND IN CONTEXT OF METABOLIC DISEASE AND SYNDROME, SOME YEARS AGO SIX, SEVEN YEARS AGO GIVE BRIEFLY METABOLIC INFORMATION ACTIVATED ASPECT OF ALSO SHARED AGING CONDITION, SO I THINK WE STILL DON'T HAVE ENOUGH UNDERSTANDING DETAIL CHARACTERISTICS OF THIS PARTICULAR INFORMATION UNDER AGE OR METABOLIC DISEASE CONDITION BUT WE BEGIN TO APPRECIATE THE CONSEQUENCES OR THE IMPACT OF THIS TYPE OF INFORMATION T REGULATORY SITE IN THE HYPOTHALAMUS, I LIST A FEW HERE. WHEN THIS INFORMATION IS INDUCED IN CONTEXT OF OVERNUTRITION OR AGE INCREASE, IT CAN AFFECT DIFFERENT NEURAL CELL TYPES INCLUDING NEURONS SO THE CHANGE IN NEURONAL SIGNAL AND CONTROL OF BODY PHYSIOLOGY CAN GLIAL CELLS GLIAL FUNCTIONS THEREFORE EFFECT GLIAL CROSS TALK AND INTERACTION. AND CAN AFFECT VASCULATURE. MAYBE AS WELL AS THE BRAIN DYSFUNCTIONS BECAUSE CHANGE BLOOD SUPPLY OF THE BRAIN. APPRECIATE ANOTHER LEVEL NEW STEM CELL LEVEL. TALK AN ADULT NEW STEM CELLS, WE HAVE A SESSION THIS AFTERNOON ON THIS, NOT TRY TO TALK TOO MUCH BUT THIS SUBJECT BUT BRIEFLY OVER THE LAST DECADE THE LITERATURE HAS BEEN CONVINCED THAT SEVERAL TYPES OF NEURAL STEM CELLS IN THE BRAIN AND IN THE RODENTS. SO TWO IMPORTANT REGIONS IDENTIFIED, ONE IS DEN TSAI GYRUS HIPPOCAMPUS, THE OTHER IS VENTRICAL ZONE, THE LEVEL VENTRICAL IN THE BRAIN SO THESE TWO REGIONS HAVE HIGH LEVEL FOR NEURAL STEM CELLS. AND LAST FEW YEARS WE HAVE SEEN THE HYPOTHALAMUS IS PERHAPS A THIRD IMPORTANT BRAIN REGION THAT WITHOUT NEW STEM CELLS. AND SOME ADDITIONAL EXPERIMENTS FOR EXAMPLE (INAUDIBLE) FOR SYSTEM CELLS HAVE BEEN DONE SUGGESTING THOSE CELL POPULATION, THESE CELL POPULATION DOES HAVE PHYSIOLOGICAL ELEMENTS AND I JUST PUT THIS IN I GUESS THE IMAGE (INAUDIBLE) CAN GIVE A LITTLE BIT ILLUSTRATION SO THIS IS THE HYPOTHALAMUS STEM CELL ISOLATE MICE AND CULTURALLY RICH IN FORM OF -- DIFFERENTIATED INTO NEURAL CELLS NEURONS, ASTROCYTE IN GREEN, YOU CAN APPRECIATE THE RATIO DIFFERENCIALLY IN NEURONS AND ASTROCYTE AND WE'RE STRUCK BY OBSERVATION THAT UNDER CHRONIC OVERNUTRITION THIS IS A SIGNIFICANT COMPROMISE NOT ONLY DEPLETION OF NEURAL STEM CELLS BUT ALSO THEY HAVE POOR DIFFERENTIATION THAT YOU CAN SEE HERE. THEY BARELY DIFFERENTIATE IN NEURONS THOUGH THEY CAN DIFFERENTIATE INTO ASTROCYTE IN QUITE A LIMITED MANNER. IT SUGGESTS THAT THE MODEL HERE, IN A NORMAL PHYSIOLOGY, THE HYPOTHALAMUS STEM CELLS CAN CONTRIBUTE TO A FRACTION OF NEURAL CELLS INCLUDING NEURONS, PERHAPS TO E REPLACE CERTAIN NUMBER OF DYSFUNCTIONAL NEURONS. AND THIS POPULATION, THIS TURN OVER IS PROBABLY SMALL IN THE SHORT TIME WINDOW, IS NOT DETECTED OF JUST WITHIN THE SHORT TIME WINDOW. BUT IT COULD BE VERY SIGNIFICANT OVER TIME. AND DISEASE CONDITION I MENTION DEGENERATION, AND ALSO INDUCE NEUROGENETIC FUNCTION OF THOSE CELLS. ALTOGETHER IN THE PARADIGM WE THINK CHRONIC OVERNUTRITION ACTIVATE THE NEW INFORMATION LEAD TO NUMBER OF EVENTS INCLUDING IMPAIRED NEUROGENESIS AND PROMOTE NEURODEGENERATION AND PROBABLY SHOULD CONTRIBUTE THE DEVELOPMENT OF METABOLIC SYNDROME AS WELL AS AGE DISEASE, PERHAPS COMMON BASIS LIST A FEW POINTS FOR YOU TO CONSIDER AS POTENTIAL DISCUSSION. THAT'S IT. >> THANK YOU VERY MUCH. [APPLAUSE] >> THANK YOU, DONGSHEN. FINAL TALK OF THIS SESSION WILL BE GIVEN BY ROMAN KONDRATOV FROM CLEVELAND STATE. HE WORKS ON MOLECULAR CLOCKS AN CIRCADIAN RHYTHM AND WILL PARTICULARLY SPEAK ABOUT CIRCADIAN DESYNCHRONIZATION AS A KEY PLAYER IN METABOLIC DISEASE OF AGING. ROMAN. >> OKAY. THANK YOU ORGANIZERS FOR THE INVITATION AND THIS OPPORTUNITY TO SPEAK HERE. BEFORE I ADDRESS QUESTIONS, I HAVE BEEN ASKED VERY INFORMED QUESTION AND PROBABLY ACTUALLY MULTIPLE QUESTIONS. I WILL START FROM THIS SLIDE JUST TO TELL YOU CIRCADIAN CLOCK IS A LITTLE BIT MORE COURSE OF JET LAG. FOR AGES WE KNOW CIRCADIAN DYSFUNCTION IS ASSOCIATED WITH SLEEP DISORDERS AND PSYCHIATRIC DISORDERS. WHAT WE KNOW NOW FROM EPIDEMIOLOGICAL STUDIES THAT CIRCADIAN MISALIGN. IN HUMANS WILL INCREASE RISK OF CARDIOVASCULAR DISEASES, AND INCREASE RISK OF METABOLIC SYNDROMES OBESITY, DIABETES AND OTHER PATHOLOGIES. WHAT IS CIRCADIAN CLOCK? WE KNOW RHYTHMS IN SLEEP WORK CYCLE WRITTEN IN THE AREA, THIS IS A FUNCTION OF CENTRAL CLOCK WHICH IS SITUATED IN HYPOTHALAMUS, IN THE BRAIN. BUT WHAT IS PROBABLY LESS KNOWN THAT CIRCADIAN CLOCKS OPERATE IN EVERY CELL IN EVERY TISSUE. SO AND THIS CIRCADIAN CLOCK FROM CIRCADIAN POINT OF VIEW, THIS CLOCK GENERATE RHYTHM IN GENE EXPRESSION SO ABOUT 10% OF GENES IN EVERY TISSUE SHOW CIRCADIAN RHYTHM IN GENE EXPRESSION. AND THIS RHYTHM GENE EXPRESSION TRANSFORM INTO CIRCADIAN RHYTHM AND PHYSIOLOGY AND METABOLISM. FIRST QUESTION NEED TO BE ANSWERED WHAT IS EFFECT OF AGING ON CIRCADIAN CLOCK? AN ANSWER WILL BE NOT SIMPLE ONE, BECAUSE AN AFFECT ON CENTRAL CLOCK WILL EFFECT SLEEP, WILL AFFECT RESPONSE THE LIGHT AND BEHAVIOR, ALL CAN CONTRIBUTE TO DEVELOPMENT OF DIFFERENT METABOLIC DISEASES AND OTHER DISEASES, BECAUSE FOR EXAMPLE, IT IS VERY WELL KNOWN SLEEP DISRUPTION IS ASSOCIATED WITH METABOLIC DISEASES AND PROBABLY CONTRIBUTE TO THIS. WHAT IS MUCH LESS APPRECIATED FROM MY POINT OF VIEW IS AFFECT OF AGING ON CLOCK BUT ACTIVITY OF ACTIVITY OF PERIPHERAL CLOCKS IS EXTREMELY IMPORTANT FOR CRIMINAL OF METABOLISM AND FOR CONTROL OF PHYSIOLOGY. WHAT WE KNOW ABOUT CHANGES IN THE PERIPHERAL CLOCK THE AMPLITUDE OF CIRCADIAN RHYTHMS REDUCED WITH AGE. AND WHEN WE HAVE REDUCTION IN AMPLIMY TUESDAY IT CAN BE ACHIEVED IN TWO WAYS. ONE WAY THE CLOCK IN ALL TISSUES OR IN CELLS, WILL STOP. WE WILL SEE THE (INAUDIBLE). ANOTHER POSSIBILITY IS DESYNCHRONIZATION OF INDIVIDUAL CLOCKS IN THE TISSUE. IF EVERY CELL HAVE A FACE OF ISOLATION, WHEN WE MEASURE RHYTHM AS A GENE EXPRESSION PARAMETER WE WILL SEE AMPLITUDE RHYTHM DECREASE IF CLOCK STOPPED WITH AGE OR SAY UPON DISEASE, THE QUESTION IS, AT WHAT TIME POINT CLOCK WAS STOPPED BECAUSE THIS IS AN EXAMPLE OF TWO TYPE OF CIRCADIAN MUTANTS SO IT'S DEFICIENT ONE IN CIRCADIAN CLOCK. SO BOTH OF THESE GENES EXPRESSION AND ACTIVITY ISOLATE UNDER NORMAL CIRCUMSTANCES. BUT UPON MUTATION, ISOLATION WILL DISAPPEAR. BUT YOU WILL SEE THAT ISOLATION WILL STOP AT EXACTLY OPPOSITE TIMES. AND THEY AFFECT METABOLIC EFFECT AND PHYSIOLOGICAL AFFECT IN THESE TWO MUTANTS WILL BE DIFFERENT. SO WHY IS IT IMPORTANT TO UNDERSTAND IS IT STOP OR DESYNCHRONIZATION? TO CONTINUE PARALLEL WITH DRIVING CAR, IF YOU HAVE CLOCK WHICH IS STOPPED, WITH TRANSMISSION, YOUR GEAR IS LOCKED OR IN GEAR, IF YOU ARE DEALING DESYNCHRONIZATION YOU'RE DEALING WITH CAR TRANSMISSION IS ACTUALLY REACHED IN RANDOM WAY, CONSTANTLY. SO THEREFORE IN ORDER TO FEED THE PROBLEM, YOU NEED TO KNOW WHAT THE PROBLEM. THE SECOND QUESTION IS HOW THIS CIRCADIAN MISALIGNMENT, HOW THE CIRCADIAN DYSFUNCTION COULD AFFECT AGING. SPECIFICALLY CHRONIC DISEASE OF AGING. AGAIN, ANSWER IS NOT SIMPLE, AND MY FEELING IS IT WILL BE TISSUE AND CELL TYPE SPECIFIC BECAUSE CIRCADIAN CLOCK CONTROL DIFFERENT SET OF GENE IN DIFFERENT TISSUES. AGING WOULD AFFECT CENTRAL CLOCK AND PERIPHERAL CLOCK AND MANY THINGS WILL BE CHANGED. BUT IF WE'RE TALKING ABOUT METABOLISM AND P EFFECT OF DISRUPTION ON METABOLISM, I THINK THERE ARE SEVERAL PATHWAYS WHICH ARE -- COULD BE UNIVERSAL AND NEED TO BE CONSIDERED. SO ONE THING CRIMINAL OF CELLULAR RHYTHM STATE AND OXIDATIVE STRESS RESPONSE, THERE IS A BIG BODY OF DATA ON THIS SUBJECT. ANOTHER IMPORTANT PATHWAY IS CELL PATHWAY. IT WAS DEMONSTRATED THAT CIRCADIAN CLOCK SIGNALING PATHWAY ARE INTERLINKED AND IT WAS DEMONSTRATED BY LABORATORY (INDISCERNIBLE) AND JOE BUTTS P AND I THINK THIS IS IMPORTANT. MY LAB HAS INTEREST IN CIRCADIAN CLOCK DEPENDENT REGULATION OF MTOR SIGNALING PATHWAY. I DON'T THINK I HAVE TO INTRODUCE THIS IN THIS MEETING. WHAT WE FOUND TOR SIGNALING PATHWAY UNDER NORMAL PHYSIOLOGICAL CONDITION IS CONTROLLED BY CIRCADIAN CLOCK. THIS CONTROL IS ACTUALLY FEELING UNDERSTOOD PENDENT, AND THIS CONTROL IS DISRUPTED IN CIRCADIAN MUTANTS AND THAT CAN CONTRIBUTE TO METABOLIC DISORDERS OF AGING. FINAL SLIDE WHY IT IS IMPORTANT TO TARGET CIRCADIAN CLOCK AS THERAPEUTIC TARGET. ONE ADVANTAGE IS WHEN YOU TARGET THE CLOCK YOU TARGET MULTIPLE PHYSIOLOGICAL SYSTEMS. THE SECOND ADVANTAGE AND PROBABLY NOT ADVANTAGE FROM ECOLOGICAL POINT OF VIEW. YOU CAN TARGET CLOCK. BEHAVIORAL DIETARY INTERVENTION BECAUSE YOU DON'T HAVE TO USE DRUG AN THERE ARE A FEW EXAMPLES OF THIS. RESTORATION OF CIRCADIAN RHYTHM DELAY DEVELOPMENT OF PATHOLOGY AND ANIMAL MODEL SO CELLULAR DEMONSTRATED OBESITY AND METABOLIC SYNDROMES IN MICE HIGH FAT DIET. SO I'M DONE. [APPLAUSE] >> INVITE PANELISTS TO COME UP TO THE STAGE AND WHILE THEY'RE DOING THAT I'LL MAKE SOME COMMENTS. I THINK FROM THIS DISCUSSION, IT'S VERY BROAD AND VARIETY AREAS METABOLISM, U THINK AS EVERYBODY CAN SEE, METABOLISM EFFECTS MANY PARTS OF AGING P AND AGING RELATED DISEASE PROCESSES. I THINK THAT IS A PRIMARY TAKE HOME MESSAGE. AS CHRIS SAID AT THE BEGINNING CAUSE AND EFFECT. WE HAVE NO IDEA. THIS IS AN ENTERRELATED COMPLEX PATHWAY. WHETHER DNA METHYLATION, HISTONE MODIFICATION, PROTEIN ACELATION ALL REQUIRE METABOLIC SUBSTRATES AND PRODUCTS AND ANY DISREGULATION OF ONE EFFECTS THE OTHER ONE. SO YOU THINK ABOUT PROTEINS ACETATE, ACETATE DOESN'T COME ON TO A PROTEIN AS ACETATE, IT COMES THROUGH ACETYL COA THROUGH METABOLISM, YOU HEARD ABOUT NAD CYCLES. THE OTHER POINT TO MAKE IS THE CYCLING PART FUNDAMENTAL ASPECT OF METABOLISM WHICH IS NOT APPRECIATED AND DECK BY DID A GOOD JOB DESCRIBING HOW FUTILE CYCLES ARE POISESSED TO CHANGE VARIOUS FUNCTIONS. THINK OF CIRCADIAN RHYTHMS WHICH ARE IMPORTANT IN AGE PROGRESS SYCES, AS ROMAN DESCRIBED, THAT IS A CYCLIC PATHWAY, A FETAL CYCLE WHERE THINGS ARE CYCLING AROUND. SO THOSE COMMENTS I WILL LET THE PANEL ANSWER QUESTIONS. >> >> LEFT SIDE MIC PLEASE. THERE WE GO. >> THANK YOU. I'M BOB JOKE FROM AMERICAN SOCIETY OF BONE AND MINERAL RESEARCH. I HAVE A COMMENT A LOT OF SPEAKER WHOSE DEALT WITH AGE RELATED BONE LOSS RELATED TO SARCOPENIA PORTRAYED IT A SARCOPEDIA BEING CAUSE OF BONE LOSS BUT IN FACT THERE IS A LOT OF CONFUSION ABOUT THAT IN THE LITERATURE NOW AND -- THERE'S EVIDENCE BONE LOSS TAKES PLACE INDEPENDENTLY OF SARCOPENIA ESPECIALLY IN THE MOUSE. I ADMIT THE DATA IN THE HUMANS IS NOT WELL ESTABLISHED. IN MY OWN LAB WE HAVE SHOWN AND FIELD IN GENERAL HAS ACCEPTED THE IDEA THAT BONE LOSS IS -- WITH AGING IS DUE TO A DEFICIT IN OSTEOBLAST THAT SEEMS TO OCCUR INDEPENDENTLY OF CHANGES IN MUSCLE FUNCTION SO I INVITE DR. EVANS OR OTHERS ON THE PANEL TO COMMENT ON THAT. >> HOPEFULLY I DIP SUGGEST SARCOPENIA IS DRIVING LOSS OF BONE. WHAT IS CLEAR IN THE LONGITUDINAL OBSERVATIONAL STUDIES IS WEIGH LOSS RESULTS IN BONE LOSS. THE GREATER THE AMOUNT OF WEIGHT LOSS THE GREATER BONE LOSS. THAT'S FOR SURE. WE ALSO KNOW THERE IS A PRETTY GOOD RELATIONSHIP SHOWN DIETARY PROTEIN INTAKE IS ASSOCIATED WITH BONE HEALTH AS WELL. I HAVE NO DOUBT THERE IS ALWAYS A STRONG RELATIONSHIP BETWEEN BONE MASS AND MUSCLE MASS. WHAT DRIVES THE OTHER IS WHAT IS LESS WELL UNDERSTOOD. I APPRECIATE THAT. >> I EACH RICHARD MILLER UNIVERSITY OF MICHIGAN. DR. CAI'S STUDIES INDICATED THAT THE GONADOTROPIM RELEASING HORMONES PLAY A ROLE IN HYPOTHALAMIC INFLAMMATION IN MOUSE MODELS SO IT WOULD BE GOOD TO HEAR YOUR IDEAS BUT ANYONE ELSE IDEAS WHETHER THIS MAY OR MAY NOT BE A MAJOR DRIVING FORCE BEHIND THE SEXUAL DIMORPHISM IN RESPONSES TO MALE AND FEMALE MICE TO DIET OR DRUGS SOME WHICH HAVE SPECIFIC EFFECTS ON ONE SEX OR THE OTHER TO WHAT EXTENT TO THESE ANTI-AGING INTERVENTIONS INTERACT WITH MALE SPECIFIC OR FEMALE SPECIFIC PATHWAYS AND IS THE HYPOTHALAMUS THE PLACE TO LOOK FOR THE ANSWER. >> THANK YOU, DR. MILLER FOR THIS. NICE COMMENT. GNIH IS ONE MOLECULE DOWN STREAM HYPOTHALAMIC INFORMATION, I BELIEVE THERE ARE OTHERS BUT WE HAVE STUDIED NIH RECENTLY REPORTED. I THINK IT IS AN INTERESTING POINT THE LINK THE CONNECTION BETWEEN PHYSIOLOGY AND BUYOLOGY ON AGING SO THERE'S ALWAYS A RELATIONSHIP BETWEEN THIS AND THE AGENT. THERE'S MANY MODELS SHOWING THAT SEPARATION OR ACTIVE CONTROL MOLECULES CAN AFFECT THE AGING AND LIFE SPAN IN A POSITIVE SOME CASE NEGATIVE MANNER. SO IT'S VERY POSSIBLE THAT GNIH LEVEL MAY EXPLAIN DIMORPHISM IN FEMALE IN TERMS OF LIFE SPAN EXPECTANCY. SO THERE'S SOME -- EVEN SOME EXAMPLES IN A SOCIETY, FOR EXAMPLE IN MALE IN LOYAL FAMILY HAVE BEEN SHOWN INCREASED -- HIGH INCIDENCE OF (INAUDIBLE) FROM THAT POPULATION. AND IN ANIMALS HAS BEEN ALSO SEEN THE SITUATION. MAYBE IN TERMS OF MALE AND FEMALE POST (INAUDIBLE) CHANGE SEXUAL HORMONES AND INFLUENCE OF GNIH MAY CONTRIBUTE TO THE DIFFERENT (INAUDIBLE) MALE AN FEMALES. >> I WANT TO PUSH ON THAT. JUST A LITTLE BIT. ONE OF THE THINGS I SAID IN MY REMARKS SOME OF US COMING FROM OUTSIDE EVERY DAY WORK IN THE AGING FIELD. I WAS STRUCK BY THE CAR BOASE OBSERVATION THE BLOCKER OF INTESTINAL CARBOHYDRATE IN GLUCOSE ABSORPTION LEADS TO A SEX SPECIFIC EFFECT ON AGING. SO HAS ANYONE LOOKED AT IN THE STUDIES WHETHER THERE ARE CHANGES IN HORMONES WE VIEW SEX SPECIFIC OR SEX DRIVING. IF SO IT HAS A CONNECT. I'M NOT AWARE OF AN AGENT THAT BLOCKS GLUCOSE ENTRY HAVING ANY PARTICULAR EFFECT ON SEX HORMONES. JUST NOT AWARE OF IT. >> (INDISCERNIBLE) INSTITUTES. THE QUESTION IS STAGE 4 CANCER AN DIET. IT IS A TUG OF WAR BETWEEN DIFFERENT ACADEMIES CARBOHYDRATE METABOLISM INTAKE, METABOLISM INTAKE. SOME OF THEM RECOMMEND GENIC DIET IS THE ONLY WAY TO FIGHT CANCER. THIS PROMOTES SARCOPENIA, SO WHAT'S THE BALANCE, WHAT KIND OF DIET WOULD YOU RECOMMEND TO BALANCE CANCER IN SARCOPENIA AT THE SAME TIME. >> THE ONLY THING I WOULD SAY IS IN CANCER WE NEED TO INCREASE FOOD INTAKE PERIOD. ACCELERATED PROTEIN DEGRADATION IT IS UNRESPONSIVE TO NUTRITION. HALL HARK OF CACHEXIA IS NUTRITIONAL STRATEGIES HAVE NO EFFECT ON MUSCLE LOSS. SO WHETHER YOU COMBINE WITH ANABOLIC AGENT INTERVENTION LIKE PHYSICAL ACTIVITY, NUTRITION PER SE AFFECTS MUSCLE LOSS. WE KNOW FROM SEPSIS DATA. YOU CAN MAINTAIN THEIR WEIGHT ABSOLUTELY BUT THEY WILL CONTINUE TO LOSE MUSCLE MASS. >> HOW DO YOU CORRECT ANOREXIA? >> THESE PEOPLE HAVE SEVERE ANOREXIA. >> I DON'T KNOW. >> I ENJOYED THE TWO TALKS IN THE HYPOTHALAMUS. IF YOU LOOK AT THE REGULATORY THERMOSTATS IN THE HYPOTHALAMUS, THEY'RE A YIN YANG SYSTEM, WE TALKED OBESITY AND AGING. IF YOU ENTER LIFE AS SOMEBODY SUGGESTED YESTERDAY THAT HE DID, WITH INCREASED DRIVE MORE PROTEIN DRIVE IN NUCLEUS, IF WE'RE LOOKING AT THINGS LIKE EFFECT OF INFLAMMATION AM I RIGHT IN THE GENIC SYSTEM, ONE WOULD HAVE TO GO UP. AND THE AN REXIC SYSTEM THEY HAVE TO GO DOWN. SO PERHAPS WE NEED TO LOOK MORE CAREFULLY AT SPECIFITY WITHIN THESE SYSTEMS BECAUSE IF INFLAMMATION IS AFFECTING BOTH THE SAME, CHANGE THE DRIVE. >> DO YOU KNOW TO TAKE THAT ONE? >> I THINK THIS IS AN INTERESTING POINT. THERE ARE A VARIETY OF DIFFERENT TYPES OF CELLS IN THE HYPOTHALAMUS, MAYBE CELLS ALSO CRITICAL. THE INFLAMMATION HAS IMPACT ON THEM. I GUESS OFF STREAM CHANGES PROBABLY SHARED BUT DOWNSTREAM CONTEXT DEPEND ON SIGNALING CASCADE AND FUNCTIONS. IT COULD HAVE DIFFERENT OPPOSITE FUZZ OWE LOGICAL IMPACT DEPEND ON WHAT KIND OF -- FOR EXAMPLE PROMOTE FOOD INTAKE, THE FUNCTIONS, REGULATORY FUNCTION OF THOSE NEURONS ARE IMPAIRED THEIR RESPONSE TO OFF STREAM SIGNAL, CAN HAVE THE SAME DIRECTION BOTH CONTRIBUTE TO THE NET BOLLIC SYNDROME LIKE PHENOTYPE. SO I GUESS QUITE DYNAMIC, THEY HAVE TO CONSIDER THE CONTEXT OF SPECIFIC CELL TYPES AN UPSTREAM DOWNSTREAM EVENTS. >> DID YOU WANT TO SAY SOMETHING ALSO? >> I CAN HEAR YOU. >> WE AGREE WE HAVE TO TAKE INTO CONSIDERATION SPECIFIC CELL TYPES AND PROBABLY RESPONSE WILL BE DIFFERENT IN DIFFERENT CELL TYPES. FROM CIRCADIAN POINT OF VIEW. QUESTION WOULD BE WHAT TIME OF DAY RESPOND WOULD BE DIFFERENT IN DIFFERENT TIME OF THE DAY SET OF GENES REGULATED IN DIFFERENT PART OF THE BRAIN. I CANNOT ANSWER THIS QUESTION IN ONE WAY OR ANOTHER BEFORE WE HAVE ALL THIS. >> I WONDERED IF THERE WAS ANY EVIDENCE. Q. I CAN MAKE A COMMENT. THE NF KAPPA B SIGNALING PATHWAY IS DIFFERENT IN DIFFERENT CELL TYPES. IT'S NOT PUBLISHED BUT WE HAVE DATA NF KAPPA B ADIPCYTES GIVES A DIFFERENT OUTPUT THAN IMMUNE CELL SO YOU GET DIFFERENT CYTOKINE PRODUCTION AND DIFFERENT RESPONSES. SO SOME ARE SIMILAR SOME ARE DIFFERENT. SO YOUR QUESTIONS ARE A GOOD VALID ONE, YOU MIGHT HAVE DIFFERENT SIGNALING OUTPUTS AND EGRP -- YOU COULD HAVE A DECREASE IN INFLAMMATORY SIGNALING BUT PERHAPS AN INCREASE, YOU GET A DIFFERENCE OUTPUT. THOSE THINGS YOU'RE ABSOLUTELY RIGHT ARE VERY IMPORTANT TO SORT OUT. >> (INAUDIBLE) FROM THE LIFE EXTENSION FOUNDATION. DR. NEWGARD BRANCH CHAIN AMINO ACIDS INCREASE INSULIN RESISTANCE AND DIDN'T DISCUSS THAT AT THIS PRESENTATION. DR. EVANS HAS SAID IMAGING RESTRICTION INCREASES THE NEED FOR PROTEIN BUT SPOKE ONLY OF ENERGY RESTRICTION RATHER THAN SOMETHING LIKE CARBOHYDRATE RESTRICTION. SO I'M WONDERING IF WE COULDN'T GET MORE SPECIFIC DIETARY RECOMMENDATIONS TO ADDRESS THESE PROBLEMS. >> FIRST OF ALL, I'M RIGHT. WHAT WE ON SEVERED IS A VERY STRONG ASSOCIATION OF INCREASE IN BRANCH CHAIN AMINO ACIDS AN RELATED METABOLITES IN MULTIPLE HUMAN DISEASE SETTINGS. AS WE HAVE BROUGHT THAT OBSERVATION BACK TO THE LABORATORY, WE PROMOTE AMINO ICEYS IN HIGH CONTEXT DIETS SO BRANCH CHAIN AMINO ACIDS BEGIN TO EXERT NEGATIVE EFFECTS NOT NECESSARILY IN THE OLDER SEDENTARY SUBJECTS BILL WAS SPEAKING ABOUT EARLIER. IN THE CONTEXT OF HIGH ENERGY DIETS, THAT BECOME A BAD ACTOR WHEN OTHER NUTRIENTS ARE ABUNDANT. THAT'S WHEN THEY EXERT EFFECTS ON METABOLIC HOMEOSTASIS IN OUR EXPERIENCE. CONVERSELY IF WE RESTRICT BRANCH CHAIN AMINO ACIDS, IN OBESE ANIMALS, WE CAN RESOLVE A SIGNIFICANT AMOUNT OF UNDERSTAND LYNN RESISTANCE. THAT'S THE DATA WE HAVE TO THIS POINT. >> I DIDN'T DISCUSS IT BECAUSE I DIDN'T HAVE TIME ENERGY OUR BEDREST STUDIES, IF WE SUPPLEMENT HEALTHY OLDER PEOPLE WITH AIMEE MOW ACIDS, 15 IN THE MORNING AN EVENING IT OBLITERATES THE LOSS OVERMASTING. WHILE THERE IS AN BOLL UK RESISTANCE IN OLDER PEOPLE, YOU CAN OVERCOME IT WITH ESSENTIAL QUANTITIES OF AMINO ACIDS WHICH INCLUDE THE BRANCH CHAIN. ONE OF THE CHALLENGES THAT WE HAVE IS TO PROVIDE ADEQUATE PROTEIN NUTRITION. AMOW KNOW ACIDS THERE ARE IN THE FOOD THE GREATER THE STIMULATION OF PROTEIN SYNTHESIS IN OLDER PEOPLE. HIGH QUALITY LOW FAT PROTEIN IS MY RECOMMENDATION. >> >> IN HONOR OF (INAUDIBLE) THAT BROUGHT US TOGETHER NIDDK, NIA, OTHERS, I WANT TO DO A (INAUDIBLE) THAT HAS TO DO WITH GEROSCIENCE, IT'S TO YOU GUY WERE FUNNED BY THE NIDDK. FROM THE FIRST DAY OF ME COMING FROM NIDDK SWITCHING FROM NIDDK TO NIA, THE CONCEPT IN DATA NIDDK DOES INSULIN RESISTANT WAS ESSENTIAL TO DISEASES. AND IN AGING, INSULIN RESISTANCE IS CENTRAL TO LONGEVITY. THERE WERE TOTALLY DIFFERENT VIEWS. IT WAS INSULIN RESISTANT IN NIDDK BECAUSE WE HUMANS HAVE POOLED BETTER CELLS AND DEVELOPED DIABETES. SO THAT WAS THE PROBLEM. WHILE USING MODELS EVERY TIME YOU CAUSE INSULIN RESISTANCE YOU HAVE LONGEVITY. LIEU LUIGI FERRUCCI AND I PUBLISHED IN JOURNAL OF ONTOLOGY ALL STUDIES AT END POINT OF LIFE SPAN. IN ANIMALS THAT ARE INSULIN RESISTANTS OR INSULIN SENSITIVE. THERE'S NO RELATIONSHIP TO LONGEVITY. MAYBE POINTING TO NEWGARD'S POINT IT'S UPSTREAM SOMEWHAT ALSO. BUT THERE ARE LOTS OF MODELS THAT ARE INSULIN RESISTANTS LIKE THE MORRIS WHITE IRS, THAT CLEAVE LONGER OR MAUREEN SHOWN GLUTE 4 OVEREXPRESSION SENSESIVE AND A LITTLE BIT SHORTER. WHY IS IT SO IMPORTANT FOR US TO DISCUSS THOSE THING? AND IT HAS THE TO DO WITH HUMANS HEALTH SPAN. BECAUSE WHEN WE DECIDE TO TREAT PEOPLE FOR INSULIN RESISTANCE KNOWING IT IS A STRESS RESPONSE ALSO, WHY ARE ARE WE SO SURPRISED SURPRISED IT DOES CARDIO VASCULAR DISEASE, WE REMOVED A STRESS RESPONSE, THE APPROACH THAT COMES FROM AGING MOLECULAR APPROACH THAT COME TO NIDDK ARE REALLY EXEMPLIFIEDED BY WHAT YOU SAID MORRI AND SAID AT THE END. I DON'T UNDERSTAND THE INSULIN RESISTANCE, BUT THERE ARE THINGS TO CONSIDER ABOUT INSULIN RESISTANCE VERY RELEVANT TO THE INSTITUTIONS. >> THEY'RE LOOKING AT ME. WHICH DOESN'T P SURPRISE ME. BEFORE I DO THAT WE HAVE A REAL PROBLEM, THAT IS THE DEFINITION OF INSULINS RESISTANCE. THE PROBLEM IS THAT PHRASE CEASED TO MEAN ANYTHING WE CAN AGREE ON. SO WHEN YOU SAY, WHEN YOU REFER TO INSULIN RESISTANCE, FROM POPULATION OF OBESE INDIVIDUALS INCREASING PREVALENCE TODAY, NOT SURE WHAT THAT MEANS. IF YOU TAKE THE CLASSICAL AS WELL AS BIOLOGICAL ACTIONS OF INSULIN. OR IN MANY MANY WAYS TYPE ONE DIABETIC BECAUSE THEY DON'T HAVE SIGNALING. THE THING THAT I THINK ALL OF US KNOW IS OBESE PATIENT WITH INSULIN RESISTANCE DOES NOT FEE KNOW COPY A LOSS OF INSULIN SIGNALING. THERE ARE REASONS YOU CAN ARGUE THAT, YOU CAN ARGUE THAT'S THE CASE. ONE IS BECAUSE CONCEPT OF SELECTIVE INSULIN RESISTANCE, SOME PATHWAYS ARE RESISTANT, OTHERS AREN'T. THE OTHER OF COURSE IS FUNDAMENTAL CONCEPT OF INSULIN RESISTANCE AND ETIOLOGY OF TYPE 2 DIABETES IS WRONG. THERE'S NOT TIME TO DISCUSS REASONS WHY THAT MIGHT NOT BE THE CASE BUT THE IMPORTANT MEASURE -- MESSAGE TO GET ACROSS IS WHEN WE REFER TO QUOTE INSULIN RESISTANCE IN TYPE 2 DIABETIC PATIENTS IN OBESE INDIVIDUALS, THAT IS NOT AN EXCLUSIVE SELECTIVE -- SELECTIVE TO INSULIN SIGNALING PATHWAY BUT GENERALIZED IN TERMS OF TISSUES DECREMENT IN INSULIN SIGNAL. WHEN YOU TALK ABOUT MANIPULATING INSULIN SIGNALING TO EXTEND LIFETIME, THAT'S WHAT YOU'RE TALKING DOWN, KNOCKING DOWN AKT, AFFECTING ARCKT SIGNAL AS WELL AS OUTPUTS AND ALL TISSUES. SO UNFORTUNATELY THE TERM RESISTANCE IS UNIFORM REDUCTIONS IN SIGNALING AND SYNDROME PEOPLE GET BUT SAME TERM APPLIED TO TWO DIFFERENT SITUATIONS. >> I THINK IN MY MIND, DEFINITION OVINES HUNDRED RESISTANCE IS NOT CLEAR CUT. THERE MIGHT BE A FIZZ QUO LOGICAL INSULIN RESISTANCE PATHOLOGICAL INSULIN RHESUS TAN. I DON'T THINK IT'S BENEFICIAL A -- ETIOLOGY WE NEED A LEVEL OF INSULINS RESISTANCE, IN -- IN A STRESS CONDITION. PARTICULARLY EVEN SOME TISSUES, FOR EXAMPLE BRAIN, LITTLE BIT INSULIN RESISTANCE MAYBE HELP THE BRAIN TO COPE WITH THE STRESS RESPONSE, CONSISTENT WITH DISCUSSION YESTERDAY AFTERNOON ABOUT STRESS RESISTANCE, MAYBE CONSISTENT. SO WE MAY HAVE LANGUAGE BETWEEN TWO INSTITUTIONS. >> ONE ADDITIONAL POINT, A LITTLE BIT DIFFERENT THAN DR. CAI. PEOPLE BELIEVE THERE'S NO DIFFERENCE BETWEEN PATHOLOGICAL INSULIN RESISTANCE, THAT'S AN APPROPRIATE RESPONSE TO OVER EATING AND OVERNUTRITION AND IF PEOPLE DIDN'T HAVE THAT RESPONSE THEY WOULD BE IN WORSE SHAPE THAN WITH RESPONSE. THAT'S AN IMPORTANT CONCEPT TO HAVE OUT THERE. >> ROWDILY VINE FOR NIH. QUESTION FOR DEPUTY MUOIO. YOU POINT OUT EFFECT OF FUTILE CYCLE IS TO CONVERT ARCTPASE TO CONVERT ENERGY TO HEAT. AND ONE WAY TO DO THAT IS TO HAVE TWO ENZYME, ONE THAT CATALYZE FORWARD REACTION, ONE THE OTHER. BUT WOULDN'T IT BE SUFFICIENT TO SIMPLY TARGET ANY FORE REACTION THAT USES ARCTP? THE GLUTAMATE GLUTAMINE PAIR ON YOUR LIST, P IF YOU HAD A DRUG THAT INTERFERED WITH THE FULL CYCLE OF GLUTAMINE SYNTHASE YOU WOULD BE HYDROLYZING ATP. >> ONE POSSIBLE ANSWER TO THAT QUESTION, IF YOU LACK THE ARM OF THE CYCLE THEORETICALLY WHAT YOU'RE MISSING IS METABOLIC CONTROL ASPECT OF THE CYCLE AND NOT ENERGY WASTING OF THE CYCLE. SO THERE'S DEBATE WHY THEY EXIST AND CONCEPTUALLY IT DOESN'T MAKE SO MUCH PHYSIOLOGICAL SENSE THERE WOULD BE SOME SYSTEM TO SIMPLY WASTE THAT ENERGY. AND THE ARCHITECTURE, THE NETWORK WOULD HAVE TO BE DESIGNED SOD THAT PROCESS CAN BE TURNED OFF ON PHYSIOLOGICALLY APPROPRIATE. EXPANDING THE DEFINITION AND EWE L FILLTY OF THOSE CYCLES, TO MEAN REGULATING SIGNALING AND METABOLIC PRECURSORS, IT'S IMPORTANT TO HAVE THAT CONTROL IN VARIOUS SITES OTHER THAN JUST THE ONE ENZYME. >> I AGREE, IF YOU WANT TO INTERVENE YOU HAVE THE OTHER STRATEGY. >> I THINK FOR PURPOSES OF ENERGY WASTING THAT CUB BENEFICIAL BUT THEN AS PART OF THE DISCUSSION WE HAVE BEEN HAVING HERE, THERE WILL BE SOME DETRIMENT TO UNCONTROLLED ENERGY WASTINGING SO TURNING ON A PATHWAY THAT WASTES ENERGY UNCONTROL WOULD BE FAVORABLE AND ONE EXAMPLE OF THAT IS METABOLIC UNCOUPLERS SO TURNING ON UNCOUPLING THE PROCESS OF UNCOUPLING WITHOUT HAVING SOME REGULATORY ARM IS DETRIMENTAL. >> NICK (INAUDIBLE) FROM THE BAR INSTITUTE SAN ANTONIO. I'M ENDOCRINOLOGIST INTERESTED IN AGING SO WHEN ARLEN RICHARDS AND RANDY STRONG FOUR OR FIVE YEARS AGO ASKED WHETHER WE WOULD STUDY AFFECT OF RAPAMYCIN IN HUMAN SUBJECTS WE THOUGHT IT WAS A CRAZY IDEA, IT WAS RAPAMYCIN WAS THOUGHT TO INDUCE INSULIN RESISTANCE AND PROMOTE INTOLERANCE. AS WE HAVE BEEN LEARNING MORE IT'S WARRANTED TO STUDY IN HUMANS. ALSO IN HEALTHY SUBJECTS FOR SEVERAL REASONS, THE EFFECTS ON GLUCOSE METABOLISM ARE UNCLEAR, EVIDENCE FROM A HE ROW ROW GENIUS MOUSE IS NOT OBSERVED. SOME PEOPLE THAT HAVE DONE PILOT STUDIES, HEALTHY SUBJECTS, WE HAVEN SEEN HYPERGLYCEMIA, FASTING GLUCOSE. THE OTHER HAS TO DO WITH CARDIOVASCULAR DISEASE, SEVERAL GROUPS DEMONSTRATING RAPAMYCIN HAS A STRONG EFFECT TO PREVENT ATHEROSCLEROSIS AND APOE AND KNOCKOUT MICE AND THAT'S WHY RAP LOGS ARE USED AND CORONARY SIN. LASTLY, AS YOU POINTED OUT, IT MAYBE THE EFFECT OF RAPAMYCIN TO EXTEND LONGEVITY TO POTENTIAL EFFECT ON INSULIN ACTION. THE DIFFICULT SITUATION NOW IS THERE'S COMPELLING REASONS WHY YOU WOULD WANT TO TRY RAPAMYCIN IN PEOPLE. THERE ARE THEORETICAL BUT EQUALLY COMPELLING REASONS YOU WORRY ABOUT IT. ONE ISSUE WITH RAPAMYCIN AND INSULIN RESISTANCE IS INDIRECT. THE THOUGHT IS BASED ON DAVID SACKATINI'S WORK IS WHEN YOU HAVE RAPAMYCIN YOU SEQUESTER TOR TO INACTIVE TOR 1 COMPLEX AND TURN DOWN TORK 2 SIGNALING. SINCE IT IS IMPORTANT FOR THE ACT VISION OF AKT WHICH IS AN INTEGRAL INTERMEDIATE AND CLASSICAL CANONCAL METABOLIC SICKNALLING PATHWAY YOU GETTINGS LINS RESISTANCE. -- YOU GET INSULIN RESISTANCE. ALTERNATIVE EXPLANATIONS WHY, ONE POSSIBLY THAT'S NOT AN ISSUE, THAT IS THE ASSUMPTION IN THE COMMUNITY, THAT IS THE PREVALENT BELIEF IS TYPE TWO DIABETES, UNDERSTAND LUNES RESISTANCE IS INACTIVATION OF AKT THAT IS THE PREVALENT BELIEF. THERE ARE OTHER PLAUSIBLE SEQUENCES OF EVENTS THAT WOULD LEAD TO DECREASE IN AKT ACTIVATION AS CONSEQUENCE AND NOT CUSS OF DISEASE. IF THAT'S THE CASE RAP MYSPIN IS NOTD WITH AT ALL. IT'S GOING TO FEE KNOW COPY THE FINAL STEP. BUT IT'S NOT BAD. SO TO RESOLVE WE HAVE TO UNDERSTAND RAPAMYCIN BUT WE HAVE TO BE MUCH CLEARER ON THE SEQUENCE OF HE WANT THAT LEADS TO QUOTE INSULIN RESISTANCE TYPE 2 DIABETES. YOU CAN SUMMARIZE BY SAYING THERE'S A HUNDRED PERCENT CERTAINTY THAT RAPAMYCIN EXTEND YOUR LIFE SPAN IF YOU DON'T DIE FIRST. >> MY NAME IS CHRIS LIEU FROM BOSTON UNIVERSITY. MY QUESTION IS FOR DR. EVANS, REGARDING INTERVENTIONS FOR -- TO PREVENT NITROGEN LOSS WITH BEDREST. CURIOUS TO KNOW, YOU LOOKED AT AMINO ACID SOUPLE MINTAGE AS INTERVENTION, IF YOU LOOK AT PHYSICAL ACTIVITY AS INTERVENTION, IN PARTICULAR CYNTHIA BROWN'S WORK AT UNIVERSITY OF ALABAMA WHERE SHE DID AN INTERVENTION TO INCREASE MOBILITY AND HOSPITALIZE ELDERS. >> ONE INTERVENTION WE DID WE GOT EACH OF THE OLDER PEOPLE OUT OF BED FOR 15 MINUTES PER DAY AND THEY WALKED ON A TREADMILL. SPECIFIC PERCENTAGE OF THEIR MAXIMAL AROW BIB CAPACITY. IT HAD NO EFFECT ON LOSS OF MUSCLE. SO 15 MINUTES A DAY OF WALKING WAS INADEQUATE TO PREVENT THE LOSS. IT MAY BE STUDIES BY NASA SHOW THAT ONE OR TWO BOUTS OF RESISTANCE EXERCISE EXERT A MORE POWERFUL EFFECT THAN GIVE -- GETTING UP AND MOVING AROUND. THAT IS PROBABLY MOST APPROPRIATE WAY TO EXERCISE THE PATIENT. THERE WERE A NUMBER OF OTHER EFFECTS. THESE ARE HEALTHY PEOPLE, THEY WEREN'T SICK. THIS IS THE BEST CASE SCENARIO. THEY BECAME HYPOTENSIVE TRYING TO AMBULATE AND MOVE AROUND. SO THERE ARE A LOT OF CHANGES THAT OCCUR IN CARDIOVASCULAR SYSTEM AS WELL AS MUSCLE PROTEIN METABOLISM. THE INSULIN RESISTANCE WE SEE IMPAIRS AMINO ACID STIMULATED BLOOD FLOW TO MUSCLE. THAT MAYBE ONE OF THE SOURCES OF THE SO CALLED ANABOLIC RESISTANCE. >> WE HAVE TEN MINUTES LEFT IN THE SESSION SO TRY TO KEEP IT -- THE QUESTIONS AND THE ANSWERS BRIEF. >> DEAN JONES EMORY UNIVERSITY. BROADEN THE SCOPE OF DISCUSSION A LITTLE BIT. IT IS POSSIBLE WITH METABOLIC PROFILING TO MEASURE 20 OR 30,000 CHEMICALS IN A LIVING ORGANISM. WE HAVE DONE THAT IN ASSOCIATION WITH AGING AND FLIES AN MARMOSETS AN HUMANS. WHAT IS REMARKABLE IS THERE ARE HUNDREDS OF CHEMICALS THAT CHANGE SIGNIFICANTLY IN ASSOCIATION WITH AGE, WE DON'T HAVE REGARD TO LONGEVITY. THE QUESTION COMES FROM THAT. AND THAT IS THAT HAS ANYBODY OR DO YOU KNOW EXPERIMENTS WHERE PEOPLE HAVE TAKEN THE CALORICALLY RESTRICTED MODELS. AND ASKED METABOLIC QUESTIONS AND INTERVENTIONAL QUESTIONS YOU'RE DISCUSSING. IT WOULD SEEM IF YOU MAXIMALLY EXTEND WITH RESTRICTION, YOU CAN CONTRIBUTE TO OTHER MECHANISMS. >> I DON'T THINK -- I'M SNOT AWARE AS ONE OF THE CENTERS THAT DOES METABOLIC PROFILING WE HAVE LOOKED AT MULTIPLE MODELS OF CALORIC RESTRICTION. METABALOMIC OUTCOMES IN SUCH BUT NOT IN THE CONTEXT OF THE AGING -- THE LIFE SPAN EXTENSION TIME POINT. WE CAN TELL YOU WHAT HAPPENS METABOLICALLY WITH WEEKS OF CALORIC RESTRICTION IN A DIO HIGH FAT DIET ANIMAL, CHANGES ARE PROFOUND. AS YOU SWITCH TO A MORE CAT BOLLIC CONDITION. HOW THAT PLAYS IN TERMS OF SUBSEQUENT TEMPORAL PATTERN OF METABOLIC CHANGE IN AN ANIMAL THAT EXPERIENCES LIFE SPAN EXTENSION VERSUS ONE THAT IS MAINTAINED ON AN AD LIB REGIMEN, NOT AWARE OF DATA SETS OF THAT NATURE. IF ANYONE S SPEAK UP. LIKE SO MANY THINGS IN THIS FEEL, THINKING THACKS IMPORTANT HURTS MY HEAD. >> THERE IS A PUBLICATION FROM THE WISCONSIN NON-HUMAN PRIMATE CALORIE RESTRICTION STUDY. (INAUDIBLE) FIRST AUTHOR FROM 2009, IT'S EXPERIMENTAL DERMATOLOGY. SO THEY DID NMR PROFILING THREE TIME POINTS THROUGH AGING AS PART OF THE LONGITUDINAL STUDY. TRAJECTOR ROY CHANGE WAS DIFFERENT FOR THE CONTROL ANIMALS THAN CR ANIMALS. SIGNIFICANT DIFFERENCES IN EACH TIME POINT BUT THAT DYNAMIC OF CHANGE THROEING WAS DIFFERENCE FOR BOTH GROUPS AS WELL THAT'S THE ONLY ONE FOR THE NON-HUMAN PRIMATES I'M AWARE OF PUBLISHED. >> GEORGE MARTIN UNIVERSITY OF WASHINGTON. FIRST I WANT TO CONGRATULATE, NOT IN TO METABALOMICS BUT YOU DID A GREAT JOB. I DON'T KNOW MUCH ABOUT THE HIPPOCAMPUS, BUT WONDERING WHAT'S THE PROXIMAL STIMULUS FOR INFILTRATION OR ACTIVATION OF THE MICROGLIA. AND THE INFLAMMATION, WOULD IT BE JUDY CAMPISI'S SASS WITH GLIAL CELLS KNOWN TO UNDERGO REPLICATIVE SENESCENCE? OR IS IT SOMETHING TO DO WITH PROTEOSTASIS, PROTEIN AGGREGATES THAT TRIGGERS IT? HOW DO YOU THINK ABOUT THAT? >> EXCELLENT QUESTION. AT THIS MOMENT WE DON'T HAVE A CLEAR ANSWER. WE NEED SOME YEARS TO P DEVELOP A PICTURE. CONCEPTUALLY THINKING DEPEND WHAT STAGE YOU'RE TALKING ABOUT. INITIAL STAGE OR DURING THE DEVELOPMENT OF AGE. IF YOU TALK ABOUT DURING THE CAUSE OF AGE AND DEVELOPMENT, THERE'S CERTAINLY THE CROSS TALK BETWEEN MICROGLIA ASTROCYTE AND NEURONS. THEY INFLUENCE EACH OTHER, CLEARLY THEY'RE (INAUDIBLE) PATHWAYS CYTOKINES AND MAYBE MORE THAN CYTOKINES. IF YOU TALK INITIAL CHANGE OF INFLAMMATORY SIGNAL, INTRINSIC HOMEOSTATIC CHANGES. INTRACELLULARLY MAY AS I SAID IN A TALK MAY HAVE GOOD CONTRIBUTION BUT WE STILL DON'T KNOW EXTRA CELLULAR STIMULI WHAT KIND OF IMPORTANT FACTORS WE COULD BE MOST IMPORTANT IN THAT PROCESS. BUT THAT'S I THINK SIGNIFICANT AREA FOR THE FUTURE. >> WE DIDN'T HEAR ABOUT BROWN FAT IN THIS STORY, UNCOUPLED AND GENERATES HEAT. WE'RE LOSING IT AS WE GET OLD. LOSING BROWN FAT. >> I'M NOT AWARE OF ANY STUDY SHOWING THAT. >> ACTIVITY GOES DOWN >> DID CHANGE IN SYMPATHETIC NERVE ACTIVITY BUT THERE'S CERTAINLY I THINK A LOT OF EVIDENCE THAT MODIFYING THE ACTIVITY OF BROWN FAT CAN BE BENEFICIAL FOR HEALTH OUTCOMES THAT WE STUDY IN THE CONTEXT OF OBESITY AND DIABETES. I THINK IT'S AN INTRIGUING POSSIBILITY YOU MIGHT BE ABLE TO TARGET SOME OF THESE CYCLES IN THE BROWN FAT TO THEN GENERATE SIGNALS FAVORABLE FOR LIFE SPAN. >> THERE IS DATA IN ANIMALS THAT HAVE MORE BROWN FAT. THEY DO LIVE LONGER. WHETHER THEY'RE HEALTHY AGING OR PROTECTED AGAINST DISEASE, IT'S STILL NOT NOW. >> JIM NELSON, BAR SHOP INSTITUTE SAN ANTONIO. COMMENT AND QUESTION WITH RESPECT TO THIS PARADOX IF YOU WILL INSULIN RESISTANCE AN MAMMALS AN INVERTEBRATES JUST WANTED TO BRING TO ATTENTION A FASCINATING PAPER PUBLISHED IN JBC 7, 8 YEARS AGO. REPLACED MURINE RECEPTOR WITH A MUTATED INSULIN RECEPTOR BY KNOCKING IN ONE THAT HAD A MUTATION AT A SITE COMMON TO THE MUTANTS THAT LIVE LONG IN C ELEGANS. WHAT THEY FOUND OUT WAS THAT THIS REPLACEMENT INCREASED QUOTE UNQUOTE CLASSIC MAMMALIAN INSULIN RESISTANCE DEFINED BY GLYCEMIC RESPONSES BUT IT ALSO HAD A REMARKABLE AFFECT ON INCREASING RESINCE ANSWER STRESSORS IN VITRO AND IN VIVO AND WENT ON PUBLISHED THE LIFE SPAN DATA AND SHOWED THAT DESPITE CONTINUED INSULIN RESISTANCE, THERE WAS NO DIFFERENCE IN LIFE SPAN. IT WAS A VERY SOLID STUDY. MY QUESTION IS WITH RESPECT TO BILL EVANS. THANK YOU FOR THAT CONSCIOUSNESS, DISCUSSION OF ANABOLIC RESISTANCE, FOUR YEARS AGO WHEN THE NIA WAS FIRST FORMED THERE WAS DISCOVERY RECENTLY BEEN MADE OF THE STEROID AND PEPTIDE HORMONE RECEPTORS AND SIGNALING PATHWAYS AND THERE WAS A BIG PUSH THEN TO LOOK AT AFFECT OF AGING ON THOSE PATHWAYS. THIS IS KIND OF FALLEN BY WAYSIDE. WONDERING WITH RESPECT TO ELDERLY INDIVIDUALS WHAT WE KNOW ABOUT ANABOLIC HORMONE RECEPTOR SIGNALING PATHWAYS, IS THERE RESISTANCE IN THOSE PATHWAYS? >> THE LIMITED DATA PUBLISHED INDICATE THERE CONTINUES TO BE RESISTANCE TO STRAIGHT ANABOLIC THERAPY THOUGH NOT CLEAR WHERE THE RESISTANCE LIES. THERE ARE DATA THAT SHOW THAT IF YOU STIMULATE GLAD FLOW, MUSCLE SYMPATHETIC NEVER ACTIVITY DECREASED PRODUCTION OF NOS AN DECREASED VASODILATION ASSOCIATED WITH FEEDING. THAT IS A LARGE PART OF INSULIN RESISTANCE LIES. IF YOU CHANGE BLOOD FLOW WITH ANABOLIC THERAPIES YOU SEE ALMOST NO RESISTANCE AT ALL TO EITHER AMINO ACID FEEDING OR ANABOLIC EFFECTS OF TESTOSTERONE. >> WE ONLY HAVE TWO MINUTES LEFT. VERY BRIEF. >> MIKE ANSON, THIS WE FOR BILL EVANS IN MACRO NUTRIENT STUDY THERE'S A TIME OF ADAPTATION TO THE CHANGE IN PROTEIN LEVEL OR CHANGE IN CARBOHYDRATE LEVEL AND IN REAL LIFE WITHOUT NON-EXPERIMENTAL WHEN YOU'RE HOSPITALIZED IT HAPPENS UNEXPECTEDLY DO WE KNOW WHETHER WE LOSE THE ABILITY TO ADAPT TO SUDDEN CHANGES IN MACRO NUTRIENT, TIME COURSE CHANGE WITH AGE? DO WE KNOW THAT? >> I'M IN THE AWARE OF ANY ONE THING THAT HAPPENS IN HOSPITAL SETTINGS IS OLDER PATIENTS BECOME -- EVEN WORSE SHOW THAT IN MOST HOSPITAL SETTINGS, OLDER PEOPLE ARE GIVEN 50% OR LESS OF WHAT TRUE ENERGY REQUIREMENT IS, THAT'S WHAT THEY'RE GIVEN, NOT WHAT THEY EAT. THOSE PATIENTS DO EXTRAORDINARILY POORLY. NUMBER ONE REASON FOR THAT IS DOCTORS WERE GOOD TO TAKE NPO OFF THEIR ORDERS. THAT'S 30% OF ELDERLY PEOPLE IN THE VA SYSTEM. SO WE HAVE A LOT OF WORK TO DO. >> I THINK THIS IS A QUESTION ABOUT FETAL CYCLES BUT BECAUSE I DON'T KNOW ANYTHING ABOUT FETAL CYCLES. ME THIONEINE RESTRICTION EXTENDS RAT LIFE SPAN BY 40% THOUGH THEY EAT LIKE CRAZY, THEY'RE NOT RESTRICTED JUST THEY CAN'T GET THE BINDING RESIDUE. HOW DOES THAT WORK? HOW DOES THAT EXTEND THEIR LIFE SPAN IN >> LET ME -- >> I REFER THAT TO MORRIS. >> LET ME JUMP IN ON THAT ONE. WE HAVE BEEN STUDYING A INHIBITOR WHICH IS THE ENZYME THAT BLOCKS ME THIONEINE OFF END TERMINUS PROTEINS. IF YOU INHIBIT THAT ENZYME YOU DROP CELLULAR ME THIONEINE LEVELS BY 40%. THERE'S A VERY IMPORTANT ROLE FOR REMOVAL OF THAT IN TERMS OF HOMEOSTASIS OF ME THIONEINE POOLS. THAT INTERVENTION, A BLOCK OF THE INHIBITOR CAUSE AS DRAMATIC WEIGHT LOSS, REDUCED CALORIC INTAKE. SO I DO THINK MORE STUDY OF THIS ME THIONEINE MECHANISM IN TERMS OF LONGEVITY AND FEEDING BEHAVIOR IS VERY WELL WARRANTED. THERE'S LITERATURE OUT THERE LIKE OTHER ASPECT OF NUTRITIONAL LITERATURE CAN BE A BIT OKAY CULT AT TIMES. I THINK IT'S SOMETHING WE SHOULD HAVE A FRESH LOOK AT. >> ONE QUICK COMMENT, IN PREPARING FOR THE CONVERSATION I HAD HERE, THE RESEARCH ON THESE CYCLES GOES BACK TO THE MID 50s AND 60s AND IT WAS VERY ACTIVE AT THAT TIME AND FASCINATING IDEAS. I THINK THAT A LOT OF THE CYCLES THAT WE THINK ABOUT PROTEIN BREAKDOWN AND RESYNTHESIS IN THE CONTEXT OF THE ME THIONEINE DIET VERY WELL ACT ACTIVATED. OTHER THAN UNCOUPLING PROTONE FOR THE LAST COUPLE OF DECADES AND NOW WE HAVE THE TOOLS TO ASK THOSE QUESTIONS IN A MUCH MORE ROBUST DEEPER WAY. >> LAST QUESTION. >> (INAUDIBLE) INSTITUTE THIS IS A QUESTION ON CIRCADIAN RHYTHMS TO ROMAN. I'M SURE THEY MUST HAVE BEEN SOME STUDIES THAT HAVE BEEN DONE ON HUMANS AN ANIMALS THAT HAVE BEEN KEPT (INAUDIBLE) IN THE DARK, IS THERE A CHANGE IN THE PERIPHERAL CENTRAL CIRCADIAN GENES AND DOES THIS I WERE PACT METABOLIC SYNDROME AND INSULIN RESISTANCE FEATURE? >> MY BEST KNOWLEDGE (INAUDIBLE) -- (OFF MIC) >> MICROPHONE. >> TO BEST OF MY KNOWLEDGE, NO, BUT IT IS THIS EXPERIMENT. PEOPLE DID EXPERIMENTS UNDER MINIMAL CONDITIONS BUT NOBODY CHECK EXPRESSION. >> ANY DISEASES LINKED TO KEPT PERMANENTLY IN DISRUPTED FREE RUNNING RHYTHM? >> WHAT IS KNOWN VERY WELL, IF YOU TAKE DISEASE OBESITY OR METABOLIC SYNDROME, ANIMAL MODELS IN OWL THESE CONDITIONS IN ALL THESE CONDITIONS CIRCADIAN RHYTHM IN GENE EXPRESSION IN PRODUCTION OF CIRCULATORY MOLECULES, THEY ARE DISRUPTED. THAT IS WHAT IS KNOWN. THIS RESULT DISRUPTION GENE EXPRESENTATION, GENE EXPRESSION CONTRIBUTE TO THE DEVELOPMENT OF DISEASE IS THE QUESTION. I DON'T KNOW IF I ANSWERED THE QUESTION. >> E WE'LL TALK ABOUT IT LATER. >> SO THANKS TO ALL THE PANELISTSND THE AUDIENCE AND SURE THEY'LL BE READY (INAUDIBLE). [APPLAUSE] >> OKAY, EVERYONE. TAKE YOUR SEATS, WE'RE READY TO START THE NEXT SESSION WHICH WILL BE ON MACRO MOLECULAR DAMAGE. MY NAME IS RON JOHNSON, I'M PROGRAM DIRECTOR NATIONAL CANCER INSTITUTE. FOR TODAY'S SESSION OUR CO-CHAIRS FOR SESSION FOR MACRO MOLECULAR DAMAGE IS ARLEN RICHARDSON AND ARI SCHADT. DR. RICHARDSON IS CELLULAR AND STRUCTURAL BIOLOGY AND UNIVERSITY OF TEXAS HEALTH SCIENCE CENTER SAN ANTONIO AND SEENIER CAREER RESEARCH SCIENTIST AT THAT TIME BAR SHOP INSTITUTE. HE'LL GIVE AN INTROER OVERSUIT OF THE TOPIC FOR THIS SESSION. DR. SCHADT IS PROFESSOR CHAIR DEPARTMENT OF GENETICS AND GENOMICS SCIENCES AND DIRECTOR OF THE INSTITUTE FOR GENOMICS AND MULTI-SCALE BIOLOGY AT THE MOUNT SINAI SCHOOL OF MEDICINE IN NEW YORK CITY. HE WILL BE DIRECTING THE DISCUSSION WHICH WILL FOLLOW THE LAST SPEAKER AND SUMMARY OF THE SESSION. DR. RICHARDSON. >> THANK YOU VERY MUCH, RON. BEFORE WE START I WANT TO GIVE RON CREDIT FOR THE WORK BEHIND THE ORGANIZATION AND KEEPING US ON COURSE HE WOULD ALWAYS SENT ATONIAS SAY EMAILS AND I WOULD PLAY THE GOOD GUY AND SAY MAYBE YOU CAN GET BY WITH ANOTHER DAY OR TWO. SO THANK YOU VERY MUCH, RON. I WOULD LIKE TO START THE SESSION WITH -- I WANTED TO KIND OF CAPTURE THE IMPORTANCE OF OF DAP DAMAGE AND AGING. I APPRECIATE PHILIPPE SIERRA GIVING THIS PICTURE OF THIS CAR WHICH DEMONSTRATE DEMONSTRATES RAVAGES OF DAMAGE AND AGINGING. >> IT WOULD HAVE BEEN NICE IF YOU WOULDN'T HAVE THROWN AWAY THE HUB CAPS OF THE CAR PHILIPPE. WE ON THE OTHER HAND WHY TAKING SENATORS AND CONGRESSMAN, WE AREN'T GETTING MORE MONEY IN AGING. THE CONCEPT THAT DAMAGE WAS IMPORTANT IN AGING IS A RELATIVELY OLD CONCEPT. IT STARTED WITH (INAUDIBLE) IN 42, HE WAS A CHEMIST WITH KODAK WHO CAME ONE A CONCEPT THAT PROTEIN CROSS LINKS MIGHT BE IMPORTANT MECHANISM UNDERLYING AGING. IN '56 DEN HAM HARMON CAME UP WITH FREE RADICAL THEORY OF AGING, THE REACTIVE OXYGEN SPECIES THEORY OF AGING OR OX DAYSIVE STRESS THEORY OF AGING AND THEN CAME UP WITH CONCEPT FREE RADICALS WERE CAUSING DAMAGE AND THIS DAMAGE GAVE RISE TO AGING. SUBSEQUENTLY LEO (INAUDIBLE) FATHER OF THE ATOMIC BOMB A A PHYSICIST CAME ONE THE CONCEPT THAT DNA DAMAGE WAS A KEY IN AGING. AND SUBSEQUENTLY LESLIE CAME UP WITH THE AIR CATASTROPHE CONCEPT WHICH DAMAGE OR MISTAKES IN THOSE PROTEINS AND MOLECULES IN GENE EXPRESSION CALL MORE DAMAGE WHEN THEY WERE IN TRANS SUBSCRIPTION AND TRANSLATION. AND THESE WOULD BUILD UP AND YOU WOULD HAVE THIS CASCADE OF THIS AIR CATASTROPHE. NOW, THE QUESTION IS AT THAT TIME DOES DAMAGE INCREASE WITH AGE? THERE WAS A GREAT DEBATE FOR SUBSTANTIAL AMOUNT OF TIME BUT LATE '70s AND '80s IT WAS SHOWN YES INDEED DAMAGE DOES OCCUR. THESE ARE TWO LEADERS I REMEMBER SPEAKING AT ALMOST EVERY AGING CONFERENCE I WENT TO BACK IN THE '80s. BRUCE AIMS WAS FOCUSING ON DNA DAMAGE THE SLIDE SHOWS DATA FROM OUR STUDIES WHERE WE'RE LOOKING AT DEOXYGAIN SEEN UP WITH AGE. EARLY SATURDAYMAN'S GROUP OX DAYTIVE DAMAGE PROTEIN INCREASE WITH AGE BUT EARL'S GROUP ESSENTIALLY DISCOVERED PROTEINS DID BECOME DAMAGE OR OXIDATIVELY MODIFIED. IT'S INTERESTING ROD WAS TELLING ME THAT EARL ESSENTIALLY WAS UP UNTIL HIS '60s AS THE PERSON STUDYING REGULATION OF PROTEIN WAS IN HIS 65, LATER YEARS THAT HE GOT INCH TO AGING BECAME A KEY PACK FACTOR WE REMEMBERED HIM FOR. SO DAMAGE INCREASES WITH AGE. WHAT EVIDENCE DAMAGE PLAY AS ROLE IN AGING SOMETHING THAT'S THE THEME WITH WE HEARD ALL ALONG HERE. IN OTHER WORDS DOES IT CAUSE IT OR IS IT -- DOES IT CAUSE AGING OR DOES IT ARISE FROM AGING. EVIDENCE FOR DNA DAMAGE IS SEVERAL FOLD. DNA DAMAGE OXIDATIVE DAMMING IS REDUCED BY DIETARY RESTRICTION WHICH INCREASES LIFE SPAN, SLOWS DOWN AGING. THE OTHER INTERESTING THING IS THAT DEFICIENCIES IN DNA REPAIR, THIS WAS OBSERVED IN HUMANS EXHIBIT ACCELERATEDDED AGING PHENOTYPES. THIS IS A STUDY OF A WARNER SYNDROME PATIENT. THIS ISES ARE FOUND IN MOUSE MODELS WHERE THEY KNOCKED OUT VARIOUS DNA REPAIR GENES. IN ADDITION MICE HAVE BEEN MADE -- MAKE MUTATIONS IN THE MITOCHONDRIAL DNA AND THESE ALSO SHOW ACCELERATED AGING INDICATING THAT DAMAGE IN MITOCHONDRIAL DNA IS IMPORTANT. WHAT, HOWEVER THERE'S EVIDENCE SUPPORTING THE CONCEPT THERE'S A FEW PROBLEMS. LEVEL OF MUTATIONS OR DELETIONS WERE DAMAGED THAT YOU SEE IN MODELS OF ACCELERATED AGING, THE AMOUNT OF DAMAGE YOU SEE IS VERY HIGH COMPARED WITH DAMAGE YOU NORMALLY SEE. THE OTHER FACT IS ALTHOUGH WE SEE THIS DIFFERENCE IN DAMAGE AND ACCELERATED AGING, IT WOULD BE IMPORTANT TO SHOW REVERSE, THIS HAS NOT BEEN DONE. IF YOU REDUCE DAMAGE IN DNA WOULD THIS LEAD TO INCREASE LIFE SPAN. WITH RESPECT TO PROTEIN, PROTEIN IS OXIDATIVE DAMAGE TO PROTEIN IS INCREASED IN MANY AGE RELATED DISEASES. INCLUDING SUBJECTS WITH PROGERIA , DIETARY RESTRICTION REDUCES OXIDATIVE DAMAGE TO PROTEIN. NO STUDIES HAVE SHOWN WHERE YOU HAVE BEEN ABLE TO REDUCE DAMAGE TO INT DAP AND INCREASE LIFE SPAN AND (INDISCERNIBLE) SHOWED IN CONTRAST IF YOU WOULD PREDICT BECAUSE THESE ARE LONGER LIVED THEY ACTUALLY HAVE MORE DAMAGE DUE TO BUY LOOKCAL ACTIVITY OF A PROTEIN' BIOLOGICAL ACTIVITY OF A PROTEIN. NOW THE ARGUMENT IS THIS DAMAGE RESULTS IN OLIGOMERIZATION AND AGGREGATION AND THIS IS SOMETHING THAT'S -- THE CELL WANTS TO GET RID OF POTENTIALLY TOXIC SO WE TALK ABOUT PROTEOTOXICITY. AS AS DEAN JONES WILL TALK ABOUT THE DAMAGE TO THOSE PROTEINS IN WHAT HE TERMED THE REDOX PROTEOME, IN OTHER WORDS YOU HAVE A FREE TILE GROUP IMPORTANT IN BIOLOGICAL REACTIONS, THIS COULD HAVE A MULTITUDE OF EFFECTS NOT ONLY TRANSCRIPTION FACTORS BUT IN A VARIETY OF OTHER PLACES SO THAT YOU WOULD HAVE THIS GLOBAL EFFECT. WHAT WE'RE GOING TO DO TODAY ON OUR SESSION IS SPEAKERS IN THE FIRST TWO PRESENTATIONS WILL FOCUS ON THE IMPORTANCE OF DNA DAMAGE IN AGING. THE NEXT TWO WILL FOCUS ON PROTEIN AND ONE PROBLEM PROBLEM OR LIMITATION OF DATA IN THIS FIELD IS WE LOOK AT DNA DAMAGE OR PROTEIN DAMAGE OR MAYBE DAMAGE TO LIPIDS AND EVEN LOOKING AT DNA DAMAGE OR PROTEIN DAMAGE WE LOOK AT ONE OR TWO TYPES OF DAMAGE. NO ONE HAS REALLY LOOKED AT WHAT WOULD BE THE CONCENTRATIONS OF A LITTLE BIT OF DAMAGE EVERYWHERE. SO I THINK THIS WOULD BE IMPORTANT IN THE FUTURE. SO WITH THAT, WE'LL GO TO OUR FIRST SPEAKER AND THIS IS JANUARY VIJG, PROSEARCHER OF DEPARTMENT OF GENETICS. I AM SO OLD I REMEMBER JANUARY WHEN HE WAS A OHIO JAN AS A JUNIOR FACULTY MEMBER, A SHORT LITTLE GUY THEN AND HAD REALLY LONG HAIR. SO JAN FOR THOSE OF YOU THAT DON'T KNOW, JAN STARTED OUT HE DEVELOPED THE FIRST -- IT WASN'T CALLED BIG BLUE BUT FIRST MOUSE TO LOOK AT MUTATIONS TO STUDY POTENTIAL TO ASK THE QUESTION, DNA MUTATIONS INCREASE WITH AGE. THIS IS AN AREA HE'S WITHIN A LEADER IN SUBSEQUENTLY. JAN. >> THANKS ARLEN FOR THIS NICE INTRODUCTION. I JUST WANT TO SAY SERIOUSLY THANK YOU. I LEARNED A LOT FROM YOU. I CAN TELL ALREADY I WON'T ABLE THE ASK A QUESTION. WHY DNA? WHY DAMAGE DNA? YOU ALREADY MENTIONED THAT I THINK IT'S THE ONLY CAUSE OF AGING FOR WHICH YOU SHOW WHEN YOU ARE GOING TO LOOK TO THE (INAUDIBLE) NEUTRALIZING DNA DAMAGE MUTATIONS PREMATURE AGING, THAT'S TRUE FOR MICE. ANOTHER REASON IS SOME OF THE DAMAGE TO DNA, ALTERATION OF DNA ARE REVERSIBLE AND THAT'S ALSO LOOKED FOR MOST OTHER MOLECULES. FROM Z BECAME AWARE IMPORTANT DNA DAMAGE GOES BACK TO WORK IN 1970s WHO DEMONSTRATEDDED THAT DNA IS FAIRLY UNSTABLE MOLECULE UNDER PHYSIOLOGICAL CONDITIONS. PROBE THOUSANDS TO TENS OF THOUSANDS OF DNA LESIONS IN EACH CELL EACH DAY. THE FIRST QUESTION HERE WAS IF THAT IS THE CASE, CAN DNA DAMAGE THEMSELVES DNA DAMAGE PHYSICAL AL THE TEAR RATIONS, CROSS LINKS DEEP RUMINATIONS, (INDISCERNIBLE) SO THE IDEA WAS TO HAVE DNA DAMAGE CELL SOME POINT REACH LEVELS AND OLD AGE HIGH ENOUGH (INAUDIBLE) TRANSCRIPTION. ALSO THE QUESTION ALMOST CERTAINLY KNOW IT'S VERY UNLIKELY THAT THEY'RE NECESSARILY TRUE. FIRST OF ALL SEVERAL LABORATORIES INCLUDING MY OWN -- STEADY STATE DAMAGE LEVEL IS VERY LOW EVEN ALL AGES. THE REASON IT IS LOW AND ACTUALLY STAYS LOW, DETECTABLE IN FACT IT HAS TO DO WITH THE COMPLEX OF GENOME MATRIX SYSTEMS YOU CAN ROUGHLY DIAVIDIN TO TWO ARCS THERE'S ACTUAL REPAIR OF ACTION, DAMAGE REPLACEMENT OF DAMAGE PIECE OF DNA, DOUBLE STRAND BREAKINGS, SO ON BUT THE OTHER IS A RESPONSE TO DAMAGE. I THINK DAVID SINCLAIR ALREADY MENTIONED THAT. BECAUSE DNA DAMAGE IS RANDOM BUT SIGNALING TO SYSTEMS RARE DAMAGE TURNED THAT INTO PHENOTYPIC ENDS POINTS, APOPTOSIS AND CELLULAR SENESCENCE, YOU HAVE A NICE EXAMPLE THAT YOU DON'T NEED CONSISTENT EFFECTS OF DNA DAMAGE, SIMILAR AND ACROSS SPECIES. YOU ALREADY HEARD LOSS OF CELL ATROPHY AND SENESCENCE CELLS IS PROBABLY A MAJOR ENPOINT OF AGING, AS A RESULT OF DNA DAMAGE. THE OTHER END REPAIRTIVE DAMAGE ALSO LEADS TO CHROMATIN, DAVID MENTIONED THAT BRIEFLY FOR THE NEXT SPEAKER. I WILL BE SPECIFICALLY TALKING ABOUT ERRORS THAT ARE BEING MADE DURING THE ACTUAL REPAIR OF DAMAGE. DAMAGE REPAIR IS GOOD FOR SURVIVAL OF CELL BUT INEVITABLE ERRORS, THOSE ERRORS MUTATION, SO MUTATIONS NOT DNA DAMAGE. DNA DAMAGE IS REVERSIBLE. YOU CAN REPAIR IT. YOU LOSE THE INFORMATION DNA TEMPLATE. THE QUESTION IS HOW MANY MUTATIONS ARE THERE? AND CAN THEY EXERT AN EFFECT. RANDOM CHANGES IN THE EPIGENOME. OCCUR ALL THE TIME. I'M TALKING HERE ABOUT DNA METHYLATION OR HISTONE MODIFICATION THAT NEED TO BE RETORIED AFTER REPAIR AND DAMAGE. -- RESTORED AFTER REPAIRTIVE DAMAGE. YOU CAN PUT IT IN A MODEL THAT I TRIED THE DO HERE, THIS IS A YOUNG HEALTHY TISSUE, INDICATING INCREASED MUTATIONS POSSIBLE FUNCTIONAL DECLINE. AFTER AGING YOU SEE ACCUMULATION OF SENESCENCE CELLS. YOU SEE ATROPHY, CELLS THAT HAVE BEEN REMOVED BY APOPTOSIS AND SOMETIMES THERE'S A VERY UNFORTUNATE COMBINATION OF MUTATIONS THAT GIVE RISE TO TUMOR. SO PRINCIPLE THIS EXPLAINS A FAIR PART OF THE AGING PROCESS. MA I WILL DO NOW IS SPEND TIME POSSIBILITY MUTATIONS WILL ACCUMULATE IN INDIVIDUAL CELLS TO LEVELS THEY CAN EXERT PHENOTYPIC EFFECT. THIS IS A SITUATION WITH RESPECT TO MUTATIONS. ERRORS REPAIRING DNA DAMAGE OCCUR FROM THE BEGINNING. DNA MUTATIONS ACCUMULATE DURING THE DEVELOPMENT AND THE AGING PROCESS YOU CAN SEE HERE EACH COLOR IS DIFFERENCE MUTATION, IT'S A RANDOM PROCESS, BUT THERE'S A MUTATION OCCASIONALLY THAT SELECT -- (INAUDIBLE) THAT GROWS OUT TO A TUMOR. YOU CAN TAKE THAT TUMOR FROM TISSUE AND THE GENOME HAS BEEN DONE AND WHAT YOU WILL SEE IS PATIENTS THAT WERE OLDER, CONTAIN MANY MORE MUTATIONS THAN YOUNGER PATIENTS. THIS OF COURSE IS BECAUSE P MOST OF THE MUTATIONS THAT YOU FIND IN TUMOR ARE ACCUMULATING IN NORMAL CELLS BEFORE THE TUMOR (INAUDIBLE). OCCASIONALLY THAT'S ALSO (INAUDIBLE) PARTICULAR MUTATION THAT'S SHOWS FRACTION OF THE TISSUE CELL POPULATION. THERE ARE A NUMBER OF PAPERS BUT THE MUTATIONS YOU CANNOT DETECT, THAT'S MAJOR PROBLEM FORS TO ANSWER THIS QUESTION I MENTIONED BEFORE. IF YOU LOOK AT THE MUTATIONS, SOME PEOPLE ARGUE THAT CAN NEVER HAVE A FUNCTIONAL CONSEQUENCE BECAUSE THE FREQUENCY IS TOO LOW. FUNCTION IS NOT PROVIDED BY -- IT'S PROVIDED BY PATHWAYS AND NETWORKS. MANY OTHER PROTEASES ARE GOING TO AFFECT THOSE GENES NOT CODING REGULATORY SEQUENCES. SO TOTAL SEQUENCE CODE AFFECTED IS FAIRLY LARGE SO WHATEVER MUTATION LEFT IN THIS SYSTEM IS OF COURSE HAVEN'T HAD ADVERSE EFFECT (INAUDIBLE) SO HERE IS ANOTHER EXAMPLE HOW A RANDOM MUTATION CAN ACTUALLY HAVE -- SIMILAR INDIVIDUAL AND ACROSS SPECIES. OF COURSE WHAT YOU WILL LIKE TO DO LAST SLIDE, IS CAN YOU USE THIS ADVANCES, SEQUENCING TO LOOK AT MUTATIONS AND INITIALLY KNOW IT'S DIFFICULT. REMEMBER THIS WHOLE TISSUE MUTATIONS ARE RANDOM, THIS IS ONE OF THOSE MUTATIONS, ONE SINGLE CELL, YOU GRIND THE TUSH SHOE TOGETHER YOU START TO SEQUENCE IT, THIS IS TYPICAL WHEN YOU GET NEXT GENERATION SEQUENCING, AND HERE YOU SEE THE MUTATION. YOU KNOW THAT THIS IS CRITICAL MUTATION, THIS IS WHAT I TOLD YOU A MOMENT AGO. HOW YOU DISTINGUISH BETWEEN THIS MUTATION IN ONE READ, SEQUENCING ERRORS YOU CANNOT DISTINGUISH. SO IT'S NOT POSSIBLE TO GET INFORMATION ABOUT SOMATIC MUTATIONS BUT YOU CAN TAKE CELLS AND DO A MUTATION AND THE SAME MUTATION SHOWS IN 50% OF THE READS. BECAUSE THERE'S ONLY ONE ALLELE. A RANDOM MUTATION HIT THE SAME, SINGLE CELL SEQUENCING YOU CAN GET A HANDLE ON FINDING OUT HOW MANY MUTATIONS THERE ARE IN SOMATIC TISSUE. SEQUENCED SINGLE NUCLEI FROM DROSOPHILA (INAUDIBLE) THIS IS OF COURSE YOU COME THROUGH SINGLE CELL BUT WE -- A SINGLE NUCLEI FROM THE (INAUDIBLE) YOUNG FLIES AN OLD FLIES. THIS IS SIX NUCLEI. YOU CAN CLEARLY SEE THAT YOU SEE PARTICULAR DRAMATIC INCREASE IN MUTATION FREAK SCHISM THIS IS MUTATION AS NUCLEOTIDE VARIANT PER MEGA BASE, .25 PER MEGA BASE THIS NECESSARY NUCLEI AND IT'S HIGHER IN THE OLD ANIMALS, VARIATION IS HIGHER. THE MOUSE HEPATOCYTE, SO CONCLUSIONS YOU CAN SEE THREE HERE, WHEN YOU HAVE EVEN IF YOU HAVE LOW LEVELS THEY CAN DRIVE APOPTOSIS. YOU HAVE A CLEAR POSSIBLE (INAUDIBLE) DNA DAMAGE, RETINA MUTATION EVEN IF THEY ARE OCCURRING INDIVIDUALLY AT LOW FREQUENCY, CAN AFFECT TRANSCRIPTIONAL NETWORKS AND FINALLY WHEN YOU DO SINGLE CELL GENOMICS YOU CAN GET ACCESS NOW FINALLY TO THIS (INAUDIBLE) SOMATIC MUTATIONS. TWO LAST CONCLUSIONS ON HERE. IT IS VERY IMPORTANT DEVELOPING -- YOU'RE NOT ONLY FOCUSING ON CELL SURVIVAL, CELLS MAY SURVIVE BUT WILL CONTAIN MORE AN MORE MUTATIONS THAT'S NOT GOOD SO SUPPORTED TO HAVE A MEASURE FOR OCCURRENCE OF RANDOM SOMATIC MUTATIONS. SAME TRUE FOR STEM CELLS, FINALLY IN TERMS OF (INAUDIBLE) YOU CAN ASK YOURSELF THE IF MUTATIONS ARE INEVITABLE AND ALSO IRREVERSIBLE. AND POSSIBLE TO DO SOMETHING ABOUT AGING IN THE FIRST PLACE. WAY WAY OR THE OTHER REFER THAT. THANK YOU. [APPLAUSE] >> THANK YOU VERY MUCH, JAN. THE NEXT IS WILLIAM BONNER, LABORATORY OF MOLECULAR PHARMACOLOGY NCI. BILL WAS INVOLVED IN IDENTIFYING SEVERAL HISTONE VARIANTS SHOWING ASSOCIATION WITH DOUBLE STRAND BREAKS. BILL. >> I WASN'T PREPARED TO GO ON WITH YOUR CV. >> OKAY. THANK YOU VERY MUCH. IS THIS TOTALLY AUTOMATIC OR MY SLIDES GOING TO POP UP OR TO I NEED TO PRESS SOMETHING? I'M GOING TO TALK ABOUT CHROMATIN. I'M GOING TO CHANGE THE TITLE TO NUCLEAR CHANGES, BECAUSE WHAT I HAVE WRITTEN HERE. WE HAVE DNA AND HISTONES AND NONHISTONE PROTEINS AND RNA AND LIPIDS RESIDES IN THE NUCLEUS AND TAKES UP THE WHOLE NUCLEUS. VIRTUALLY. OF COURSE, ITS JOB IS CONTROL GENEK PRESENTATION BY TIMING AND LEVEL. OBSERVATIONS HAVE BEEN MADE IN THE LITERATURE, AGING LEADS TO TELOMERE SHORTENING. HETERO CHROMETYIZATION WITH SENESCENCE HETERO CROWMATIC FOCI WHICH CONTAIN DAMAGE TO THE X. LOSS OR CHANGES IN SOME OTHER MARKERS. FIRST WE HAVE TELOMERES AND WE KNOW THAT TELOMERES GET SHORTER WITH AGING BECAUSE OF POLYMERASE CAN'T MAKE IT TO THE DNA. THIS WAS THEORY OF AGING BUT THERE ARE MULTIPLE LINES OF EVIDENCE THAT DON'T SUPPORT IT. FOR ONE THING MICE HAVE LONG TELOMERES. EVEN WHEN OLD AND DEAD. SO WE CAN SEE NOW SENESCENCE ASSOCIATED HETERO CROWMATIC FOCI. HERE IS A PAPER THAT SAYS THEY ARE DISPENSABLE. AT LEAST IN CERTAIN CASES, HERE WE HAVE WHEN WE TAUGHT EPIGENETIC MODIFICATIONS, HERE IS A CATALOG OF MANY OF THESE EPIGENETIC MODIFICATIONS. AS YOU CAN SEE ON THE FOUR CORE HISTONES PRIMARILY THE AMINO TERMINAL ENDS. MAYBE I DONE. AS YOU CAN SEE HISTONES ARE PACKED INTO THE NUCLEOSOMES. SO OUVRIERIOUS ACETYLATIONS, E EAR FAMILIAR WITH THIS. THIS ONE HAS BEEN REMARKED IN THE LITERATURE ENRICHED DURING AGING. WE HAVE 5 METHYL C CHANGES. THIS PAPER CAME OUT LAST WEEK WHICH IS INTERESTING, TALKS ABOUT A DNA METHYLATION CLOCK. STEVE HORVATH FOUND 353 METHYLATION SITES WHEN YOU TRACK THEM OUT TRACK AGE VERY CLOSELY SO VIRTUALLY EVERY TISSUE OF THE BODY SO YOU CAN CALCULATE THE AGE OF A TISSUE BY LOOKING AT THESE METHYL GROUPS. THERE IS A RAPID INCREASE IN THESE PARAMETERS. MORE GRADUAL STRAIGHT CLOCK. SO INTERESTING OBSERVATIONS FROM THIS, THIS HAPPENS IN NEURONS AS WELL. WHAT ABOUT PROGERIA? PREMATURE AGING DISEASE, HUNTINGTONS. ACCORDING TO THIS, YOU DON'T GET -- THESE DO NOT FALL ON THESE LINES. NEIGHBOR THERE IS A QUESTION ABOUT PROGERIA MODEL OR METHYLATION CLOCK. BUT HERE IS A LITTLE AREA DISAGREEMENT. WHAT IS INTERESTING IS WHEN YOU MAKE INDUCED PLURIPOTENT STEM CELLS YOU RESET THE CLOCK. ENCOURAGING FOR THE PEOPLE IN THAT FIELD. SO NOW LET'S TALK ABOUT WHAT'S IN THE NUCLEUS BESIDES THE CHROMATIN. THERE IS A BUNCH OF LITTLE FACTORIES THAT HANG ON IN BETWEEN THE CHROMATIN. FOR EXAMPLE HERE, THIS IS THE NUCLEAR SPECKLES WHERE THEY HAVE THE SPLICE SITES. A FAULTY CRYPTIC SPLICE SITE TURNS LAMIN TO PROGERIA AND CAUSES HUNTINGTON'S DISEASE. HERE IS THE LAMIN THIS WIGLY LINE OUTSIDE THE MEMBRANE. ANOTHER THING THAT CAME UP EARLIER IN THE CONFERENCE, NUCLEAR STRESS BODIES OBVIOUSLY DESEPTEMBER NAME THAT WORK TO COUNTER ACT STRESS. YOU CAN HAVE MILD HEAT STRESS CORRECTING NASCENT PROTEINS, I THOUGHT IT WAS INTERESTING, THE TEMPERATURES HERE AND HEAT STROKE AND HUMANS HAPPENS ABOUT 105-DEGREES. CERTAINLY WE COULD BE UNDERGOING MILD HEAT STRESS. WHETHER THIS IS EQUIVALENT TO SAUTEEING NEMATODES, I DON'T KNOW. I THOUGHT WHEN THAT TALK WAS GIVEN, YOU LOOK AT PEOPLE WHO SPENT HOURS IN STEAM BATHS ALL THE TIME, DO THEY LIVE LONGER? I DON'T KNOW. TRANSCRIPTION FACTORS, WE HAVE THE SENESCENCE HETERO CROWMATIC FOCI. PML BODIES HAVE TO DO WITH TEAL MERE MAINTENANCE. SO WE HAVE A BUNCH OF OTHER THINGS IN THE NUCLEUS, SO WE HAVE A COMPLEX THING. NOW, HERE IS A LIST OF PROGERIA LIKE DISEASES AND THE DEFECTS. AS YOU CAN SEE HERE, THESE ARE ALL HELOCASES. HERE WE HAVE -- AND THE OTHER ONES, WITH ONE EXCEPTION, THESE ARE ALL NUCLEOTIDE EXCISION REPAIR PROTEINS. WE HAVE SCLERODERMA AND TRY WHATEVER IT IS. HERE WE HAVE TELOMERE ASSOCIATEDDED CONSIDER TEESIS CON GENERAL TA WHICH HAS TO DO WITH DEFECTS AT THE TELOMERE. THEN WE HAVE HUTCHINSON'S GILFORD PROGERIA, ONE WHICH THE LAMB MA IS ACTUALLY FAULTY AND THE OTHER WHICH THE PROTEASE WHICH CHOPS THE PARRELLA MINUTE INTO LAMIN IS MISSING. SO IT'S INTERESTING THESE ARE ALL SINGLE STRAND DAMAGE. DOUBLE STRAND DAMAGE REPAIR PATHWAYS OF COURSE ATM USES AND THINGS LIKE THAT ARE SERIOUS BUT THEY DON'T SEEM TO SHOW ANY AFFECTS OF ACCELERATED AGING. IT WAS ALLUDED TO BEFORE, WHEN WE TALK ABOUT SINGLE STRAND DAMAGE WE'RE TALKING 50, 100,000 EVENTS PER DAY PER CELL. SO THIS -- THESE PATHWAYS ARE ALWAYS ON. SOMEWHERE LOOKING AROUND DOING SOMETHING. ON THE OTHER HAND DOUBLE STRAND BREAKS, MAYBE TEN PER DAY. THAT MAYBE MORE IN CYCLING CELLS, THE DEMAND MAYBE LESS SO IT MAY HAVE ENOUGH TIME TO TAKE CARE OF STUFF UNDER HIGHLY SUBOPTIMAL SITUATIONS. TO FINISH, I THOUGHT IT WOULD BE INTERESTING TO PRESENT THIS AS OTHER OPTIONS FOR EMPLOYMENT AS YOU CAN SEE. THERE'S ALREADY SOMETHING AROUND HERE TO TAKE CARE OF PROJEER I DON'T KNOW PROBLEMS BY THIS COSMETIC COMPANY AND YOU CAN GET A CERTIFICATE OF TELOMERE SCIENCE IF YOU WOULD LIKE. THANK YOU VERY MUCH. >> NOW IT'S ON. OUR NEXT SPEAKER IS ROWDILY VINE, I HAVE KNOWN HIM A LONG TIME. HE IS CHIEF OF THE LABORATORY BIOCHEMISTRY, NATIONAL HEART LUNG AND BLOOD INSTITUTE. AS I SAID, I HAVE KNOWN RON SINCE HE WAS A REAL TALL PERSON AND HE HAD NO BEARD. NOW HE SHRUNK AND HE HAS A BEARD. SO BUT SERIOUSLY, RON WAS PART OF EARLE STAIDMAN'S GROUP INVOLVED IN DISCOVERY OF PROTEIN OXIDATION AND HE'S MADE MAJOR CONTRIBUTIONS IN THIS FIELD. SO WE WILL TALK ABOUT THE ROLE OF PROTEIN OXIDATION AND AGING. >> GOOD MORNING, HAPPY HALLOWEEN. THERE IS A LOT OF OFFICIAL CELEBRATION OF HALLOWEEN HERE AT NIH, NOT JUST THE CANDY PUT OUT AT THE REGISTRATION DESK, AT LUNCH, CHECK IT OUT, GO OUT THE CAFETERIA DOORS, WALK TO SIDEWALK YOU WILL SEE SCARY, THERE IS A COYOTE MOVING THERE. SOME OF YOU MAY HAVE SPOTTED THIS. IT APPARENT HI WASN'T PUT OUT TO SCARE US, IT IS PUT OUT BECAUSE WE HAVE AN INVASION OF GEESE WHICH MAKE A GOD AWFUL MESS AND THEY LOVE THIS AREA. SO THE GROUNDS PEOPLE PUT THESE OUT AND THEY LOOK REALISTIC AND THEY MOVE AND ACCORDING TO THEIR POORLY CONTROLLED STUDY ACTUALLY WORKS. THIS IS SUPPOSEDDED TO BE A HEADLIGHT TALK SO THERE'S A SIMPLE QUESTION. CAN OXIDIZED PROTEINS BE PREPARED. I WILL RAISE A COUPLE OF SIMPLE POINTS. SULFUR CONTAINING AMINO ACIDS, SISTINE AND ME THIONEINE WHEN OXIDIZED CAN BE PREPARED BUT WHAT ABOUT BEYOND THAT? THE POINT I WOULD LIKE TO MAKE FIRST, WE DON'T ACTUALLY KNOW WHAT'S REPARABLE BECAUSE MOST OF US HAVEN'T ACTUALLY HAD THE STRENGTH TO TACKLE IT. I WANT TO SUGGEST WE REALLY MIGHT BENEFIT BY DETERMINING WHICH COVALENT CAN BE AVAILABLE, CAN BE PREPARED. THERE SAKE CAVEAT THAT I WOULD LIKE TO OFFER YOU DESPITE THE FACT MY UNDERGRADUATE DEGREE IS CHEMISTRY. EVOLUTION ISN'T RUN BY ORGANIC CHEMISTS. A LOT OF OUR THINKING ABOUT WHAT'S REPARABLE HAS BEEN DOMINATED BY THAT. MY COLLEAGUES ARE MORE CERTAIN ABOUT WHAT'S POSSIBLE AND WHAT ISN'T POSSIBLE MOST BIOLOGICAL RESEARCHER. HERE IS AN EXAMPLE. SUPPOSE YOU WANT TO OXIDIZE ACETATE TO CARBON DIOXIDE? TALK TO A CHEMIST. IT'S HARD, YOU HAVE TO USE HIGH TEMPERATURE AND ELEVATED PRESSURE TO DO IT. THE CELL, NO PROBLEM. ACTIVATE THE ACETATE WITH CA, POP THE TCA CYCLE YOU'RE GOOD THE GO. IT WORKS AT ROOM TEMPERATURE. IT IS A CONDITION TRAYS IN THE WAY OF THINKING. HERE IS WHAT I THINK IS A RELEVANT EXAMPLE. PREPARING OVEROXIDIZED SISTINE IN PROTEINS THAT HAVE AN ESSENTIAL SISTINE AT THE ACTIVE SITE. IN SUCH ENZYMES CONVERSION OF THE -- IS CRITICAL FOR THE CATALYTIC CYCLE, REDUCTASE, HYDROGENASE AND OTHERS. THIS IS A FRAGILE MODIFICATION IN THAT THE ACID IS SUSCEPTIBLE TO FURTHER OXIDATION. IT'S PRETTY EASY TO GO ON TO SULFINIC ACID TO OX GENERALS ADDED. FROM THE CHEMIST POINT OF VIEW, I LEARNED THIS, MANY OF YOU DID, THAT'S SIMPLY IRREVERSIBLE. AND WE HAVE KNOWN THAT FOR YEARS, SIMPLE. BUT IT'S WRONG. (INAUDIBLE) AND COLLEAGUES DURING COURSE OF STUDYING ANTIOXIDANT DEFENSES BY ROW DOXINS SHOWED EXTRACTS FROM HELO CELLS AND LATER A NUMBER OF MAMMALIAN CELL LINES COULD REVERSE IT. THEY PUBLISHED THIS OBSERVATION IN THIS FIRST PAPER. THEY DIDN'T KNOW HOW IT WAS MEDIATED BUT IT WAS QUITE AN IMPRESSIVE DISCOVERY IT COULD HAPPEN, IT WASN'T SUPPOSED TO BE POSSIBLE. HIS COLLEAGUES DIDN'T SUCCEED IN ICE ISOLATING THE ACTIVITY, IDENTIFYING IT AND TELLING US WHAT IT WAS. BECAUSE SUBU OUTSTANDING BIOCHEMIST HAD A HANDICAP. Ph.D. WAS IN CHEMISTRY. -- CHEMISTRY. HE MADE ASSUMPTIONS. BUT MICHELLE AND COLLEAGUES DID FIND IT, THEY WERE THINKING OUTSIDE THE BOX AND THEY NAMED IT SULFUR ROW DOXIN. IT WOULD HAVE BEEN RIDICULOUS THINKING ABOUT THIS LOGICALLY. TO CONSIDER ATP MIGHT HAVE BEEN REQUIRED FOR THIS ACTIVITY. HIM AND FOLK NEVER THOUGHT ABOUT THAT. ALL SORTS OF REDUCING AGENTS TO GET THIS TO GO. I'M A GENETICIST, MOLECULAR BIOLOGIST, THROW EVERYTHING IN. ACTTP WAS THE KEY. EVOLUTION ISN'T RUN BY ORGANIC CHEMISTS, MAYBE MORE OPEN MIND. HOW FAR CAN REPAIR GO? A LOT OF MODIFICATIONS BUT THE TASK MASTER GOT HIS WHIP OUTND TOLD ME TO CUT AT LEAST ONE SLIDE IN THE LIST OF MODIFICATIONS. I MENTION NITRATION PARTICULARLY TYROSINE SINCE IT'S RATHER THAN IMPORTANT MODIFICATION MEDIATED INFLAMMATORY CHANGES INITIALLY THROW MODIFICATION NITRIC OXIDE. BUT LET ME FOCUS ON ONE THING THAT IRELAND DID INTRODUCE, THAT'S ACCUMULATION OF CARBONLATED PROTEINS. WHICH COME FROM A VARIETY OF SOURCES. LET ME JUST MENTION THIS ONE A COMMON OXIDATIVE MODIFICATION PROTEINS WITH METAL BINDING SITES. STRUCTURAL PROTEINS HAND ENZYMES IS THE OXIDATION AND IT'S BLOWN TO PIECES BY OXIDATION THIS HOCK CURS AT METAL BINDING SITE AND AFFECT IT IS ARGININES. AND YOU END UP WITH THE CARB NEAL GROUP, THE ANALOG OF GLUTAMIC ACID. I HOPE I HAVE NOT DONE THIS, I MIGHT HAVE, BUT IT IS WIDELY STATED THIS IS AN IRREVERSIBLE ACTION, THE ONLY WAY THE DEAL WITH THIS IS DEGRADE THE PROTEIN. IS THAT TRUE? LET ME SHOW YOU ANY OH HELL SLIDE BECAUSE WHEN I SHOW YOU THE NEXT PAR YOU'LL BE THINKING OH, HELL IS HE GOING TO GO THROUGH ALL THAT. I'M NOT, I WANT TO ZOOM IN ON THE ARGININE METABOLISM AND SHOW YOU SOMETHING MICROORGANISMS AND OTHER ORGANISMS TO WITH NO PROBLEM. THEY MAKE ARGININE. HOW? IT'S A SIMPLE TWO STEP PROCESS STARTING WITH GAMMA GLUTE MILL SEMIALDEHYDE. CITRULLINE, YOU POP TO THE UREA CYCLE AND YOU END UP WITH ARGININE. ALL YOU HAVE TO DO IS FOLLOW THIS. WORKS ON SMALL MOLECULES NO REASON IT COULDN'T WORK ON PROTEIN IF ACCESS WERE AVAILABLE AND ENZYMES EXISTED. NOBODY LOOKED. SO I HAVE TWO OTHER SLIDES THAT MADE ME TAKE OUT TO SAY THE SAME HAPPENS WITH LYSINE OXIDATION, TWO STEPS YEAST ARE GOOD AT IT. OXIDIZED RESIDUE YOUR BACK TO LYSINE. MY CONCLUSION IS SIMPLE. THERE'S AN OPPORTUNITY HERE TO UNDERSTAND PROTEIN REPAIR. THAT MIGHT BE IMPORTANT OBVIOUSLY IN CERTAIN ASPECTS AND MECHANISM OF AGING. OR IN OUR ABILITY TO INTERVENE IN THE RATES OF AGING. I CLOSE WITH A REMINDER FROM ALFRED EINSTEIN. THANK YOU. [APPLAUSE] >> OUR NEXT SPEAKER IS DEAN JONES FROM EMORY, PROFESSOR OF MEDICINE AND DIRECTOR OF THE CLINICAL BIOMARKERS LABORATORY. AND DEAN IS GOING TO BE TALKING WITH RESPECT -- HE'S BEEN INVOLVED IN LOOKING AT REDOX SYSTEM IN ORGANISMS AND IN THE LAST FIVE, TEN YEARS BECOME MORE IMPORTANT THIS MIGHT PLAY AN IMPORTANT ROLE PARTICULARLY IN OXIDATIVE STRESS MEDIATED PROBLEMS. >> THANK YOU. I JUST HAVE TO FOLLOW-UP WITH THE COMPLIMENTS TO PHILIPPE FOR THE -- DIRECTING US SO CLEARLY, I HAVE TO ADD AT ONE POINT, MANY YEARS AGO DECADES AGO CONVERSATION WITH MY FATHER, WHAT IS THE MEANING OF TAKING SCIENTIFIC DATA AWAY FROM A SCIENTIST? WHICH PHILIPPE DID FOR US. WHAT MY FATHER SAID, IF YOU TAKE THE SCIENTIFIC DATA AWAY YOU HAVE A PREACHER. SO I'M GOING TO BE A PREACH EVERIER. -- PREACHER. SO THE QUESTION I HAVE ASKED TO ADDRESS IS WHAT WE DO ABOUT REVERSIBLE REDOX CHANGE THAT OCCURS WITH AGING. I HAVE BROKEN THIS INTO THREE COMPONENTS, ONE TO ADDRESS BRIEFLY, WHY THIS OCCURS AND THEN DOES IT MATTER AND THEN TALK ABOUT WHAT WE MIGHT BE ABLE TO DO ABOUT IT. BUT THOSE OF YOU NOT FAMILIAR WITH THE CONCEPT, A BIT OVER A DECADE AGO WE FOUND THAT THERE IS A SLOW CHANGE IN OXIDATION REDOX POTENTIAL ASSOCIATED WITH AGE SPECIFICALLY BEYOND 50 YEARS OLD WE BECOME 1 MILIMOLE MORE OX DICED FOR YEAR. -- OXIDIZED PER YEAR. SO THE QUESTION, WHY DOES THIS OCCUR? , DOES IT MATTER? WE EXPANNED IF RESEARCH, WE HAD A PAPER COME OUT ON THE COMMON MARMOSET MODEL OF AGING, AND FOUND MARMOSET AS IN MICE AND ALSO PREVIOUSLY FOUNDED RHESUS THAT THEY SIMILARLY CHANGE IN REDOX POTENTIAL WITH AGE THE QUESTION WHY DOES THIS OCCUR I THINK TO ADDRESS THAT, WHAT I WOULD LIKE TO GO TO IS THE CONCEPT OF THE EXPOSESOME AS CHRISTOPHER DEFINED IT SEVERAL YEARS AGO. THIS CONCEPT WE HAVE A CUMULATIVE LIFE LONG EXPOSURE TO MANY DIFFERENT DIETARY ENVIRONMENTAL INFECTIOUS EXPOSURES AS WELL AS VARIOUS CONTRIBUTIONS OF THE MICROBIOME THERAPEUTIC DRUGS LIFESTYLE BEHAVIORS, SO FORTH. SO ALL OF THESE, THE CUMULATIVE NATURE THEY IMPACT THE ORGANISM. THEY IMPACT THE ORGANISM ALONG THIS AXIS BETWEEN THE PROTEOME AND METABALOME. SOME THINGS DRIBBLE THROUGH AND DO CAUSE DIRECT DAMAGE TO THE DNA BUT I THINK OUR MAJOR INTERFACE IS RIGHT HERE. WHAT WE FOUND OVER THE PAST TEN YEARS OR SO, IN STUDYING THIS, THE REDOX PROTEOME COMPONENTS WITHIN THE PROTEIN THAT UNDERGO CHANGES IN RESPONSE TO OXIDATION. FOLLOWING UP ON WHAT ROBLE VINE SAID IN TERMS OF BELIEVING THE CHEMIST, YOU HAVE TO BE CAREFUL BELIEVING THE BIOCHEMISTS. BECAUSE ATTITUDE IN BIOCHEMISTRY IS EFFECTIVELY THE (INAUDIBLE) SHOULD BE OXIDIZED. IF THEY'RE NOT OR SHOULD BE REDUCED AND IF THEY OTHER NOT ALL REDUCED, WE HAVE AN ARTIFACT FROM OUR BIOCHEMICAL ANALYSIS. WHAT WE FOUND NOW IS USING MASS SPECTROMETRY BASED REDOX PROTEOMIC METHOD IS IN FACT THAT IS REALLY COMPLETELY WRONG BUT IN FACT THE REDOX PROTEOME IS -- EXISTS AS A SPECTRUM OF OXIDATION STATES OF DIFFERENT PROTEINS AND THAT THIS -- THE CONCEPT IS THAT THIS REDOX PROTEOME IS REALLY OUR PRIMARY INTERFACE IN INTERACTING WITH THE DIET AND ENVIRONMENT AND INDEED AT A MOLECULAR LEVEL, IT'S A FOLLOW-UP TO WHAT LINDA FRIED TALKED ABOUT YESTERDAY, THIS IS A PRIMARY ADAPTIVE STRUCTURE. THIS IS A PRIMARY WAY, IT HAS EVOLVED SINCE THE EVOLUTION OF OXYGEN ATMOSPHERE. IT'S OBVIOUSLY PRE-DATED THAT, BUT IN TERMS OF THE CHARACTER IN US AND MAMMALIAN SYSTEMS, IT IS THIS ADAPTIVE STRUCTURE THE REDOX PROTEOME PROVIDE THAT IS REALLY THE KEY IN TERMS OF THE ENVIRONMENTAL ASPECTS, THE DIET ENVIRONMENT AND ALL THESE OTHER INTERACTIONS THAT WE HAVE, STRESSORS THAT WE HAVE. THERE ARE 214,000 SUSTAINS ENCODED IN THE MAMMALIAN GENOME. NOT ALL CHARACTERIZED FUNCTIONALLY. BUT AS WE DEVELOP METHODS ABOUT 4,000 NOW HAVE BEEN MEASURED IN A REDOX SENSE, ALMOST ALL WILL BE PARTIALLY OXIDIZED AS THEY EXIST IN BIOLOGIC SYSTEMS. WHAT'S VERY IMPORTANT IN LINKS -- AND LINKS BACK TO THE SESSION THIS MORNING, THE REDOX PROTEOME IS DYNAMICALLY CONTROLLED BY THE REDOX METABALOME. SO THE REDOX PROTEOME AS ARLEN INDICATED IMPACTS ALL ASPECTS OF FUNCTION OF CELLS. WHAT'S VERY IMPORTANT IN TERMS OF CLOSING THE LOOP, THE GENE DIRECTED ACTIVITIES ARE CONTROLLING THE METABALOME AND THE METABALOME ALSO IS ENTERACTING WITH THE DIET AND ENVIRONMENT. THIS INCLUDES NOTABLY THE NAD, NADH SYSTEM, THE NADP, NADPH SYSTEM, WE TALKED ABOUT IT A BIT. IT ALSO INCLUDES GLUE TA THIO, SUSTAIN, SISTINE, LIPOIC ACID WE HAVEN'T HEARD ABOUT HERE. BUT COENZYME A. THERE ARE MANY SYSTEMS THAT ARE ACTUALLY INVOLVED IN THE REDOX METABALOME. THESE ARE DIRECTLY ENTERACTING WITH THE REDOX PROTEOME. OXIDATION OF REDOX POTENTIAL REFLECTS LOSS OF ADAPTABILITY TO THIS EXTERNAL ENVIRONMENT. DOES IT MATTER? WE DISCUSSED PROBLEMS OF CORRELATION ANALYSIS. I'M IN THE CLINICAL DEPARTMENT -- I'M A BIOCHEMIST BUT I'M IN THE CLINICAL THE PRESIDENT SO INVOLVED IN MANY STUDIES THE REDOX SYSTEM. WHAT WE FIND IS THAT IN OBESE PEOPLE, MORE OXIDIZED. DIABETICS ARE MORE OXIDIZED. PEOPLE WHO HAVE AGE RELATED -- ATRIAL FIBRILLATION, OTHER ASPECTS OF CARDIOVASCULAR DISEASE, IDIOPATHIC PULMONARY FIBROSIS. THERE'S A LONG LIST OF CONDITION ASSOCIATED WITH OXIDATION OF THESE THIOL SYSTEMS. THAT DOES NOT ESTABLISH CAUSE EFFECTMENT WHAT'S THE EVIDENCE IN TERMS OF CAUSAL MECHANISM? ONE THING THAT'S BEEN DONE, WE AND OTHERS HAVE LOOKED AT CHANGING THE REDOX POTENTIAL IN SYSTEM AND ASKED WHAT DOES THAT DO? TWO VERY IMPORTANT THINGS THAT IT DOES IS IT STIMULATES PROFLAMTORY SIGNALING AND STIMULATES PROFIBROTIC SIGNAL SIGNALING. I DON'T HAVE TIME TO GO INTO ANY DETAILS ON THAT, KEY POINT IS WHAT WE CAN DO ABOUT THIS. ONE ASPECT THAT FOLLOWS FROM THE LAST SESSION THIS NEEDS TO BE PAID ATTENTION TO. AMERICAN INTAKE. THE MEDIAN AMERICAN INTAKE OF ME THIONEINE IS 2.5 TIMES RECOMMENDED DIETARY ALLOWANCE. WHAT WE -- WHAT I -- MY COLLEAGUES AND I HAVE SHOWN, IF ONE TAKES IN EXCESSIVE ME THIONEINE AND SUSTAIN, WHAT HAPPENS IS WE INCREASE OXIDIZED FORMS, IT'S VERY POORLY CLEAR. IT'S UNCLEAR AS FAR AS THE -- WHETHER OR NOT WE SHOULD DECREASE TOTAL PROTEIN INTAKE. IT IS RELATIVELY STRAIGHT FORWARD WE SHOULD AT LEAST CONSIDER THE POSSIBILITY THAT WE HAVE A PRETHETY WIDE MARGIN OF INTAKE ON EXCESS INTAKE ON ME THIONEINE WORTH LOOKING INTO THE POSSIBILITY THAT THIS IS BENEFICIAL. A SECOND IS THE DIETARY THINK WE HAD AN ANCILLARY STUDY WHICH WE EXAMINE THE EFFECT ZINC SUPPLEMENTATION, THE OXIDIZED FORM. WHAT WE FOUND INDEED, THIS WAS THE FIRST EVIDENCE THAT THE REDOX ACTUALLY CHANGES LONGITUDINALLY WITHIN INDIVIDUALS, AND THAT SHOWED THE ZINC DECREASED OR PREVENTED THE INCREASE OF -- WITH AGE AND WHAT COLLEAGUES HAVE MORE RECENTLY SHOWN IS THAT IS -- THAT ZINC MECHANISM OCCURS THROUGH ACTIVATION OF NERF 2. I DON'T HAVE TIME THE TALK ABOUT SELENIUM BUT IF WE HAD METHODS TO MEASURE HOW MUCH SELENIUM WE SHOULD BE EATING, THIS IS A GOOD TARGET. BECAUSE WE ARE OUT OF TIME I WANT TO JUMP BACK AT THE PREACHER FOR A SECOND. AND THIS PREACHER IS RIGHT HERE. I THINK IF WE HAVE MISSED THE BOAT AS FAR AS LONGEVITY RESEARCH, IT'S HERE. THAT IS THAT WE NEED TO PAY ATTENTION TO THIS CONCEPT OF THE EXPOSESOME AND MOVE FORWARD JUST AS THE GENETICS AN GENOMICS COMMUNITY SEQUENCED THE HUMAN GENOME, BENEED TO DEVELOP STRATEGIES TO SEQUENCE EXPOSERS, SEQUENCEs EXPOSEOME. THIS WILL GIVE US A TRY MEN ADVANTAGE IN UNDERSTANDING AGE RELATED DISEASE AND PROLONGING HEALTH. THANK YOU. [APPLAUSE] >> THANK YOU VERY MUCH, DEAN. THE LAST SPEAKER IN OUR SESSION I DIDN'T KNOW HIM WHEN HE WAS LITTLE, SO THIS IS MY FIRST TIME TO MEET DR. YAFFE, HE'S CO-CHAIR OF BIOLOGY AND BIOENGINEERING AT MIT. HE'S GOING TO BE BRINGING A SYSTEMS APPROACH TO LOOKING AT THIS. I THINK PARTICULARLY ON THE LAST SPEAKER YOU GET A FEELING AS WELL AS FROM JAN'S SPEECH WE HAVE LOTS OF DIFFERENT TYPES OF DAMAGE AND HOW WE LOOK AT THE END PRODUCT OF ALL THESE TOGETHER. MICHAEL. >> LET ME START BY THANKING ARLEN AND RON FOR THE OPPORTUNITY TO SPEAK AND WHAT I WAS CHARGED WITH TELLING ABOUT WAS HOW IN THE SURROUNDING MICROENVIRONMENT AFFECTS THE RESPONSE TO MACRO MOLECULAR DAMAGE FROM A SYSTEMS BIOLOGY POINT OF VIEW. I WAS INTRIGUED BY ARLEN'S ANALOGY OF TRANSPORTATION AND THE CAR, SO I'M SHOWING HERE DIAGRAM THAT LOOKS A BIT LIKE ROD'S ARGININE METABOLISM DIAGRAM, A SYSTEMS LEVEL DIAGRAM BUT WHAT I WANT TO POINT OUT HERE IS THAT THIS SYSTEMS BIOLOGY BACKGROUND IS THE REAL WAY LINES IN TOKYO. KEEP WITH THAT TRANSPORTATION MODEL SO YOU FIND COMPLEXITY IN EVERYTHING, NOT JUST A BIOLOGY FOR IT. FIRST POINT I WANT TO STRESS IS THERE'S CROSS TALK BETWEEN INNATE AND ADAPTIVE IMMUNE CELLS AND THE MICROENVIRONMENT IN WHICH DAMAGE CELLS FINE THEMSELVES AND MACRO MOLECULAR DAMAGE. SO IN THE CASE OF INFLAMMATION, NEUTRAPHILS AN T LYMPHOCYTES CONTRIBUTE TO SOME OF THE RADICALS THAT CAUSE DNA OR PROTEIN DAMAGE, THESE CELLS ALSO PLAYS A ROLE IN WHAT HAPPENS WHEN THEY'RE DAMAGED. THE BEST EXAMPLE WHICH WE UNDERSTAND THIS IS IN THE SETTING OF CANCER. WHAT I HAVE SHOWN YOU HERE IS AN SECTION OF EPITHELIUM UNDERGONE DAMAGE. AND WHEN CELLS ARE DAMAGED THEY HAVE A LIMITED NUMBER OF REPERTOIRE OF RESPONSES CELLS CAN REPAIR DAMAGE FULLY OR ONLY PARTIALLY AND MAINTAIN THAT EPITHELIUM. ALTERNATIVELY, YOU GET SOME TYPE OF REPAIR PROCESS, AND A COUPLE OF THINGS CAN HAPPEN. CELLS AND KNEW THAT PHILS AN MONOCYTES AND MACROPHAGES MOVE TO THAT AREA. I WANT TO STRESS WHAT US ALLUDED TO, THERE'S A IMPORTANT ROLE FOR PLATELETS AND FIBRIN AND THROMBIN, THESE CAN DO TWO THINGS, BOTH RESULT IN LOSS OF FUNCTION. ONE WHICH RESULTS IN LOSS OF FUNCTION. YOU CAN LEADS TO SUBSTITUTION BY A HEALING RESPONSE. YOU CAN END UP WITH FIBROTIC DEGENERATION AND THAT LIMBS FUNCTION OR ALTERNATIVELY YOU CAN STIMULATE THE DIVISION OF SOME TYPE OF PRECURSOR CELL, A STEM CELL, IF YOU WILL, TO REPOPULATE. THE PROBLEM IN CHRONIC INFLAMMATION THE CREOLIANS GOES CONTINUE DAMAGE AND REPAIR, IT CAN TURN INTO A MALIGNANT CLONE, IT SIGNALS EMERGEED FROM THE MICROENVIRONMENT THAT STIMULATE AND HAYNE TAIN UNTIL IT TURN INTO A TUMOR. I WANT TO STRESS A VERY IMPORTANT ROLE FOR THE ENDOTHELIUM. MORE ABOUT THAT IN A MOMENT. SO IT'S CROSS TALK BETWEEN WHAT'S HAPPENING TO MACRO MOLECULARLY DAMAGED CELLS, INTRINSIC PATHWAY, MACROPHAGES AND NEUTRAPHILS THAT DETERMINE WHAT RESPONSE WE SEE. EXTRINSIC PATHWAYS AN CELLS AND CYTOKINES AND GROWTH FACTORS THAT MODULATE WHAT IT IS THAT THE DAMAGE CELL ITSELF DOES. WE HAVE UNDERSTANDING OF CYTOKINES AN GROWTH FACTORS THAT PLAY ROLE IN THIS. YOU HER ABOUT THIS OVER THE LAST TWO DAY, IL-6 AND IL-1, IR-8 AND TNF L FA. IGF 1 INCLUDED EARLIER, ALL PLAYED AN IMPORTANT ROLE. ALL OF THIS STARTS OF COURSE WE THINK IN PART BY ROS GENERATION RESPONSE TO STRESS, TUMOR CELLS THEMSELVES WHEN THEY UNDERGO THE CHANGE IN METABOLISM WE'RE AWARE OF, GENERATE ROS AND THIS LEADS TO A CYCLE WHERE TUMOR CELLS MODIFY MODULATE THE MICROENVIRONMENT AT THE SAME TIME THE MICROENVIRONMENT MODULATES THE TUMOR CELLS BUT THERE'S ONE CELL WE HAVEN'T HEARD MUCH ABOUT THAT PLAYS AN IMPORTANT ROLE AND THAT'S THE ENDOTHELIAL CELL BECAUSE WE KNOW THAT SOME OF THE THINGS THAT TUMOR CELLS SECRETE STIMULATE ENDOTHELIAL CELLS AND THEY SECRETE CYTOKINES AN CHEMOKINES THAT CONTROL WHAT HAPPENS TO THE DAMAGE CELLS WHETHER WHETHER THEY PROLIFERATE OR INDUCE FURTHER ANGIOGENESIS. SO REMINDED IN THE CASE OF LYMPHOMASTOR EXAMPLE, WHEN YOU TREAT WITH CHEMOTHERAPY A SUBSET OF CELLS BECOME RESISTANT AND LEAD TO RECURRENCE OF DISEASE AND THEY EXIST IN THE THYMUS. THE REASON THEY'RE RESISTANTS ISN'T BECAUSE WE HAVEN'T DAMAGED DNA BUT IL-6 IN THE THYMUS MAKES A NICHE WHERE THE MACRO MOLECULARLY DAMAGED LYMPHOCYTES CAN BE MAINTAINED. THE CELL TYPE IN THE THYMUS IS NOT AN IMMUNE CELL, ENTHELIAL CELLS IN THE THIGH MOUSE SO SIGNALING MECHANISMS INVOLVED IN WHAT HAPPENS TO MANY MACRO MOLECULAR DAMAGED CELLS WE NEED TO KEEP THEM IN MIND. WE HAVE THE THINK ABOUT SIGNALING PATHWAYS INVOLVED IN THESE GROWTH FACTOR SIGHKINE RESPONSES. FROM A SYSTEMS POINT OF VIEW, THERE'S THE TALK ON SYSTEMS BIOLOGY. WHAT YOU GET OUT DEPENDS ON THE LEVEL OF GRANULARITY YOU WANT TO LOOK. WE HEARD EARLIER FROM LINDA THE TYPES OF THINGS WE LOOK AT EPIDEMIOLOGICALLY IN PATIENTS ARE THINGS LIKE NUTRITION AND INFECTIOUS STATUS AND WHAT HAPPENS TO THE PATIENT, WHOLE BODY DYNAMICS BUT IN FACT WHERE SYSTEMS BIOLOGY IS BEST SUITED IS DOWN HERE AT THE LEVEL OF INDIVIDUAL CELLS WHERE WE LOOK AT CERTAIN QUEUES THAT CELLS SEE, CERTAIN RESPONSES AND ASSAY THING WE KNOW HOW TO ASSAY IN ORDER TO BUILD A MODEL WE TRY TO CORRELATE THE QUEUE THAT THE CELL SAW TO THE RESPONSE THAT THE CELL PROVIDED BASED ON THE SIGNALS THAT IT SAW. QUICK EXAMPLE TWO WAYS YOU CAN DO THIS. WHAT I SHOW HERE IS CROSS TALK. BETWEEN FOR INSTANCE MODIFIERS OF METABOLISM LIKE INSULIN AND CYTOKINES THAT RESULT IN CELL DEATH. P IF WE WANT TO UNDERSTAND HOW IT IS SIGNALING PATHWAYS TELL WHETHER CELL IN RESPONSE TO INSULIN AND TNF SURVIVES OR REPLICATES. WE HAVE TO START TO LOOK AT PATHWAYS THAT CONTROL EACH OF THESE RESPONSES AND HOW THESE PATHWAYS TALK TO ONE ANOTHER SO WE CAN BREAK IT DOWN INTO PATHWAYS THAT WE THINK INSULIN DRIVE SURVIVAL PATHWAYS LIKE ERK AND MTOR AND AKT, STRESS PATHWAYS TNF ALPHA DRIVE, JUNK AND P-38 AND OUTCOMES LIKE DNA DAMAGE RESPONSES AND CASPASE ACTIVATION. WE HAVE TO COME UP WITH ASSAYS THAT ALLOW US TO MEASURE WHAT'S HAPPENING DYNAMICALLY IN THIS ENTIRE NETWORK. AND THERE ARE A SUBSET OF PROTEINS WHICH YOU SEE MARKED WITH P WHICH WE HAD MEDIUM HIGH THROUGH PUT ASSAYS THAT ALLOW US TO DO THIS. USING REVERSE PROTEIN ARRAYS, IN VITRO KINASE ASSAYS. FROM THIS WE CAN GATHER FAIR AMOUNT OF DATA. THIS IS A QUICK SHOW YOU ONE EXAMPLE HOW THE DNA DAMAGE PATHWAY CROSS TALKS WITH GROWTH FACTOR RECEPTOR PATHWAY, EVERYTHING IS SHOWN IN WHITE. AND THIS IS THE TYPE OF DATA THAT EMERGES FROM THE SYSTEMS BIOLOGY STUDIES. I WANT TO STRESS IF YOU'RE GOING TO DO SYSTEMS BIOLOGY YOU CAN'T DO IT WITH ONE CELL TYPE AND ONE STIMULUS AND ONE RESPONSE. YOU HAVE TO TAKE THE SAME RESPONSE, SAME STIMULUS IN A BUNCH OF CELL TYPES OR EVEN BETTER, ONE CELL TYPE AND LOT OF STIMULI BEST EVER AND MULTIPLE STIMULI AND MEASURE DENSELY, YOU HAVE TO MEASURE A BUNCH OF THING, SAMPLE AS MANY PATHWAYS AS YOU CAN AND YOU WANT TO SAMPLE THEM TWO ASPECTS OF IT. FIRST DYNAMIC. YOU WANT TO MEASURE THE RESPONSES IN SOME SORT OF WAY, THIS DYNAMIC, YOU CAN'T PICK ONE MEASUREMENT SO MEASURE IN THIS CASE WE DID 12 MEASUREMENTS THEN THE MEASUREMENTS ARE RIGOROUSLY QUANTITATIVE BECAUSE YOU WANT TO USE MATH TO EXPLAIN RESULTS. THIS MEANS YOU HAVE TO DO SOME KIND OF A TECHNIQUE WHERE WHAT COMES OUT ARE NUMBERS THAT ARE MEANINGFUL. WESTERN BLOTS. BUT YOU HAVE TO GET MEASUREMENTS TO TELL YOU WHAT'S HAPPENING. THEN YOU ALSO HAVE TO MEASURE IN A QUANTITATIVE WAY THE RESPONSES, YOU HAVE TO MEASURE FOR EXAMPLE, WHAT HAPPENS TO THE CELLS IN RESPONSE TO MACRO MOLECULAR DAMAGE, DO THEY DIE BY APOPTOSIS? PROLIFERATE? STUCK IN THE CELL CYCLE? DO THEY UNDERGO AUTOPHAGY? AND MULTIPLE STEM CELL TYPES TO HAVE A DATA SET AND USE MATHEMATICS TO CONNECT SIGNALS TO THE RESPONSES. WHEN THEY THINK ABOUT SYSTEMS BIOLOGY MOST THINK ABOUT DIFFERENCIAL EQUATION BASED MODELS BUT WHAT I WANT TO STRESS IS THAT'S JUST ONE EXTREME OF A BROAD RANGE OF KNOWLEDGE, BROAD RANGE OF MODELS THAT YOU CAN USE. YOU CAN GET MORE INSIGHT ABOUT THE LARGER SERIES OF NETWORKS INVOLVED. USE A REGRESSION MODEL. ACQUIRE LESS KNOWLEDGE, EASIER MODELS TO CALIBRATE, BROAD SAMPLING OF MANY PATHWAYS. THE DOWN SIDE IS NOT AS MUCH MECHANISTIC INSIGHT. YOU GET CORRELATIONS RATHER THAN CAUSATION BUT THE RESULTS THAT COME OUT ARE QUITE IMPORTANT. THE IDEA WITH ALL OF THESE MODELS IS YOU SOMEHOW WANT TO RELATE THE RESPONSES TO THE SIGNALS. KEY IS WHETHER THE FUNCTION IS MECHANISTIC FROM DIFFERENCIAL EQUATIONS OR CORRELATIVE, COMES FROM THESE REGRESSION MODELS. THERE ARE SIGNALS IMPORTANT IN AGING AND MACRO MOLECULAR DAMAGE, WE DON'T MEASURE. IF YOU BELIEVE PATHOLOGICAL REPAIR THAT CONTROLS WHAT THE RESULT MILLION OUTCOME YOU HAVE TO THINK ABOUT THE PATHWAYS WE CAN MEASURE WHICH I HAVE SHOWN YOU, AND THEN REALLY IMPORTANT PATHWAYS WE DONE DO A GOOD JOB MEASURING OR AT THE MOMENT MEASURING ACTIVATION OF THE NOTCH PATHWAY OR BETA CATENIN WINT PATHWAY. THERE AREN'T POST TRANSLATIONAL MODIFICATIONS EASY TO SET UP TO MEASURE. WE HAVE TO LOOK AT INDIVIDUAL CELLS AND PROTEIN TRANSLOCATION HERE. SO TO SUM UP, I WANT TO LEAVE YOU WHAT I THINK ARE CHALLENGES AND UNMET NEEDS. WHAT ARE CELL TYPES IMPORTANT IN CONTROLLING THE RESPONSE OF CELLS TO MACRO MOLECULAR DISCHARGE DAMAGE? DAMAGE CELLS THEMSELVESES OR SPECIFIC IN THE MICROENVIRONMENT THAT WE SHOULD MEASURE. WHETHER SIGNALING PATHWAYS ARE THE MOST IMPORTANT TO MEASURE? I THINK THIS IS A CASE WHERE IT'S A BIT LIKE THE DRUNK UNDER THE LAMP POST, WE HAVE GREAT TOOLS MEASURING SOME PATHWAYS AN POOR TOOLS FOR OTHERS. WHAT SHOULD WE MEASURE AS A RESPONSE? PHENOTYPES LIKE SENESCENCE OR APOPTOSIS OR SOMETHINGEESIER TO MEASURE QUANTITATIVELY LIKE IL-6 OR TNF OR FOCUS ON CELL MARKERS, MODIFICATIONS OF HISTONES FOR EXAMPLE, OR SOME LEVELS OF OX GEYSER DIEED PROTEINS HOW DO WE MEASURE IN VIVO SUBMIT'S EASY IN ACTUALTURE BUT NO WAY TO MEASURE MULTIPLE PATHWAYS IN CELLS WITHIN A TISSUE AT MOMENT. HOW IMPORTANT IS THIS PATHOLOGICAL REPAIR PROCESS AND SIGNALING REPAIR MODEL? HOW GRANULAR SHOULD MATHEMATICAL MECHANISTIC MODELS BE? LINDA TALKED ABOUT A MODEL OF OF FRAILTY WHERE GRANULARITY IS THE WHOLE PERSON LEVEL. GRANULARITY AT THE LEVEL OF INDIVIDUAL MOLECULES AN PATHWAY. HOW CAN WE USE FIX PATIENT SAM LES TO COLLECT IN ORDER TO GET AT THIS DATA AND HOW CAN WE MAKE THE APPROPRIATE MOUSE MODELS THAT ALLOW US TO TEST SPECIFIC PREDICTIONS OF SIGNALING IN VIVO. THANK YOU VERY MUCH. >> I'LL TURN THIS TO ERIC. I WANT TO THANK THE SPEAKERS FOR KEEPING ON TIME. >> SO SPEAKERS COME TO THE PODIUM, NO SHORTAGE OF THINGS TO BE THINKING ABOUT GIVEN THOSE GREAT MANY LECTURES. SO PEOPLE WHO WANT TO ASK QUESTIONS COME UP TO THE MICROPHONES. I PARTICULARLY LIKE THE LAST PART OF MICHAEL'S TALK ON HOW TO -- ALL THE DIFFERENCE QUESTIONS AND IT IS GOING TO COME TOWN TO HOW CAN WE AGGREGATE CHECKING AND AGGREGATING THIS INFORMATION OVER SYSTEMS TO RECEIVE INTEGRATE AND TAKE A DATA DRIVEN VIEW PERHAPS OF AGING. SO WE'RE READY TO ENGAGE? >> SO PICK A QUESTION. THE OXIDATION OF THE ARGININE AND LYSINE, YOU SUGGESTED THAT HAPPENS TO METAL PROTEINS. IS THAT ABSOLUTELY THE CASE THE OXIDATION ARGININE AND LIE SEEN ONLY HAPPENS TO METAL PROTEIN? >> NOT AT ALL. IT CERTAINLY HAPPENS WITHOUT METAL PROTEINS WITH THE MANY LYSINES ON THE SURFACE OF PROTEINS. WHAT I DIDN'T MAKE CLEAR IS THE FUNCTIONAL EFFECTS WHAT I WAS EMPHASIZING. IF IT'S AN ARGININE FORMING THE METAL BINDING SITE OR NEARBY, IT BECOMES DYSFUNCTIONAL OR NON-FUNCTIONAL. WHEREAS SURFACE EXPOSED, IT IS UNCLEAR HOW MUCH DIFFERENCE IT MAY MAKE. LYSINE ON THE SURFACE MIGHT BE ANTIOXIDANT PROTECTION MECHANISM. >> THANK YOU. RICH MILLER UNIVERSITY OF MICHIGAN. I HAVE THE SAME QUESTION FOR JAN AND ALSO FOR ROD. I WOULD LIKE TO ASK YAN IF THERE'S EVIDENCE OF LIFE SPAN OF NORMAL HEALTHY ANIMAL OF SAY A MAMMAL COULD BE IMPROVED BY FIXING DNA MUTATIONS OR SLOWING THEIR ACCUMULATION. FOR ROD, IS THERE ANY EVIDENCE THE LIFE SPAN OF HEALTHY NORMAL MAMMAL, IN THE A MUTANT, NOT SICK LITTLE THICK, CAN BE IMPROVED BY PREVENTING OXIDATION OF PROTEINS OR REPAIRING THEM. >> OKAY. SO FIRST OF ALL, OF COURSE, THERE HAVE BEEN MANY THOUGHTS ABOUT TRYING TO IMPROVE DNA REPAIR. BUT THAT TURNS OUT TO BE DIFFICULT IF YOU OVEREXPRESS FOR EXAMPLE PARTICULAR COMPONENTS OF BASIC EFFECTS OUR -- MERELY TOXIC. I DON'T THINK THERE'S EVIDENCE AT ALL. HOWEVER, FROM OUR WORK SHOWS MUTATION ACCUMULATION, AGE RELATED ACCUMULATION MUTATION SLOWS DOWN FOR EXAMPLE, IN DWARF MICE, MUCH SLOWER IN DWARF MICE THAN CONTROL MICE. SAME DROSOPHILA, SAME AGE LEVEL. Q. THAT'S THE ANSWER TO A DIFFERENT QUESTION. THE QUESTION I ASKED IS WHETHER YOU CAN ACCOMPLISH THE GOAL OF LIFE SPAN EXTENSIONS SPECIFICALLY BY IMPROVING DNA HYGIENE. >> >> NOBODY HAS TRIED TO IMPROVE IT, IN SENSE YOU SLOW DOWN, THAT'S THE IDEA, WHEN IGF BY LOWERING IGF SIGNALING IS IN EFFECT IMPROVING YOUR CELLULAR DEFENSE SYSTEMS STRESS RESPONSE AND GENO TOXIC ALSO I THINK THAT IS EVIDENCE THAT AT LEAST CORRELATION BETWEEN IGF LEVEL SIGNALING. >> MY ANSWER IS NO, THERE ISN'T ANY EVIDENCE AT THIS POINT PRIMARILY BECAUSE PEOPLE HAVEN'T LOOKED. THAT'S YOUR POINT, HAVEN'T LOOKED IN THE HEALTHY NORMAL ANIMAL, THOUGH STAY TUNED WHAT THE PROTEOSTASIS PEOPLE HAVE TO SAY ABOUT THAT. >> MY QUESTION IS FOR JAN. AND ALSO RICH -- >> I THINK WE HAVE ONE OTHER COMMENT. >> ONE COMMENT TO THE LAST ANSWER, FOR EXAMPLE, NON-HOMOLOGOUS ENJOINING IS INHERENTLY MUTAGENIC. THIS IS THE ONLY TYPE OF DOUBLE STRAND BREAK REPAIR YOU HAVE FOR EXAMPLE, IN CELLS. THE QUESTION ABOUT CAN YOU COMPLETELY PREVENT A GENOME FROM CHANGING IN SPACE SEQUENCE DURING ITS LIFETIME, I HAVE A FEELING NO BECAUSE INHERENT PROCESSES LEAD TO THAT. >> GEORGE MARTIN UNIVERSITY OF WASHINGTON. JAN MOSTLY. IN RELATION TO RICH MILLER'S QUESTION, BRING US UP TO DATE WHAT COMPARATIVE GEROSCIENCE TELLS US ABOUT INTRINSIC RATE OF SEMATIC MUTATION AS FUNCTION OF LIFE SPAWN COMPARING SPECIES FIDELITY AND DNA REPLICATION FOR EXAMPLE, I HAD ONE OTHER FOLLOW-UP QUESTION ABOUT SOMATIC MUTATION. WE NEVER HAD METHODS TO ANALYZE DNA SOMATIC MUTATION RATE IN DIFFERENCE SPECIES, THEY'RE LIKE REPORTER GENES SO COMPARE FOR EXAMPLE DROSOPHILA MOUSE BUT NOT FOR ALL SPECIES, THERE ARE REPORTS OF DISEASE IN MOUSE FREQUENCY LOWER. GERM LINE MUTATION IS A DIFFERENT METHOD, GENERALLY LOW SOMATIC MUTATIONS AND THEY SEEM TO BE A FUNCTION IN A SENSE, YEAST GERM LINE MUTATIONS SOMATIC MUTATIONS IS MUCH LOWER THAN IN HUMANS. YOU SEE A CLEAR RELATIONSHIP MORE COMPLEX ORGANISMS IS GENERALLY HIGHER MUTATION RATE. THAT IS THE ANSWER TO THAT QUESTION. >> I'M GLAD YOU BROUGHT UP SOMATIC STORY. WHEN I READ THE FAMOUS PNAS PAPER MORE LIKE WHAT HE'S TALKING ABOUT IS INACTIVATION OF ENTIRE CHROMOSOMAL ARM. IT'S LIKE A LINE HYPOTHESIS. NOW WITH ANDY CHES STORY, WHEN HE FINDS EVIDENCE THAT'S SOMATIC MOW SAY SCHISM FOR AUTOSOMAL GENES THE BETA AMYLOID PRECURSOR GENE SO PATCHES OF TISSUES HE SAY, OPERATING ALLELICLY EITHER ON YOUR -- ON THE PATERNAL OR THE MATERNAL. SOMETIMES DIPLOID, SOMETIMES HOMOZYGOUS. SO WHAT TEASE FOLLOW-UP ON THAT? >> THIS IS INTERESTING, THERE'S NOT A LOT OF VERY RECENT EVIDENCE FOR SOMATIC MOSAICISM, MOSAICISM, THIS OF COURSE BECAUSE THERE'S SEQUENCING AND ALL SORTS OF CHROMATIN ANALYSIS SO YOU CAN SEE NOW THAT THE POPULATIONS OF CELLS, THEY ARE SHOWING UP SOMETIMES THEY CLONAL DEVELOP MUTATIONS YOU CAN DETECT IT. VERY LOW ONE IS TIP OF THE ICEBERG DISCOVERED SLOWLY BUT IMPLICATIONS IN AUTISM DISEASE PATHOGENESIS IS BECOMING CLEARER AND CLEARER. THE ONLY WAY SO TAKE A SINGLE CELL APPROACH TO SEQUENCE WHOLE GENOMES OF MANY SINGLE L CELLS. >> THERE'S A NICE PAPER THAT CAME OUT IN SCIENCE TODAY ON THE SOMATIC MOSAICISM IN NEURONS. THESE COPY NUMBER VARIATION ALTERATIONS, AVERAGING TWO MEGA BASES TO 70, HALF THE CELLS THEY SEQUENCED INDIVIDUALLY HARBORED AT LEAST ONE OF THESE RATHER LARGE COPY NUMBER VARIATIONS MUTATIONS. A FAIRLY NATURAL THING THE JOELS ARE SHUFFLED AROUND. >> THANK YOU. >> (INAUDIBLE) FROM NIDDK. MY QUESTION IS FOR DR. JONES STIMULATED BY THIS NICE SLIDE BEHIND YOU. ARE THERE INTERACTIONS BETWEEN PHYSICAL ACTIVITY AND REDOX PROTEOME? YOU TALKED ABOUT DIET BUT DO YOU KNOW OF ANYTHING? >> CERTAINLY THERE'S LITERATURE ON THAT. IT FALLS TWO PHASES, IMMEDIATELY AFTER EXERCISE THERE IS AN OXIDATION RESPONSE, OXIDATIVE RESPONSE AND THEN PROLONGED RESPONSE FROM THAT IS A MORE REDUCED STATE. AS FAR AS FOLLOWING THAT OVER IN TERMS OF LONG TERM INDIVIDUALS, I DON'T KNOW OF STUDIES THAT HAVE DONE THAT. CERTAINLY THE STUDY IN YOUNGER INDIVIDUALS AND MORE IN TERMS OF CONDITIONING AND ENDURANCE ATHLETES, IT'S VERY CLEAR THE EXERCISE DOES IMPACT REDOX STATE. >> (INDISCERNIBLE) DRUG DISCOVERY FOUNDATION. THERE'S A LOT OF USE IN GENERAL PUBLIC IN TERMS OF ASSAYING TELOMERE LENGTH FOR AN IDEA OF THE AGENT. THIS WAS CLEAR FROM THE TALKS TODAY THAT'S NOT NECESSARILY WHAT ANYBODY CAN BE USING. BUT IT'S BEING USED. I'M CURIOUS THE TYPES OF MACRO MOLECULAR DAMAGE DISCUSSED IN THE PANEL. WHAT MIGHT BE FEASIBLE FOR TESTING HUMANS IN BIOMARKER AN PERSPECTIVE OF PEOPLE BUYING THIS AND MAKES THE DANGERS. WHAT MIGHT BE USEFUL, THERE IS DATA TELOMERE LENGTH MIGHT BE ASSOCIATED WITH CARDIOVASCULAR DISEASE. SO WITH THESE VARIOUS APPROACHES, WHAT'S PRACTICAL FOR TESTING IN HUMANS? >> I'LL START BY MAKING THE POINT THAT WE NEED TO DISTINGUISH BIOMARKERS THAT ARE USEFUL IN A SCREENING POPULATION OR GROUP TO GIVE US A CLUE ABOUT MECHANISM AND THOSE THAT ARE USEFUL FOR A SPECIFIC INDIVIDUAL. IN THE KISS OF PROTEIN OXIDATION A REMARKABLE NUMBER OF DISEASES THAT HAVE A STRONG CORRELATION WITH PLASMA, OXIDIZED PROTEIN LEVEL OFTEN OBSERVED BEFORE CLINICAL ONSET OF DISEASE. BUT I DON'T KNOW OF A SINGLE ONE OF THOSE WHOSE VARIABILITY IS LOW ENOUGH TO MAKE IT LOW ENOUGH FOR INDIVIDUAL USE. >> I WAS GOING TO MENTION THERE HAVE BEEN STUDIES FOR EXAMPLE, FOR GAMMA H 2AX THIS MIGHT BE INFLAMMATION RATHER THAN AGING SPECIFICALLY. IN ITALY THEY SCREENED A BUNCH OF CHILDREN AND FOUND THE GAMMA H 2AX LEVELS INCREASED WITH BMI OF THE INDIVIDUAL. THERE ARE SOME THINGS YOU CAN MEASURE. >> I THINK IT'S NOT WISE TO FOCUS ON ONE PARTICULAR ASPECT. LIKE TELOMERE EROSION IS ONLY ONE MANY FORMS OF GENOME INSTABILITY. I WAS WONDERING WHY -- I CAN GUESS WHY THERE'S SO MUCH INTEREST BUT NEVERTHELESS YOU SHOULD LOOK AT GENOME INSTABLE ANY MORE BROADER SENSE. AND OTHER MARKERS, CYTOGENETIC ARRAY GCAs, AS I HOPEFULLY TRY TO CHOLEDOCHO IN THE FUTURE, SEQUENCING TO TRY TO CAPTURE ALL THESE VARIATIONS. >> FROM -- IT'S PROBABLY GOING TO TURN OUT USEFUL TO MEASURE NOT JUST MARKERS OF DAMAGE IN A PARTICULAR CELL, I AGREE, GAMMA H 2AX IS A USEFUL MARKER TO START. BUT ALSO MEASURE CIRCULATING CYTOKINES OR CIRCULATING COAGULOPATHY PROFILES AND SEE IF THERE'S A WAY TO PUT THE DATA TOGETHER TO STRATIFY RISK. I THINK THAT IF WE FOCUS ON DAMAGE AND IGNORE REPAIR COMPONENT AND WHAT IT IS THAT CONTROL MR. THE DAMAGE RESULTS IN FUNCTIONAL LOSS WE'RE ONLY LOOKING AT ONE PART OF A BIGGER EQUATION. >> I DON'T KNOW ABOUT HUMANS BUT CERTAINLY IN MICE WE USED ASSAY OF PROSTATES, THAT'S NOT TOO BAD BECAUSE IT GIVES YOU TISSUE OUT TO THE BLOOD. I AGREE WITH ROD, THERE WOULD BE SO MUCH VARIATION. I WAS GOING TO ASK DEAN, HE'S DONE THINGS LOOKED AT THE GLUTATHIONE OXIDIZED REDUCED RATIO AND SEEM TO ME THAT THAT WOULD BE AN INTEREST THING. YOU HAVE OPPORTUNITY THAT IN HUMANS. >> WE LOOKED AT GLUTATHIONE REDOX POTENTIAL A VARIETY OF OTHER BIOMARKERS, I THINK THAT THE EMERGING VIEW IS THE CONCEPT OF A GALCION TYPE DISTRIBUTION OR THAT IS MONOTONIC IS REALLY PROBABLY ERRONEOUS WAY TO THINK ABOUT THINGS AS WE GO FROM A POPULATION TO AN INDIVIDUAL BASED HEALTH ASSESSMENT. WE'RE PROBABLY GOING TO AS WE GET LARGE ENOUGH SYSTEMS IN A SYSTEMS BIOLOGY SENSE, WE WILL HAVE SUBCLASSIFICATIONS OF PEOPLE SO WE'RE GOING TO BE ABLE TO IDENTIFY BUCKET. AND DIFFERENT BUCKETS WITH DIFFERENT RESPONSES. SO WE'LL BE BETTER ABLE TO USE BIOMARKERS WHEN WE FIGURE OUT THAT OVERALL STRUCTURE HOW WE CAN DEFINE WHICH BUCKET TO PUT A PERSON INTO IN TERMS OF METABOLIC GENETIC CHARACTERISTICS AND USEFULLY USE THOSE BIOMARKERS THAT ARE CURRENTLY NOT ACCURATE. I THINK THE PRECISION WILL BE MUCH BETTER WHEN WE GET INTO -- IMPROVED SUBCLASSIFICATION STRUCTURES. >> INSTITUTE ON AGING. I HAVE A QUESTION FOR JAN MOSTLY. THIS IS AGING AND RELATIONSHIP TO DNA REPAIR. IN THE CONTEXT OF PROLIFERATING AND NON-PROLIFERATING TISSUE. WHAT IS THE -- IS THERE ANY DIFFERENCE IN CORRELATION BETWEEN AGING RATE AND MUTATION? THOUGH CELLS DIVIDE COMPARED TO CELLS THAT NOT DIVIDE. MUTATIONS BECAUSE HA CARRIES ON. >> THAT'S A GOOD QUESTION. IF WE LOOK AT MUTATION ACCUMULATION AGING ORGAN TISSUE, FOR EXAMPLE WE SEE A DRAMATIC INCREASE IN MUTATION, THEY'RE POINT MUTATION. ALSO AN ORGAN, A LOT OF TOUGHLY PROLIFERATING CELLS, WE DO NOT SEE THAT AT ALL. I EXPLAIN THIS BUT I HAVE NO REAL EVIDENCE, APOPTOSIS IS MUCH HIGHER. SO THAT'S WHY YOU SEE MUTATIONS. YES OF COURSE, UNDER CERTAIN CIRCUMSTANCES IT'S VERY IMPORTANT TO REPAIR QUICKLY, BUT END OF THE DAY THE PRICE YOU PAY MUTATIONS. WE DON'T KNOW THAT BUT THAT'S A POSSIBILITY. HAVING SAID THAT YOU CAN INDUCE MUTATIONS IN BOTH DIVIDING CELLS AND IN CELLS DIRECTLY. CELLS ACTIVELY PROLIFERATING, ULTRAVIOLET LIGHT YOU SEE POINT MUTATIONS REPLICATION ERRORS. YOU WILL DO SAME CELLS CRY QUIESCENT NOT DIVIDING. REPLICATION ERRORS BUT ON THE OTHER HAND (INAUDIBLE) YOU SEE LOTS OF STRUCTURAL VARIATIONS. STEM CELLS IN CERTAIN SITUATIONS. >> THANKS. >> HI, BRIAN KENNEDY, BUCK INSTITUTE. I WANT TO GO BACK AND P COME BACK TO RICH'S POINT A LITTLE BIT. BROADEN IT. BECAUSE I THINK THIS REALLY ONE OF THE BIG PROBLEMS IN THE FIELD RIGHT NOW IS WE HAVE HAD TWO INTERESTING GREAT LINES OF INVESTIGATION, ONE OF THEM IS THIS MUTATION CONSERVED THAT EXTEND LIFE SPAN. MORE OR LESS UNBIASED. THE OTHER IS, LET'S LOOK AT PARTICULAR MODELS WE HAVE FOR WHAT MIGHT BE DRIVING AGING DNA DAMAGE, PROTEIN MISFOLDING. THE PROBLEM IS, IT'S BEEN HARD TO CONNECT THOSE TWO THINGS. THE CONSERVED AGING MUTANTS WE FIND CAN BE LINKED TO MULTIPLE OF THESE PATHWAYS. AND SO I GUESS THE QUESTION IS, IS THE ONLY WAY, I DON'T KNOW THE ANSWER, HOW DO WE CONNECT THOSE TWO LINES OF INVESTIGATION ONLY WAY TO EXTEN LIFE SPAN HIT MULTIPLE FORMS OF THESE EVENTS THAT ARE DRIVING AGING? AND HITTING THESE NODES? OR IS IT THAT THERE ARE JUST A COUPLE OF ONES THAT ARE DRIVING THE PROCESS AND WE DOPE HAVE THE TOOLS TO TWEAK THOSE TO GET THE EFFECTS. THIS IS SOMETHING I THINK ABOUT A LOT. YOU OF AN ANSWER, I WONDER WHAT YOUR THOUGHTS ARE ON THAT. SYSTEMS LIKE RAPAMYCIN -- DEFINITELY IGF -- YOU REALLY IN ONE WAY OR THE OTHER BEGIN TO AFFECT A HOST OF DEFENSE SYSTEMS. AND I THINK ALTHOUGH THERE'S IN THE A LOT OF EVIDENCE FOR THAT, SOME EVIDENCE AT LEAST IS THERE. WHEN WE KNOCK DOWN (INAUDIBLE) AND WE GET -- WITH PARTICULAR DNA DAMAGE AGENT WE SEE MANY MORE MUTATIONS THAN WHEN FULLY ACTIVE. THAT'S EVIDENT AND I THINK ALREADY SHOWS THAT IT IS IMPORTANT IN DNA DAMAGE RESPONSE. THE SAME I TOLD RICH WE SEE EFFECT OF ICF MUTANT MUTATION, SO MUTATION RATE IS LOWER IN THOSE ORGANS. SO AGAIN I'M SURE THERE WAS OTHER EFFECTS A HOST OF DIFFERENT CELL DEFENSES SO I REALLY THINK WHAT THEY DO IS REALLY AFFECT BROAD RANGE OF CELLS DEFENSES, I BELIEVE GENERALLY FELT IN THE FIELD. BUT THE RESPONSE DEMONSTRATES FOR EVERYTHING. >> NOT QUITE ON YOUR QUESTION BUT CERTAINLY LOOKING TRYING TO TEST OXIDATIVE STRESS AGING, IT WAS VERY FRUSTRATING BECAUSE WE MADE ANIMAL MODELS WHERE WE HAD INCREASED OXIDATIVE STRESS BECAUSE THERE WAS ALWAYS A PRETTY TIGHT CORRELATION BETWEEN DAMAGE AND DWARF MICE AND THIS SORT OF THING. I THINK IT'S REALLY IMPORTANT WHICH BOTH YOU AND RICH RAISED, TO BE GOING AND MANIPULATE THE SYSTEM. WE DID MANIPULATE THE SYSTEM SO WE GOT MORE DAMAGE OR LESS DAMAGE. THE UPSHOT IS WE DIDN'T SEE DIFFERENCE IN LIFE SPAN. ON THE OTHER SIDE I'M SKEPTICAL WHETHER MODEL SYSTEMS MAYBE THE MOST RELEVANT BECAUSE IF WE DO THE SAME GENETIC MANIPULATIONS THAT HAD NO EFFECT ON LIFE SPAN, AND WE DO IT IN A DISEASE MODEL ALZHEIMER'S OR CARDIOVASCULAR DISEASE OR SO ON, WE SEE A DIFFERENCE. AS PREDICTED FROM THE STANDPOINT OF LESS DAMAGE OR MORE DAMAGE BAD FOR YOU, LESS DAMAGE -- I'M WONDERING IN THE MOUSE MODEL SYSTEM, WE USE THE LONGEST LIFE STRAINED UNDER PERFECT CONDITION, AND I'M WONDERING UNDER THOSE CONDITIONS IF WHEN YOU DO CHANGE THE OXIDATIVE DAMAGE OR DNA DAMAGE AROUND THAT YOU'RE NOT GOING TO -- IT WON'T BE A MAJOR PLAYER IN WHAT'S AFFECTING LIFE SPAN. WE'RE VERY -- I HAVE NEVER BEEN ABLE THE TO HIT ON THIS, BUT I THINK THAT IT WOULD BE VERY INTERESTING TO LOOK AT A MODEL THAT WAS AT LEAST SLIGHTLY STRESSED AND SEE IF MANIPULATIONS THAT MAYBE NOT WORK IN LIFE SPAN UNDER THIS CONDITION. THE OTHER THING IS ALSO POSSIBLE WITH TOR, WOULD THESE MANIPULATIONS WORK IN A STRESS POPULATION, MIGHT NOT. >> TWO POINTS RELATED TO WHAT ARNOLD SAID. WE HAVE TO KEEP IN MIND GENETIC MODELS AND ACUTE INHIBITION BY DRUGS ARE NOT THE SAME THING. MAYOR MAY NOT SEE AN EFFECT IN ONE THAT YOU CAN REPRODUCE WITH THE OTHER. I THINK THE OTHER THING TO KEEP IN MIND IS MANY OF THESE -- THERE'S A LOT OF CROSS TALK BETWEEN PATHWAYS SO YOUR POINT OF TRYING TO ASK SHOULD WE HIT ONE PATHWAY OR MULTIPLE, AND HOW TO FIGURE THAT OUT IS THE BIG QUESTION. IF YOU INHIBIT ONE PATHWAY YOU GET COMPENSATORY UP REGULATION OF SIGNALING THROUGH OTHER PATHWAYS SO WE HAVE TO THINK ABOUT THIS IN A DINE MAKE WAY. PROBABLY EASIER ON AGING THAN CANCER BECAUSE CELL TYPES AREN'T HYPERMUTAGENNIC SO LESS LIKELY TO EVOLVE A BYPASS MECHANISM. I WOULD ARGUE IT IS PROBABLY A COMBINATION THERAPY OF TREATMENT LIKELY TO BE THE MOST EFFECTIVE. THAT OF COURSE ALSO RAISES THE QUESTION WHAT HAPPENS TO THE OTHER CELLS THAT AREN'T DAMAGES. >> I AGREE. TRYING TO FIGURE OUT A NEW WHICH OF THINKING ABOUT HOW TO SOLVE THIS PROBLEM IS CRITICAL ISSUE. I DON'T KNOW THE ANSWER. THERE ARE NO THING AS PATHWAYS ONLY NETWORKS AND TO USE THE CONSTRUCTS AND SAY WE UNDERSTAND HOW THINGS ARE INTEGRATING WITHOUT LOOKING AT THE SYSTEM MORE HOLISTICALLY, NOT JUST LOOKING AT NO SIGNALING PATHWAYS, TEN YEARS AGO WE DIDN'T REALLY KNOW WHAT MICRORNAs WERE AND ONCE DISCOVERED HOW IT CHANGED THE REGULATORY LANDSCAPE WE HAVEN'T SEASONED 10% GENOME IS NOT SEQUENCED, THE ABILITY THE GET STRUCTURAL VARIATIONS JUST NOW STARTING TO COME TO LIFE. SO ALL THESE MISSING PIECES HAVE TO BE ORGANIZED IN THE CONSTRUCTS THAT MICHAEL WAS TALKING ABOUT IN A MORE SYSTEMS LEVEL WAY AND GET AWAY FROM THIS IDEA THESE THINGS ARE SIMPLE SIGNALING PATHWAYS DETERMINING THINGS BECAUSE THAT'S NOT TRUE. THESE ARE OPERATING AT VERY, VERY COMPLEX ENVIRONMENTS, VERY -- THOUSANDS, HUNDREDS OF THOUSANDS OF VARIABLES AT PLAY. WE HAVE TO GET OUR HEADS AROUND THAT IF WE WANT TO BE SUCCESSFUL. >> LET ME SAY ONE OTHER THING. THE POINT YOU BROUGHT UP, ONE OF THE FEW THINGS AGING IS BEING OLDER IS GOOD FOR, GEORGE PROBABLY REMEMBERS IF YOU GO BACK 30, 40 YEARS PEOPLE WOULD ARGUE YOU WOULDN'T BE ABLE TO DO ANYTHING WITH AGING BY A SINGLE DRUG OR SINGLE GENETIC MANIPULATION. AND I THINK IT IS AMAZING THESE GENE MANIPULATIONS HAVE BEEN -- THERE'S SO MANY THAT ALTER AND THAT WE HAVE SOMETHING WHEN YOU HAVE RAPAMYCIN, ONE DRUG THOUGH PROBABLY AFFECTING MANY TARGETS. >> I WANT THE ADD AN EXAMPLE HOW CROSS TALK IS SURPRISING CONCERNS ATM, DOUBLE STRAND BREAKS DNA DAMAGE, TANYA PAUL FOUND IT IS ALSO INVOLVED IN ROS LEVELS IN THE CELL. ATM HAS SULFIDE IN IT WHICH IS OXIDIZED OR REDUCED UNCAN DEER CONDITION, YOU CAN KNOCK OUT THE RESIDUE THAT HAS TO DO WITH DNA DAMAGE, IT STILL WORKS SO HERE YOU HAVE THE SAME PROTEIN DOING TWO DIFFERENT THINGS. >> JIM NELSON, BAR SHOP INSTITUTE SAN ANTONIO. I THINK RICH'S QUESTION AND FOLLOW-UP BY BRIAN GETS TO MAJOR ISSUE WITH REASON TO THE QUESTION DOES MACRO MOLECULAR DAMAGE SEEN DURING AGING REALLY -- HOW DO YOU FIND OUT WHETHER THAT REALLY HAS AN IMPACT ON EITHER AGE LONGEVITY OR SPECIFIC ASPECTS OF AGE RELATED LIFE SPAN. I THINK THERE ARE TWO EXAMPLES THAT HAVE BEEN -- THAT HAVE ADDRESSED THIS QUESTION SUCCESSFULLY. FIRST ONE HASN'T BEEN RAISED HERE, IS ONE CONCERNING THE AGE RELATED INCREASE IN DNA DAMAGE THAT OCCURS IN THE GERM LINE. CHRIS WALTER HAS STUDIED THIS VERY BEAUTIFULLY FOR THE LAST 15, 20 YEARS AND I HAVE BEEN USING A MOUSE MODEL, A MALE, AND IDENTIFIED A SPECIFIC ENDONUCLEASE THAT SEEMS -- THAT HAS DOWN-REGULATED WITH AGE AN CORRELATES WITH THE INCREASE DNA DAMAGE THAT YOU SEE IN SPERM. SHE WHEN BEYOND THAT AS ONE HAS TO TO FINE OUT WHETHER THIS IS SIGNIFICANT WITH INTERVENTIONAL APPROACHES. FIRST WAS TO MAKE A MOUSE HETEROZYGOUS FOR A INCOME OUT OF THIS ENZYME AND SHE SHOWED SHE ACCELERATED THE DNA DAMAGE IN THE SPERM THEN SHE DECELERATED THE DAMAGE. SO THAT IS THE APPROACH NEEDED. THIS IS PERHAPS A LOW HANGING FRUIT. THIS IS AN EXAMPLE OF THE SITUATION WHERE ONLY A SUBSET OF SPERM CELLS WERE DAMAGED AKIN TO WHAT JAN IS SUGGESTING WITH RESPECT TO SOMATIC MUTATIONS BEING VERY ANECDOTAL OR CELL SPECIFIC. I WOULD ASK THE QUESTION, PERHAPS ONE CAN THINK OTHER SYSTEMS THIS THE BODY SIMILARLY AFFECTED AND ONE MIGHT BE THE STEM CELL LINEAGE AND RAMIFY IN DELETERIOUS WAYS AND LOOK AT THAT PARTICULAR SYSTEM WITH RESPECT TO SUCH INTERVENTIONS. >> STEP CELLS, THERE ARE A A NUMBER OF PAPERS SHOWING STEM CELLS HAVE A PARTICULARLY LOW SPONTANEOUS MUTATION FREQUENCY. THERE VERY LOW RATE MITOTIC RECOMBINATION SO LOW ADIPOSITY EVENT, THAT MAY OF COURSE BE RELATED TO THE FACT THAT YOU ARE -- PRESERVATION OF GENOME INTEGRITY IS PARTICULARLY IMPORTANT. THERE'S SIMILAR EVIDENCE FOR GERM CELLS, FOR EXAMPLE FROM CHRIS WALTERS GROUP. THIS IS A QUESTION HOW TO IMPROVE OR DEMONSTRATE THIS CONSTELLATION OF DAMAGE COULD HAVE AN EFFECT. MORE SUBSEQUENT FOR MICHAEL, AAGREE THIS IS A SYSTEMS BIOLOGY APPROACH, THERE'S A LOT OF MODELING INVOLVED BUT IT'S ESSENTIAL WE GET A GOOD HANDLE WHAT HAPPENS IN DIFFERENCE ORGANS AN TISSUES OF A LIVING ANIMAL DURING -- WE HAVE MADE A SYSTEMATIC -- WHAT'S GONE WRONG IN TERMS OF ACCUMULATION, THEN I THINK YOU CAN MOVE INTO MODELING AND APPROACH MICHAEL WAS TALKING ABOUT. SURE HE WANTS TO COMMENT ON THAT. STEM CELLS L HAVE HUGE REPAIR CAPABLE AS WELL SO SOMETHING IS DIFFERENT ABOUT THEM. SOME DATA MIGHT BE HIGHER LEVELS OF ATM ACTIVATION OR HIGHER ACTIVITY CERTAIN ENDOW KNEE COLLIEEASES BUT -- ENDONUCLEASES. BUT YOU HAVE TO COME ONE A WAY TO LOOK AT TISSUE AT THE EXTENT OF DAMAGE OVER TIME AND TO MY KNOWLEDGE THERE'S NO GOOD WAY TO DO THAT NOW. COME UP WITH BETTER MODEL ORGANISMS OR IN VIVO ASSAYS. >> GEORGE MARTIN UNIVERSITY OF WASHINGTON. TRYING TO THINK ABOUT PUTTING TOGETHER ASPECTS OF THE SESSION WE HEARD ON EPIGENETICS EPIMUTATION. NICE PRESENTATION HE SHOWED WHAT SEEMED TO BE SYSTEMATIC CHANGES IN GENE EXPRESSION WITH AGING. ONE INTERPRETATION WITH AGING CONCERTED COMPENSATION LARGE SCALE COMPENSATION FOR PHYSIOLOGICAL DECLINE UP REGULATE THIS, DOWN REGULATE THAT. SO IT MAYBE PROTECTIVE. DOING THAT THERE'S REMODELING GOING ON, WOULD YOU THINK MAYBE INCREASE IT IS PROBABILITY OF EPIMUTATION? IS HE GETTING A LOT OF REASSESSMENT AND REMODELING OF THE CHROMATIN? >> YES. I THINK -- I DIDN'T EXPRESSLY MENTION IT BUT THERE'S ENORMOUS. OF EPIGENOMIC REMODELING GOING ON BEFORE THE ACTUAL ENZYMES HAVE DONE THE DNA REPAIR WORK. MANY OF THOSE RESPONSES ARE SHOWING UP AS CONSISTENT CHANGE, HAD BEEN FOR EXAMPLE AT GAMMA H 2AX THIS LAYER OF GAMMA H 2AX IS WHOLE PAINING THIS IS LARGE AREA. AS JUDY SHOWED, SOME OF THIS -- SOME EVENT, FOCI THEY STAY AROUND IN SENESCENCE CELLS. THAT WOULD BE A CONSEQUENCE OF EPIGENOMIC CHANGES AND THOSE WILL SHOW IN A FAIRLY CONSISTENT WAY AS PART OF THE THE SENESCENCE CELL HOUSEHOLD. THERE ARE PROBABLY MANY, DAVID ALSO MENTIONED THIS RELOCATION OF SORE THANK YOU WINS. PURELY STOCHASTIC DNA DAMAGE RESPONSE SO THERE'S A TARGETED REACTION THERE. >> I HAD A QUICK COMMENT. THESE EPIGENETIC MARKERS, THIS ASPECT I DON'T REMEMBER IT COMING UP BUT CAN GO TO THE NEXT GENERATION TOO. STUDIES OF WOMEN SMOKING, HAVING GRANDCHILDREN WITH HIGHER LEVELS OF ASTHMA. EVIDENTLY THIS -- SO THIS COULD HAPPEN WITH AGING TOO. SENSITIVITY TO VARIOUS TYPES OF DAMAGE. >> IF I CAN COMMENT. THIS IS ONE OF THE ARGUMENTS THAT I FEEL IS STRONG GOING BACK SYSTEMATICALLY UNDERSTAND HUMAN EXPOSURES BECAUSE WE HAVE EXPOSURES VARIABLE IN TERMS OF CHILDHOOD INFECTION, DIETARY CHANGES, ACTIVITY LEVELS, ENVIRONMENTAL EXPOSURES TO PESTICIDES AND SO FORTH. THESE ARE HIGHLY VARIABLE AND IF IN FACT MODIFY THE EPIGENOME AS THE EVIDENCE REALLY IS ACCUMULATING TO SHOW THEY DO, WE REALLY HAVE TO THINK ABOUT THIS, WE HAVE TO SYSTEMATICALLY THINK ABOUT THIS IN TERMS OF ALL OF THE -- ALL OUR AGING AN AGE RELATED DISEASE INVESTIGATION. >> IMPORTANT POINT. I ABSOLUTELY AGREE. >> GEORGE'S COMMENT AND QUESTION PROM MESS TO MENTION ANOTHER KIND OF EPIGENETIC CHANGE THAT ALSO OCCURS ON PROTEINS BUT THE OXIDATIVE ONE. IF A CELL OR ANIMAL IS EXPOSED TO ELEVATED OXIDATIVE STRESS, CHRONIC INFLAMMATION THE MOST PROTEINS SUSCEPTIBLE TO OXIDATIVE MODIFICATION HAVE INCREASED FREQUENCY OF OXIDATION. AS I MENTIONED IF THAT OXIDATION IS SITE SPECIFIC SAY ACTIVE SITE A METAL IS FOUND, YOU HAVE A DYSFUNCTIONAL ENZYME OR STRUCTURAL PROTEIN WHOSE DYSFUNCTION IS EXACTLY THE SAME AS YOU GET FROM INHERITED DNA MUTATION. >> SO LET ME JUST -- IT'S KIND OF LITTLE FOLLOW-UP BUT BACK TO NELSON'S QUESTION, WE'RE TALKING ABOUT DAMAGE. I THINK JAN MADE A GOOD POINT HE WAS ABING ABOUT WHERE YOU HAD A LOT OF DAMAGE YET PREPARED. BUT THE CO-WAS GOING INTO MUTATIONS. I TALKED TO ROD ABOUT THIS, BUT I THINK FROM THE STANDPOINT OF OXIDATIVE DAMAGE, WHEN WE TALK ABOUT DAMAGE MOST OF IT IS PREPARED AND ROD POINTED OUT A LOT OF THINGS IN PROTEINS MAY ACTUALLY BE REPARABLE. WHAT I THINK NOW BASED ON STUDIES NEGATIVE THE KEY MAYBE TO DAMAGE IS NOT THE LEVEL OF DAMAGE BECAUSE IT'S REPARABLE BUT WHEN YOU HAVE ENOUGH DAMAGE YOU TIP AND MAKE A CELL, THIS IS WHERE JUDY'S STUDY ON CELL SENESCENCE ARE VERY ATTRACTIVE BECAUSE ONCE YOU GET ENOUGH DAMAGE YOU CAN TIP A CELL TO BECOME SENESCENCE YOU CAPTURED IF CELL JUST LIKE YOU CAPTURE WITH THE MUTATION. SO IN OTHER WORDS, IT MAY BE THE DAMAGE WE'RE MEASURING IS NOT THE KEY THING, IT WOULD BE THOSE THINGS THAT WOULD MAKE ATE KEY, WHETHER CELL SENESCENCE OR CHANGE IN CHROMATIN METHYLATION, THIS SORT OF THING. >> I WANT TO STRESS WHEN YOU LOOK AT EPIGENETIC CHANGES AND EPIGENOMIC MODIFICATION IN THE SETTING OF DNA DAMAGE YOU HAVE TO DISTINGUISH WHETHER SEEING DIFFERENCE MT. AMOUNT OF DAMAGE OR DIFFERENCE IN THE RESPONSE. IF YOU TREAT CELLS WITH HDAC INHIBITORS OR BROMODOMAIN MODULATORS THAT RECOGNIZE ACETYLATION, IF YOU TREAT CELLS WITH HDAC INHIBITORS YOU SEE A MORE PROFOUND GAMMA RESPONSE, BUT GEL ELECTROPHORESES THERE'S NO INCREASE IN BREAKS. YOU SEE ENHANCED RESPONSE BECAUSE THE CHROMATIN IS OPEN AND BETTER ABLE TO LOAD LARGER STRETCHES OF H 2AX RATHER THAN INCREASE THE AMOUNT OF DAMAGE PER SE. >> CHRIS SELF, FILL DELL FEE Y I WANT TO INSERT INTO THE DISCUSSION HERE YOU GUYS TALK ABOUT THE ISSUE OF TIMING OF REPAIR. SO TIMING WE HAVEN'T TALKED ABOUT AT ALL IN THIS SESSION AND I THOUGHT EARLIER IN THE METABOLIC SESSIONS WE KNOW DEVELOP MENTAL RATE TIDE TO LONGEVITY, WE KNOW SELECTION FOR DELAYED DEVELOPMENT, INCREASES LONGEVITY. THE CELLULAR LEVEL, INTERESTING TO THINK ABOUT TIMING OF REPAIR RELATIVE TO CELL CYCLE PROGRESSION AND HOW THAT HAS TO BE COORDINATED AND ITS RELATIONSHIP TO SPECIES LONGEVITY AND INDIVIDUAL LONGEVITY AS WELL. >> I THINK IT'S AN EXCELLENT POINT. WE HAVEN'T THOUGHT ABOUT IT ENOUGH. TAKING AS AN EXAMPLE WHAT ARLEN MENTIONED, IF YOU CAN REPAIR THE DAMAGE MACRO MOLECULES FAST ENOUGH YOU MY NOT BE TIPPED INTO SENESCENCE, IF YOU CAN YOU END UP WITH A SENESCENCE CELL. >> I AGREE BUT YOU HAVE TO THINK ABOUT THE DIFFERENCES BETWEEN DIFFERENT SPECIES IN TERMS OF THEIR REPAIR, IN TERMS OF THEIR AN TO ARREST, TO REPAIR. WE DON'T THINK ABOUT CELL CYCLE REPAIR IN TERMS OF MACRO MOLECULES, OR THE RELATIONSHIP BETWEEN MACRO MOLECULE DAMAGE AND CELL CYCLE PROGRESSION BUT WE SHOULD, WE ALWAYS THINK ABOUT IT IN TERMS OF DNA REPAIR, IT'S IMPORTANT TO DO CONSIDER ALL THESE NEEDS TO BE PREPARED BEFORE WE CAN GO THROUGH THE CELL CYCLE IN ORDER TO AVOID ON AN UNWANTED CONSEQUENCES. I THINK SYSTEMS BIOLOGY IS THE PLACE TO START, SYSTEMS BIOLOGY OF CELL BIOLOGY REPAIR, WITH WE SEE NETWORKS YOU LAID OUT, SYSTEMS BUY I DON'T IMAGINE EXPERM. CAN YOU MODEL THIS FOR US. >> YOU'RE COMPLETELY CORRECT. PART MAY HAVE NO DO WITH THE CELLS WE CHOOSE TO LOOK AT. IT'S POSSIBLE THE BULK OF THE CELLS WE STUDY ARE TERMINALLY DIFFERENTIATED. THOUGH HAY MAY ACCUMULATE DNA AND PROTEIN DAMAGE, THAT MAYBE RELATIVELY IRRELEVANT TO WHAT HAPPENS. IT MAYBE A PRECURSOR POPULATION OR STEM CELL POPULATION TIMING BETWEEN REPAIR AND WHERE THEY ARE IN THE CELL CYCLE IS CRITICAL FOR PREDICTING WHETHER WE GET SUCCESSFUL REPAIR, PATHOLOGICAL E REPAIR SO I AGREE WITH YOU BUT WE ALSO HAVE THE THINK ABOUT WHETHER WE SHOULD BE STUDYING THE CELL TYPES, BECAUSE THE BULK OF THE POPULATION OR MINORITY RESPONSIBLE FOR THE AGING PHENOTYPE. >> TWO MORE QUESTIONS. AND I THINK WE'RE GOING TO BE OUT OF TIME. >> MY NAME IS JOE FROM THE UNIVERSITY OF AKRON. I'M A RESEARCH ENGINEER. SO I REMEMBER THE SLIDE WE KIND OF STARTED OFF WITH PHILIPPE'S CAR AND I THINK THAT FROM THE ENGINEERING PERSPECTIVE, THIS TYPE OF THING, I'M INTERESTED IN THE CLASSES OF DAMAGE AND PERHAPS THIS IS WHAT DEAN WAS HINTING T LITTLE BIT WITH EXPOSESOME, HAS ANYBODY ENUMERATED ALL OF THE CLASSES OF DAMAGE AND THEN POTENTIAL THERAPIES FOR INTERVENING THIS THOSE CLASSES OF DAMAGE FOR WHERE THE BODY IS -- THE NATURAL REPAR MECHANISMS ARE INSUFFICIENT AS WE'RE SEEING SO WE SEE -- AND THEN ALSO MAYBE THE FAILURE MODE. SO SENESCENCE, CELLULAR SENESCENCE LOOKS LIKE THE MAIN FAILURE MODE AS JUDY CAMPISI HAS SHOWN AND THIS TYPE OF THING THAT LENDS TO SYSTEMS BIOLOGY. AT THE SAME TIME I WANT TO CAUTION THAT WE DON'T NEED TO LOOK FOR COMPLEXITY, IT WILL FIND US WITHIN THAT TYPE OF THING SO WE DONE NECESSARILY NEED TO LOOK AT NEIGHBOR THE BALL BEARINGS APPROXIMATE TYPES OF THINGS IN ORDER TO TO KNOW WE NEED A NEW BALL BEARING. PRIORITIZATION, WHAT WOULD BE THE MOST IMPORTANT DAMAGE IN THAT TYPE OF THING. THANKS. >> THIS IS AN IMPORTANT P QUESTION. PART OF IT IS LIMITED THE TECHNOLOGY. MEASURING DAMAGE IS VERY DIFFICULT TO DO. MAINLY BECAUSE A LOT OF THE DAMAGE IS VERY LOW LEVELS. SO WHAT YOU HAVE IN MOST CASES IS INDIVIDUAL MEASURES ONE KIND OF DAMAGE WHETHER DNA OR PROTEIN CARBNEL HEALTHCARE,S WERE A BIG DEAL TO MEASURE. WE DOPE HAVE THAT. AS I MENTIONED BEFORE, IT COULD BE THAT IT'S NOT WHEN WE'RE MEASURING ONE TYPE OF DAMAGE, WE'RE NOT GETTING A GOOD SNAP SHOT. THE ASSUMPTION IS THAT WE MEASURE ISOPROSTAINE OR CARBNELLES OR DNA OXIDATION, WE GET THE SAME RESULTS. THAT'S JUST THREE ASSAYS. I DON'T KNOW ANYBODY THAT'S DONE THAT THOUGH WE HAVE HAD THE ABILITY TO DO IT, SOME ARE JUST SO DIFFICULT THAT WE HAVEN'T DONE IT. THIS COMES BACK TO A QUESTION I RAISED TO RICH BEFORE ABOUT TALKING ABOUT STRESS RESISTANCE. AND WHY WE SEE IN MANY OF OUR LONG LIVED ANIMALS THAT THIS RESISTANCE TO STRESS BUT YET YOU HAVE ANIMALS THAT YOU MAKE RESISTANT TO STRESS THEN YOU DONE SEE ANY DIFFERENCE. AND I THINK THE POSSIBILITY IS THERE'S TYPES OF DAMAGE THAT MAYBE IMPORTANT THAT WE'RE NOT PICKING UP. SO MY FEELING IS WE NEED TO DO THIS BUT IT'S GOING TO BE VERY DIFFICULT. AND IT IS NOT PARTICULARLY INEXPENSIVE. >> SO I WILL COMMENT. IT'S A HUMBLING EXPERIENCE. A STUDY THAT MARIA KADISKA AT NIEHS CONDUCTED LOOKING AT OXIDATIVE STRESS IN A MOUSE MODEL AND IT WAS REALLY A BIOBRILLIANTLY DESIGNEDDED EXPERIMENT TO GIVE CARBON TETRA CHLORIDE, KNOWN FREE RADICAL INDUCING OXIDANT AND FOLLOW THAT WITH COLLECTING SAMPLES AND SENDING THEM OUT TO AT LEAST A HALF DOZEN DIFFERENCE LABORATORIES WHO WERE ABSOLUTELY EXPERT IN ALL THE DIFFERENT MEASUREMENTS THE GLUTATHIONE, ISOPROS STAIN, SO FORTH. WHAT HAPPENS IS TRULY REMARKABLE. WHEN WE GOT THE DATA BACK OF EVERYTHING BLINDED AND SENT -- REALLY DONE RIGOROUSLY. WHEN WE GOT THE RESULTS BACK, WHAT THEY SHOWED WAS THAT THE MOST SENSITIVE MARKER WAS INCREASE IN GLUTATHIONE IN THE BLOOD. IT TURNED OUT THE DAMAGE TO THE LIVER WAS SUCH IT KILLED THE LIVER, RELEASING THE ANTIOXIDANTS INTO CIRCULATION THOUGH ABSOLUTELY MAJOR OXIDATIVE I WERE SHUT THE MOST SENSITIVE INDICATOR WAS THE OPPOSITE OF WHAT YOU WOULD GET. IT ILLUSTRATES THE POINT NO MATTER HOW CLEVER WE ARE IN TERMS OF DESIGN AND SOPHISTICATION IN TERMS HOW WE MEASURE THINGS, THAT WE'RE STILL LIMITED IN THIS IS THE ANALYTICAL MODELS. THAT'S WHERE WE -- THAT'S WHY I ARGUE THAT WE REALLY IMMEDIATE TO COME BACK TO HUMANS, AS BEST WE CAN DEVELOP METHODS FOR DEALING WITH THE COMPLEXITY IN HUMANS AND BEING ABLE TO EVALUATE THAT IN TERMS OF THE AGING PROCESS. >> WOULD IT BE FAIR TO SUMMARIZE ITS MEASUREMENT IN HUMAN BEINGS THAT WITH OTHER LOOKING TO GO FORWARD ON ON THAT? >> MY ARGUMENT IS THE BEST WAY WE'RE GOING TO DEAL WITH AGE ASSOCIATED DISEASE AND HEALTHY LONGEVITITY IN HUMANS. IF >> OUR TIME IS UP. RON IS GOING TO TAKE OVER. >> I WOULD LIKE TO THANK CO-CHAIRS AND PANELISTS AND THE AUDIENCE FOR A FINE SESSION. [APPLAUSE] >> SO NOW WE WILL BREAK FOR LUNCH. AND BE BACK AT 1 P.M. FOR THE NEXT SESSION ON PROTEOSTASIS. GOOD AFTERNOON TO EVERYONE, WELCOME TO THE FIRST SESSION IN THE AFTERNOON, WHICH IS GOING TO FOCUS ON PROTEOSTASIS. MY NAME IS HENRY RODRIGUEZ, I'M THE DIRECTO DIRECTOR OF PROTEOMICS AT THE NATIONAL CANCER INSTITUTE. MY COORGANIZER FOR THIS SESSION IS BRADLEY WEISS, THE CHIEF OF NEUROBIOLOGY OF AGING BRANCH AT THE NATIONAL INSTITUTE ON AGING. SO FOR THE SESSION ON PROTEOSTASIS, WE HAVE TWO CO-CHAIRS. ONE IS RICHARD MORIMOTO AND THE OTHER IS ANA MARIA CUERVO. SO DR. MORIMOTO HAPPENS TO BE THE DIRECTOR OF THE WRITES INSTITUTE FOR BIOMEDICAL RESEARCH AT NORTHWESTERN UNIVERSITY. HE'S ALSO THE BILL AND GAIL COOK PROFESSOR OF BIOLOGY WITHIN THE SCHOOL'S DEPARTMENT OF BIOCHEMISTRY, MOLECULAR BIOLOGY, AND CELL BIOLOGY. SO RICK IS GOING TO FIRST GIVE AN ENT DUC INTRODUCTORY OVERVIEW OF THE TOPIC FOR TODAY'S SESSION. THE OTHER CO-CHAIR IS DR. ANNA MARIA CUERVO, SHE'S THE CO-DIRECTOR OF THE INSTITUTE FOR AGING RESEARCH AT ALBERT EINSTEIN COLLEG COLLEGE OF MEDICINE, AND ALSO HER DISTINGUISHED COLLEAGUE, MR. MORIMOTO, SHE IS ALSO A CHAIR FOR THE DI STUDY OF NEURODEGENERATIVE DISEASES AND IS A PROFESSOR IN THE DEPARTMENT OF DEVELOPMENTAL AND MOLECULAR BIOLOGY. SO ANA MARIA IS GOING TO DIRECT THE DISCUSSION THAT FOLLOWS AFTER THE SPEAKERS AND SHE'S GOING TO GIVE A SUMMARY OF THE OVERALL OF THE SESSION. SO AT THIS POINT, PLEASE WELCOME DR. MORIMOTO. >> GOOD AFTERNOON, WELCOME BACK. WE HAVE A WONDERFUL SESSION FOR YOU ON PROTEOSTASIS. WE'RE GOING TO HEAR FROM FRED GOLDBERG, ON HOW DOES AGING IMPAIR PROTEOSTASIS, ROBERTA GOTTLIEB ON MITOCHONDRIAL QUALITY CONTROL, JUDITH FRYDMAN ON PROTEOSTATIC MECHANISMS, RANDY KAUFMAN ON THE UNFOLDING PROTEIN RESPONSE AND JEFF KELLY ON THE NOVEL THERAPEUTICS. A LITTLE BACKGROUND SO WE CAN ALL COME TOGETHER. ONE OF THE FUNDAMENTALS IN ALL OF LIFE, OF COURSE, IS THE MAKING OF A PROTEIN. A LOT OF WHAT YOU ALREADY HEARD FROM OTHERS ARE HOW DIFFERENT TYPES OF O DAMAGE IMPINGE ON PROTEINS. WHAT WE'RE GOING TO BE TALKING ABOUT ARE A POWERFUL CELL STRESS MECHANISM THAT DETECT BALANCE AND THE IMPORTANCE OF THE PROTEOSTASIS MECHANISM TO MAKE SURE THAT EVERY PROTEIN THAT'S MADE IS INTACT. SO EVERY PROTEIN STARTS OFF ON YOUR RIBOSOMES AS UNFOLDED, THEY GO TOWARDS THE NATIVE STATE. THE PROBLEM IS THE ENVIRONMENT, MUTATIONS, SINT SYNTHESIS, SHIFT THE EQUILIBRIUM TOWARDS DAMAGE. FROM A BIOPHYSICAL PERSPECTIVE THIS, IS AN ENERGY LANDSCAPE DIAGRAM, PROTEINS ENTER THE POOL AS UNFOLDED. IF HE ONLY HAVE INTRAMOLECULAR CONTACTS, THE PROTEIN FOLDS TO ITS LOWEST FREE ENERGY STATE, THAT'S GOOD. THE PROBLEM IS MUTATIONS, ENVIRONMENT, EXPANSIONS OF POLYQUEUE, ALZHEIMER'S DISEASE, HUNTINGTON, THESE TYPES OF PROTEINS, WHEN THEY SHIFT INTO AN ALTERNATE CONFORMATION, FORM MORE FIS AGGREGATES OF LIG MERES AND AN LLOYD FIBERS. THE PROBLEM IS, FOR EVERY PROTEIN IN OUR DREL CELL, THERE IS A RISK FOR A SHIFT BETWEEN INTRAMOLECULAR NORMAL NATIVE STATES AND INTERMOLECULAR. THE PROBLEM FOR THE CELL IS HOW TO DEAL WITH THIS. FORTUNATELY, WHAT COMES TO THE RESCUE ARE MOLECULAR CHAPERONS, AS YOU'RE GOING TO HEAR ABOUT, AND CLEARANCE MECHANISMS. THE CHAPERONS PLAY THE CRITICAL ROLE TO MAKE SURE THAT ALL OF OUR PROTEINS ARE PROPERLY FOLDED FOLDED, TOGETHER WITH THE CLEARANCE MECHANISMS, THE PROTEOSOME, THEY MAKE SURE THESE OFF PATHWAY INTERMEDIATES THAT CAN ACCUMULATE AND INTERFERE DON'T CAUSE DAMAGE TO THE CELL. OF COURSE THE PROBLEM OVER TIME IS THAT WE ACCUMULATE, BECAUSE OF MUTATION, YOU ALREADY HEARD ABOUT THAT, BIOSYNTHETIC ERRORS IS HUGE, THERE'S DOZENS OF STEPS FROM THE TIME THAT DNA IS TRANSCRIBED TO THE POINT OF THE PROTEIN HAS ACTUALLY FOLDED AND IN EVERY ONE OF THESE STEPS, THERE IS ERRO RFLT-PRONE EVENTS THAT OCCUR. SO THE CONSEQUENCES, THE ACCUMULATION OF PROTEIN DAMAGE, AGING, OF COURSE, IT'S A MAJOR ISSUE. PROTEOSTASIS REPRESENTS THE INTEGRATION, IT'S THE NETWORK OF MOLECULAR INTERACTIONS, THE STRESS RESPONSES, THAT DETERMINE THE BALANCE BETWEEN FUNCTION IN OUR CELLS AND DYSFUNCTION. WHEN THINGS GO AWRY, FOR EXAMPLE, IN AGING OR IN DISEASES OF PROTEIN CONFORMATION, THE NEURODEGENERATIVE DISEASES, ADULT ONSET DIABETES, MANY CANCERS, AND METABOLIC DISEASES, YOU NOW HAVE AN IMBALANCE, NOW PUSHING THE SYSTEM. FROM A CELLULAR LEVEL, THIS EXAMPLE OF A CELL, THIS PROTEOSTASIS NETWORK THEN PLAYS A CRITICAL ROLE TO TAKE OUR PROTEIN, DESPITE THE POLYMORPHISMS THAT WE ALL ENCODE AND ALLOW PROTEINS TO BE MADE. AGING, WITH MISTRANSLATION, ACCUMULATION OF PROTEIN DAMAGE AND PROTEOTOXIC STRESS, OF COURSE CHALLENGES THIS SYSTEM. IN FACT, IT SETS UP COMPETITION FOR THE PROTEOSTASIS NETWORK. YOU NOW HAVE AN INFLUX OF MISMAN ANLEMISMANAGED PROTEINS. YOU THEN HAVE ANOTHER CHALLENGE HA OCCURS WITH DISEASE-ASSOCIATED MUTATIONS. THESE ARE THE DISEASES WHICH NOW CAUSE PROTEOSTATIC ILLNESSES. THEY LEAD TO -- PROTEINS WHICH THEN GENERATE TOXIC SPECIES. ONE NOW HAS AN ENTIRE SHIFT IN THE EQUILIBRIUM. THE BALANCE IN OUR CELLS, OUR TISTISSUES AND THE ORGANISM IS ALTERED, LEADING TO CELLULAR DYSFUNCTION. AND IT'S NOT GOING TO BE JUST ONE PROTEIN. IT'S LITERALLY HUNDREDS OF PROTEINS THAT BECOME MISMANAGED AND THEY COLLECT OVER TIME, AND AS YOU'LL HEAR, THEY CHALLENGE BOTH THE FOLDING AND CLEARANCE APPARATUS. GENETIC INTERACTIONS THEN PLAY A CRITICAL ROLE BECAUSE OBVIOUSLY ALL WE NEED TO DO IS LOOK AROUND ANY POPULATION OF HUMANS TO REALIZE SOME PEOPLE ARE WELL PROTECTED, OTHERS ARE INCREASINGLY AT RISK. SO THE CRITICAL THING IS PROTEIN FUNCTION. AROUND THIS ARE PATHWAYS, WE'VE ALREADY HEARD ABOUT MANY OF THESE PATHWAYS, SYNTHESIS, POST TRANSLATIONAL MODIFICATIONS, OXIDATION, FOR EXAMPLE, DEGRADATION PATHWAYS. AROUND THIS ARE SIGNALING PATHWAYS. YOU'VE ALREADY HEARD ABOUT SOME OF THESE SIGNALING PATHWAYS. YOU HAVE THE INSULIN SIGNALING PATHWAY, WHICH WAS DISCOVERED TO HAVE A CRITICAL ROLE IN REGULATING PROTEOSTASIS. ORGANIZE NE ALREADY CALCIUM, INFLAMMATORY PATHWAYS, CALORIC RESTRICTION, SIGNALING PATHWAYS SUCH AS THE HEAT SHOCK RESPONSE, AND YOU'LL HEAR ABOUT THE UNFOLDED PROTEIN RESPONSE. THES THEN SIT AROUND THE GENETIC MODIFIERS, GENETIC, EPIGENETICS, METABOLITES AND PHYSIOLOGICAL STRESS LITERALLY DICTATE WHICH PROTEINS ARE MADE AND THEIR FUNCTION AT ANY PARTICULAR MOMENT. JUST TO REMIND YOU THIS, IS A REMARKABLY CONSERVED SYSTEM THAT'S PRESENT IN EVERY ORGANISM IN LIFE. THIS ABILITY TO TAKE A POLYPEPTIDE AND PARTITION IT TO PREVENT AGGREGATION. THIS REPRESENTS THE CHAPERON NETWORKS, THE DIFFERENT CHAPERON PROTEINS THAT THEN IN BLUE PREVENT AGGREGATION FROM OCCURRING OR ALLOW FOLDING. THEN FINALLY, THEY LITERALLY DETECT CHANGES IN THE ENVIRONMENT. THE CONSEQUENCE IS YOU HAVE A SINGLE CELL THAT FUNCTIONS AND SENSES STRESS. SOMEWHERE BETWEEN 3 AND 4 BILLION YEARS AGO, THE CELL CHANGED, SOMEWHERE AFTER THAT, YOU HAVE A SIMPLE MET ZONE. WHAT BECOMES IMPORTANT HERE IS THAT THIS REPRESENTS THE EVOLUTION AND ADAPTATION OF CELL AUTONOMOUS NETWORKS TO CELL NON-AUTONOMOUS, BECAUSE IN AN ORGANISM WITH 959 CELLS, YOU STILL HAVE A SPECIFICATION OF THE SENSORY STRESS APPARATUS. SO IN -- IT'S THE NEURONS THAT ESSENTIALLY SENSE THE HEAT SHOCK STRESS BUT OTHER TISSUES RECOGNIZE AN EMBALANCE IN PROTEOSTASIS. MISMANAGEMENT OF PROTEIN IN AN INTESTINE OR THE MUSCLE OR THE NEURON SENDS SIGNALS BETWEEN THE TISSUES TO RESTORE BALANCE. SO THIS CRITICAL POINT WE HEARD EARLIER ABOUT INFORMATION FLOW, HORMONES AND SIGNALING MOLECULES ALSO OCCURS TO MAINTAIN PROTEOSTASIS TO ENSURE FUNCTIONALITY. WHAT WE'LL ALSO HEAR ABOUT IS HOW THE CELLULAR CHALLENGE AND CELLULAR TOXICITY REALLY HAS A THERAPEUTIC OPPORTUNITY. IF ONE UNDERSTANDS THE THRESHOLD, THE BANDWIDTH BETWEEN CHALLENGE AND TOXICITY, IT GIVES US A REMARKABLE SPACE AND MANY OPPORTUNITIES TO THINK ABOUT THERAPEUTICS. WITH THIS, I'D LIKE TO NOW MOVE TO THE FIRST SPEAKER IN THE SESSION. ANA MARIA, YOU WANT TO INTRODUCE THEM? HE DOESN'T NEED INTRODUCTION, BUT IT'S DR. FRED GELD BERG, AGING AND SELECTIVE DEGRADATION. >> AS RICK MORIMOTO'S ELOQUENT INTRODUCTION EMPHASIZED, EVOLUTION OF HIGHER ORGANISMS AND LOWER ORGANISMS, OF ALL ORGANISMS, HAS REQUIRED THE DEVELOPMENT OF MAJOR SYSTEMS TO ELIMINATE MISFOLFOLDED POTENTIALLY TOXIC PROTEINS. AND THE MOST FUNDAMENTAL ONE THAT'S SHOWN IN THE FIRST SLIDE -- HOW DO I DO THIS? HOPE THAT WASN'T AN I.Q. TEST. TODAY WE'RE GOING TO FOCUS ON REALLY THE EFFECTS AND THE ROLE OF THE INTRACELLULAR SYSTEM IN ELIMINATING SUCH PROTEINS. AND BY MISFOLDED PROTEINS, WE MEAN A LIST OF -- WELL, A LITTLE BIT OF PROTEIN SOCIOLOGY INDICATING THE KINDS OF PROTEINS THAT ARE RAPIDLY DEGRADED IN BASICALLY ALL CELLS, AND EVEN WITHIN ORGANELLES, WITHIN THE MITOCHONDRIAL MATRIX. WHEREVER IT'S BEEN LOOKED AT, INCOMPLETE PROTEINS SUCH AS ARISE BY NONSENSE MUTATIONS, MISTAKES IN RNA PROCESSING, ALSO -- TRIGGERS RAPID DEGRADATION. ABOUT 20% OF THE -- MUTATION AFFECTING, SAY, HEMOGLOBIN, GENERATE MI SENSE PROTEINS THAT ARE RAPIDLY DEGRADING. IT ONLY LOOKS LIKE THOSE PROTEINS THAT MOST -- OVERALL CONFORMATION TRIGGER THIS SELECTIVE DEGRADATION. A BIG SOURCE OF SUBSTRATES FOR THIS SYSTEM ARE SUBUNITS OF MU TIEMERIC COMPLEXES THAT MAY BE FREE. WE USUALLY THINK OF MULTOMERS BEING EXPRESSED IN EXACT -- IN FACT, THAT'S NOT TRUE. THE FREE EXCESS SUBUNITS GET RAPIDLY ELIMINATED AN IN ALL CELLS WHERE IT'S BEEN EXAMINED. YOU'VE HEARD A LOT -- AGING HAS LONG BEEN FOCUSED ON DAMAGE TO PROTEINS. WE STILL DON'T KNOW HOW OFTEN THIS HAPPENS. CERTAINLY THE FORMS THAT YOU'VE HEARD ABOUT THIS MORNING THAT GENERATE STABLE PROTEINS THAT ORGANIC CHEMISTS CAN ASSAY, THESE ARE NOT THE PROTEINS THAT EVOLUTION IS WORRIED ABOUT ELIMINATING RAPIDLY. AND MANY FREE RADICALS TRIGGER RAPID DEGRADATION. IN ADDITION, OF COURSE, THERE'S INTRACELLULAR -- HAPPENING IN A SOCASTIC MANNER ALL THE TIME. THE COMMON FEATURE SEEMS TO BE THE MISFOLDED STATE, WHICH IS A VERY ILL DEFINED PHENOMENA I COULD HAVE PUT THIS SLIDE UP 30 YEARS AGO AND PROBABLY DID. [LAUGHTER] >> AT THAT TIME, UNTIL TODAY, WE REALLY DO NOT KNOW THE DETAILS ABOUT HOW THESE PROTEINS -- IN ANY DETAILED MANNER HOW THESE PROTEINS ARE RECOGNIZED. THIS PATHWAY IS A MAJOR DEFENSE AGAINST SUCH PROTEINS LEADING TO THEIR RAPID ELIMINATION. THIS IS THE UNDERGRADUATE VERSION OF THE PATHWAY. AGAIN, HERE'S ANOTHER TEST I'M FAILING. WHAT THIS PATHWAY IS MEANT TO INDICATE, THERE'S DEGRADATION THROUGH THEIR SELECTIVE UBIQUINATION, THE UIQUININE GETS ACTIVATED AND LINKED TO SUBSTRATE MOLECULES THROUGH ISOPEPTIDE BONDS. AND WHEN THERE'S MORE THAN FOUR IN A ROW, THE PROTEINS ARE RAPIDLY DEGRADED BY THE PROTEOSOME. NOW THIS IS A VERY PROTEOSOMOCENTRIC VIEW OF THE CELL. UNFORTUNATELY I DON'T HAVE TIME TO DO SO, BUT SPEND A LOT OF TIME TALKING ABOUT THE UBIQUINATION ENZYME. THERE ARE ABOUT 700 DIFFERENT ENZYMES IN THIS PROCESS IN THE HUMAN GENOME, ABOUT 20 TO 40 DIFFERENT CON JEW GATING ENZYMES AND PROBABLY 600 THAT RECOGNIZE SUBSTRATES AND CAN ATTACH UBIQUITINE TO THEM. WHICH OF THESE ENZYMES TARGETS MISFOLDED PROTEINS IN MAMMALIAN CELLS IS STILL NOT CLEAR. WE KNOW ABOUT TWO ON THE E.R. THAT GET PROTEINS IN THE SECRETORY PATHWAY AND LEAD TO THEIR DEGRADATION, PROTEOSOMES. THERE'S ONE WELL CHARACTERIZED UBIQUITINE LIGASE BECAUSE THEY HAVE CHAPERONS BOUND TO THEM. THERE HAVE GOT TO BE MANY OTHER SYSTEMS, QUITE A FEW KNOWN IN YEAST. WE STILL DO NOT UNDERSTAND OR KNOW ABOUT THE QUALITY CONTROL MECHANISMS. THEREFORE, WE CAN'T STUDY THE EFFECTS OF AGING. NOW ABOUT TWO PERCENT OF THE CELL PROTEINS IN MOST OF OUR CELLS ARE THE STRUCTURE CALLED THE PROTEOSOME 26S AND THIS IS A 60 SUBUNIT MOLECULAR MACHINE THAT BINDS THE UBIQUINATED PROTEIN AND DEGRADES THEM TO SMALL PEPTIDES, 2 TO 20 RESIDUES LONG. AND 99% OF THESE PEPTIDES WILL GET CONVERTED TO AMINO ACIDS WITHIN 10 SECONDS. PEPTIDES IN THE CYTOSOL ARE VERY DANGEROUS AND ARE QUICKLY ELIMINATED. A SMALL FRACTION, PERHAPS 1%, IS TAKEN UP INTO THE E.R. AND GET DISPLAYED TO THE CELL SURFACE ON MHT CLASS 1 MOLECULE. THIS IS THE SOURCE OF INFORMATION THAT THE IMMUNE SYSTEM, THE CYTOTOXIC T CELLS, USES TO FIND VIRUS INFECTED AND CANCEROUS CELLS, SO THE PROTEOSOME -- THE SYSTEM SCREENS INTRACELLULAR SPACE JUST LIKE THE NYSOSOME AND ENDOSEW MALL SYSTEM SCREENS EXTRA CELLULAR SPACE FOR NON-NATIVE PROTEINS. IT'S AN IMPORTANT ROLE WITH THE SYSTEM ALSO WHEN WE THINK ABOUT AGING. LET ME FOCUS BECAUSE THIS IS GOING TO BE A PROTEOSOMEIC CENTRIC VIEW, BECAUSE OUR VIEW OF ITS FUNCTION, ESPECIALLY CLEARANCE OF MISFOLDED PROTEINS, HAS CHANGED DRAMATICALLY IN THE PAST COUPLE YEARS. THIS IS A MORE REELING REALISTIC BUT NOT QUITE UP TO DATE FIGURE, IT EMPHASIZES THAT THE CORE 20TH CHROMOSOME IS THE DEG DAYTIVE STRUCTURE. INSIDE ITS CENTRAL CHAMBER, PROTEINS ARE CUT UP BY SIX ACTIVE SITES, SO THE PROTEOMI TICK ENZYMES ARE NOT FREE IN THE CYTOSOL, THEY'RE ISOLATED, AND PROTEINS TO BE DEGRADED GET BOUND TO THE 19S REGULATORY PARTICLE AND GET INJECTED INTO THE CENTRAL CORE THROUGH A SMALL OPENING AT EITHER END, AND THIS PROCESS IS -- BY SIX ATPA -- ARE INVOLVED IN THE DECISION OF WHETHER A PROTEIN GETS DEUBIQUINATED AND LIVES LONGER OR GETS TRANSLOCATED IN AND GETS DESTROYED. THIS IS DEFINITELY A LIFE/DEATH DECISION FOR EACH PROTEIN, AND IT'S NOW CLEAR THAT IT IS REGULATED. THIS STEP OF DEGRADATION IS ONE THAT HAS HAD A BIG MEDICAL APPLICATION RECENTLY, AS MANY OF YOU KNOW, PEOPLE STUDYING THIS PATHWAY ARE NOW ABLE TO USE PROTEOSOME INHIBITORS SUCH AS MG132 AS REAGENTS TO DISSECT HOW IMPORTANT THIS PROCESS IS IN DIFFERENT HOMEOSTATIC MECHANISM MECHANISMS. ABOUT 20 YEARS AGO, WE PUT IN SOME EFFORT TO DEVELOP SUCH MOLECULES FOR CLINICAL APPLICATIONS, AND NOW TWO PROTEOSOME INHIBITORS ARE WIDELY USED DRUGS. AVELCATE IS NOW A $2 BILLION DRUG USED BY AT LEAST 400,000 PATIENTS. IT BLOCKS THE DEGRADATION OF MISFOLDED PROTEINS, AND IN CERTAIN CANCERS, SUCH AS MULTIPLE MYELOMA, SENDS THE CELLS INTO A -- THE OPPOSITE OF WHAT WE WANT TO DO IN CONSIDERING AGING, IT'S PROBABLY WHAT YOU WANT TO DO TO ELIMINATE CERTAIN CANCERS WHICH ARE ASSOCIATED WITH LARGE AMOUNTS OF MISFOLDED PROTEINS. A CHALLENGE IN THINKING ABOUT AGING IS TO ENHANCE THE PROTEOSOME'S EFFICIENCY. I THINK THAT IS NOW TURNING OUT TO BE PRACTICAL, AND LET ME SHOW YOU A MORE SOPHISTICATED VIEW THAT'S JUST REALLY EMERGED IN, SAY, THE LAST YEAR OR SO ABOUT PROTEOSOME FUNCTION. IT TURNS OUT THE PROTEOSOME WAS MORE INTELLIGENT THAN IT LOOKS, MORE INTELLIGENT THAN WE HAD LONG BELIEVED, EVEN THOSE OF US WORKING ON IT, AND IT'S MAKING DECISIONS OF A KEY KIND. 19S REGULATORY PARTICLE RECOGNIZES SUBSTRATES IN TWO STEPS. ONE, IT RECOGNIZES THE UBIQUITINE CHAINS AND BINDS IT TO TWO RECEPTORS. THAT, ITSELF, IS THE INITIAL AND EASILY REVERSIBLE REACTION. THEN THE UBIQUITINE DEGRADING ENZYME, THE DEUBIQUINATIG ENZYME START CHOPPING IT UP SO IT CAN BE RECYCLED. TURNS OUT THE MAJOR ENZYME DOING THIS, USP14, IS ALSO A KEY REGULATOR OF THE REST OF THIS MACHINERY. IN FACT, THIS MACHINERY IN THE CELL IS IN AN OFF STATE, UNTIL SUBSTRATES BIND AND THEY INTERACT WITH USP14, WHICH ACTS VAITS THE PROTEOSOME. IT OPENS A GATE HERE, IT CHANGES SEVERAL DIFFERENT REACTIONS, BUT MOST IMPORTANTLY, THE PROTEINS WILL CONTINUE TO GET DEUBIQUINATED UNLESS THEY HAVE A LEWIS END ON THE SUBSTRATE. THIS COMES FROM WORK FROM MATUSEK AND FINDLY. THE MAIN POINT IS THAT THIS IS A TWO-STEP REACTION, AND ONLY PROTEINS ARE DESTROYED IF THEY HAVE A LOOSE END. AND THIS SEEMS TO BE AN IMPORTANT POINT IN QUALITY CONTROL. IF THEY ARE AGGREGATED, THE PROTEOSOME HAS A MAJOR CHALLENGE, AND WE DON'T KNOW IF IT REALLY CAN OR CANNOT DISAGGREGATE THE KINDS OF PROTEINS WE ASSOCIATE WITH DISEASE. SO LET ME END WITH A CRITICAL QUESTION THAT YOU'RE PROBABLY ASKING, AND I WISH I COULD ANSWER. FIRST OF ALL, WE HAVE MANY STEPS IN THIS PATHWAY. HOW DOES AGING AFFECT THEM? WELL, BASICALLY FOR ALL THE CRITICAL FIRST STEPS WHICH ARE RATE LIMITING FOR MANY, MANY -- FOR MOST PROTEINS, WE SIMPLY DON'T KNOW. WE DON'T KNOW THE AFFECTS OF AGING ON THE CYTOBOLIC LI LIGASES. WE DON'T KNOW WHAT THEY ARE OR HOW THEY CHANGE. THE PROCESS OF QUALITY CONTROL IN THE SECRETORY PATHWAY, THE EFFECTS OF AGING HAS NOT BEEN STUDIED. OUR RIBOSOME ASSOCIATED LIGASES THAT CARRY QUALITY CONTROL -- OF THE NASCENT PROTEINS AS SHOWN BY DR. FRYDMAN, THE EFFECT OF AGING IS UNKNOWN. FOR THE PROTEOSOME, IN THE LAST FEW YEAR PROBABLY, CRITICAL INFORMATION HAS ARISEN THAT REALLY IS CONVINCING IN MY VIEW THAT IN THE BRAIN WITH NORMAL AGING, THERE ARE DECREASES IN THE CAPACITY OF THE UBQUITIN PROTEINS TO BE DEGRADED BY THE PROTEOSOME. PATHOGENIC PROTEINS LIKE -- AS WELL AS PRP SCRAPIE, CAN DEFINITELY INTERFERE WITH THE 26S REGULATORY COMPONENT. THIS IS NOT AN ESSENTIAL PART OF AGING, IT'S MAYBE AN ESSENTIAL PART OF THE DISEASE. IF YOU LOOK IN THE MUSCLE, THE PROTEOSOMES ARE WORKING FINE, ACCELERATED IN SARCOMA PENA, THE WASTING OF MUSCLE THAT WE'VE TALKED. THERE'S ONE OTHER STEP BE, THAT'S THE P97ATPA COMPLEX WHICH HELPS IN THE DISASSEMBLY OF MANY LARGE COMPLEXES, PROBABLY AGGREGATES TOO, AND NO ONE HAS STUDIED ITS PRECISE FUNCTION AS IN DISEASES OR AGING. >> THANK YOU VERY MUCH, FRED. [APPLAUSE] SHE'S GOING TO ADDRESS THE INTERPLAY BETWEEN PROTEOSTASIS IN THE CYTOSOL AND ORGANIZEELLES USING AS A MODEL HER FAVORITE IN THE MITOCHONDRIA AND HOW IT IS REMOVED BY MY TO MITOPHASY. >> I'D LIKE TO THANK THE ORGANIZERS FOR INCLUDING ME. I'M GOING TO OFFER IN THE SPIRIT OF THE PREACHING THAT WE HEARD EARLIER, I'M GOING TO OFFER THE CONCEPT THAT MITOCHONDRIA ARE THE CENTER OF THE ISSUE AND CLEARANCE BY AUTOPHAGY IS THE KEY TO THIS PROCESS. LET ME PUT THIS FIRST IN AN ORGAN CONTEXT, THE MOST PROFOUNDLY AFFECTED BY AGING ARE THOSE THAT HAVE LONG LIVED CELLS, SO THE HEART AND THE BRAIN, I GUESS ANOTHER ORGAN THAT YOU WOULD CONSIDER WOULD BE THE RETINA. THESE ARE ALL CELLS THAT HAVE A HIGH METABOLIC DEMAND BUT THE CELLS THEMSELVES TURN OVER WITH A VERY SLOW RATE IF AT ALL. SO IT'S IMPORTANT FOR THE CELL TO BE ABLE TO MAINTAIN ALL IT'S ORGANELLES IN OPTIMAL WORKING CONDITION. SO THIS IS QUITE A CHALLENGE FOR LONG LIVE CELL. YOU CAN'T REPLACE THAT CELL, YOU NEED TO REPLACE THE COMPONENT PARTS. UP TO NOW, WE HAVEN'T HEARD MUCH ABOUT AUTOPHAGY AS A MECHANISM OF CLEARING THE ORGANIZE NELS, BUT THIS IS THE PRIMARY IN THE CASE OF THE MITOCHONDRIA, PROBABLY THE ONLY MEANS BY WHICH YOU CAN ELIMINATE DAMAGED MITOCHONDRIA OR OTHER DAMAGED ORGANELLES. HERE I'VE LISTED KIND OF ALL THE BAD THINGS THAT CAN HAPPEN WHEN DYSFUNCTIONAL MITOCHONDRIA OR EVEN OLD BEAT UP TIRED MITOCHONDRIA ARE NOT CLEARED IN A TIMELY FASHION. SO OBVIOUSLY M MITOCHONDRIA ARE THERE TO PROVIDE ATP FOR THE CELL AND AS THEY DETERIORATE IN THEIR FUNCTIONAL CAPACITY, THEN ATP LEVELS DRIFT DOWN. THIS CAN IMPACT, FOR INSTANCE, THE PANCREATIC BETA CELL, WHICH REQUIRES VERY HIGH HP LEVELS FOR INSULIN SECRETION, BUT IT HAS ITS IMPACT IN MANY OTHER CELL TYPES AS WELL. AND THE LIMITATION OF ATP IS GOING TO IMPACT THE FUNCTION OF OTHER ATP-DEPENDENT ENZYME SYSTEMS WITHIN THE CELL, INCLUDING THE PROTEASOME. SO THE ADDITIONAL EFFECT OF ACCUMULATING DAMAGED MITOCHONDRIA IS THE PRODUCTION OF -- OXYGEN SPECIES AND ALTHOUGH THIS BU BY ITSELF MAY NOT BE A PROBLEM BECAUSE OF THE ABUNDANT DETOXIFICATION SYSTEM IT DOES PUT ADDITIONAL BURDEN ON THE CELL THAT MEETS THE DEMANDS OF DETOXIFICATION. FINALLY, ONE THING THAT WE HAVEN'T TALKED ABOUT VERY MUCH BUT WHICH IS REALLY IMPORTANT IS THE CONNECTION TO INNATE IMMUNITY AND INFLAMMATION. MITOCHONDRIA ESSENTIALLY ARE FREE SWIMMING BACTERIA LIVING WITHIN THE HOST CELL. AND THEY'RE REALLY RECOGNIZED BY THE CELL AS FOREIGNERS, AND PARTICULARLY THE MITOCHONDRIAL DNA, NOTABLY WHEN IT'S OXIDIZED MITOCHONDRIAL DNA, IS RECOGNIZED BY THE NRLP3 -- CELL AND BY TLR9 AND THIS CAN ACTUALLY DRIVE INFLAM -- ASSEMBLY AND PRODUCTION OF CYTOKINES INCLUDING IL1 PAY TA AND IL18. SO THIS IS A VERY IMPORTANT ASPECT, THEN THE REACTIVE OXYGEN SPECIES ONLY FACILITATE THIS BY CONTRIBUTING TO -- SIGNALING. FINALLY IN CELLS WHERE IT'S REALLY IMPORTANT FOR THOSE CELLS TO STAY ALIVE, A DAMAGED MITOCHONDRIA HAVE A LOWER THRESHOLD FOR RELEASING CYTO -- WHICH WOULD TRIGGER PROGRAM CELL DEATH, SO IT'S IMPORTANT TO ELIMINATE THOSE MITOCHONDRIA BEFORE THEY REACH THE THRESHOLD OF RELEASING THEIR -- AND TRIGGERING CELL DEATH OR HAVING PERMEABILITY TRANSITION -- OPENING AND TRIGGERING NECROSIS. MITOCHONDRIA ARE VERY IMPORTANT IN DEALING WITH CALCIUM -- THE CALCIUM PLEXUS, SO IMPAIRMENT OF MITOCHONDRIAL FUNCTION WILL HAVE ITS IMPACT ON A VARIETY OF CALCIUM-DEPENDENT SIGNALING PATHWAYS. SO YOU CAN IMAGINE THEN THAT THE MITOCHONDRIA MIGHT BECOME DAMAGED AND DEPO DEPOLARIZED AND BE A COSTIMULATORY SIGNAL FOR NLRP3. AS THE MITOCHONDRIA FURTHER DETERIORATES, MITOCHONDRIAL DNA MAY BE RELEASED INTO THE CYTOSOL WHERE IT CAN INTERACT WITH EITHER NRLP3 OR TLR9 AND THIS DRIVES IN THE PRODUCTION OF INFLAMMATORY CYTOKINE. NOT ONLY IS IT THIS CELL AT RISK BUT NEIGHBORING CELLS THEN WOULD BE IMPACTED BY THE PRESENCE OF THESE INFLAMMATORY CYTOKINES. THE CURE FOR THIS IS MITOPHAGY. SO THE USE OF AN AUTO PHAGOSOME TO -- BEFORE THESE BAD THINGS HAPPEN IS A KEY CELLULAR RESPONSE THAT'S ENGAGED IN A VARIETY OF DIFFERENT CELLS IN RESPONSE TO CELLULAR STRESS AND AS PART OF THE NORMAL HOUSEKEEPING PROCESS IN ANY CELL TO TIGHTLY CONTROL OF NUMBER OF MITOCHONDRIA AND THEIR FUNCTION ANIN RECRUITMENT OF A -- THAT SIGNALS RECRUITMENT TO P62 -- SO MITOPHAGY IS A SUBSET OF THE AUTOPHAGY SYSTEM, MACRO YOU A TO FA GEE, AND THAT SYSTEM IS INDUCED BY STARVATION BUT IS SUPPRESSED BY OVERNUTRITION, AND YOU'VE HEARD A LITTLE ABOUT THE IMPACT OF METABOLIC SYNDROME AND CALORIC EXCESS, AND SO THERE ARE A VARIETY OF SITUATIONS WHERE AUTOPHAGY IS IMPAIRED, INCLUDING ADVANCED AGE AND ALSO THE MUTATIONS IN THE -- PATHWAY THAT ARE ASSOCIATED WITH PARKINSON'S AND OTHER NEURODEGENERATIVE DISORDERS. SO THE IMPORTANCE OF CLEARING THE MITOCHONDRIA THEN IS REALLY THIS ESSENTIAL ELIMINATION OF THESE DAMAGED ELEMENTS, AND METABOLIC SYNDROME IS SOMETHING THAT'S VERY IMPORTANT IN TERMS OF SUPPRESSING AUTOPHAGY. THERE ARE A NUMBER OF STUDIES THAT HAVE SHOWN THAT METABOLIC SYNDROME DECREASES AUTO PHAGIC FLUX AND CLEARANCE OF TARGET PROTEIN. WE'VE USED TRANSIENT ISCHEMIC STRESS OR ISCHEMIC CONDITIONING IN THE HEART TO LOOK AT THE CLEARANCE -- TO LOOK AT THE PROTECTIVE EFFECTS, AND THIS DEPENDS ON MITOPHAGY AND AUTOPHAGY, AND THE PROTECTIVE EFFECT IS LOST IN ANIMAL MODELS OF METABOLIC SYNDROME WHERE THERE IS NO LONGER THE ABILITY TO CLEAR THOSE DAMAGED ORGANELLES AND TO REPLACE THEM. I THINK THE OTHER IMPORTANT ASPECT OF THIS IS THAT NOT ONLY IS MITOPHAGY REQUIRED BUT THIS IS COUPLED TO BIOGENESIS, AND THIS MAY BE RELEVANT IN, FOR INSTANCE, SKELETAL MUSCLE WHERE -- IS IMPORTANT FOR OPPOSING THE EFFECTS OF SARCOM SAR COPENA. STAYING RIGHT ON TEAM HERE, SO THE KEY THEN IS, HOW CAN WE ENHANCE OR HARNESS THE AUTOPHAGY, MITOPHAGY PATHWAY? OBVIOUSLY THERE ARE MEN BIT BENEFITS LISTED HERE, BUT THERE ARE A NUMBER OF AGENTS IDENTIFY TO ACTIVATE MI TOE FA GEE. THES INCLUDE -- IN PART BECAUSE THEY ELICIT THE MITOCHONDRIAL UNFOLDED PROTEIN RESPONSE. THERE HAVE BEEN A VARIETY OF DIFFERENT AGENTS -- HAS SHOWN THAT THOSE AGENTS THAT PROLONG LIFE ALSO INDUCE MY TOE DONE DREEL -- PROTEIN RESPONSE, GUILT BY ASSOCIATION ALSO INDUCED MITOPHAGY. THIS IS GOING TO HAVE BENEFITS NOT ONLY FOR THE TARGET CELL BUT ALSO FOR ASSOCIATED CELLS, AS REACTIVE OXYGEN SPECIES AND CYTOKINES ARE PRODUCED, THIS IS GOING TO IMPACT ENDOTHELIAL FUNCTION BY CONSUMING BIOAVAILABLE NITRIC OXIDE AND ALSO BY INCREASING INFLAMMATORY SIGNALING OF THOSE ADJACENT TARGET CELLS, SO YOU CAN SEE THAT IN ANY TISSUE WHERE MITOCHONDRIAL DYSFUNCTION IS ONGOING T WILL HAVE IMPACT ON ADJACENT CELL TYPES. SO THERE'S A CAVEAT TO THIS THAT EXCESS MITOPHAGY MIGHT NOT ALWAYS BE A GOOD THING, BECAUSE YOU HAVE TO HAVE A BALANCE BETWEEN DEGRADATION OF THE MITOCHONDRIA AND THEIR REPLACEMENT BY BIOGENESIS, AND IT MAY BE IN CERTAIN SETTINGS, FOR INSTANCE, NUTRITIONAL DEPRIVATION OF THE VERY ELDERLY, WHERE BIOGENESIS CAN'T MATCH THE MITOPHAGY, AND ONE POSSIBLE SETTING OF THAT IS IN THE ELDERLY, PATIENTS RECEIVING STATIN THERAPY WHO ACTUALLY DON'T DO VERY WELL WITH THAT BECAUSE WE HYPOTHESIZE BECAUSE THEY'RE UNABLE TO REGENERATE THEIR MITOCHONDRIA. SO WITH THAT, I WILL THANK YOU FOR YOUR ATTENTION. [APPLAUSE] >> SO OUR NEXT SPEAKER IS GOING TO BE DR. JUDITH FRYDMAN, THAT IS COMING FROM STANFORD UNIVERSITY, AND SHE'S GOING TO TRY TO ANSWER THE QUESTION OF HOW THESE NETWORKS ARE REGULATED BY ILLUSTRATING THE GAIN OR THE INTERPLAY BETWEEN THE DIFFERENT CHAPERONS. >> THANK YOU VERY MUCH FOR INVITING ME TO THIS REALLY EXCITING SUMMIT, SO AS WE'VE HEARD FROM THE PREVIOUS SPEAKERS AND FROM MANY OTHER SPEAKERS, IN THIS MORNING AND YESTERDAY, PROTEINS ARE IMPORTANT, AND IN ORDER FOR PROTEINS TO FUNCTION, THEY HAVE TO BE FOLDED INTO THEIR UNIQUE STATE. WHEN THEY ARE NOT FOLDED IN THEIR UNIQUE -- THEY CAN EITHER LOSE FUNCTION OR GAIN AN ADDITIONAL TOXIC FUNCTION AND THIS IS VERY DLEE TIER JUST FOR THE CELL. IT HAS BECOME VERY CLEAR FROM A NUMBER OF LABS, PARTICULARLY MORIMOTO, JEFF KELLY, THAT PROTEIN HOME STASIS, THE MACHINERY THAT HELPS PROTEINS FOLD, IS IMPAIRED PROGRESSIVELY WITH AGING IN WAYS THAT WE DON'T QUITE UNDERSTAND, AND I'LL TRY TO GIVE YOU A FLAVOR OF THIS PROBLEM TODAY. SO A NUMBER OF DISEASES AS CHARACTERIZED AS INCREASING IN INCIDENCE OF AGING ARE ASSOCIATED WITH PROTEIN FOLDING FOLDING -- TO THE AMYLOID -- ALZHEIMER'S, ALS, HUNTINGTON, THESE ARE PROTEINS THAT HAVE NOTHING TO DO WITH EACH OTHER, THEY DON'T RESEMBLE AT ALL EACH OTHER WHEN THEY ARE FOLDED, BUT THEY FORM THESE VERY STEREOTYPICAL AGGREGATES THAT ARE THE HOMEWORK OF AFFECTED NEURONS IN OLD AGE. MUTATIONS, WE HEARD TODAY THAT WITH AGE, THERE IS AN INCREASING INCIDENCE OF MUTATIONS THAT CAN BE TO CANCER, METABOLIC DISEASES, IT'S VERY OFTEN ALSO IMPAIRED TO FOLD OR TO REMAIN STABLE, WE'VE ALSO HEARD OF DAMAGE SUCH AS THAT OCCURS -- AND STROKE. SO WITH AGING, THERE IS A DECLINE IN PROTEIN FUNCTION, INCREASED PROTEIN AGGREGATION, INCREASED -- TO DISEASE AND WE CAN ASK, WHY IS THIS HAPPENING WITH AGE? THE CULPRIT HERE IS MOST LIKELY THE PROTEIN HOMEOSTASIS MACHINERY. THIS IS A VERY COMPLEX SET OF ENZYMES THAT ALL TOGETHER MAINTAIN THE HEALTH AND FUNCTIONALITY OF THE PROAM YOAM. PROAT YOAM. THERE ARE ALMOST 2,000 COMPONENTS IN THE MACHINERY. SO THE QUESTION IS, WHY IS IT SO COMPLICATED, WHAT ARE ALL THESE COMPONENTS DOING, AND WHICH AND HOW ARE THEY AFFECTED WITH AGE? SO WHAT IS THE PROTEOSTASIS MACHINERY? IT'S IN A WAY A COLLECTION OF ENZYMES, MANY OF THEM ARE CHAPERONES, MANY ARE COMPONENTS OF THE UBIQUITIN PATHWAY THAT CAN RECOGNIZE NON-NATIVE PROTEINS, AND THEY RECOGNIZE DIFFERENT ASPECTS OF NON-NATIVE PROTEIN, AND THEN SOMEHOW FACILITATE FOLDING, ASSEMBLY AND DISAASSEMBLY, STABILIZATION AND DEGRADATION. SO CHAPERONS HAVE A BIOCHEMICAL CYCLE THAT THEY USE, FOR INSTANCE, FOR FOLDING, AND SOME CHAPERONES INTERACT -- VERY INTIMATELY LINKED TO THE PROTEIN HOMEOSTASIS MACHINERY TO PROMOTE THE FOLDING OF AGGREGATION AGGREGATION-SENSITIVE PROTEINS. -- IN HIS VERY NICE TALK AND ACTUALLY HERE WHERE I'M LOOKING AT THE PATHWAYS OF FOLDING AND QUALITY CONTROL, TUMOR SUPPRESSOR -- WHERE MUTANTS LEAD TO CANCER, YOU CAN SEE THE DIFFERENT -- LEAD TO FOLDING SUCH AS HSB70 -- OR TO DEGRADATION SUCH AS -- 70 AND OTHER DOWNSTREAM -- SO WHERE DOES THE COMPLEXITY COME FROM, WHY IS THERE SUCH COMPLEXITY AND WHY DO DIFFERENT CHAPERONS DO DIFFERENT THINGS TO PROTEINS? A KIND OF VERY ELEMENTARY SCHOOL LEVEL WAY OF THINKING ABOUT THIS IS HERE. SO THE CHAPERON MACHINERY IS STRUCTURED IN SUCH A WAY WHERE THERE IS A MOTOR CHAPERON, FOR INSTANCE, HSB70, AND THERE IS A LONG LIST OF COFACTORS AND COCHAPERONS THAT REGULATE THE ACTIVITY OF 70. AND WHEN WE WANT TO THINK ABOUT TARGETING THESE MACHINERY, IT WILL BE MUCH MORE EFFECTIVE TO TARGET THE SPECIFIC FACTORS, LET'S SAY, FOR AGGREGATION OR FOR REFOLDING, THAN TO TARGET THE MOTOR THAT MAY HAVE OTHER -- MORE -- TROPIC FUNCTION. SO WHAT DO WE NEED TO DO TO UNDERSTAND THE PROTEIN HOMEOSTASIS MACHINERY AND ITS REGULATION DURING DISEASE? WE NEED TO UNDERSTAND THE FUNCTION, HOW ARE CHAPERONS ORGANIZED IN THE CELL, WHAT'S THEIR DIVISION OF FLAVOR, WE NEED TO UNDERSTAND THEIR PHYSICAL LABOR. I MENTIONED THAT SOME CHAPERONS INTERACT WITH RYE BO WITH RYE BOW RIBOSOME S, THEY COMMUNICATE -- THE PROTEINS TRANSCRIBED IN THE NUCLEUS, THEY TAKE THEM TO THE MITOCHONDRIA, AND THEY'RE A BIG PART IN THE COMMUNICATION. WE NEED TO UNDERSTAND THE REGULATORY NETWORK SO THERE'S BEEN A LOT OF TALK ABOUT STRESS, BUT DIFFERENT TYPES OF STRESSES INDUCE DIFFERENT TYPES OF CHAPERONES, AND THESE DIVERSITY AND NUANCED NATURE OF THE STRESS RESPONSES MAY EXPLAIN SOME OF THE PARADOX THAT WE HAVE HEARD ABOUT HERE, AND WE NEED TO REALLY UNDERSTAND HOW STRESS REGULATES CHAPERONE NETWORKS, AND AN EMERGING CONCEPT THAT HAS ACTUALLY BEEN QUITE SURPRISING IS THAT SHA CHAPERONES DON'T OPERATE LIKE THEY WOULD IN A TEST TUBE, THEY'RE NOT FLOATING AROUND AND ENCOUNTERING THEIR SUBSTRATES. THERE IS A LOT OF SPATIAL ORGANIZATION TO PROTEIN HOMEOSTA CYST AND QUALITY CONTROL, AND THAT RAISES THE DETERMINE OF WHAT DETERMINES THE LOCALIZATION AND WHAT IS THE CELL BIOLOGY IN A WAY OF PR PROTEIN HOMEOSTASIS. AND OF COURSE A VERY IMPORTANT CHALLENGE IS WHAT ARE THE ASSAY READOUTS AND APPROACHES THAT WE NEED TO ADDRESS THIS. SO VERY QUICKLY GIVE YOU A FLAVOR OF HOW PEOPLE IN THE FIELD INCLUDING MY LAB ARE TRYING TO ADDRESS THESE ISSUES AND I WANT TO MAKE SOME POINTS THAT I IT THINK ARE IMPORTANT IN MOVING FORWARD TO UNDERSTAND THIS MACHINERY. FOR INSTANCE, WHEN WE TALK ABOUT CHAPERONE NETWORKS. FOR A NUMBER OF LABS INCLUDING MY OWN, FOUND THAT -- CELLS, THERE IS DISTINCT CHAPERONE NETWORKS, PROTEIN BIOSINT CYST -- WE CALL THIS CLEAVE -- CLING TO PROTEIN SYNTHESIS, AND THERE IS A STRESS INDUCIBLE CHAPERONE NETWORK COMPOSED OF THE SO CALLED HSB -- PROTEINS THAT PROTECTS THE PROTEIN FROM STRESS. INTERESTINGLY, WHILE STRESS -- MANY STRESSES, ALL STRESSES ACTUALLY, INDUCE CHAPERONES OF THIS FAMILY TO VARYING DEGREES, STRESS REDRESSES THE CHAPERONES OF THIS NETWORK, AND THEREFORE, STRESS WILL ALSO HAVE A NUANCED EFFECT ON THE AVAILABLE CHAPERONES TO THE CELL. ONE IMPORTANT THING IS THAT THESE TWO NETWORKS ARE CONNECTED AND -- HAD A VERY NICE PAPER SHOWING THAT ONE OF THE KEY CHAPERONES IN THE COTRANSLATIONAL ACTING NETWORK IS SEQUESTERED BY AMYLOID AGGREGATES, WHICH REALLY WOULD SUGGEST THAT HAVING AGGREGATES CAN AFFECT A VERY UPSTREAM PROCESS THAT IS UNRELATED TO STRESS AND MISFOLDING. SO WE THINK ABOUT THIS NETWORK HAVING SEVERAL HIERARCHICAL LEVELS THAT -- RETAINING -- WE WANT THE NETWORK THAT IS ROBUST AND IN FACT ONE OF THE PROBLEMS WITH AGING IS THAT IT STOPS BEING ROBUST AND IT BECOMES VERY SENSITIVE TO MISFOLDING. NOW THE OTHER ASPECT OF THE ORGANIZATION IS WHAT HAPPENS ONCE YOU HAVE A PROTEIN IN THE CELL AND IT IS MISFOLDED, AND FRED HAD A VERY NICE SUMMARY OF ALL THE BAD THINGS THAT CAN HAPPEN TO PROTEINS IN THE CELL. SO NOW THE CELL HAS TO DECIDE WHETHER DEFOLD IT, DEGRADE DID, OR SEQUESTER IT, AND WE REALLY NEED TO UNDERSTAND IT'S VERY IMPORTANT TO UNDERSTAND THE LOGIC OF THIS DECISION. IT TURNS OUT THAT IT IS EMERGING THAT THERE ARE PATHWAYS THAT CONNECT THE -- AND SPECIFIC SUBCELLULAR LOCATIONS. SO IT HAS BEEN KNOWN FOR MANY TIMES THAT AMYLOIDS FORMING A GATES -- IN THE CELL THAT WORK FROM A NUMBER OF LABS, INCLUDING MY OWN, HAVE SHOWN THAT QUALITY CONTROL PATHWAYS ARE SPATIALLY LOCALIZED. ONE INTERESTING ASPECT OF THIS LOCALIZATION IS THERE IS THIS DISTINCT QUALITY CONTROL COMPARTMENT WHERE DIFFERENT PROTEINS ARE CONCENTRATED. SO WE'VE CHARACTERIZED SYSTEM -- HAPPENS IN MAMMALIAN CELLS AND WORMS, SO ONE OF THE PATHWAYS THAT WE -- NUCLEAR QUALITY CONTROL CONCENTRATES SOLUBLE MISFOLDED PROTEINS, YOU CAN THINK OF IT AS A HOLDING CELL THAT'S CONCENTRATE -- PROTEINS UNTIL THE CELL DECIDES WHETHER TO REFOLD THEM OR DEGRADE THEM, WHETHER AS THE INSOLUBLE PROTEIN DEPOSIT SEQUESTERS -- INSOLUBLE AGGREGATES SUCH AS HUNTINGTON AND AMYLOID PROTEIN. THE INTERESTING THING IS THAT ALTHOUGH CHAPERONES ARE INVOLVED IN THESE TWO PATHWAYS, THEY ARE INVOLVED IN DIFFERENT WAYS IN THESE TWO PATHWAYS, UBIQ -- INTERESTINGLY, THE DIFFERENT DEGRADATION PATHWAYS INTERFACE DIFFERENTLY WITH THESE COMPARTMENTS, THE PROTEASOME SEEMS TO ACT PRIMARILY ON JUNK DIRECTED PROTEINS WHEREAS THE -- SEEMS TO BE LINKED TO AUTOPHAGY. OKAY? THE OTHER INTERESTING THING IS THAT WE HAVE SHOWN RECENTLY THAT THE JUNK PATHWAY DECLINES WITH AGING, IN SIMPLE MODELS SO THAT THE -- THROUGH THE JUNK IS ACTUALLY IMPAIRED IN AGED CELLS AND IT'S ALSO REGULATED BY SIGNALING PATHWAYS AND IN PARTICULAR TREATMENT WITH -- INCREASES THE -- OF PROTEINS OF MISFOLDED PROTEINS THROUGH THESE PATHWAYS. SO -- DOESN'T ONLY WORK ON AUTOPHAGY OR INITIATION OF TRANSLATION. IT SEEMS TO AFFECT GENERALLY THE PROTEIN HOMEOSTASIS MACHINERY. OKAY. SO I JUST WANT TO MAKE TWO ADDITIONAL POINTS. I'LL MAKE THEM VERY BRIEFLY. ONE ADDITIONAL POINT. [LAUGHTER] TWO HALF POINTS. OKAY. ONE OF THEM IS THAT POST TRANSLATIONAL MODIFICATIONS ARE ALSO VERY IMPORTANT FOR REGULATING CHAPERONE ACTIVITY, AND SO WHEN WE THINK ABOUT THE REGULATION OF PROTEIN HOMEOSTASIS, WE ALSO NEED TO THINK BEYOND TRANSCRIPTIONAL RESPONSES. OKAY. THE OTHER POINT I WANT TO MAKE IS THAT WE HEARD ABOUT MUTATIONS, VERY BRIEFLY, SO WHEN WE THINK ABOUT MUTATIONS, THOSE THAT ACCUMULATE WITH AGE -- THAT HAPPENED DURING -- INFECTIONS, WE ALSO NEED TO THINK ABOUT THE CHAPERONE MACHINERY, BECAUSE CHAPERONES ARE VERY IMPORTANT IN EITHER REFUNCTIONALLIZING OR MANAGING MUTATIONS SUCH AS THOSE THAT DRIVE THESE TWO DIFFERENT TYPES OF PROCESSES. SO THAT'S MY POINT. AND OF COURSE WE WANT TO UNDERSTAND HOW THESE MODIFY -- ARE MODIFIED WITH AGING. [APPLAUSE] >> OKAY, THANK YOU VERY MUCH, JUDY. OUR NEXT SPEAKER IS RANDY KAUFMAN, COMING FROM THE BURNHAM INSTITUTE. HE'S GOING TO BE ILLUSTRATING GOING BACK TO ORGANELLE QUALITY CONTROL, BUT HOW THAT PLAYS IN THE INTERPLAY AMONG CELLS AND IN THE WHOLE ORGANISM, AND HE'S GOING TO BE MOSTLY FOCUSING ON ER STRESS. >> THANK YOU VERY MUCH, AND I'D LIKE TO THANK THE ADVISORS AND MARIA AND RICK FOR LETTING ME TALK TO YOU TODAY. WHAT I'D LIKE TO REALLY DESCRIBE TO YOU IS THE PROCESS OF PROTEIN FOLDING IN THE ENDOPLA MIC RE TICK LUM AND DESCRIBE HOW THAT IS DIFFERENT. IT'S QUITE A CHALLENGING ENVIRONMENT FOR PROTEIN FOLDING. SO BASICALLY THE ENDOPLASMIC RE TICK LUM IS A SITE FOR CALCIUM STORAGE. THE MAJOR SITE FOR CALCIUM STORAGE IN THE CELL. AND IT'S RELEASED IN RESPONSE TO GROWTH FACTOR STIMULATION, AND HORMONAL STIMULATION. OBVIOUSLY THAT'S TELLS YOU IT'S GOING TO BE SOMEWHERE IMPORTANT IN SIGNALING. CALCIUM FORMS A BUFFER FOR PROTEIN FOLDING IN THE ER AND ON MOST CHAPERONES IN THE ER REQUIRE CALCIUM FOR THEIR FUNCTION IN PROMOTING PROPER FOLDING. PROTEIN FOLDING IS OXIDATIVE. THAT MEANS THAT SULFIDE BONDS ARE FORMED AND IT'S VERY DEPENDENT ON THE -- POTENTIAL OF THE ER. SO ANYTHING THAT CHANGES REDOCKS POTENTIAL ALTERS BY SULFIDE BOND FORMATION AND DISRUPTS PROPER PAIRING OF THE SULFIDE BONDS. THERE'S QUALITY CONTROL WHERE ONLY THOSE PROPERLY FOLDED PROTEINS ARE ABLE TO TRAFFIC TO THE GOLGY, AND THOSE THAT ARE MISFOLDED ARE VERY -- ARE RETAINED IN THE ER AND EVENTUALLY DEGRADED THROUGH EITHER ER ASSOCIATED PROTEIN DEGRADATION BY THE PROTEASOME OR THROUGH AUTOPHAGY. THIS IS A VERY TIGHT SURVEILLANCE SYSTEM. IT'S BASICALLY THE SITE THAT DETERMINES HOW MUCH LIPIDS THE CELL HAS. YOU CAN SEE HERE IS A PANO CELL, EXTENSIVE AMOUNTS OF ER. THINGS IMPORTANT TO NOTE, THE ER ARE VERY INTEGRATED WITH THE STRUCTURE OF MITOCHONDRIA. YOU FIND THAT THE MEMBRANES ARE BASICALLY JUXTAPOSED. AND THAT BASICALLY FROM EARLY STUDIES SUGGESTED THAT MITOCHONDRIA BEING A SOURCE OF ATP, THAT MAYBE THE PROTEIN FOLDING OR PROTEIN SYNTHESIS AT THE ER MEMBRANE REQUIRES AN ENERGY. PROTEIN FOLDING IN THE ER IS PROBABLY ONE OF THE MOST ENERGY-REQUIRING STEPS IN THE CELL. PROTEIN FOLDING ITSELF IS THE MOST ERROR-PRONE STEP IN GENE EXPRESSION. WHAT'S THE IMPACT OF PROTEIN MISFOLDING IN ONE CELL VERSUS ANOTHER CELL IN THE BODY? WELL, YOU CAN IMAGINE. ALL PROTEINS THAT GO THROUGH THE SECRETORY PATHWAY HAVE SOME ASPECT, ONE-THIRD OF OUR GENOME ENCODES PROTEINS THAT GO THROUGH THE SECRETORY PATHWAY, AND MANY ARE -- ENDOCRINE OR EX-OWE KRIN FUNCTIONS, THE SYNTHESIS OF STEROIDS, ALL THE WAY FROM CORD COSTEROID SYNTHESIS OF SYNAPTIC TRANSMITTERS, SO YOU CAN IMAGINE THAT PROTEIN MISFOLDING CAN AFFECT A LOT OF INTRACELLULAR AND INTERCELLULAR FUNCTIONS WITHIN THE ORGANISM. SO WHEN PROTEINS COME IN AN UNFOALED STATUNFOLDED STATE -- VERY IMPORT ANT FOR INTERACTION WITH PROTEIN CHAPERONES THAT RECOGNIZE SPECIFIC STRUCTURES TO PROMOTE THE PROPER FOLDING AS WELL AS TRAFFICKING OUT OF THE CELL. -- BONDS FORM AND THEY REARRANGE AND IT'S THE PROPERLY FOLDED FORM THAT'S RECOGNIZED FOR TRAFFICKING OUT ANYTHING THAT DISRUPTS THE ER HOMEOSTASIS. THIS COULD BE A WIDE VARIETY OF INDUCERS SUCH AS CALCIUM DEPLETION ALTERED -- AS IF YOU HAVE A GLUCOSE DEPRIVATION, NUTRIENT DEPRIVATION, AS WELL AS NUTRIENT EXCESS, LIPID EXCESS, REDOCKS STRESS -- INCREASE THE AMOUNT OF PROTEIN MISFOLDING. SO IN THE ACTIVATION OF -- PATHWAY WILL RESULT IN MORE ER MISFOLDED PROTEIN. THE MISFOLDED PROTEIN ITSELF HAS TWO BASICALLY -- SEVERAL DESTINATIONS, IT CAN BE REFOLDED. WE DON'T UNDERSTAND THE MACHINERY INVOLVED IN THIS, BUT IT'S VERY COMPLEX AND IT DOES HAPPEN, BUT WHAT ARE THOSE ENZYMES? WE REALLY DON'T KNOW TO DATE. ALTERNATIVELY IT'S DEGRADED THROUGH AUTOPHAGY OR ERAD. IF YOU REMOVE THESE FUNCTIONS FROM THE CELL, THE ACCUMULATION OF THAT PROTEIN WILL KILL THE CELL, SO IT'S THE TOXIC ACCUMULATION OF THESE PROTEINS. THERE'S AB ADAPTIVE RESPONSE, CALLED THE UNFOLDED PROTEIN RESPONSE, AND THAT IS BASICALLY AN ADAPTIVE RESPONSE TO RESOLVE PROTEIN MISFOLDING. IT'S CARRIED OUT AT THREE LEVELS. ONE IS YOU DECREASE PROTEIN SYNTHESIS BECAUSE YOU DON'T WANT TO SYNTHESIZE PROTEIN IF YOU CAN'T FOLD THE PROTEIN. YOU ACTIVATE TRA TRANSCRIPTION OF SEVERAL THOUSAND GENES. AT LEAST 50% OF THEM PROVIDE FUNCTIONS THAT ARE ALL DEPICTED HERE. ASSOCIATED DEGRADATION, TRAFFICKING, TRANSLOCATION, FOLDING, AND FINALLY IF THESE ARE NOT SUFFICIENT TO RESOLVE THE PROTEIN FOLDED STRESS -- RESPONSE. ONE OF THE MAJOR SIGNALS IN THE APOPTIC RESPONSE IS REACTIVE OXYGEN SPECIES. SO WHAT SIGNALS THE RESPONSE? THERE ARE BASICALLY THREE TRANSDUCERS THAT HAVE PRIMARILY BEEN DISCOVERED AND CHARACTERIZED QUITE WELL THROUGH DELETION STUDIES AND OVEREXPRESSION STUDIES. JUST BRIEFLY, TWO OF THEM ARE PROTEIN KINASE, THEY GET ACTIVATED BY DIMERIZATION, AND THEY GO THROUGH BASICALLY A SPECIFIC LINEAR PATHWAY, VERY SPECIFIC. I -- INITIATE SPLICING OF 1RMA TRANSCRIPTION FACTOR THAT ACTIVATES A VAST OF FUNCTIONS, PRIMARILY ERAD, PROTEIN CHAPERONES AND LIPID BIOSYNTHESIS. ATF6 IS A TRANSCRIPTION FACTOR THAT ACTIVATES -- CLEAVAGE AS IT TRAFFICS TO THE GOAL GEE UPON RELEASE FROM A STRESS ER AND THAT ACTIVATES A COMPLEMENTARY SET OF GENES. THESE ARE PRIMARILY ADAPTIVE. BUT IMPORTANTLY IS THAT THE -- PATHWAY IS VERY IMPORTANT FOR DIFFERENTIATION OF HIGHLY SEE KREE TRI CELLS. THIS PATHWAY IS REQUIRED FOR BETA CELL DIFFERENTIATION AND FOR A NUMBER OF OTHER HIGHLY SEE KRE TRI CELL TYPES. IF YOU KNOCK THIS OUT, IT'S -- ATF6 IS IN CONTRAST HAS NO REAL -- IN THE MOUSE WHEN YOU KNOCK IT OUT. BUT IT'S VERY IMPORTANT FOR ADAPTATION TO AN ACUTE STRESS. IN CONTRAST, PERC IS BASICALLY THE SIGNAL THAT BASICALLY DETERMINES WHETHER THE CELL WILL SURVIVOR DIE. WHEN IT GETS ACTIVATED, YOU BASICALLY SHUT DOWN PROTEIN SYNTHESIS TRANSIENTLY. THIS IS A REVERSIBLE PROCESS, AND THAT BASICALLY ENSURES THAT PROTEIN SYNTHESIS IS COUPLED WITH PROTEIN FOLDING. PARADOXLY -- TRANSLATED, ONE OF THEM IS ATF4 THAT ACTIVATES A LARGER SET OF GENES, IMPORTANTLY, IT ACTIVATES AUTOPHAGY, BIOSYNTHESIS, METABOLISM, AND ACTIVATES THIS TRAN SCRIMTION DESCRIPTION FACTOR -- A MAJOR APOPTIC FACTOR. THIS IS THE PATHWAY THAT PRIMARILY DETERMINES WHETHER THE CELL WILL SURVIVOR DIE. IT'S THE EXPRESSION OF CHOP IMPORTANT IN THAT DECISION, EXPRESSION OF CHOP CORRELATES WITH PRODUCTION OF REACTIVE OXYGEN SPE SPECIES. SO HOW DOES THIS OCCUR? IF WE ENVISION THE ER JUXTAPOSED TO MITOCHONDRIA, PROTEINED FOLD IN THE ER AND THEY AGGRAVATE, THEY BIND THEIR PROTEIN CHAPERONE CALLED BIF. THIS IS PROBABLY ONE OF THE MOST ATP-DEPENDENT STEPS IN PROTEIN FOLDING IN THE ER. ATP HIGH DROL CYST AND BINDING ARE REQUIRED FOR RELEASE BY SULFIDE BOND FORMATION REQUIRES TRANSPORT ON TO OXYGEN WHICH FORMS HYDROGEN PROX SIDE AS A REACTIVE OXYGEN SPECIES. AT THE SAME TIME, WHEN UNFOLDED PROTEINS ACCUMULATE, THERE'S CALCIUM LEAK OUT OF THE ER. THE MECHANISM FOR THIS IS REALLY NOT WELL-DEFINED. IT'S EITHER THROUGH THINGS LIKE IP 3 RECEPTORS, BUT THE CALCIUM LEAK RESULTS IN A MITOCHONDRIAL LOADS, AND IF THE MITOCHONDRIAL LOADING OF THIS CALCIUM THAT ACTUALLY ACTIVATES THE HYDROJE NAISES AND ACTIVATES ITP SYNTHESIS, SO IN A WAY, TO SAY IT'S USING THE ATP IN THE ER, YOU'RE STIMULATING ELECTRON TRANSPORT AND OXIDATIVE PHOSPHORYLATION IN MITOCHONDRIA. WE THINK IN THIS WAY THAT THE ER IS EVOLVED TO BE AN ATP SENSOR. HOWEVER, WHEN ECK SE CYST PROTEIN ACCUMULATES, YOU GET EXCESSIVE LOADING OF THE MY TOE MITOCHONDRIA, THAT RESULT IN SUPER OXIDE PRODUCTION. IN ADDITION, ATP IS REQUIRED FOR OTHER ASPECTS AND THAT'S FOR RELOADING CALCIUM INTO THE ER. SO YOU CAN SEE THAT THERE ARE A NUMBER OF STEPS THAT ARE ALL VERY INTIMATELY RELATED THAT WOULD HAVE REVOLVED AROUND ATP UTILIZATION, CALCIUM TRAFFICKING IN AND OUT, AND PRODUCTION OF ATP AND PRODUCTION OF SUPER OXIDE. WHAT ARE THE REASONS THE CELLS MAKE THE SUPER OXIDE? INDEED IF WE PUT ANTIOXIDANTS ON CELLS, WE CAN PREVENT THE CELL DEATH, WE CAN IMPROVE THE PROTEIN FOLDING, SO OBVIOUSLY THOSE EVENTS RELATED. WHERE ARE THE SUPER OXIDES COMING FROM? WE THINK MOST OF THEM ARE COMING FROM MITOCHONDRIAL FUNCTION. I THINK MOST EVIDENT FROM THAT IS STUDIES IN WHICH WE'VE LOOKED AT INSULIN RESISTANCE IN THE MOUSE AND BASICALLY INSULIN RESISTANCE BY EITHER DIET-INDUCED OBESITY, CAUSES THE BETA CELL TO INCREASE INSULIN PRODUCTION THROUGH BIOSYNTHESIS INCREASING TRANSLATION OF INSULIN, BUT -- ON THE EVIDENT R, THAT ACTIVATES THE UPR. 800 MILLION PEOPLE IN THE WORLD THAT ARE INSULIN-RESISTANT, TWO THIRDS OF THOSE WILL GO ON AND COMPENSATE THEIR BETA CELLS WILL MAKE MORE INSULIN, AND THEY'LL NEVER DEVELOP A DIABETES. BUT ONE-THIRD THAT DO DEVELOP DIABETES DEVELOP -- YOU CAN SEE THE INSULIN IS MISFOLDED, THEY HAVE FEWER GRANULES AND THEY ACTUALLY ACTIVATE ALL EVENTS OF OXIDATIVE STRESS, APOP PTOSIS, AND YOU HAVE HIGH -- INSUFFICIENCY OF SIGNALING YOU'RE PRONE TO THIS PARTICULAR BETA CELL -- AND AN ANTIOCTS DANANTIOXIDANTCAN PROTECT ALL OF THAT. BASICALLY THE INTERACTION BETWEEN PROTEIN MISFOLDING IN THE ER AND HOW BOTH PROTEIN AGO DATES AND OXIDATIVE DAMAGE ARE ASSOCIATED WITH A WIDE VARIETY OF DAMAGES FROM METABOLIC DISEASE, IMMUNE INFLAMMATORY RESPONSES, CANCER, AS WELL AS NEURODEGENERATIVE DISEASES. THANK YOU FOR YOUR ATTENTION. [APPLAUSE] >> OUR LAST SPEAKER OF THE SESSION IS GOING TO BE JEFF KELLY FROM THE SCRIPS INSTITUTE. HE'S GOING TO ADDRESS THE POSSIBILITY OF TARGETING PROTEOSTASIS WITH THERAPEUTIC -- AS A THERAPEUTIC APPROACH. SO HE'S GOING TO BE TELLING US ABOUT ALL THE DRUGS THAT THEY ARE DEVELOPING. INTO >> I'LL START MY TIMER. IT'S A REAL PRIVILEGE TO BE AT THIS SUMMIT, AND TO TALK ABOUT A SUBJECT THAT'S NEAR AND DEAR TO MY HEART. UNDERLYING HYPOTHESIS BEING THAT IT'S NOW POSSIBLE TO ADAPT PROTEOSTASIS THAT YOU'VE HEARD A LOT ABOUT IN THIS SESSION TO APPEAL YOUR ATE DISEASES LINKED TO AGING AND AGGREGATION. IN THE FIRST SHORT PART OF THIS, I'LL TALK ABOUT DECIDEDLY CHEMICAL STRATEGIES THAT HAVE BEEN AT LEAST IN ONE CASE REALIZED AND IN THE SECOND PART, I'LL TALK ABOUT WHAT I THINK IS A MUCH MORE GENERAL STRATEGY OF BIOLOGICAL APPROACH TO ADAPT PROTEOSTASIS THAT SHOULD BE MORE GENERAL. SO WORK FROM A NUMBER OF INVESTIGATORS' LABS HAVE NOW TAUGHT US THAT WHEN POLYPEPTIDE CHAINS COME OFF THE RIBOSOME, THERE'S A KINETIC -- ALSO EXHIBITING UNO MOLECULAR KINETICS AND CONCENTRATION-DEPENDENT AGGREGATION. OF COURSE FROM AGGREGATION DOMINATES, THAT CAN LEAD TO DEGENERATIVE DISEASES, LIKE ALZHEIMER'S AND PARKINSON'S AND HUNTINGTONS AND THE -- AMYLOID DISEASES THAT I'M GOING TO TALK ABOUT SOME MORE. THE OTHER THING THAT'S IMPORTANT IS THAT WHEN PROTEINS FOLD IN THE ER THAT YOU JUST HEARD ABOUT FROM RANDY, AND WHEN THEY'RE SECRETED, THEY CAN STILL, IN THE EXTRA CELLULAR SPACE, SAMPLE UNFOLDED STATES THAT CAN DRIVING A GAITION. THAT'S THE BIG PROBLEM WITH NEURO DEGENERATIVE DISEASES, BECAUSE THE PROTEOSTASIS NETWORK WE KNOW VERY LITTLE ABOUT BUT IT'S CLEAR IT'S NOWHERE NEAR AS SOPHISTICATED AS THAT IN THE CELL. SO OUR FIRST CHEMICAL STRATEGY, IT'S PRETTY SIMPLE. THE IDEA IS THAT YOU CAN POPULATE A FOLDED CONFORMATION, SAY IN A MUTE TATED OR AGGREGATION-PRONE PROTEIN, AND IF YOU CAN CRAFT THE SMALL MOLECULE THAT BINDS WITH HIGH AFFINITY TO THAT MISFOLDING PRONE PROTEIN, THEN YOU CAN PREVENT CONFORMATIONAL EXCURSIONS THAT DRIVING A GAITION. SO WE DECIDED TO TEST THAT CONCEPT IN THE CONCEPT OF THE -- AND AMYLOID DISEASE SE THAT ARE QUITE REPRESENTATIVE OF HUMAN AMYLOID DISEASES. THE HUGE ADVANTAGE HERE IS THAT YOU CAN DO A CLINICAL TRIAL WITHOUT WORRYING ABOUT THE BRAIN, BECAUSE THESE DISEASES DESTROY THE -- NERVOUS SYSTEM AND THE HEART. SO THIS PROTEIN IS SECRETED FROM THE LIVER AS A AT THE TROA MER. WHEN IT'S IN THE EXTRA CELLULAR SPACE, IT AGGREGATES INTO A WHOLE BUNCH OF STRUCTURES, INCLUDING AMYLOID FIBRILS, AND THIS PROCESS OF AGGREGATION DRIVES THE ULTIMATE DEGRADATION OF THESE TISSUES THAT DON'T EASILY REGENERATE. WE NOW HAVE, I THINK, INCREDIBLY STRONG DATA THAT THE PROCESS OF AGGRAVATION DRIVES THESE DISEASES. WHAT WE DON'T UNDERSTAND IS HOW DOES THAT COUPLE TO THE LOSS OF POST MY TO TICK TISSUE? THAT'S A BIG UNANSWERED QUESTION. SO WE KNOW THAT THE AT THE TROA MER -- IN ORDER TO DRIVING A GAITION. SO WE REASON THAT IF WE COULD FASHION SMALL MOLECULES THAT COULD BIND TO THESE SMALL MOLECULE BINDING SITES, THEN WE COULD STABILIZE ENOUGH -- THE PROTEIN ENOUGH SO THAT THE PROTEIN WOULD NO LONGER AGGREGATE. REALIZING THAT WHEN PEOPLE BECOME SYMPTOMATIC, THEY HAVE ALL THESE STRUCTURES IN THE PLASMA. SO THE IDEA IS THAT IF YOU CAN STOP FURTHER AGGREGATION, YOU CAN STOP THE PATHOLOGY. WHILE I DON'T WANT YOU TO SPARE US THE DETAILS OF THE DATA, THE BOTTOM LINE IS THE PATIENTS GET WORSE WHEN THESE -- WHEN THE DATA GO UP THE Y AXIS. THE TREATED GROUP IS IN THE FILLED SYMBOLS AND THE UNTREATED GROUP IS IN THE UNFILLED. IT'S CLEAR, VERY CLEAR, THAT THIS DRUG WORKS. AND, THEREFORE, IT'S BEEN APPROVED IN EUROPE AND IN JAPAN AND HOPEFULLY SOON BY THE FDA. WHAT'S THE PROBLEM? THE PROBLEM IS YOU HAVE TO DEVELOP A SMALL MOLECULE FOR EVERY -- PROTEIN. THERE ARE 50 OF THEM. THIS TOOK US 15 YEARS TO DO. WE CAN PROBABLY DO IT IT IN 10 NOW, BUT STILL A BIG PROBLEM, RIGHT? WE NEED A MORE GENERAL STRATEGY. SO THE MORE GENERAL STRATEGY IS CLEAR. WE SHOULD BE ABLE TO TAP THE PROTEIN HOMEOSTAY TIS NETWORK TO ADAPT IT TO KEEP US FREE FROM DISEASE WHEN WE GET OLDER. THIS SYSTEM IS AMAZING AT PREVENTING DEGENERATIVE DISEASES WHEN WE'RE YOUNG, RIGHT? NOBODY GETS PARKINSON'S DISEASE WHEN THEY'RE 25. NO ONE GETS THE AMYLOIDOSIS WHEN THEY'RE 25. THEY CARRY ALL THE MUTATIONS. SO WHAT PROTECTS THEM, IT'S THIS SYSTEM. SO HOW DO WE ADAPT THE SYSTEM? NOW THE WAY THAT I THINK ABOUT THE PROTEOSTASIS NETWORK IS QUITE SIMPLE MINDED, PERHAPS. BUT AS THE POLYPEPTIDE CHAIN COMES OFF THE RIBOSOME, THE CRITICAL DECISION IS, DO YOU FOLD THE PROTEIN OR DO YOU DEGRADE IT? AND WE'RE GOING TO TAKE ADVANTAGE OF THAT CENTRAL DECISION THAT THE PROTEOSTASIS NETWORK MAKES IN COMING UP WITH A STRATEGY FOR THESE DEGENERATIVE AGGREGATION-SOART AGGREGATION-ASSOCIATED DISEASES. OF COURSE WHAT HASN'T BEEN MENTIONED BUT HAS BEEN IMPLIED IS THAT THERE'S A PROTEOSTASIS NETWORK FOR EVERY SUBCELLULAR COMPARTMENT. THEY'RE DIFFERENT, RIGHT? EACH ONE OF THEM IS REGULATED BY ANOTHER STRESS RESPONSIVE SIGNALING PATHWAY. I'M GOING TO TALK MORE ABOUT THE UNFOLDED PROTEIN RESPONSE WHICH REGULATES THE PROTEOSTASIS CAPACITY IN THE ENDOPLASMIC RETICULUM. WE'RE GOING TO DO THAT AGAIN IN THE CONTEXT OF THE TRAN SIRE TIN AMYLOID DISEASES. I WANT TO POINT OUT SOMETHING NEW ON THIS SLIDE YOU'VE ALREADY SEEN. SO IN THE FAMILIAL DISEASES, OF COURSE, THE PATIENTS ARE HETEROZYGOUS. SO ONE OF THE ALLYL CODES FOR THE WILD TYPE PROTEIN, THE OTHER ALLYL FOR THE MEU TENT. THEREFORE, THE AT THE TROA MERES THAT ARE SECRETED FROM THE LIVER ARE MIXTURES OF WILD TYPE AND MUTANT ALLYL. SO WON'T IT BE WONDERFUL IF WE COULD SOMEHOW DEPLETE THE TET MERES THAT WERE -- OF THE MISFOLDING PRONE MUTANT SUBUNIT? SO OUR THINKING IS AS FOLLOWS, IF WE COULD TUNE UP THE ENDOPLASMIC RETICULUM TO HAVE A HIGHER LEVEL OF QUALITY CONTROL, WHEN MUTANT PROTEINS ARE INJECTED, THE DESIRE WOULD BE TO HAVE THEM TO BE DEGRADED BY THE PROTEOSOME WHILE PROCESS OF ERAD THAT ANNIE MENTIONED, RANDY, WHILE THE WILD TYPE PROTEIN COULD STILL BE SECRETED. OF COURSE THE SITUATION WOULD BE NOW THE TET MERES WOULD BE MOSTLY WILD TIEF, WHICH WOULD BE MUCH MORE STABLE THAN THOSE INCORPORATING MUTANT. SINCE THESE STRESS RESPONSIVE SIGNALING PATHWAYS THAT REGULATE THE PROTEOSTASIS PATHWAY OF THE ER ALL END IN TRANSCRIPTION FACTORS, WE SIMPLY DECIDED TO EXPRESS THESE TRANSCRIPTION FACTORS AT PHYSIOLOGIC LEVELS USING A TRICK THAT WE JUST PUBLISHED IN "CELL REPORT" TO ASK WHETHER ACTIVATION OF ONE OR MORE OF THESE PASTWAYS WOULD AFFECT THE KIND OF QUALITY CONTROL WE WANT SO THAT A MORE STABLE PROTEIN WOULD BE SECRETED SECRETED. TURNS OUT THIS REALLY WORKS FANTASTICALLY, BOTH FOR THE TRANSTHYRETIN -- WHEN YOU ACTIVATE -- IN THIS EXPERIMENT, YOU GO FROM HERE TO HERE. SO ABOUT 50% OF THE MUTANT PROTEIN IS ELIMINATE, AND SIMILARLY WITH -- CHAIN. SO I WOULD JUST CLOSE IN SAYING THAT WE HAVE AN AWFUL LOT TO LEARN ABOUT THE PROTEIN HOMEOSTASIS NETWORK, BUT I THINK IT'S CLEAR THROUGH FAIRLY SUBTLE ALTERATIONS OF THE PROTEOSTASIS NETWORK THAT WE CAN ALTER PROCESSES OF AGGREGATION THAT LEAD TO DEGENERATION, PROCESSES OF MISFOLDING THAT LEAD TO LOSS OF FUNCTION DISEASES LIKE CYSTIC FIBROSIS AND SO ON. SO I THINK I'LL STOP THERE. HOPEFULLY I'VE GIVEN YOU SOME FOOD FOR THOUGHT AND SOME OPTIMISM THAT PERHAPS WE CAN DISCUSS FURTHER IN THE DISCUSSION PERIOD. THANK YOU VERY MUCH. [APPLAUSE] >> THANK YOU. WE ARE GOING TO MOVE INTO THE TABLE FOR THE DISCUSSION. SO IF YOU GUYS CAN PLEASE COME TO THE TABLE, AND WHILE YOU DO THAT, I'M JUST GOING TO SUMMARIZE A LITTLE WHAT WE HEAR TODAY. BASE KRI THE WHOLE IDEA THAT YOU SAW IS THERE ARE DIFFERENT LEVELS IN WHICH YOU CAN MODULATE PROTEOSTASIS, WE ARE LEARNING A AS A GOOD SITUATION AS YOU HEAR FROM THE SPEAKERS, THERE ARE MANY THINGS WE STILL NEED TO KNOW. SO WHEN YOU THINK ABOUT THE CHAPERONES AND SYSTEM, IT'S CLEAR WE HAVE A GOOD UNDERSTANDING OF THE MOLECULAR -- WE CAN HEAR HOW YOU -- AND MAKE THEM WORK BETTER BUT WE STILL HAVE MANY QUESTIONS REGARDING -- WITH PROTEIN REPAIR THAT WE HEAR THIS MORNING IN THE SESSION, AND ALSO HOW -- INTERPLAY WITH DAMAGES MOLECULES, WHAT ABOUT THE DNA DAMAGE THAT WE SEE THIS MORNING. NOW WHEN YOU MOVE OUT -- INTO THE ORGANELLES, I THINK WE GOT A GOOD FEELING OF HOW MUCH WE KNOW ABOUT THE ER -- THE MITOCHONDRIAL STRESS, BUT HOW THESE PATHWAYS CROSSTALK, BUT WE STILL DON'T HAVE A VERY CLEAR IDEA HOW THIS COMPARTMENTS, ONE OF THEM -- WE HEAR ABOUT DNA DAMAGE AND HOM HOMEOSTASIS -- NOT VERY WELL UNDERSTAND, IT IS NOT CLEAR HOW THESE COMPARTMENTS -- THE GARBAGE FROM ONE TO EACH OTHER AND WHICH IS THE STRATEGY TO DO THAT, AS WELL AS WHEN YOU START MODULATING CHEMICALLY ONE OF THESE COMPARTMENTS OR THE -- WHAT IS GOING TO HAPPEN TO THE OTHERS. THEN AS WE MOVE TO CELL TO CELL COMMUNICATION, IT'S VERY CLEAR THAT THE PROTEOSTASIS IN ONE CELL IS GOING TO AFFECT -- IN THE DIFFERENT SYSTEMS, BUT WHAT IS NOT VERY CLEAR IS HOW HA COMMUNICATION OCCURS. IS THIS FROM CELL TO CELL INTERACTION, IS THIS -- WE DIDN'T HEAR ABOUT EX-OWE SOMES BUT WE HEAR YESTERDAY, IS THIS A WAY TO COMMUNICATE -- AND IS THIS SOMETHING THAT CONTINUOUSLY COMMUNICATING OR ONLY HAPPENS DURING STRESS. THIS REALLY LEADS INTO THE IMPORTANCE OF PROTEIN HOMEOSTASIS IN THE -- PROTEINS, USING THE PROTEINS SEEMING TO GENERATE AGGREGATES -- ANOTHER PROTEIN IS SOMETHING THAT -- WE HEAR A LOT. AND THEN THE LAST PART AND THIS IS THE ONE THAT WE UNDERSTAND -- HOW THE -- IN ONE ORGAN AFFECTS THE OTHER. WE HEAR -- HOW WHEN YOU ARE AFFECTING ONE SYSTEM, IT'S GOING TO ALTER IN A DIFFERENT ORGAN THE BRAIN -- VICE VERSA, BUT IT'S SOMETHING THAT WE CAN USE, INTERFER WITH THE PROTEOSTASIS ONE PERIPHERAL ORGAN TO AFFECT THE BRAIN AND WE HOPE THAT THAT'S GONNA HAPPEN, AND THEN ALL THESE COMMUNICATION IS HAPPENING THROUGH HORMONAL AND HAPPENING THROUGH THE BLOOD, SO HOW IT'S AFFECTED BY ALL THE THINGS WE HAVE HERE, SUCH AS THE INFLAMMATION AND NUTRIENTS, HOW IS ALL THIS PUT IN PLACE. SO WE ARE GOING TO OPEN THIS FOR DISCUSSION, AND PLEASE IDENTIFY YOURSELF BEFORE ASKING THE QUESTION. >> LET'S START OVER HERE. THANK YOU. >> WE HAVE THE MICROPHONES. >> BILL EVANS. GSK. FRED, MAYBE YOU DIDN'T GET TO IT IN YOUR TALK. BUT SOME YEARS AGO WE DID A STUDY WHERE WE BIOPSIED 90-YEAR-OLD PEOPLE, BUT THE PREEXERCISE SAMPLES SHOWED A TREMENDOUS AMOUNT OF DISORGANIZATION IN MUSCLES, D LINE STREAMING, IT LOOKED LIKE THERE WAS A TREMENDOUS AMOUNT OF DAMAGE GOING UNREPAIR IN THE MUSCLE OF THESE EXTREMELY OLD PEOPLE. IF YOU CAN SAY SOMETHING ABOUT WHY THAT MIGHT BE, AND WHAT IS IT ABOUT SENESCENT MUSCLE THAT MAY NOT ACTIVATE THE PROTEASOME APPROPRIATELY? >> I WISH I COULD ANSWER THESE QUESTIONS. THERE'S NO QUESTION -- IF YOU EVER SAW ME ON THE TENNIS COURT, AGE DECREASES MUSCLE PERFORMANCE, AND ANYONE WHO HAS LOOKED AT IT, AND I GUESS YOUR DATA IN HUMANS IS SIMILAR TO DATA WE'VE SEEN IN MICE, I MEAN -- WE BEING THE COMMUNITY, IS THERE IS A LOSS OF CLASSIC ARCHITECTURE, YOU WOULD SEE SPLITTING OF -- FIBERS, SO THE QUALITY CONTROL MECHANISMS ARE REDUCED. NOW WHETHER THAT'S IN UBIQUINATION, THE ASSEMBLY, THE RESPONSE TO, LET'S SAY, LITTLE BIT OF EXERCISE THAT A 92-YEAR-OLD PERSON DOES, ALL OF THOSE QUESTIONS, WE DON'T KNOW ABOUT. ALL I CAN SAY, THE TIME WE HAD, IS CERTAIN OF THE COMPONENTS, WE CAN'T -CAN'T MEASURE BECAUSE WE DON'T KNOW ENOUGH ABOUT UBIQUINATION AND -- FOR METHODS IN THE LAST COUPLE YEARS, IT SHOULD BE STRAIGHTFORWARD AS A PROTEASOME FUNCTION AND ALSO P97 IS THIS CRITICAL COMPLEX THAT PULLS SUBUNITS OUT OF LARGER STRUCTURES, OUT OF THE ER AND NOW WE KNOW OUT OF THE MYOFIE BRAL. WHETHER THEY'RE FUNCTIONAL, WE DON'T KNOW ABOUT, WE DON'T KNOW ABOUT THEIR PRODUCTION. IT IT IS INTERESTING, THERE IS A -- IF YOU INHIBIT THE PROTEASOMES AS SAY WITH THE PHARMACOLOGICAL DRUGS, CELLS DO UPREGULATE, PRODUCE NEW PROTEASOMES THROUGH A TRANSCRIPTION FACTOR CALLED NRF1, THEY UPREGULATE P97, AND THEY ALSO TO SOME EXTENT UPREGULATE -- 62 AND FACTORS THAT FEED -- INTO AUTOPHAGY. BE VERY INTERESTING TO SEE IF THAT TRANSCRIPTIONAL RESPONSE IS ALTERED IN THE AGED POPULATIONS. I THINK THE ANSWER TO YOUR QUESTION IS, W WE ONLY NOW KNOW THE TERMS IN WHICH TO ASK THE QUESTIONS THAT YOU'VE GOT A VERY SPECIFIC EXAMPLE. >> JUDITH WILL GIVE YOU ONE METHOD. >> EVEN LESS THAN THAT. SO I WANTED TO COMMENT ON WORK THAT MAYBE YOU'RE NOT FAMILIAR WITH, ALL THE PROTEINS IN THE MYOFIBRALS ARE VERY LARGE AND VERY COMPLICATED TO FOLD. THERE IS A NUMBER OF PEOPLE THAT HAVE LOOKED AT THE DISTRIBUTION OF CHAPERONES AND THE FOLDING OF THESE PROTEINS IN MUSCLES, AND THE WORK FROM -- IS PARTICULARLY NICE BECAUSE HE'S FOUND THAT PROTEINS ARE FOLDED ON THE Z LINE AND THERE ARE A NUMBER OF CHAPERONE COFACTORS THAT HELP THOSE PROTEINS FOLD THAT ARE LOCALIZED, THEY'RE MUSCLE-SPECIFIC AND THEY'RE LOCALIZED TO THE NASCENT MYOFIBRALS -- AND WHEN HE KNOCKS OUT THIS CHAPERONE COFACTORS, THE Z LINE IS COMPLETELY DISORGANIZED, THE MUSCLES -- LOOK LIKE WHAT YOU DESCRIBE FOR 90 -- SO THESE COULD ALSO AB FOLDING ISSUE. >> THIS MONTH WE HAVE HEARD QUITE A BIT ABOUT POW AND NFL INJURY TO THE HEAD FROM REPEATED EXPOSURES. TWO QUESTIONS. WHAT'S THE MERCK NISM OF FORMATION OF -- AND QUESTION NUMBER TWO, WHAT CAN WE DO TO PREVENT OR REVERSE SUCH TRAUMA? >> SO TAO, THE MAIN PROBLEM WITH THE PROTEIN IS THE PROTEIN THAT IS GOING TO BE ALWAYS IN YOUR BRAIN, IT'S A PROTEIN OF THE CYTOSKELETON, BUT IT UNDERGOES -- AND MODIFICATIONS AND THIS IS ONE OF THE EXAMPLES OF HOW -- MODIFICATIONS CAN ALTER THE PROTEOSTASIS, SO WHEN IT GETS HYPER -- YOU START HAVING THESE AGGREGATES OF THESE FIBERS, BUT THE INTERESTING THING AND THAT'S SOMETHING THAT HAS BEEN VERY RECENTLY UNDERSTOOD NOW, IS THAT THESE PROTEINS PROPAGATE, SO THIS IS THE IDEA THAT YOU CAN PROAP GATE PROPAGATE THE DISEASE FROM ONE CELL TO ANOTHER AND NOBODY -- ALL THE -- THAT HAS BEEN REPRODUCED IN VITRO -- INCREASING THEY SOMEHOW EXROAS MIEZING INTRACELLULARLY AND IT HAS TO PASS THROUGH -- AND THIS IS ACCELERATING, SO IT'S PROBABLY A COMBINATION OF WHAT WE HEAR ABOUT INFLAMMATION -- DAMAGE, THEY GO TO THESE REGIONS AND MAKE THEM VERY VULNERABLE -- >> I THINK THE PHYSICAL EFFECTS OF TRAUMA THAT INITIATE WHAT ANA MARIA HAS JUST MENTIONED IS A VERY OPEN QUESTION. WE'RE USED TO STRESSES OF HEAT AND CHEMICAL NATURE, BUT PHYSICAL TRAUMA TO THE BRAIN IS A LITTLE CRUDER THAN CELL BIOLOGISTS AND BIOCHEMISTS WANT TO DO. SO BASICALLY EVEN TOXICOLOGISTS DON'T WANT TO HIT MICE IN THE HEAD WITH HAMMERS. NOT SURPRISINGLY, WE CAN'T SAY ANYTHING VERY INTELLIGENT ABOUT WHAT INITIATES THAT EXCEPT THAT IT SOUNDS LIKE AN INFLAMMATORY STIMULUS, AND ONCE YOU START MA GROW FAIJS, NEUTROPHILS, GLEE YA, GENERATING FREE RADICALS, ALL THESE SEQUELAE WILL HAPPEN. >> TO SOME EXTENT, TRAUMATIC BRAIN INJURY IS AN ISCHEMIC INJURY BECAUSE WITH THE SWELLING AND THE TRANSIENT CLOTTING THAT TAKES PLACE, THERE'S ISCHEMIC INJURY. WITH THAT, YOU'RE GOING TO HAVE DAMAGE TO MITOCHONDRIA. THIS IS GOING TO LIMIT THE ATP AVAILABLE FOR PROTEIN REFOLDING OR PROTEIN REPAIR, SO THAT COULD ALSO BE AN INCITING TRIGGER. >> WHAT WOULD YOU DO PREVENTTIVELY? >> HELMETS. DON'T PLAY FOOTBALL. >> DON'T PLAY FOOTBALL. [LAUGHTER] >> THIS IS ANOTHER QUESTION FOR FRED. I THINK YOU MENTIONED IF YOU HAVE -- APPROPRIATE -- YOU HAVE -- MULL TOE AMERICA PROTEINS WITH EXTRA I SUBUNITS HANGING AROUND. IS THAT THOSE IN THE PROTEASOME, I WAS WONDERING IS THERE ANY INFORMATION ABOUT THOSE -- PRO PROTEINS WITH STOIK YOM TRI WAS ALTERED WHERE IT DOESN'T WORK SO WELL? I WAS THINKING MITOCHONDRIAL PROTEINS WHERE MOST OF THE GUYS ARE COMING FROM THE NUCLEUS AND A FEW FROM MITOCHONDRIA, I'D MANL YOU GET INTO TROUBLE ABOUT STOIK YOM TRI, AND IS THAT -- ANYTHING KNOWN ABOUT THAT, THAT THAT IS A PROBLEM IN DEALING WITH THAT? >> ABSOLUTELY GOING BACK TO BEST CHARACTERIZED CASES MIGHT BE THAT WILL SEEM YAS. IF YOU HAVE -- THEN THEY TEND UNLIKE NORMAL AT THE TROA MERES TO PRECIPITATE OUT -- RED CELLS, SO THAT'S ONE EXTREME RELATIVELY -- EXAMPLE. IN MITOCHONDRIA, THIS IS AN EXPERIMENT WE DID IN THE EARLY 80s, IF YOU PUT IN AN INHIBITOR OF MITOCHONDRIAL GENE EXPRESSION SUCH AS -- YOU WILL FIND THE IMPORTED SUBUNITS ARE RAPIDLY DEGRADED, OR IF YOU TAKE ISOLATED MITOCHONDRIA AND FOLLOW THE FATE OF THE GENE PRODUCTS MADE THERE, WHEN THEY DON'T GET THE SUBUNITS FROM THE CYTOSOL, THEY'RE ALL RAPIDLY DEGRADED, WITHIN THE MATRIX. SO THERE ARE MANY, MANY EXAMPLES YOU CAN SHOW IN VITRO, IF YOU STOP R RIBOSOME RNA PRODUCTION, ALL OF THE PROTEASOME SUBUNITS ARE RAPIDLY DEGRADED. SO I THINK THIS IS A VERY COMMON EVENT. THE COMMENTS THAT YOU HEARD FROM SOME OF THE OTHER SPEAKERS, CHAPERONES WORKING AT THE RIBOSOME ON NEWLY SYNTHESIZED PROTEINS, AND THEY MAY BE DIFFERENT FROM THE ONES THAT MAINTAIN THE PROAT YOAM OF THE MATURE PROTEINS, THAT JUST EMPHASIZES THAT THE CHALLENGE OF NEWLY SYNTHESIZED PROTEINS, UNTIL THEY GET FOLDED AND FIND THEIR OTHER MATES, IN LARGER STRUCTURES, THEY'RE VERY SUSCEPTIBLE TO PROBLEMS -- INTERFERES WITH THAT PROCESS AS OTHERS ON THIS PANEL HAVE SHOWN. SO I THINK THAT'S A VERY IMPORTANT ASPECT OF THE CHALLENGES FOR SPACES THAT HASN'T BEEN EMPHASIZED ENOUGH. JUDITH? >> SO OF COURSE BECAUSE OF THE DANGER OF -- THE GENOME HAS TRIED TO BALANCE TRANSCRIPTION AND TRANSLATION OF SUBUNITS OF -- COMPLEXES. AND PERHAPS THE MOST DRAMATIC EXAMPLE -- OF CHROMOSOMES BECAUSE THERE YOU DISRUPT THE NATURAL BALANCE OF -- AND THERE IS NICE WORK FROM -- SHOWING THAT RIEB SOMOL PROTEINS ARE ALL IN DIFFERENT CHROMOSOMES, IF YOU HAVE ONE MORE OR ONE LESS OF ONE OF THE CHROMOSOMES, THEN YOU HAVE AN IMBALANCE OF ALL THE OTHER SUBUNITS. I THINK THIS IS WHY -- ONE OF THE REASONS WHY CANCER CELLS ARE SO DEPENDENT ON HAVING A ROBUST -- RESPONSE FOR SURVIVAL. >> SO THAT'S HAPPENING IN DOWN SYNDROME PRESUMABLY. >> EXACTLY. SAME REASON. >> THAT'S INTERESTING. >> IN THAT CONTEXT, I DIDN'T GET A CHANCE TO MENTION IT, ONE OF THE EXCITING NEW DEVELOPMENTS IN THIS AREA ON THE PROTEASOME WORK IS WORK FROM FINLEY AND KING THAT'S SHOWN THAT IF YOU ALTER THE PROAT SOMOL REACTIONS THAT I QUICKLY WENT THROUGH, YOU CAN ACTIVATE BREAKDOWN OF PROTEIN, AND AND IN ANGELICA AMEN'S WORK, THIS IS PARTICULARLY -- CANCERS THAT ARE POLYEMPLOYEDY THAT'S INAPPROPRIATE, SO I THINK THIS IS A VERY INTERESTING CONNECTION BETWEEN EVERYTHING THAT'S GOING ON HERE AND HASN'T BEEN STUDIED ENOUGH IN CANCER. >> THIS QUESTION IS ALSO TO FRED GOLDBERG. FINDINGS BETWEEN THE BRAIN AND THE MUSCLE IN TERMS OF PROTEASOME FUNCTION, I'M QUITE FASCINATED BY ONE BEING INNOVATEED WITH AGING AND THE OTHER BEING DECREASED WITH AGING, AND I WONDERED IF THAT HAD ANYTHING TO DO WITH THE INCIDENCE OF MAYBE INFLAMMATION OR OXIDATIVE STRESS BEING ACTIVATED AND PHENOTI PICK PLASTICITY OF THE PROTEASOME AND IN PARTICULAR THE IMMUNOPROTEASOME. WOULD YOU LIKE TO COMMENT? >> YES. WE'RE FASCINATED BY THE SAME PHENOMENA, BUT I HAD A FOOTNOTE WHICH HE DIDN'T GO INTO SAYING EVERYTHING I WAS SAYING WAS BASED ON ONE PAPER THAT IS REALLY RIGOROUS, BECAUSE ONLY RECENTLY HAVE PEOPLE BEEN ISOLATING THE -- PROTEASOME AND REALLY STUDYING IT WITH PSYCHOLOGICAL SUBSTRATES. ASSUMING THOSE TWO STUDIES ARE VALID, I THINK WHAT I WAS TRYING TO EMPHASIZE IS THAT NOT ALL ORGANS ARE RESPONDING THE SAME WAY. WHEN YOU THINK ABOUT IT, THERE IS A MUSCLE WASTING PHENOMENA, THERE'S AN INCREASE IN UBIQUITIN CONJUGATES IN THE MUSCLES OF ELDERLY ANIMALS AS WELL AS HUMANS CALLED SARCOMA PENA. PROTEASOME -- HAVING PROT SOME IN THE UBIQUITIN PATHWAY HAS A VERY DIFFERENT ROLE MAINLY INVOLVED IN QUALITY CONTROL -- THAT'S VERY IMPORTANT, IN ADDITION TO QUALITY CONTROL. SO THE ANSWER TO YOUR QUESTION IS, I THINK THERE ARE BIG DIFFERENCES. WE HAVE NO REASON TO BELIEVE THAT WHAT WE TALKED ABOUT HAS ANYTHING TO DO WITH IMMUNE RESPONSE. I RESUME IT'S MORE RELATED TO AGING AND INDEPENDENT TO THE PHENOMENON OF IMMUNOPROTEASOME. IMMUNOPROTEOSOMES GO UP WITH AGING BUT THEY DO NOT AFFECT THE RECOGNITION OF MISFOLDED PROTEIN. THAT OCCURS THROUGH UBIQUINATION IN THE 19S. THE IMMUNOPROTEOSOMES, WHICH IS A VARIANT IN THE ACTIVE SITE INFLUENCES THE EFFICIENCY OF GENERATING PEPTIDES THAT ARE APPROPRIATE FOR ANTIGEN PRESENTATION. THAT'S RATHER IMPORTANT BUT IT'S A LITTLE DIFFERENT FROM THE PHENOMENON THAT I WAS REFERRING TO. >> MELISSA HARRIS, NIH, NATIONAL GENOME RESEARCH INSTITUTE. I'VE GOTTEN THE IDEA THROUGH THE SYMPOSIUM THAT WE HAVE A LOT OF GLOBAL DISREGULATION ON A NUMBER OF PROCESSES SO PROTEOSTASIS, AND MAYBE IT'S BECAUSE OF THE IDEA OF THE TED TOX, BUT I WAS STRUCK BY A COMMENT I THINK FROM STEWART KIM YESTERDAY WHERE HE TALKED ABOUT GENE EXPRESSION CHANGES CAN BE VERY DRAMATIC WITH AGE FROM YOUNG TO OLD, BUT THERE ARE SOME TISSUES WHERE YOU DON'T SEE THESE DRAMATIC CHANGES. SO WHAT I WANTED TO POS POSE TO THIS PANEL IS, HOW MUCH DO WE KNOW ABOUT DISREGULATION OF PROAT STASIS AND ALL DIFFERENT CELL TYPES, VERSES TERMINALLY DIFFERENTIATED, AND IS IT REALLY AN ORGANIZISMAL PHENOMENON OR ARE THERE CERTAIN TISSUES THOO ARE MORE DRIVING -- THAT HAVE -- THAT CONTRIBUTE TO THE AGING PHENOMENON? >> THERE'S WORK THAT'S LOOKED AT THE INDUCTION OF AUTOPHAGY AND THE ABILITY TO RESPOND TO A STARVATION STIMULUS IN DIFFERENT TISSUES AS A FUNCTION OF AGE. AND ALTHOUGH IN MANY TISSUES, THE PROCESS OF AUTOPHAGY DIMINISHES, THIS ISN'T CONSISTENT ACROSS EVERY TISSUE, AND THE RESPONSE TO STARVATION IS NOT CONSISTENT ACROSS DIFFERENT TISSUES. I THINK WHEN WE THINK ABOUT AUTOPHAGY OR MANY OF THESE CELLULAR PROCESSES IN A WHOLE ANIMAL, THERE ARE A NUMBER OF CAVEATS THAT WE HAVE TO TAKE INTO CONSIDERATION. IF WE'RE USING MOUSE MODELS -- CAN IMPACT VERY SIGNIFICANTLY THE FUNCTIONALITY OF THE AUTOPHAGY PATHWAY AND LIKELY OTHER CELLULAR HOMEOSTATIC MECHANISMS. TIME OF DAY, SEX OF THE MOUSE, AND FEEDING STATUS. SO ALL OF THESE DIFFERENT FACTORS CAN IMPACT THE RATE OF PROTEOSTASIS IN DIFFERENT TISSUES. >> LET ME ADD A SPECIFIC -- THAT WILL HIGHLIGHT QUITE DRAMATICALLY HOW THE PROTEOSTASIS CHANGES IN AN AGING MODEL SYSTEM. IF ONE TAKES THESE ANIMALS AND THEY HAVE A SERIES OF FOLDING SENSORS, PROTEINS IN DIFFERENT COMPARTMENTS, THAT YOU CAN MONITOR IN REALTIME AND THEY ALL HAVE A VERY STRICT PHENOTYPE, MIACIN, PARAMIACIN, RATS, FOR EXAMPLE, AZITO COLEAN,, UNFOLDING OCCURS BETWEEN -- THEY'RE NOT MASSIVELY ALTERED. IF YOU THEN LOOK AT THE HEAT SHOCK RESPONSE THAT YOU'VE HEARD ABOUT OR THE UNFOLDED PROTEIN RESPONSE, THEY SHUT DOWN PRECISELY BETWEEN 8 AND 12 HOURS OF ADULTHOOD. THE REASON THIS IS INTERESTING IS THE ANIMAL IS REALLY SET IN STAGE -- THEY MAKE THE EGGS AND THE SPERM, AND SO ESSENTIALLY THAT'S ACCOMPLISHED. AND THIS IS A VERY PRECISE GENETIC REPROGRAMMING THAT'S OCCURRING EARLY IN LIFE THAT THEN DICTATES WHAT'S HAPPENING IN THE OUTYEARS. SO AS TO WHETHER ONE TISSUE IS AFFECTED MORE THAN ANOTHER, WE DON'T KNOW AT THIS POINT. BUT IT'S NOT RANDOM, IT OCCURS AT A PRECISE MOMENT, AND ALL THE DIFFERENT STRESS RESPONSES ARE BEING SHUT DOWN SIMULTANEOUSLY. AND AT THIS POINT, WE THINK IT'S ALMOST ENTIRELY EPIGENETIC CONTROL. SO I THINK IT'S HIGHLY SPECIFIC AND CATASTROPHIC. I'VE HEARD THERE'S OBSERVATIONS IN -- THAT ARE VERY SIMILAR TO SUGGEST AT LEAST THE INVERTEBRATE MODEL, THERE'S GOING TO BE A SIMILAR PROCESS. >> SO I'M INTERESTED IN THE OPINION OF THE PANEL REGARDING WHAT IS THE EFFECT OF AGING ON THE EXPRESSION AND ACTIVITY ON UBIQUITIN -- LIGASES AND IF THEY'RE AFFECTED WHERE WHAT ARE THE -- LOGIC CONSEQUENCES? >> YES, THE -- MAYBE I SHOULD FILL IN THE PACK GROUN THE THE BACKGROUND. I MENTIONED THERE ARE 600 OR SO UBIQUITIN LIGASES. THERE A GROUP OF THEM ASSOCIATED WITH MUSCLE WASTING, IF YOU -- FAST, HAVE SEPSIS, CANCER -- AND WE CALL THESE -- GENES, INCLUDING MRF WHICH TAKES APART THE -- FILAMENT AND -- WHICH AFFECTS NUCLEAR PROTEINS AND PROTEIN SYNTHESIS. THOSE ARE DEFINITELY INDUCED MUCH MORE DRAMATICALLY IN YOUNG ANIMALS THAN IN OLD ANIMALS, THE WORK OF OTHERS, IF YOU GIVE A SPECIFIC STIMULUS SUCH AS GLUCOCOURGLUCOCOURTGLUCOCORTICOI DS, IT'S N OT INDUCED AS DRAMATICALLY. ON THE OTHER HAND, WORK FROM OURSELVES AND DAVID GLASS FINDS THAT MRF IS HIGH IN SARCOPENIC ANIMALS. A LOT OF THAT MAY BE POST TRANSDESCRIPTIONAL. WHAT THAT MEANS, WE'RE NOT CERTAIN OF, BUT IT PROBABLY MEANS EXCESSIVE -- UBIQUINATION AND BREAKDOWN. BUT THERE ARE ONGOING EXPERIMENTS ADDRESSING THEM. IN THIS BUSINESS, IN TERMS OF THE PREVIOUS QUESTION, WHAT YOUR QUESTION RAISES IS THE IDEA THAT UBIQUITIN LIGASES, THAT EARLY PART OF THE PATHWAY, IS NOT UBIQUITOUS, UBIQUINATION IS NOT UBIQUITOUS. THERE ARE ORGAN-SPECIFIC DIFFERENCES DIVIDING CELLS A DIFFERENT FROM NON-DIVIDING CELLS, MUST ELSE ARE DIFFERENT THAN NEURONS, THERE ARE A WHOLE GROUP OF UBIQUITIN SPECIFIC -- SO THE BIOLOGY OF THAT PART OF THE PATHWAY -- OF THE PROTEASOME IS VERY SPECIFIC AND TO THE PHYSIOLOGICAL CONTEXT OR THE PATHOLOGICAL CONTEXT. >> JOE, UNIVERSITY OF AKRON. MY QUESTION IS FOR ROBERTA. SO COULD YOU TALK A LITTLE BIT MORE ABOUT MAYBE A STRATEGY FOR ENHANCING THE AUTOPHAGY? I IMAGINE, YOU KNOW, HOW -- WHERE MAYBE THE FIRST PLACE IS TO LOOK WHETHER YOU'D THINK, YOU KNOW, HUMAN CELLS VERSUS THE MOUSE MODELS, THOSE SORT OF THINGS, BUT HOW YOU WOULD GO ABOUT THAT? >> I LOOKED AT YO AUTOPHAGY AND MITOPHAGY FROM ANIMAL MODELS TO HUMAN MODELS, THIS IS RESPONSE TO ISCHEMIC STRESS AND IS INDUCED BY NUTRITIONAL DEPRIVATION SUPPRESSED BY NUTRITIONAL EXCESS. IN THE SETTING OF HUMANS, WE'VE LOOKED AT AUTOPHAGY IN THE HUMAN HEART, DURING SURGICAL -- THE STRESS OF ISCHEMIA DURING A CARDIAC SURGERY. AND THEIR AUTOPHAGY IS MOUNTED AS PART OF A PROTECTIVE RESPONSE RESPONSE. IN ANIMALS, WE KNOW IT'S PROTECTIVE RESPONSE BECAUSE THE MAGNITUDE OF INJURY IS MUCH GREATER, SO THERE ARE -- I THINK THE OTHER QUESTION WAS WAYS OF INDUCING IT. >> YES. >> SO THE QUESTION IS ON A CHRONIC BASIS, CERTAINLY THERE ARE LOTS OF PHARMACOLOGIC AGENTS THAT INDUCE AUTOPHAGY AND PROBABLY INDUCE THE WHOLE -- A WHOLE HOST OF OTHER PROTEOSTATIC PATHWAYS. AND MAYBE THAT'S A GOOD THING TO DO ALL OF THEM IN CONCERT SO THAT THEY CAN KEEP EACH OTHER IN BALANCE. BUT I'LL TELL YOU A STORY FROM WORK DONE -- WHERE MICE WERE SUBJECTED TO FASTING TWICE A WEEK, STARTING AT THE AGE OF 10 MONTHS AND CONTINUING FOR A YEAR. AT THE END OF THAT POINT, WHAT WE FOUND WAS THEIR AUTOPHAGY CAPACITY WAS MUCH MORE INTACT THAN THEIR AGED LITTERMATE AND THEIR CARDIAC RESERVE IN TERMS OF RESPONSE TO ISCHEMIA WAS MUCH BETTER AND THEIR IMMUNOLOGIC FUNCTION WAS BETTER, SO THAT'S ONE MECHANISM, THIS INTERMITTENT FASTING, POTENTIALLY EXERCISE WOULD ELICIT THE SAME SET OF PATHWAYS TO UPREGULATE -- ON A PHYSIOLOGIC CHRONIC BASIS. >> SO -- ABOUT -- BECAUSE THAT'S SOMETHING THAT HAS -- ENOUGH EVIDENCE THAT IN CONDITIONS LIKE ALZHEIMER'S, PARKINSON'S, HUNTINGTON, YOU HAVE COMPROMISE OF THE AUTOPHAGY SYSTEM, SO IN THAT CASE, SOMETHING THAT WE HAVE DISCUSSED MANY TIMES, IT'S VERY IMPORTANT TO KNOW WHAT IS THE PROBLEM, SOMETIMES THAT YOU MIGHT NOT WANT TO PUT MORE OF THIS OUT OF -- DESCRIBED VERY NICELY BECAUSE YOU HAVE A -- SO I THINK IN THOSE CASES, IT'S VERY IMPORTANT -- TO EVERYBODY IT'S TIME TO SEE WHAT IS THE PROBLEM AND DO MUCH MORE SPECIFIC TREATMENT THAT I THINK WE ARE REACHING AT THIS STAGE THAT IS GOING TO BE POSSIBLE. >> GREAT. THANK YOU. GORDON? >> GORDON -- INSTITUTE. I'D LIKE TO GO BACK TO LARGE ASSEMBLIES. I'M CURIOUS WHAT WE KNOW AND DON'T KNOW ABOUT REPAIR OR REMOVAL THE SINGLE SUBUNITS IN ASSEMBLIES. SO IF THERE'S IRREPARABLE DAMAGE TO A SINGLE SUBUNIT IN THE -- COMPLEX, CAN YOU REPAVER OR REMOVE THAT SUBUNIT? DO YOU HAVE TO TAKE DOWN THE ENTIRE COMPLEX, OR IN FACT DO YOU HAVE TO TAKE DOWN THE ENTIRE MITOCHONDRIA? >> I THINK WE DON'T HAVE GOOD ANSWERS TO THAT YET, PARTLY BECAUSE OF THE LIMITATION OF THE TOOLS. THERE ARE TRIPLE A PROTEASES THAT ARE ABUNDANT IN THE MITOCHONDRIA AS WELL AS PROTEASOMES, SO THERE ARE MECHANISMS FOR DEGRADING INDIVIDUAL COMPONENTS, BUT WHETHER THEY CAN ACCESS THE COMPONENTS THAT ARE ALMOST IN THESE CRYSTALLINE ARRAYS ISN'T CLEAR, BUT IF WE IMAGINE THESE STRUCTURES KIND OF BREATHE AND REASSEMBLE DYNAMICALLY, THEN YOU HAVE A CHANCE FOR MISFOLDED COMPONENTS TO NOT BE REINCORPORATED AS THESE STRUCTURES REASSEMBLE. YOU CAN IMAGINE THEY GET SHOVED ALONG THE MEMBRANE TO SOME REGION THAT'S NOT SO -- THAT MIGHT BE THE PLACE WHEREAS THE -- THAT JUNK GETS BUTTED OFF AND IT'S REMOVED FOR AUTOPHAGY BECAUSE WHEN MITOCHONDRIA GOES THROUGH FISSION, YOU GET ONE THAT'S NOT SO GOOD AND THAT'S TARGETED FOR DISPOSAL. >> WE HAVE ABOUT FIVE MINUTES SO WE HAVE TO HAVE SHORTER QUESTIONS AND VERY SUCCINCT ANSWERS. >> I HAD A COMMENT MORE THAN A QUESTION. I WOULD LIKE TO DRAW THE ATTENTION OF THE PANELISTS TO ANOTHER PROTEIN MODIFICATION THAT HAS NOT BEEN MENTIONED, AND THIS IS RELATED TO THE GLYCOMIC, SO THE ALTERATION OF ENGLIE KIENS AND -- THESE ARE -- THIS ALTERATION ARE INVOLVED IN INFLAMMATION AND OTHER PATHWAYS, BUT THIS -- FOR EXAMPLE, THE ACCUMULATION OF -- GLIE CANS IN THE PROTEINS IN IGG AND ANOTHER LIVER PROTEINS -- SOME OF THE MOST POWERFUL BIOMARKERS OF AGING. SO I THINK THAT THIS SHOULD BE ADDED TO THE LEAST OF THE ALTERATIONS. >> WE'LL ASK RANDY AND JEFF BECAUSE IT TURNS OUT THAT GLYCO -- THE ADENGS OF GLIE CANS IS A MAJOR SIGNAL IN THE UNFOLD UNFOLDED PROTEIN RESPONSE. >> THE N GLYCANS ARE PUT ON IN THE ER AND THEIR MODIFICATIONS INVOLVE TRIMMING AND THEN REBUILDING UP THROUGH THE GOLGY STACKS. THOSE GLYCANS IN THE ER ARE ESSENTIAL FOR INTERACTION WITH LEK TIN CHAPERONES AND FOR TRAFFICKING THROUGH -- MEDIATED TRAFFIC THROUGH THE GOLGY WHERE THERE ARE OTHER GLYCOSEAL -- EVENTUALLY IS A LICK ACID IS PUT ON AND THEY'RE SECRETED. FOR THE OLD GLYCANS, THOSE ARE PRIMARILY PUT ON IN THE GOAL GEE PARTMENT, AND THEY TEND TO BE PARTICULARLY ENLARGED IN TUMOR CELLS, IN PARTICULAR MEU SANTS OF THE EPITHELIAL -- EP THEEL YAM TRACT. WHAT YOU FIND ARE THE TUMORS IN THOSE TISSUES, THEY HAVE EXCESSIVE LARGE AMOUNTS -- A WAY TO EVADE THE IMMUNE SYSTEM. BUT IN TERMS OF AGING, I ACTUALLY LOOKED FOR THIS BEFORE COMING HERE AND I REALLY COULDN'T FIND ANY EVIDENCE THAT THE EXPRESSION OF ANY OF THESE GLYCOSEAL TRANSPHRASES IS ALTERED WITH AGING. >> WE PUBLISHED SEVERAL PAPERS THAT THERE IS THIS ALTERATION OF N GLYCANS, SO -- AND THERE ARE MANY PAPERS. >> THANK YOU. >> YOU CAN GIVE A LOOK. >> RICH MILLER FROM UNIVERSITY OF MICHIGAN. THE NIA INTERVENTION TESTING PROGRAM IS ALWAYS INTERESTED IN TRYING TO FIND DRUGS THAT MIGHT SLOW AGING AND EXTEND LIFESPAN BY IMPROVING THE ABILITY OF CELLS WITHIN THE MICE TO DEAL WITH SOME PROTEIN MISFOLDING. WHENEVER I HEAR A TALK FOR THE LAST FIVE YEARS ABOUT THIS TOPIC, THERE IS ALWAYS ONE SUCH COMPOUND THAT DOES IT. IT'S CALLED COMPOUND 1. SO I WAS WONDERING IF THERE ARE DRUGS THAT ARE NOT COMPOUND 1 THAT COULD ANY OF YOU RECOMMEND FOR ADMINISTRATION TO MICE THAT FOR TESTING TO SEE WHETHER LIFESPAN AND HEALTH SPAN CAN BE IMPROVED BY A FARM ENCLOSE CAL IMPROVEMENT OF THE PROTEIN FOLDING APPARATUS. >> THE ANSWER IS YES. >> SO THERE ARE CHEMICAL CHAPERONES THAT BASICALLY BUFFER PROTEIN AGGREGATION, AND THESE MOST CLASSICAL -- THEY'RE IN THE CLINIC ACTUALLY FOR PROBABLY OVER A DOZEN DIFFERENT DISEASES NOW, ALL ASSOCIATED WITH PROTEIN MISFOLDING. -- ACID IS APPROVED FOR -- DISORDERS AND -- HAS BEEN APPROVED FOR BILE OBSTRUCTION DISEASES. BUT THEY'RE IN THE CLINIC FOR PARKINSON'S AND THEY'VE BEEN SHOWN TO IMPROVE INSULIN SENSITIVITY IN HUMANS, THE NUMBER OF STUDIES IN ANIMAL MODELS SHOW THAT MOST DISEASES OF PROTEIN FOLDING ASSOCIATED WITH MISFOLDING IN THE END PLASMIC PRE TICK LUM CAN BE PROTECTED BY THESE EXTREMELY HIGH DOSES, GRAMS, PER DAY. IN SOME OF OUR EXPERIMENTS, WE CAN ACTUALLY DELETE A MOLECULAR CHAPERONE AND COMPLETELY CORRECT THE PHENOTYPE BY ADDING A CHEMICAL CHAPERONE TO THEIR DIET. >> SO THERE WAS JUST A PAPER FROM A GROUP IN FINLAND SHOWING THAT ABOUT 1% OF HUMANS HAVE A MUTATION IN TRANSPYRO TIN THAT WORKS STRICTLY ANALOGOUS TO THE DRUG WE INTRODUCED THAT PFIZER NOW SELLS. WHAT'S INTERESTING ABOUT THOSE PEOPLE, THAT THEY LIVE FIVE TO 10 YEARS LONGER THAN THE OTHER 70,000 PEOPLE IN THAT POPULATION THAT STUDIES FOR A MEDIAN OF 32 YEARS, AND THE AUTHORS ATTRIBUTED THAT LIFESPAN EXTENSION TO A DECREASE IN VASCULAR DISEASE, AND THAT WAS VERY CLEAR FROM THE DATA. SO I THINK THERE ARE DRUGS THAT WILL EXTEND LIFESPAN BY AMELIORATING DISEASE. AND VASCULAR DISEASE IS A BIG ONE. >> TO THE EXTENT THAT INDUCING HIGH TOMY TOE GEE TO HAVEINDUCE --THERE AR E ALREADY DRUGS AVAILABLE ON THE MARKET AND THAT THESE INCLUDE RAH PA MY SIN -- BUT I THINK THAT BECAUSE WE HAVE THIS BALANCE OF STIMULI THAT ARE BOTH PRO AND ANTI-AUTOPHAGY GOING ON IN EACH PERSON, IT'S NOT CLEAR. FOR INSTANCE, SYMPATHETIC STIMULATION IS GOING TO TEND TO SUPPRESS AUTOPHAGY BETA-BLOCKERS, SO MANY OF THESE BENEFITS MAY ALREADY BE ACTIVE IN THE PATIENT POPULATION. >> I WOULD JUST ADD, THERE ARE ENOUGH AGES BEING STUDIED AT THE MOMENT THAT YOUR QUESTION IS A LITTLE PREMATURE, WHEN THE PHARMACODYNAMICS ARE READY, I'M SURE SEVERAL PEOPLE WILL BE PHONING TO YOU TEST THEM, BUT WHEN YOU THINK ABOUT IT, RAP MY SIN, ASIDE FROM ACTIVATING AUTOPHAGY, WE HAVE NOW FOUND IT ACTIVATES UBIQUINATION GENERALLY, SO THE TWO MAIN DEG DAYTIVE PATHWAYS ARE ACTIVATED, AND ANYTHING THAT AFFECTS THE IGF PATHWAY IS AFFECTING FOXO, AND ASIDE FROM METABOLIC EFFECTS, FOXO IS INDUCING THE TRANSCRIPTION OF ALL THE AUTOPHAGY GENES I KNOW AND ALSO IS INDUCING MAJOR COMPONENTS OF THE UBIQUITIN PROTEASOME PATHWAY NND OTHER PROTEOSTATIC MECHANISMS. SO WITHOUT KNOWING IT, YOU MAY BE THE LEADER IN INTRODUCING MANIPULATIONS OF THE UBIQUITIN PROTEASOME PATHWAYS. >> IF THERE'S NO OTHER QUESTIONS, I'M BRAD WEISS FROM THE NATIONAL INSTITUTE ON AGING, AND TO CLOSE THE SESSION, I'D LIKE TO THANK THE CO-CHAIRS, THE PANELISTS, AND THE AUDIENCE FOR A VERY INTERESTING AND INFORMATIVE SESSION. WE WILL BREAK NOW, AND RECONVENE AT 3:15 FOR THE LAST SESSION OF THE DAY. THANK YOU. >> GOOD AFTERNOON. WELCOME TO THE SESSION ON STEM CELLS AND REGENERATION. WE'RE NOW ON THE HOME STRETCH. I AM FROM THE NATIONAL INSTITUTE OF DENTAL AND CRANIOFACIAL RESEARCH. MY COORGANIZERS FOR THIS SESSION ARE YUNG O AND RON FROM THE NATIONAL INSTITUTE ON AGING. OUR CO-CHAIRS FOR THIS SESSION ARE THOMAS RAN DOUGH AND TONY WEISS-CORE RAY. DR. RAN DOUGH IS A PROFESSOR IN THE LABORATORIES FOR BIOLOGY OF AGING AND STANFORD UNIVERSITY SCHOOL OF MEDICINE. HE WILL GIVE AN INTRODUCTORY OVERVIEW OF THE TOPIC FOR THIS SESSION. DR. WEISS-CORAY IS PROFESSOR IN NEUROLOGY AND NEUROLOGICAL SCIENCES OF STANFORD UNIVERSITY SCHOOL OF MEDICINE AND ASSOCIATE DIRECTOR OF THE CENTER FOR TISSUE REGENERATION, REPAIR AND REGENERATION AT THE VETERANS' ADMINISTRATION PALO ALTO HEALTHCARE SYSTEMS. HE WILL BE DIRECTING THE DISCUSSION WHICH FOLLOWS THE LAST SPEAKER AND GIVING A SUMMARY OF THE SESSION. THANK YOU. >> THANKS, LESLIE, AND THANKS, ALL OF YOU FOR STICKING AROUND. I AM FROM STANFORD BUT I GREW UP IN NEW ENGLAND AND I DID MOST OF MY SCHOOLING IN BOSTON, WHICH I JUST WANTED TO MENTION BECAUSE IT'S ALSO HOME OF THE WORLD CHAMPION BOSTON RED SOX. THIS IS THE LAST SESSION OF THE SUMMIT, AND THIS IS TITLED STEM CELLS AND REGENERATION SO YOU'LL HEAR ABOU -- BUT I WANTED TO POINT OUT THIS IS REALLY A KIND OF DIFFERENT SESSION IN A WAY THAN THE OTHERS THAT WE'VE HEARD ABOUT SO FAR. THIS IS JUST THE TITLES OF THE OTHER SESSION, AND TEASE ARE EITHER REALLY KIND OF EXTRINSIC INFLUENCES ON ANY CELLS, ALL RELATED TO AGING, WHEREAS IN THIS SESSION, WE'RE TALKING ABOUT A PARTICULAR CELL TYPE AND WE CAN TALK ABOUT ANY CELL, SO REALLY THE QUESTION IS, WHY ARE STEM CELLS IN A WAY UNIQUE OR SPECIAL IN TERMS OF THE BIOLOGY OF AGING, WHEN REALLY WE HEAR ABOUT ALL OF THESE ASPECTS OF PROCESSES THAT GO ON IN AGING IN STEM CELLS. I WOULD THROW OUT A COUPLE IDEAS OF WHY WE WOULD TALK ABOUT STEM CELLS AS OPPOSED TO ANY OTHER CELL THAT MIGHT BE INTERESTING TO THE IMMUNE SYSTEM, AND CLEARLY THERE'S A RELATIONSHIP CONCEPTUALLY IN TERMS OF STEM CELLS BEING IMPORTANT FOR TISSUE HOMEOSTASIS IN THE SAME WAY A LOT OF THESE PROCESSES ARE IMPORTANT FOR CELLULAR HOMEOSTASIS, SO CLEARLY YOU HAVE TO HAVE CONTROL OF PROTEINS TO MAINTAIN A CELL, WE HAVE TO HAVE -- OF THE TISSUE COMPONENTS OF A TISSUE TO MAINTAIN THAT TISSUE. SO THERE'S REALLY THE HOMEOSTATIC COMPONENT. THERE'S ALSO OBVIOUSLY KIND OF A THERAPEUTIC ASPECT OF STEM CELLS PLAYING A ROLE IN AGING THINKING ABOUT USING STEM CELLS BUT IN TERMS OF THE BIOLOGY OF AGING, THE STEM CELLS I THINK HAVE KIND OF A UNIQUE ROLE IN THINKING ABOUT HOW CELLS IN THE BODY AGE OR DON'T AGE. IN ONE SENSE, THE KIND OF CELLS WE'LL TALK ABOUT, IN IN A WAY INTERMEDIATE BETWEEN THE KIND OF IMMORTAL GERMLINE CELLS AND THE -- SOMOL CELLS. SO THERE THESE ASPECTS OF WHAT ARE UNIQUE ABOUT SOMATIC STEM CELLS THAT GIVE THEM THE ABILITY THROUGHOUT THE LIFE OF AN ORGANISM TO MAINTAIN THAT TISSUE. SO I THINK THERE'S THIS INTERESTING INTERMEDIATE -- THERE ARE ALSO, I THINK, PROBABLY IMPORTANT UNIQUE ASPECTS OF STEM CELLS THAT PLAY A ROLE IN THINKING ABOUT THE PROCESSES OF AGING, AND WE'LL MENTION MANY OF THEM DURING THIS SUMMIT, I WANT TO MENTION TWO IN PARTICULAR, ONE IS THIS IDEA THAT STEM CELLS CAN UNDERGO ASYMMETRIC CELL DIVISION TYPICALLY, GIVING RISE TO DAUGHTERS WITH TWO FACE, OFTEN IS -- THE OTHER DAUGHTER GOES ON TO BECOME A MORE DIFFERENTIATED CELL. THERE'S THE POSSIBILITY AT LEAST OF ASYMMETRICALLY SEGREGATING ASPECTS OF THE BIOLOGY OF AGING, WHETHER IT'S DEFECTIVE PROTEINS OR DAMAGED DNA, SO I THINK THERE'S SOMETHING IMPORTANT IN AGING, THE OTHER ASPECT I THINK IS VERY IMPORTANT IS THAT IDEA THAT MANY STEM CELLS CAN RESIDE IN THE QUIESCENT STATE. THIS IS AN ILLUSTRATION OF SOMETHING INTERESTED ABOUT STEM CELLS, WE'VE TALKED ABOUT DIFFERENT ASPECTS OF AGING IN THIS SUMMIT IN TWO WAYS. WE TALKED ABOUT CHRONOLOGICAL AGING IN TERMS OF CELLS THAT EXIST THROUGHOUT THE LIFE OF AN ORGANISM AND THEREFORE EXPERIENCE THE CHALLENGES OF PROLONGED EXISTENCE WITHOUT REPLENISHMENT. WE TALKED A LOT ABOUT -- AGING THAT OCCURS AS CELLS UNDERGO MULTIPLE CELL DIVISIONS AND MANY STEM CELLS ACTUALLY EXIST IN BOTH STATES, SO THEY WILL STAY IN A QUIESCENT STATE FOR A LONG TIME AND THEN DIVIDE, SO THEY EXPERIENCE BOTH CHRONOLOGICAL AGING AND RE PLEKTIVE AGING. SO CLEARLY THERE ARE A LOT OF DIFFERENT TYPES OF STEM CELLS, I THANK RON FOR STIMULATING THE IDEAS FOR THIS SLIDE, THINKING ABOUT STEM CELLS FROM DIFFERENT TISSUES REALLY HAVING DIFFERENT ROLES, SO THERE ARE STEM CELLS IN THE TISSUES THAT TURN OVER CONTINUOUSLY LIKE SKIN, INTESTINES, BLOOD, WHERE THE CELLS ARE ALWAYS REPLENISHING THE MORE DIFFERENTIATED CELLS, THEN THERE ARE CELLS IN TISSUES REALLY THERE FOR MUCH MORE REPAIR. THEY HAVE THE ABILITY TO ACTIVATE, PROLIFERATE, DIFFERENTIATE IN RESPONSE TO STIMULATE AND SO FORTH. SO THERE ARE REALLY DIFFERENT TYPES OF STEM CELLS, SO IT'S HARD TO TALK ABOUT THIS ALMOST AS A SINGLE CELL TYPE. THESE REALLY HAVE VERY DIFFERENT BIOLOGY. I'M GOING TO POSE AT LEAST ONE THEORETICAL QUESTION THAT WILL COME UP IN DIFFERENT WAYS IN THE TALKS. THIS IDEA IS KIND OF A DICHOTOMY OF HOW STEM CELLS PLAY A ROLE IN AGING IN THE SENSE DO STEM CELLS CAUSE AGING OR DO STEM CELLS PREVENT AGING. I THINK ONE CAN REALLY THINK OF THESE IN EITHER WAY, AND IT'S IMPORTANT FOR US TO THINK WHICH OF THESE THE DATA SUPPORTS. I'M GOING TO START WITH THIS IDEA THAT STEM CELLS CAUSE AGING, AND THIS IS FROM NED AND RHONDA FROM A FEW YEARS AGO IN WHICH THEY ACTUALLY TITLED THE REVIEW SOMETHING LIKE HOW STEM CELLS AGE AND WHY THIS MAKES US GROW OLD OR SOMETHING LIKE THAT. SO REALLY THE IDEA THAT STEM CELLS ARE CONVEYING THE AGING INFORMATION TO THE REST OF THE BODY, AND THE WAY THEY'VE ILLUSTRATED THAT IS YOU HAVE THE YOUNG STEM CELLS WHICH GIVE RISE TO HEALTHY ARRAY OF DIFFERENTIATED CELLS, THEN WITH AGE, THESE STEM CELLS CHANGE AND WHEN THEY DIFFERENTIATE, THEY GIVE RISE TO A DIFFERENT ARRAY OF CELLS THAT AREN'T QUITE NORMAL, THEY'RE NOT THE SAME AS YOUNG. SO IN THIS SENSE, IT'S REALLY THE PROBLEM, IS THAT THE STEM CELLS ARE AGING, BUT IS THIS REALLY TRUE. I THINK WE HAVE TO ASK IF THIS IS REALLY SUPPORTED BY THE DATA. I'LL JUST PRESENT A COUPLE OF IDEAS HOW WE MIGHT THINK OF THIS. DERRICK ROSSI WHEN HE WAS A POSTDOC, THEE ARE HEMATOCRIT STEM CELLS FROM YOUNG OR OLD INDIVIDUALS, AND WHAT HE'S LOOKING AT IS EVIDENCE OF DNA DAMAGE. SO IN THE YOUNG CELLS, THERE'S VERY LITTLE EVIDENCE OF DNA DAMAGE SHOWN BY THESE GAMMA H26789X STAINING WHEREAS IN THE OLD CELLS, THERE'S A LOT OF EVIDENCE OF DAMAGE. SO THIS WOULD SUGGEST CONSISTENT WITH THIS MODEL THAT BASICALLY THE STEM CELLS THEMSELVES ARE EXPERIENCING SOME KIND OF CHRONOLOGICAL AGING AND ARE CARRYING INFORMATION, DNA DAMAGE, WHICH IS DIFFERENT THAN YOUNG CELLS. I'LL JUST POINT TO THIS, THIS MAY NOT BE THE SIMPLEST ANSWER BECAUSE WHAT MANY GROUPS IS SHOWN, EVEN IF YOU SEE THESE FOCI OF DAMAGE, HERE'S ACTUALLY A SKELETAL MUSCLE FIBER WITH FOUR CELLS SHOWN HERE, AND THESE ARE STANFORD GAMMA H2X, HERE'S 123, BUT THIS FOURTH STEM CELL WHICH IS A PROJECT NATE OF ONE CELL INTO TWO, ONE EXHIBITS A LOT OF DAMAGE, THE OTHER ALMOST NONE. SO SOMETHING HAPPENED HERE ASYMMETRICALLY IN TERMS OF CELL DIVISION WHERE THIS DAMAGE THAT WAS OBSERVED IN THE STEM CELL ACTUALLY IS AMELIORATED IN THE PROCESS OF THIS DIVISION. SO ON THE OTHER END OF THE SPECTRUM IS THE IDEA THAT MAYBE STEM CELLS ARE ACTUALLY THE GUARDIANS OF YOUTHFULNESS THAT WHAT THE STEM CELLS ARE ARE THESE PRISTINE CELLS WHICH HAVE VERY LITTLE DNA DAMAGE, VERY LITTLE PROTEIN AGGREGATION AND WHAT THEY DO IS THEY LIVE IN THIS ENVIRONMENT, SYSTEMIC INFLUENCE, EXTERNAL INFLUENCES, THESE ARE THE CELLS THAT ARE ACTUALLY MAINTAINING THE HEALTH OF THE TISH TEU BY MAINTAINING THEY WILL SEFS IN THIS PRISTINE STATE, AND IS IT POSSIBLE IN FACT THAT OLD STEM CELLS ARE REALLY PERFECTLY FINE AND JUST AS GOOD AS YOUNG STEM CELLS, BUT WHEN THEY EXIST IN AN AGED ENVIRONMENT, THEY BEHAVE LESS EFFECTIVELY. I'LL PRESENT ONE PIECE OF DATA IN SUPPORT OF THIS, IT THIS WAS FROM MY CO-CHAIR TONY WEISS-CORAY'S LAB, TO THINK ABOUT HOW THE AGING INFLUENCE, THE E TERNL INFLUENCE -- IT'S KNOWN FROM A YOUNG TO OLD MOUSE NEUROGENERAL SUS DECLINES PRO SIP TUSLY, SO A MARKED DECREASE IN NEUROGENESIS WITH AGE, BUT WHEN MICE ARE OLDER EXPOSED TO THE YOUNG ENVIRONMENT OF A YOUNG ANIMAL, THERE'S KIND OF A REJUVENATION OF THE AGED STEM CELLS SO THEY BEHAVE LIKE YOUNG STEM CELLS. SO MAYBE THE AGED ARE PERFECTLY FINE AND WHEN GIVEN THE RIGHT ENVIRONMENT, THEY NOW BEHAVE PERFECTLY FINE JUST AS WELL AS YOUNG STEM CELLS. SO THAT'S SORT OF THE OTHER END OF THE SPECTRUM, CLEARLY IT DOESN'T HAVE TO BE BLACK AND WHITE, THERE COULD BE AN INTERMEDIATE BETWEEN STEM CELLS MAY AGE BUT THE ENVIRONMENT IN WHICH THEY LIVE ALSO HAN -- IN HOW WELL THEY FUNCTION. IT'S A QUESTION WITH THIS PARABUY TICK TECHNIQUE AND OTHER APPROACHES, CAN OLD STEM CELLS IN A SENSE BE REPROGRAMMED, AND THIS IS SIMPLY PUT UP AS AN IDEA THAT THE REPROGRAMMING WE KNOW VERY WELL IN TERMS OF CELLULAR DIFFERENTIATION, GOING TO A DIFFERENTIATED CELL TO A LESS DIFFERENTIATED CELL, VERY PROMINENT IN THE STEM CELL FIELD, IS THIS ALSO TRUE OF AGING, CAN WE THINK ABOUT WHAT HAPPENS WHEN WE'VE HEARD ABOUT SOME REJUVENATION STUDIES AND REJUVENATION INTERVENTIONS, THINGS THAT CAN REALLY MAKE AN OLD CELL LOOK LIKE A YOUNG CELL, IS IT REALLY SOMETHING AKIN TO A KIND OF EPIGENETIC REJUVENATION. SO WE'LL GET TO THIS IDEA OF EPIGENETICS DURING THE TALK TOO. I JUST WANTED TO END WITH THIS TAKEOFF, IF YOU HAVE THIS METAPHOR WHERE THE GENOME CARIES THE INFORMATION THAT HAS TO BE -- THAT PASS THROUGH A LENS WHICH IS THE EP GENOME TO GIVE YOU A CLEAR PICTURE OR A HEALTHY CELL, AND WITH AGE, YOU HAVE DAMAGE TO DNA, YOU HAVE DISRUPTION OF THE CHROMATIN, AND YOU END UP WITH A CELL THAT IS NOT AS EFFECTIVE, EFFICIENT OR HEALTHY, BUT WITH THESE REJUVENATION INTERVENTIONS, WHAT ARE WE DOING? WE'RE PROBABLY ACTING AT THE LEVEL OF THE EPIGENOME, RESTORING THE LENS THROUGH WHICH THIS INFORMATION IS PASSED TO ATTAIN CELLS THAT LOOK PARTICULARLY EFFECTIVE AND WORK PARTICULARLY EFFECTIVE IN TERMS OF THEIR FUNCTION. SO I'LL END THERE. THESE ARE THE SPEAKERS WHO WILL COVER THESE TOPICS. THE FIRST TWO REALLY SPEAK TO THIS IDEA OF WHAT HAPPENED INSIDE THE CELL, THE LAST THREE ADDRESS THE ISSUE OF WHAT HAPPENS OUTSIDE THE CELL IN THE STEM CELL COMPARTMENT THAT CAN REALLY INFLUENCE STEM CELL FUNCTION. SO WITH THAT, I'LL TURN THE MIC OVER TO TONY WEISS CORAY. >> OUR FIRST SPEAKER IS DANICA CHEN, FROM U.C. BERKELEY. SHE'S INTERESTED IN SIGNALING PATHWAYS THAT CONTROL STEM CELL ACTIVITY ESPECIALLY WITH AGING AND SHE HAS WORKED ON HOW -- MIGHT BE RESPONSIBLE FOR SOME OF THE CALORIC RESTRICTION EFFECT, AND SHE WILL ADDRESS THE QUESTION HOW STEM CELLS CHANGE INTRINSICALLY WITH AGE AND HOW THIS COULD BE EXPLOITED FOR THERAPEUTICS. >> THANK YOU, TOM, THANK YOU, TONY. TOM HAS ALREADY INTRODUCED THE STEM CELL THEORY OF AGING, AND I WILL JUST GO AHEAD AND PRESENT SOME EVIDENCE THAT SUPPORTS THIS THEORY. HOW DOES STEM CELL FUNCTION CHANGE WITH AGE? NOW THE -- SYSTEM AS AN EXAMPLE, THE DEGENERATIVE CAPACITY OF A HEMATOPOE TICK STEM CELL DOCTOR DECREASES WITH AGE. AND BROADLY SPEAKING, HEMATOPOETIC STEM CELLS CAN DIFFERENTIATE INTO THE MYELOID -- AND THE LYMPHOID LINEAGE, AND WITH AGE, THEY TEND TO DIFFERENTIATE TOWARDS THE MYELOID AGE AND -- AND THIS IS CONSISTENT WITH THE PHYSIOLOGICAL CHANGES IN THE AGING HEMATOPOETIC SYSTEM. THIS IS ALSO CONSISTENT WITH THE PATHOLOGICAL CHANGES IN THE AGED SYSTEM. CHILDHOOD LEUKEMIA TEND TO BE IN THE LYMPHOID REGION AND -- TEND TO DEVELOP LEUKEMIAS OF THE MYELOID REGION. STEM CELL FUNCTIONAL CHANGES WITH AGE IN OTHER TISSUES SUCH AS THE BRAIN AND THE MUSCLES ARE ALSO CONSISTENT WITH THE -- AND DYSFUNCTION IN AGED TISSUES. THE STEM CELL DEGENERATION ACCELERATES AGING. A GOOD EXAMPLE IS THE PREMATURE AGING DISEASE. THIS PATIENT HAS A VERY SHORT LIFESPAN ABOUT 10 TO 20 YEARS. -- IN THIS PATIENT ARE COMPROMISED AND THEY HAVE A REDUCED DIFFERENTIATION TOWARDS -- AND INCREASE DIFFERENTIATION TOWARDS OSTEOBLAST. CONSISTENTLY IN THIS PATIENT, THE MAIN TISSUES AFFECTED ARE IN THE -- REGION. FOR EXAMPLE, THEY HAVE AN INCREASED TURNOVER IN THE BONE AND A LOSS OF SUBACUTE -- -- MANIPULATION LEADS TO LIFESPAN EXTENSION. THERE'S SOME EMERGING EVIDENCE TO SUGGEST THAT STEM CELL MANIPULATION CAN EXPAND THE LIFESPAN. FOR EXAMPLE, IN FLIES, MANIPULATION OF SEVERAL MOLECULAR PATHWAYS IN -- STEM CELLS LEADS TO LIFETIME EXPANSION. HOW ABOUT MAMMALS? IN THE PROGERA MOUSE MODEL, INJECTION OF MUSCLE STEM CELLS ALMOST DOUBLED THE LIFESPAN. WE HAVE LEARNED A GREAT DEAL ABOUT ORGANIZE MANISMAL AGING AND STEM CELL AGING AT THE MOLECULAR LEVEL. IF STEM CELL DETERIORATION IS INDEED A MAJOR CONTRIBUTOR TO ORGANISMAL AGING, THEN THEY SHOULD SHARE COMMON MOLECULAR REGULATORS. REEBRECENT DATA IS ALSO CONSISTENT WITH THE IDEA THAT LONGEVITY PATHWAYS REGULATE STEM CELL FUNCTION, AND I LISTED HERE THE THREE COMMON CON -- LONGEVITY PATHWAYS AND THEY HAVE ALL BEEN TESTED. IT'S IMPORTANT TO POINT OUT THAT FOXO, A MAJOR COMPONENT IN THE IGS PATHWAY AND 33, A MITOCHONDRIAL -- HAVE BEEN SHOWN TO -- IN THE CELLS AND PREFERENTIALLY REGULATE STEM -- CELLS BUT NOT THE DIFFERENTIATED PROGENY. THE ENRICHMENT OF LONGEVITY FACTORS SPECIFICALLY IN THE STEM CELL COMPARTMENT PROVIDE FOR THE SUPPORT TO THE STEM CELL THEORY OF AGING. NEXT I WILL DISCUSS HOW DO STEM CELLS AGE. STEM CELLS MOSTLY RESIDE IN THE QUIESCENT STAGE, WHICH IS THE METABOLICALLY INACTIVE, AND IT'S THOUGHT THAT THIS IS TO REDUCE THE PRODUCTION OF METABOLIC BYPRODUCTS -- HOWEVER -- LEVELS STILL INCREASE WITH AGE IN STEM CELLS. THERE ARE A NUMBER OF OF ANIMAL MODELS WITH DEFECTIVE MANAGEMENT OF OXIDATIVE STRESS AND THEY ALL HAVE A COMPROMISE OF STEM CELL FUNCTIONS. CONVERSELY -- REVERSES THE REGENERATIVE CAPACITY OF STEM CELLS. THIS IS LIKELY TO BE A CONSERVATIVE MECHANISM THAT HAS BEEN TESTED IN DIFFERENT SPECIES AND IN MAMMALS HAS BEEN TESTED IN DIFFERENT TISSUES. ANOTHER CONTRIBUTING FACTOR IS -- MOST HUMAN CELLS DO NOT EXPRESS -- THE ENZYME THAT REPLENISHES -- TILLAMERES, HOWEVER, STEM CELLS EXPRESS HIGH LEVELS. DESPITE THE PRESENCE OF -- STILL SHORTENS WITH AGE IN STEM CELLS. -- IN MICE HAVE A DEFECT IN STEM CELL FUNCTION BUT ONLY OF AFTER THREE GENERATIONS, AND THIS IS LIKELY BECAUSE MICE HAVE EXTENDED TELOMERES. IN HUMANS -- ATREETION IS ALSO CONSISTENT WITH COMPROMISE OF STEM CELL FUNCTION. DISCARE PTOSIS -- DISEASE IN MANY WAYS RESEMBLES PREMATURE AGING AND PATIENTS HAVE A SHORTENED -- AND REPAIRED REGENERATION OF PROLIFERATIVE TISSUE SUCH AS BONE MARROW AND THE SKIN. SO FINALLY, CAN STEM CELL AGING BE TARGETED TO COMBAT AGING-ASSOCIATED DEGENERATIVE DISEASES? AS I JUST DISCUSSED, STEM CELL AGING IS IN A WAY VIEWED AS ACCUMULATION OF CELLULAR DAMAGE DUE TO VARIOUS CELL ACTIVITY THROUGHOUT THE LIFE. IN RESPONSE TO CELL DAMAGE -- SUPPRESSORS ARE SUPPOSED TO BE ACTIVATED RESULTING IN APOP PTOSIS, IN ESSENCE, CELL CYCLE ARREST. SO INDEED, A STEM CELL AGING IS REGULATED BY TUMOR SUPPRESSORS, P16, RESCUES STEM CELL AGING. SO IN A WAY, STEM CELL AGING IS VIEWED AS AN EVOLVED MECHANISM TO -- CANCER AND I THINK ANN ALSO BROUGHT UP THIS POINT YESTERDAY. SO THE IDEA IS THAT TARGETING TUMOR SUPPRESSORS TO PREVENT STEM CELL AGING IS POSSIBLE TO INCREASE -- GENESIS. SO IDEA OF STEM CELL BASED THERAPIES SHOULD AIM TO REDUCE THE ACCUMULATION OF CELLULAR DAMAGE IN ORDER TO PREVENT BOTH STEM CELL AGING AND CANCER. HOWRVE, IF THE ACCUMULATION OF CELLULAR DAMAGE IS INTRINSIC TO CELLULAR ACTIVITIES IT THROUGHOUT THE LIFE, THE QUESTION REALLY IS, IS IT POSSIBLE TO REDUCE CELLULAR DAMAGE? FOR EXAMPLE, THE PRODUCTION, THE ACCUMULATION OF -- IN AGED CELLS IS THOUGHT TO BE A RANDOM -- PROCESS. SO FREE RADICALS ARE PRODUCED DUE TO RESPIRATION, SO IT'S A BYPRODUCT OF RESPIRATION, AND THIS -- MUTATION AND DEFECTIVE MITOCHONDRIA -- AND THIS FURTHER INCREASES THE PRODUCTION OF LOF. HOWEVER, A RECENT STUDY SUGGESTS THAT -- RATHER A REGULAT REGULATED PROCESS WITH -- PLAYING A ROLE, SO -- 3 REGULATES A STRESS RESPONSE IN HEMATOPOIETIC STEM CELLS TO REDUCE OXIDATIVE STRESS AND THIS PROTECTIVE MECHANISM IS SUPPRESSED IN AGED HEMATOPOIETIC STEM CELLS AND THIS MAY CONTRIBUTE TO THE -- IN AGED STEM CELLS. INTERESTINGLY, REINTRODUCTION OF -- 3 TO AGED HEMATOPOIETIC STEM CELLS IMPROVE THE FUNCTIONAL CAPACITY OF AGE HEMATOPOIETIC STEM CELLS, SO IN A WAY, WE CAN VIEW IT AS A REJUVENATION OF AGED STEM CELLS. SO I THINK IT'S IMPORTANT TO UNDERSTAND WHICH TYPES OF CELLULAR DAMAGE IS A REGULATED PROCESS AND WHICH TYPES OF CELLULAR DAMAGE HAS THE EFFECT OF STEM CELL FUNCTION, THAT IS, THE REVERSIBLE, AND THIS MAY PROVIDE THE BASIS TO REJUVENATE AGED STEM CELLS IN ORDER TO COMBAT AGING-ASSOCIATED DEGENERATIVE DISEASES. THANK YOU. >> OUR NEXT SPEAKER IS HARTMUT GEIGER, FROM THE CINCINNATI CHILDREN'S HOSPITAL AND FROM THE UNIVERSITY OF -- IN GERMANY. HE'S INTERESTED IN HEMATOPOIETIC STEM CELLS AND SIGNALING PATHWAYS THAT CONTROL FATE AND PARTICULARLY IN HOW CHANGES IN THE LOCAL ENVIRONMENT IN THE SO CALLED NEES NICHE AFFECT THE FUNCTION OF THESE STEM CELLS WITH AGE. >> AS WAS SAID, MY LAB IS INTERESTED IN STEM CELL BIOLOGY, STEM CELL AGING, STEM CELL REJUVENATION AND ALSO NICHE INFLUENCES ON AGING. >> THIS IS MY INTRODUCTORY SLIDE, IT WAS ON "NATIONAL GEOGRAPHIC" THIS MONTH, AND IT SHOULD JUST REMIND US THAT WE'RE TALKING ABOUT A GLOBAL PROBLEM IN TERMS OF AGING. I THINK INITIALLY IT WAS SHOWN BUT I'D LIKE TO EMPHASIZE THIS ONE MORE TIME. IT'S NOT JUST RESTRICTED TO WESTERN COUNTRIES. SO YOU'VE HEARD A LOT ABOUT STEM CELLS AND AS TOM SAID, THERE IS A -- I CAN'T EVEN PRONOUNCE IT, THAT CAN BE GOOD OR BAD, WHETHER STEM CELLS AGES AND WHETHER THEY PROTECT US FROM BECOMING AGED. STEM CELLS -- AS SINGLE UNITS. WHICH IS GOOD IN TERMS OF CELL BIOLOGY. BUT LIKE HUMAN BEINGS, THEY ARE NOT ALONE. SO THEY LIKE TO EACH OTHER, TO TALK TO THEMSELVES, AND OTHER PEOPLE, SO THEY HAVE OTHER CELLS AND SYSTEMS AROUND IT -- AND WE CALL THIS -- AS TOM SAID, OR TRAUMA CELLS OR SYSTEMIC FACTORS. SO WE SHOULD NEVER FORGET THAT. IT'S IMPORTANT TO REMEMBER. IT'S ALWAYS CROSSTALK FOR THE REST OF THE TALK, THIS MEANS NICHE CAN BE ALSO FOR LARGER SYSTEMS, THIS LOOKS LIKE CELLS, FOR SOME OF THE -- AND THEY INTERACT WITH THE STEM CELLS AND THIS REGULATES THEIR BEHAVIOR. SO WHY IS STEM CELL AGING INTERESTING? THERE IS THE ASSUMPTION THAT HIGHLY PROLIFERATIVE TISSUES, WHICH DEPEND ON STEM CELL ACTIVITY ALL OF THE TIME, LIKE BLOOD, INTESTINE, MUSCLE, SKIN, THE GERM LINES, THAT STEM CELL AGING, ON TOP OF THIS HIERARCHY OF TISSUE MAINTENANCE, AS THEY AGE, THE TISSUE AGES. IT'S 10 DAYS, IN THE BLOOD SYSTEM, TWO OR THREE MONTHS, BUT THEY DEPEND ON THAT. SO IT'S VERY IMPORTANT TO UNDERSTAND STEM CELL AGE AND WHAT ACTUALLY CONTRIBUTES TO -- STEM CELLS. ALL OF THE DIFFERENT TYPE OF BLOOD CELLS. THIS HAPPENS IN THE BONE MARROW USUALLY, THERE'S A GOOD SLIDE FROM A LONG TIME AGO, STILL VERY NICE TO SHOW THAT THERE'S CERTAIN NICHES, CELLS IN THE BONE MARROW, OR BONE LINING CELLS, AND THE ASSUMPTION IS THESE STROMA CELLS INTERACT WITH -- STEM CELLS AND DETERMINE AT LEAST IN PART THE OUTCOME OF CELL RENEWAL AND DIFFERENTIATION. AGAIN STEM CELLS ARE NOT DOING THAT IN THE MIDDLE OF NOWHERE. THEY'RE ALWAYS THE -- YOU CANNOT -- CANNOT TAKE THEM AND WAIT. SO THE QUESTION IS, THERE'S A LOT OF STEM CELL INTRINSIC COMPONENT -- SYSTEM WHERE THE STROMA CELLS DO ACTUALLY CHANGE WITH TIME TOO. THIS IS FROM OUR LABORATORY, OTHER LABORATORIES WITH SIMILAR DATA THAT'S ACTUALLY UNPUBLISHED. WE JUST MADE A VERY SIMPLE EXPERIMENT. WE TOOK NICHE CELLS OUT OF A YOUNG AGE MOUSE AND -- WHAT KIND OF CYTOKINES DO THEY SEE CRETE. SECRETE. THEY GO UP UPON AGING SHE COULD ACTUALLY SHOW THAT THIS IS MOST LIKELY ALSO INVOLVED IN -- LYMPHOID SHIFT AND IT MIGHT BE CAUSATIVE CHANGE BY -- WE HAVE NOT CHECKED ALL OF THE OTHER ONES, THIS IS JUST TO EMPHASIZE A VERY SIMPLE ASSAY -- THE INTERESTING PART IS -- GO UP, ALSO TALK ABOUT -- INHIBITORS SO MIGHT BE INTERESTING HERE. WE DON'T KNOW THE FUNCTION OF -- ON THE CAUSATIVE RELATION SHOULD -- CHANGES. THE OTHER QUESTION WAS DISEASES. SO AS YOU ALL KNOW, LEUKEMIA -- THIS IS FOR MDS, IT'S PROBABLY MYELOID DYSPLASTIC SYNDROME, INCIDENCE RATE, PROBABLY ONE OF THE MOST AGING-ASSOCIATED LEUKEMIA KIND OF THING THAT HAS BEEN FOUND. SO THE QUESTION IS -- THOUGHT TO BE A DISEASE THAT INITIATED BY STEM CELLS, PLI MALIGNANT STEM CELLS IN THE BONE MARROW. THE QUESTION IS, TO MAKE ONE SHORT EXAMPLE, AN -- NICHE MIGHT ACTUALLY SUPPORT THAT. SO FAR THE THINKING IS THAT STEM CELL AGES AND THAT WILL LEAD TO -- MUTATIONS, AND THAT WILL DRIVE THE LEUKEMIA TYPE DISEASES. I THINK PEOPLE SHOULD DO MORE OF THESE, NOT BECAUSE -- BUT BECAUSE SOMEBODY TALKED YESTERDAY ABOUT YOU SHOULD TEST IN AGED MICE WHAT'S GOING ON BECAUSE A LOT OF THIS LEUKEMIA -- YOUNG MICE -- WHICH ALLOW US TO CHANGE NICHES AND ALL OF THAT. THIS WOULD ALSO BE SOMETHING STEM CELL SYSTEMS NEED TO DEVELOP A LITTLE BIT MORE TO BE ABLE TO DISSECT AGING, STEM CELLS FROM AGING FROM NICHES, BUT WHAT WE CAN DO IS WE CAN TRANSPLANT PRELOU KEY MIC CELLS. I NEED TO TAKE THE DATA OUT TO EXPLAIN IT TO YOU. TRANSPLANT OF PRELEUKEMIC SELLS SELLS -- THIS GIVES YOU CERTAIN PRELEUKEMIA IN A YOUNG MOUSE. WE ASK IF YOU TAKE THE SAME CELLS, YOUNG CELLS, JUST PUT THEM IN AN AGED MOUSE, AS A RECIPIENT. WHAT DO YOU SEE IN TERMS OF PROGRESSION IN AN AGED MOUSE VERSUS A YOUNG MOUSE? THE DATA WAS PRETTY CLEAR THAT YOU CAN SEE THAT IN THIS AGED MOUSE, YOU HAVE A FAST EXPANSION OF THE LOU KEY MIC CLONE. IT DOESN'T CONVERT IT TO DISEASE YETS BUT CAN YOU CLEARLY MEASURE A FAST EXPANSION, WHICH MEANS THIS AGED NICHE SYSTEM IS LESS SUPPRESSIVE, I GUESS, FOR PROMOTING THIS DISEASE CLONE. SO WHAT CAN HAPPEN IT WILL PROBABLY NOT CHANGE THE DISEASE ITSELF BUT THE PROGRESSION ALLOWS FOR EXPANSION OF THESE CLONES THAT THEY HIT EARLIER OR FASTER WITH THIS CLINICAL DIAGNOSIS WHICH IS BASED ON -- NUMBERS, LEUKEMIC CLONES AND PERIPHERAL BLOOD. SO SIMILAR EXAMPLE HAS BEEN -- WHERE THEY LOOKED AT AGING OF SKELETAL MUSCLE AND NICHES, WHICH IS -- YOU SEE THE STEM CELLS ARE QUIESCENT COMING BACK TO TOM'S PARADIGM, THEY ARE HARDLY EVER ACTIVATED, IF THEY ARE ACTIVATED, A DIFFERENT SHADE. THIS IS DONE BY -- 2 AND THERE'S A NOTCH INHIBITOR THAT ALLOWS NOT TO DIFFERENTIATE TOO MUCH -- IN THE AGED ONE -- THE NOTCH WHICH NORMALLY INHIBITS THAT IS NOT DOING THAT ANYMORE, AND WHAT YOU CAN SEE IS ACTIVATION AND DIFFERENTIATION BUT IT'S NOT -- WE GOT DIFFERENTIATION TYPE OF THING BECAUSE THEN THE MUSCLE FIBERS BECOME MORE FIBROTIC. SO THAT WOULD SUMMARIZE AND THERE'S OTHER DATA FROM OTHER SYSTEMS AGAIN THAT SYSTEMIC FACTORS, WE WILL HEAR ABOUT THAT MORE -- DIFFERENT FACTORS, AND THAT CAN IFN FLEUNS HOW STEM CELLS PERFORM. THREE POINTS TO MAKE, THAT'S WHAT I WANTED TO SAY. IF YOU WANT TO UNDERSTAND AGING OF STEM CELLS BETTER, WE SHOULD NOT FORGET ABOUT -- STEM CELLS ARE NOT ALONE, THEY'RE ALWAYS EXPOSED TO THESE FACTORS. -- PARTIALLY AGING OF STEM CELLS. THE INTERESTING PART IS THAT THEY HAVE A LOT OF SUPPRESSIVE ACTIVITY AT THE STEM CELL OR A LOT OF ACTIVATION ACTIVITY. IT SEEMS THAT -- LOSE SUPPRESSION ACTIVITY SO THAT WOULD MEAN STEM CELLS IN GENERAL LIKE TO GO FORWARD, DO SOMETHING, DIFFERENTIATE OR REPLICATE, AND THAT NICHES DO SUPPRESS THAT, AND -- YOU HAVE HYPERACTIVITY IN CERTAIN DISEASES ASSOCIATED WITH THAT. -- WHICH WOULD LEAD TO HYPERACTIVITY, AND CHRONIC WASTING OF CERTAIN TISSUES. SO THAT WILL BE IMPORTANT TO DIFFERENTIATE BECAUSE THERAPEUTIC VAT GEES WOULD BE VERY, VERY DIFFERENT IN ONE OR THE OTHER SETTING. WITH THAT, I WOULD LIKE TO CONCLUDE, AND THANK YOU FOR YOUR ATTENTION. >> THANK YOU, HARTMUT. OUR NEXT SPEAKER IS EMMANUELLE PASSEGUE. HER LAB IS INTERESTED IN HOW CELL INTRINSIC DAMAGE IN STEM CELLS CAN AFFECT STEM CELL SURVIVAL AND POSSIBLY LEAD TO DYSFUNCTION AND TUMOR REGENESIS, AND SHE WILL ASK THE QUESTION SPECIFICALLY HOW DNA DAMAGE CAN LEAD TO CELL DYSFUNCTION. >> THANKS, TONY. AS THE PREVIOUS SPEAKER, I'M A STEM CELL BIOLOGIST AND I WANT TO LOOK AT ONE OF THE MOST OLD AND -- PRINCIPLE OF AGING, DNA DAMAGE, FROM THE LENS OF STEM CELL BIOLOGY, AND THE EFFECT ON THE -- CELL POPULATION -- STEM CELL, WHICH MAINTAINS TISSUE -- SO STEM CELLS ARE LIKE EVERY OTHER CELL IN THE BODY, EXPOSED TO A WIRLD WIND OF DAMAGE -- OR COMING FROM THE CELLS THEMSELVES THAT ARE INTRINSIC -- THIS IS CUMULATIVE DAMAGE WHICH AFFECTS THE CELL WHICH ARE MAINTAINED OVER TIME. TO -- STEM CELL -- TO THE DNA. I'M NOT GOING TO GO OVER THIS MECHANISM, JUST TO POINT OUT THAT WHILE THE MAJORITY OF THEM HAVE A FAIRLY HIGH -- ONE OF THOSE MECHANISMS IS VERY WELL-KNOWN TO BE HIGHLY -- MEANING -- WHICH CAN ACT ON DNA ACTUALLY CAN BE SICK, ESPECIALLY IN NON-CYCLING CELL WHICH ARE -- GLUE BACK TOGETHER THE BROKEN STRAND OF DNA OR YOU SEE -- OF VALUES -- INCLUDING TRANSLOCATION, THEREFORE, BEING BY ITSELF A VERY -- EVENT. SO WHY IS THAT IMPORTANT? IT'S BECAUSE THE CONSEQUENCE OF THE TYPE OF REPAIR, THE CONSEQUENCE OF THE RESPONSE TO DNA DAMAGE WHICH IS SURVIVAL THAT -- TOGETHER WITH -- IS WHAT WILL DICTATE AT THE END OF THE DAY THE BIOLOGY OF THE TISSUE. IF A STEM CELL ENCOUNTERS DNA DAMAGE AND SURVIVES THIS DNA DAMAGE, AND WE REPAIR THE DAMAGE -- THERE IS NO CONSEQUENCE, THE STEM CELLS KEEPS -- AND PRODUCING DIFFERENTIATED PROTEIN ENSURING TISH YOE HOMEOSTA HOMEOSTASIS. IF THE SAME STEM CELL SURVIVES THE DNA DAMAGE BUT THEN REPAIR IT IT WITH A NARROW -- AND A -- THIS CAN LEAD TO THE -- WILL LEAD IT TO THE PREDICTION OF PRO -- WHICH CARIES ITS MUTATION, AND THEREFORE, CAN CONTRIBUTE TO BOTH CANCER DEVELOPMENT OR AGING IN THE -- CHRONIC DISEASE AND AGE RELATED PLAGUE NANCY. ASPECTS CAN BE AS DAMAGING FOR TISSUE MAINTENANCE AND FOR AGING, WHICH IS THAT WHEN THE STEM CELL ENCOUNTERS DNA DAMAGE, IT DOESN'T SURVIVE. IT DIES. AND YOU LOSE YOUR STEM CELL POPULATION, YOU LOSE YOUR DIFFERENTIATED -- AND YOU LOSE MAINTENANCE OF THE TISSUE, LEADING TO ORGAN FAILURE, WHICH IS AS WELL ONE COMPONENT OF AGING. SO THAT'S WHY WE CARE ABOUT THE DNA DAMAGE RESPONSE WITHIN THE STEM CELL POPULATION. SO MY -- IS THE BLOOD SYSTEM AND THERE IS NO MYSTERY OF THAT BECAUSE IN HEMATOPOIETIC STEM CELL, THE MOTHER OF -- IS THE MOST KNOWN STEM CELL POPULATION, WE CAN TAKE IT OUT, TRANSLATE, MANIPULATE, FUNCTIONALLY TEST THE BIOLOGY OF THE HEMATOPOIETIC STEM CELL POPULATION, AND YET BE ON THE FOREFRONT OF A LOT OF DISCOVERY IN THE STEM CELL FIELD. SO WHAT I JUST WANT TO SAY ABOUT THIS HEMATOPOIETIC BLOOD FORMING STEM CELL, FOR HUMAN AND MOUSE WITH A SLIGHT DIFFERENCE BUT THE GENERAL PRINCIPLE ALL TRUE IN BOTH SPECIES IS THAT THEY ARE ESSENTIAL FOR PRODUCTION OF ALL THE -- ARE VERY IMPORTANT IN THE CONTEXT OF -- AND WHAT'S VERY IMPORTANT IS IN -- ORGANISM, I'M NOT TALKING ABOUT THE DEVELOPMENT OR ESTABLISHMENT OF THE BLOOD SYSTEM, I'M TALKING ABOUT THE MAINTENANCE IN THAT -- THE STEM CELL ARE KEPT IN A QIE QUIESCENT STAGE OF THE CELL CYCLE, WHICH IS ENFORCED BY THE BONE MARROW NICHE IN THE PREVIOUS TALK, BUT -- A VERY -- SITUATION, READY TO FIRE AND READY TO DIFFERENTIATE, READY TO PROLIFERATE AND PRODUCE PROGENIES, A LITTLE BIT LIKE THE RUNNER IN THE STARTING BLOCK. -- BY READY TO PROLIFERATE -- QUIESCENT IS -- AND DNA DAMAGE. AND THAT'S A WORK WE PUBLISH A FEW YEARS BACK FROM MY LAB -- SURVIVE IRRADIATION IN THIS CASE WE USE -- FORGOT TO PUT IT IN THE SLIDE -- RADIATION AND USING -- LOW DOSES BECAUSE THE BLOOD IS ONE OF THE MOST RADIO SENSITIVE TISSUE IN THE BODY. -- TO FIX IT AND THIS IS ASSOCIATED WITH -- OF MUTATION AND WHAT WE SHOW LOTS OF FUNCTION AND -- UPON -- ACCUMULATION OF MUTATION -- FORMATION AND IT'S -- BECAUSE -- QUIESCENT AND WE -- THEM TO PROLIFERATE, THEY STILL SURVIVE. THE SURVIVAL MECHANISM OUR LEVEL OF SURVIVAL FACTOR WHICH -- MACHINERY BY -- 3 PATHWAY IS THE SAME -- BUT NOW THEY CAN USE -- COMBINATION TO REPAIR THE DOUBLE STRAND BREAK, WHICH IS MUCH MORE -- AND -- CYCLE OF ACCUMULATION OF MUTATION. AND VERY SPECIFIC TO THE STEM CELL COMPARTMENT, MORE DIFFERENTIATED -- THEY DON'T DO THAT, BECAUSE -- SO THESE PRINCIPLES, TO MUTATION DNA DAMAGE QI QUIESCENT STEM CELL -- TO A LOT OF OTHER CONTEXT, MIGHT BE GENERAL -- OF -- WHICH ARE NOT EVERY STEM CELL YOU HAVE IN YOUR BODY, BUT -- STEM CELL IN THE -- IN THE -- CANCER STEM CELL WHICH ARE DRIVING CANCER DEVELOPMENT. IT MIGHT EXPLAIN THE STEM CELL ORIGIN OF A LOT OF CHRONIC DISEASE OR CANCER BECAUSE THERE ARE -- IT'S A LITTLE BIT COUNTERINTUITIVE TO THE DOGMA OF CANCER DEVELOPMENT, BUT THESE QIQUIESCENT CELLS THAT DO NOT PROLIFERATE -- DNA DAMAGE AND THAT CLEARLY CAN CONTRIBUTE TO THE LOSS OF FUNCTION WITH AGE. AND THAT A SUMMARY OF WHAT HAS BEEN ALREADY SHOWN BY THE PREVIOUS SPEAKER, THE STEM CELLS, HEMATOPOIETIC STEM CELL IS PRETTY MUCH THE ONLY BLOOD CELL THAT AGES BECAUSE ITS PURPOSE OVER THE LIFETIME AND ALL ORGANISMS, BOTH HUMAN AND MICE, THERE IS A WHOLE ARRAY OF PROBLEM ASSOCIATED WITH THIS AGING STEM CELLS, AND THE MOST STRIKING FEATURE IS THAT THEY ACCUMULATE WITH AGE. 'NOTHER TISSUE, STEM CELL POPULATION DECREASE IN THE BLOOD SYSTEM, A ALL STEM CELL ACCUMULATES BUT THEY ARE HIGHLY DYSFUNCTIONAL AS ALREADY MENTIONED BY THE PREVIOUS SPEAKER. WHICH IS SIMILAR TO WHAT IS UP THERE DURING PHYSIOLOGY -- AGING. RELATED AS WELL TO -- IN THE LAST SESSION, THERE IS VERY STRONG ARGUMENT COMING FROM HUMAN SEQUENCING DATA WHICH SHOWS THAT -- ON RELAPSE LEUKEMIA PATIENT OVER TIME. WE SHOW THAT IN ADDITION OF DRIVING MUTATION, WHICH REALLY ARE AND WE ARE TALKING BETWEEN 500,000 MUTATION ACCUMULATING IN THIS -- OVER TIME, AND WHAT -- ALREADY SHOW ACCUMULATION OF -- IN THIS --DNA DAMAGE, THESE ARE COMING FROM INCREASED -- AND DONE IN MY LAB SHOWING THAT -- DUE TO REPLICATION ERROR. SO HOW CAN WE PREVENT THAT ACCUMULATION OF DNA DAMAGE IN A YOUNG CELL, IT'S EXACTLY THE PRINCIPLE OF -- STRATEGY -- IT'S A -- SURVIVAL -- AND THEN -- WHICH I CHECKED THIS SURVIVAL -- OF A YOUNG STEM CELL, WE SEE ACCUMULATION OF -- STEM CELL -- AND WHICH CAN REPAIR USING THIS -- TO GO ON AND CONTRIBUTE TO CHRONIC DISEASE. WITH VERY -- MAYBE VERY NICE WAY TO TRY TO INTERVENE WITH THIS ACCUMULATION OF DNA DAMAGE BY PREVENTING SURVIVAL IN ACCUMULATION -- AND TRYING TO REDUCE THE RECURRENCE OF THIS ERROR PRONE -- THANK YOU. [APPLAUSE] >> THANK YOU. OUR NEXT SPEAKER IS RICHARD LEE FROM HARVARD MEDICAL SCHOOL. HE'S STUDYING CARDIOVASCULAR DISEASE AND HOW TARGETING STEM CELL MAY BE USEFUL IN REGENERATING OR POSSIBLY REJUVENATING THE HEART TISSUE AND HE WILL ASK THE QUESTION HOW DOES REPAIR OF THE WHOLE ORGAN BE ACHIEVED IN AGING. >> THANK YOU, TONY. SO I KNOW I WAS COMING AT THE END OF A LONG TWO-DAY MEETING AND I'M GOING TO SPEAK VERY CONCEPTUALLY WITH ESSENTIALLY NO DATA AND ALL AND IT MAY BE A LITTLE BIT INFLAMMATORY SO HOPEFULLY YOU'LL STAY AWAKE. THE QUESTION IS WHAT DO YOU WANT TO DO? WE WOULD LIKE TO GROW KIDNEYS, LIMBS, HEARTS, THINKS FOR ORGAN IT IS THAT WE REALLY HAVE GREAT DISEASE PROBLEMS FOR. SO I'M GOING TO START BY MAKING A COMPARISON THAT YOU'RE GOING TO THINK IS RIDICULOUS, BUT IF YOU'D BEAR WITH ME FOR A SECOND. LET'S JUST SAY THAT YOU WANTED TO GO TO THE - MOON, WE ALREADY DID THAT HALF A CENTURY AGO, BUT LET'S JUST CONSIDER FOR A SECOND YOU DIDN'T KNOW WE'D DONE THAT 50 YEARS AGO, 45 YEARS AGO, AND YOU GO OUT TONIGHT AND YOU WERE LOOKING OUT AT THE MOON AND THINKING, WE CAN GO THERE. IT'S A RIDICULOUS CONCEPT, RIGHT? I MEAN, IT REALLY IS ALMOST MIND BOGGLING THAT THEY DID THIS. NOW HOW CAN THEY DO THIS? IT TOOK A LOT OF DETERMINATION, IT TOOK A LOT OF TEAM WORK, IT TOOK A LOT OF COURAGE, IT TOOK A FUNCTIONING GOVERNMENT. [LAUGHTER] I'M PROBABLY NEVER GOING TO BE INVITED BACK HERE AGAIN. IT ACTUALLY WASN'T THE BEST TECHNOLOGY, IT WASN'T ANYTHING NEAR WHAT WE HAVE AVAILABLE TO US TODAY. SO THIS IS MY IPHONE. IT HAS 2,000 TIMES THE COMPUTING CAPACITY OF THE APOLLO 11 SPACECRAFT. THAT'S THIS IPHONE. THIS ISN'T EVEN THE UP TO DATE IPHONE BECAUSE MY CELLULAR PLAN WON'T LET ME UPGRADE FOR LIKE ANOTHER YEAR. SO OUR TECHNOLOGY THESE DAYS REALLY IS NOT THE PROBLEM. HOW DID THEY ACTUALLY DO THIS? IT'S ACTUALLY PUN IF YOU GO BACK AND YOU LOOK AT THE FLIGHT PLANS FOR THE APOLLO 11 SPACECRAFT BECAUSE THEY WERE REMARKABLE. THIS IS JUST A SNAPSHOT HERE, BUT THEY DID THIS BY HAVING THE HUMILITY TO UNDERSTAND EVERYTHING. THEY REALLY UNDERSTOOD THE PROBLEM. THEY UNDERSTOOD EVERY LITTLE DETAIL. IF YOU GO BACK THROUGH ALL OF THE THINGS IN THE NASA WEBSITE, YOU CAN SEE THEY KNEW EVERYTHING THAT WAS GOING TO HAPPEN AT EVERY SECOND. THAT'S REMARKABLE. THAT'S ACTUALLY HOW THEY PULLED IT OFF, WITH WHAT WE WOULD CALL STONE AGE TECHNOLOGY. FROM 45 YEARS AGO. SO THE COMPARISON I WOULD LIKE TO MAKE IS THAT IF WE WANT TO DO SOME OF THESE DRAMATIC THINGS WITH REGENERATIVE MEDICINE, WE HAVE TO APPROACH IT THE SAME WAY. WE CAN'T ACTUALLY GO INTO THESE THINGS, NOT KNOWING WHAT THE HECK IS GOING TO GO ON. THIS IS A SLIDE I USE A LOT. WONDERFUL NATURE REVIEW BACK IN 2006, BUT I THINK THAT IT'S STILL VERY MUCH UP TO DATE. TOM MADE THE POINT THAT THERE ARE MANY CELLS THAT WE UNDERSTAND QUITE WELL THAT HAVE A LOT OF CELLULAR REGENERATIVE POTENTIAL AND THESE ARE PLACES WHERE WE HAVE THINGS IN CLINICAL USE FOR REGENERATION. ON THE OTHER HAND, A LOT OF THE TISSUES THAT WE'RE INTERESTED IN, LIKE THE HEART OR THE RETINA OR THE BRAIN OR THE LUNG, ARE AREAS WHERE THEIR CONTROVERSIAL BUT IT SEEMS TO BE RELATIVELY LOW REGENERATIVE POTENTIAL AND WE ACTUALLY DON'T UNDERSTAND THOSE TISSUES VERY WELL. SO JUST IN MY TISSUE, THE HEART, THERE HAVE BEEN MANY THOUSANDS OF PATIENTS TREATED WITH CELL THERAPY BASED ON THE CONCEPT THAT IT THERE'S STEM CELLS IN THE HEART BUT MANY STUDIES WERE SUGGESTING THOSE CONCEPTS WERE COMPLETELY ERRONEOUS, SO WE'RE THOUSANDS OF PATIENTS INTO HUMAN TRIALS WITHOUT UNDERSTANDING THE FUNDAMENTALS. CAN YOU JUST IMAGINE IF THE NASA GUYS SAID, WOW, YOU GUYS DIDN'T EVEN THINK ABOUT HOW FAR AWAY THE MOON WAS. SO I BELIEVE WE REALLY NEED TO UNDERSTAND THE FUNDAMENTALS, THAT THE BASIC INVESTIGATION WITH WHAT'S GOING ON IN THESE TISSUES IS CRUCIAL TO THE TRANSLATION. SO THIS IS SOMETHING THAT YOU ALL KNOW, THIS IS U.S. LIFE EXPECTANCY OVER THE PAST CENTURY. YOU SEE IT'S GONE UP, ALMOST DOUBLING, AND THE CONCEPT IS SOMETHING WE ALL KNOW, WHICH IS THAT FOR MOST OF US WHO ARE PLAYING ON THE BACK NINE OF LIFE NOW, WE ARE DEALING WITH AN ENVIRONMENT THAT IS NEW. THAT IS BIOLOGICALLY NEW. JUST TO MAKE A POINT, THIS BIG DIP AROUND 1917, THIS WAS NOT WORLD WAR I, IF THIS WERE -- IN THE UNITED STATES THIS WAS SPANISH FLU, SO THAT'S JUST A REMINDER TO GET YOUR FLU SHOT THIS YEAR. YOU MIGHT HAVE NOTICED SOME DATA THAT CAME OUT JUST A FEW DAYS AGO THAT FOR REASONS NOT CLEAR, IF YOU GET A FLU SHOT, IT DECREASES YOUR RISK OF A HEART ATTACK AND STROKE. SO IF IT WASN'T ENOUGH TO INCITE YOU TO PREVENT FROM GETTING SICK AS ALL GET OUT WITH INFLUENZA, THAT IT MAY ACTUALLY SEEM TO PREVENT A STROKE. BUT THE CONCEPT I WANT TO POINT OUT HERE IS THAT THE AGING ENVIRONMENT IS A NEW PROBLEM, IT'S SOMETHING WE REALLY DON'T UNDERSTAND. YOU HEARD OVER AND OVER TODAY THAT THERE ARE MANY STUDIES PERFORMED AND THEY'RE MOSTLY DONE ON 10 WEEK OLD MICE. THE NUMBER OF STUDIES WHERE WE LOOK AT THESE PROBLEMS IN MICE THAT ARE TWO YEARS OLD, WHICH IS REFLECTING THE SORT OF ENVIRONMENT THAT WE REALLY ARE DEALING WITH MOST OF THE TIME, ARE ACTUALLY RELATIVELY SMALL. EXCEPT FOR THE LABS SPECIFICALLY STUDYING AGING. SO WHAT SHOULD WE DO? WHAT ARE THE KINDS OF THINGS THAT I BELIEVE WE NEED TO DO? SO YOU ALL KNOW YOU CAN TAKE CELLS OUT, WE CAN MAKE -- CELLS, WE CAN REALLY FASHION REGENERATIVE ORGAN, WE CAN DO 3D PRINTING, WE HAVE SCAFFOLDS, WE HAVE LOTS OF MATRIX TOOLS, WE HAVE WAYS OF DOING REGENERATION IN THE LABORATORY, AND I THINK THE CAPACITY TO GROW LIMBS IS GOING TO ACTUALLY COME ABOUT. IN BOSTON, THAT'S A BIG TOPIC THIS YEAR. WHAT WE NEED DO IS THINK ABOUT THESE THINGS NOT IN TERMS OF JUST DOING THEM IN 10 WEEK OLD MICE. WE NEED TO THINK ABOUT THE CONTEXT OF THE PROBLEM. WE NEED TO THINK ABOUT THEM IN THE CONTEXT OF THE FUNDAMENTALS. AND THAT'S DISEASES LIKE DIABETES, WHERE THE MILL EU IS TOTALLY DIFFERENT, AND A LOT OF REGENERATIVE MEDICINE, FOR EXAMPLE, IN LIMBS AND IN THE HEART, THEY HAVE TO BE CONSIDERED IN THE CONTEXT OF DIABETIC PATIENT BECAUSE THAT'S WHAT WE'RE FACING. AND THE AGING ENVIRONMENT. THESE STUDIES ARE HARD BECAUSE THEY'RE EXPENSIVE, BUT ALL OF THOSE CLINICAL TRIALS THAT I WAS TELLING YOU ABOUT IN THE HEAR, THEY WERE BASED LARGELY ON 10, 12 WEEK OLD MICE AND SOME YOUNG RATS, VERY, VERY RARELY ARE THEY BASED UPON ANIMAL MODELS THAT ARE REFLECTIVE OF THE TYPE OF PATIENT THAT WE'RE ACTUALLY TRYING TO TAKE CARE OF. SO ULTIMATELY WHAT WE NEED TO DO IS RECOGNIZE THAT ALTHOUGH WE WANT TO TAKE REGENERATIVE MEDICINE FORWARD AS FAST AS WE CAN, IF WE DO THAT BUT WE DON'T UNDERSTAND THE SIMPLEST FUNDAMENTALS LIKE HOW CELLS ARE ACTUALLY BEING REPLACED, WHAT'S MAINTAINING HOMEOSTASIS, WHAT DOES THE SYSTEM ACTUALLY LOOK LIKE, THEN WE'RE PROBABLY GOING TO MAKE A LOT OF MISTAKES. SO THANK YOU VERY MUCH. [APPLAUSE] >> THANK YOU, RICHARD. OUR LAST SPEAKER IS IRINA CONBOY, FROM THE UNIVERSITY OF CALIFORNIA IN BERKELEY. HER LAB STUDIES, TISSUE AGING AND HOW STEM CELL ACTIVITY MAY HELP REGENERATE TISSUES, AND SHE'S GOING TO ASK WHETHER THIS WILL BE A USEFUL APPROACH FOR THE TREATMENT OF CHRONIC DISEASES. SO AGING IS -- WITH THE DISEASE, AGING OR DISEASE. AGING IS BASICALLY JUST A GENERIC TERM FOR ALL OF THESE, AND MANY MORE HORRIBLE DISEASES, AND THERE IS NO SUCH THING BY THE WAY AS HEALTHY AGING. HEALTHY AGING SIMPLY MEANS THAT THE PERSON DOES NOT SUCCUMB TO ALL OF THESE HORRIBLE DISEASES, BUT ONLY TO SOME OF THEM. MOST OF THESE DISEASES ARE DEGENERATIVE. THE SLIDES WERE GIVEN TO ME BY JUDY CAPIZZI BECAUSE IT WAS DIFFICULT FOR ME TO MAKE A 10 MINUTE TALK -- IN THE SAME WAY, SO JUDY USUALLY EXCLUDES CANCER FROM THE LIST OF DEGENERATIVE DISEASES AND MAKES VERY NICE ELEGANT CONNECTIONS TO -- BUT FROM MY STEM CELL SCIENTIST LABS, I PERCEIVE THAT CANCER ACTUALLY, DEVELOPMENT OF CANCER IS -- BY IMMUNE DYSFUNCTION, WHICH -- IS NOW WORKING ON VERY SUCCESSFULLY AT STANFORD, AND SO CANCER ALSO -- PERCEIVED AS THE DEGENERATIVE DISEASE OF THE IMMUNE SYSTEM. SO -- RISE EXPONENTIALLY WITH AGE. THIS IS THE BREAKING POINT. SO WHY IS THAT? WELL, THE LOGICAL SELF -- EXPLANATION OR INTUITIVE EXPLANATION IS THAT IT CAUSES DAMAGE -- ACCUMULATE WITH AGE. BUT ON 2-YEAR-OLD HUMAN AND 2-YEAR-OLD MOUSE IS SOMEWHERE THERE, SO WHY IS THAT? WHY IS THERE MORE DNA DAMAGE IN 2-YEAR-OLD MOUSE AS COMPARED TO 2-YEAR-OLD HUMAN, AND WHY DOES 2-YEAR-OLD MOUSE HAVE -- AND OTHER HORRIBLE THINGS THAT 2-YEAR-OLD HUMAN DOES NOT? SOME PEOPLE SAY BECAUSE MOUSE IS SMALL AND HUMAN IS BIG AND ALSO MICE ARE VERY METABOLICALLY ACTIVE, BUT 3-YEAR-OLD RATS STILL HERE, YOU HAVE MANY AGE RELATED DISEASES, AND 3-YEAR-OLD SQUIRREL, WHICH IS BASICALLY THE SAME RAT WITH BUSHY TAIL, OVER HERE. [LAUGHTER] >> AND SQUIRRELS ACTUALLY LIVE 14 YEARS. SO -- TISSUES OR ANYTHING, CANNOT REALLY EXPLAIN THE AGING. WHAT CAN EXPLAIN THE AGING IS THE ABANDONMENT OF TISSUE MAINTENANCE AND REPAIR. -- AND IN MY LAB, THEY HAVE IDENTIFIED SOME OF THESE BIOCHEMICAL PROCESSES THAT CAUSE ABANDONMENT OF ORGAN MAINTENANCE AND REPAIR OR COLLECTIVELY, AGING. -- COMMON COURSE AGING OF MULTIPLE TISSUES, WHY THEN SOME PEOPLE WHEN THEY GROW OLD -- BEAUTIFUL HEAD OF HAIR. SO JUST TO GIVE YOU AN EXAMPLE EXAMPLE -- REPAIR OF OUR AUTOMOBILES, IT DOES NOT MEAN THAT TRANSMISSION WILL FAIL IN ALL OF THEM ON THE SAME DAY, BUT WHAT IT MEANS IS THAT SOONER OR LATER, SOME OF THE KEY PARTS WILL FAIL AND THE CAR WILL DIE. >> -- AGING PROCESS CAN PREVENT MANY OR ALL AGE-RELATED DISEASES, ALL OF THEM. WHAT I CANNOT UNDERSTAND IS WHY IS THERE NOT MORE INVESTMENT FROM SOCIETY AND FUNDING INTO AGING BECAUSE WE CAN THEN ELIMINATE DISEASES AT THEIR CAUSE AND IT WILL BE BILLION DOLLARS LESS COSTLY THAN TRYING TO CURE DISEASES AS THEY DEVELOP, SOME OF THEM ARE INCURABLE, YOU DON'T KNOW WHAT TO DO WITH THEM, AND -- IMPRACTICAL. SO WHAT I WAS THINKING IS THAT WHY PEOPLE DO NOT REALLY INVEST -- PARACHUTE WHEN THEY KNOW THEY'RE GOING TO JUMP OFF THE PLANE SOONER OR LATER IS THAT THEY SIMPLY CANNOT BELIEVE THAT THERE COULD BE ONE FUNDAMENTAL CAUSE OF AGING OF MULTIPLE TISSUES AND ONE FUNDAMENTAL CURE. SO TO GIVE YOU AN EXAMPLE, IF -- 150 YEARS AGO, YOU TOLD SOMEBODY THAT COMPLETELY DIFFERENT DISEASES SUCH AS FOR EXAMPLE TUBERCULOSIS AND MENINGITIS COULD BE CURED BY FUNDAMENTALLY SIMILAR PROCESS, PEOPLE WOULD NOT HAVE BELIEVED YOU. THEY WOULD SAY NO -- THEY CAN AFFORD IT TO -- AND -- WELL, YOU CANNOT DO ANYTHING, WE CAN JUST HOPE THAT OUR DIAGNOSIS IS WRONG. BUT THEN WHEN YOU DISCOVER -- BY BACTERIA, AND BACTERIA AND -- HAVE FUNDAMENTALLY DIFFERENT TRANSLATIONAL MACHINERY AND -- NOW THIS WHOLE CLASS OF DISEASES COULD BE ELIMINATED. SO WHAT -- THE SAME FUNDAMENTAL UNDERLYING PROCESS FOR TISSUE AGING EXISTS, AND -- ALL OF THESE DISEASES COULD BE ELIMINATED BEFORE -- SO IN MY LAST SLIDE, I'M GOING TO GIVE YOU AN ILLUSTRATION OF THAT. THEY TRIED TO GLOBAL -- AGING BY -- MOLECULES, AND WHAT YOU CAN SEE HERE IS THE SKELETAL -- THAT HAS BEEN --. IN THE YOUNG MOUSE OR HUMAN -- BY HEALTHY TISSUE, BUT IN THE OLD MOUSE -- AND FIBROSIS, WHICH CAUSES, OF COURSE, LACK OF FUNCTION AND ALSO OTHER DISEASES OF AGING SUCH AS INFLAMMATION, DEVELOPMENT OF -- AND SO FORTH. HOWEVER -- OF THE SAME -- THE YOUNG ONE. -- CAME FROM EXPERIMENTS SUCH AS -- OR HIG HYDROCHRONIC TISSUE TRANSPLANT WHICH TOLD US TWO MAIN THINGS. ONE IS THAT IT OLD STEM CELLS DO NOT -- AGE VERY MUCH, AND WHEN THEY FIND THEMSELVES IN A YOUNG ENVIRONMENT, THEY ARE COMPLETELY COMPETENT AND CAN REGENERATE YOUNG, HEALTHY TISSUE. THE SECOND LESSON WAS THAT THE BEHAVIOR OF STEM CELLS REGULATED BY -- PATHWAYS AND -- IS THE DEFENSE TO MAKE OLD -- CELLS TO REGENERATE -- BY THE RESIDENT OLD STEM CELLS. WHAT I WOULD LIKE TO EMPHASIZE IS THAT HYDRO -- OF ANIMALS ARE SHARED IS NOT THE SAME AS -- BAG OF WATER WITH -- BAG OF WATER. ALL -- BENEFITS FROM YOUNG SET OF LUNGS, YOUNG HEART, YOUNG IMMUNE SYSTEM AND OTHER ORGAN SYSTEMS, EACH COLLECTIVELY -- STEM CELL MAY HAVE YOU BEHAVIOR, SO I ASSU ME IT WILL BE ONE -- IN THE CALCULATION -- TO GROW OLD OR ONE -- GROW YOUNG. BUT THE IMPORTANT POINT IS THAT -- STEM CELLS -- AND -- HEALTHY TISSUES, AND THAT SPECIFIC BIOCHEMICAL SIGNALING SIGNALING -- SO THAT BRINGS ME TO THE PART WHICH IS -- PARABIOCYST, IF YOU KNOW THAT ALL YOU HAVE TO DO IS CALIBRATE -- PATHWAYS TO USEFUL -- STRENGTH, WHY NOT DO JUST THAT AND NOT TORTURE MICE BY -- EACH OTHER AND ACTUALLY BY THE WAY, IN 1957, THEY CONNECTED ONE POOR OLD RAT TO SIX YOUNG ANIMALS, AND AGING PREVAILED LONG TERM AND THERE WAS NO EXTENSION IN LIFESPAN. SO PARABIOSIS JUST TELLS US SOMETHING -- NOW IN REVIEW -- MY COLLABORATOR -- INTRODUCED ONE SMALL MOLECULE -- OLD MOUSE, AND IN ABOUT COUPLE OF WEEKS -- ENHANCED MYELOGENESIS -- GENESIS BY -- CALIBRATION OF ONE AND THE SAME -- SO THIS MOUSE -- AND WILL NOT HAVE LACK OF -- DUE TO OLD AGE. THIS IS -- THERE ARE MANY DISEASES WHICH ARE NOT ASSOCIATED WITH AGING, FOR EXAMPLE, JUVENILE DIABETES, BUT INTERESTINGLY TISSUE AGING BECOMES PREMATURELY -- NOT ONLY BY -- BUT ALSO BY INHIBITORS OF THE -- SO ONCE AGAIN, AGING AND DISEASE ARE -- AND NORMALLY -- AGING IS DEVELOPMENT OF MANY DISEASES, AND WHEN DISEASE -- YOUNG -- HUMAN TISSUE DEGENERATION BECOMES -- THANK YOU SO MUCH. [APPLAUSE] >> THANK YOU. MAYBE CAN I ASK ALL THE SPEAKERS TO COME TO THE PODIUM AND ALSO THE CHAIR, AND I MAKE A FEW COMMENTS. AS TOM ALLUDED ALREADY IN HIS INTRODUCTORY REMARKS, THE AGING OF STEM CELLS CAN REALLY BE SEEN AS AN INTEGRATIVE HALLMARK OF AGING. AS WE'VE SEEN IN ALL THE PREVIOUS SESSIONS, THE FUNDAMENTALS OF THE BIOLOGY OF AGING APPLY VERY MUCH TO THE AGING OF OF STEM CELLS. TO JUST SUMMARIZE A FEW OF THE MAIN POINTS THAT WE WANTED TO MAKE IN THIS SESSION, STEM CELLS COME IN DIFFERENT FLAVORS. THERE ARE STEM CELLS WITH HIGH TURNOVER THAT PRODUCE CELLS CONTINUOUSLY SUCH AS IN THE BLOOD, THERE ARE STEM CELLS THAT GIVE FUNCTION IN TISSUE REPAIR, AND THEN THERE'S STEM CELLS WITH VERY SPECIALIZED FUNCTION SUCH AS THOSE FOR EXAMPLE IN THE ADULT BRAIN THAT GIVE RISE TO NEURONS INVOLVED IN MEMORY PROCESSES. WE'VE ALSO LEARNED THAT STEM CELL ACTIVITY DECLINES DRAMATICALLY WITH AGE IN ALMOST ALL TISSUES AS FAR AS WE KNOW. WE KNOW THAT THE STEM CELL ACTIVITY IS -- BOTH BY CELL INTRINSIC AS WELL AS EXTRINSIC EFFECTS, AND THE INTRINSIC EFFECTS CAN BOTH LEAD TO DYSFUNCTION OF STEM CELLS OR TUMOR FORMATION, BUT THEY CAN ALSO PROMOTE AGING OF TISSUES, AND ON THE UPSWING, BENEFICIAL EFFECTS, FOR EXAMPLE, IN THE ENVIRONMENT BUT ALSO WITHIN THE CELLS, CAN HELP IN REPAIRING TISSUES AND POSSIBLY REJUVENATING THEM. AND I THINK THIS IS ONE OF THE EXCITING ASPECTS OF THIS FIELD, AND SO WHY DON'T WE TAKE QUESTIONS NOW. >> I'D LIKE TO ASK IF THERE'S ANY TISSUE FOR WHICH STEM CELL FUNCTION DECLINES BECAUSE OF DECLINING TELOMERES ACTIVITY. >> I CAN TELL YOU THAT STEM CELL FUNCTION DOES NOT DECLINE -- ACTIVITY, AND THAT YOU PUBLISHED THE PAPER A KUS COUPLE OF YEARS BACK SAYING THAT AT LEAST -- ACTIVITY IN STEM CELLS REMAINS HIGH, DOES NOT SIGNIFICANTLY DECLINE, BUT STEM CELLS FAIL IN TISSUE REGENERATION. SO THE AGE IS -- DECLINE -- ACTIVITY. >> I CAN ADD TO THAT AGAIN, IN THE MOUSE SYSTEM WHERE TRANSPLANTATION -- WHICH HAVE -- TELOMERE ACTIVITY, WHEN YOU -- FOR MORE THAN FIVE TIMES, AND FOR THEM TO UNDERGO A LOT OF -- STRESS, YOU SEE A VERSION OF -- AND A CRASH OF -- ACTIVITY, SO IT DOES -- BUT IT DOES -- IN THE -- CONTEXT COMPARED TO -- OF AGING AND THE FUNCTION OF THE STEM CELLS IN AN OLD ORGANISM. >> I JUST WANT TO ADD ONE THING. YOU CAN CAN OBVIOUSLY DEPLETE STEM CELLS BY INHIBITING -- YOU CAN MAKE STEM CELLS WORSE. THE QUESTION YOU'RE ASKING, REALLY, IS, IF YOU WERE TO BE ABLE TO MAINTAIN TELOMERES IN HUMANS, WOULD THERE BE A STEM CELL POPULATION THAT WOULD BE -- WE OBVIOUSLY CAN'T DO THAT SEW WNT DO KNOW THE ANSWER TO THAT, BUT AT LEAST THE EVIDENCE SO FAR AND CERTAINLY IN MICE WHERE THE TELOMERES ARE LONG IS THAT THERE IS A DEPLETION OF STEM CELLS BECAUSE OF A SHORTENING OF THE TEAL TELOMERES. >> WHEN YOU RE-ESTABLISH -- YOU DO NOT REALLY MAKE THE STEM CELLS GROW STRONGER IN A MOUSE SYSTEM. PLAWIN -- PLAYING A MAJOR ROLE IN STEM CELLS. >> OKAY. CAN >> I UNDERSTAND THAT YOU ARE ALL INTERESTED IN STEM CELL BIOLOGY. BUT I AM ALSO INTERESTED IN THE OTHER PART OF THE BODY, SO THE QUESTION FOR YOU IS, THE SYSTEMIC MILLEAU CAN IMPINGE UPON THE AGING PHENOTYPE OF SOMATIC CELLS, SO IN OTHER WORDS, AN AGED ENVIRONMENT CAN HAVE NOT ONLY EFFECT ON YOUNG STEM CELLS BUT ALSO ON OTHER SOMATIC CELLS SO THAT THIS COULD BE IN FAVOR OF WHAT I PRESENTED THE FIRST DAY THIS CRAZY IDEA THAT AGING IS PROC PROPAGATING AMONG THE CELLS AND THE TISSUES OF THE BODY >> I CAN TAKE THIS FROM THE POINT OF THE BRAIN. WE KNOW THAT CELLS IN THE BRAIN, IMMUNE CELLS, MICRO GLIA, ARE AIIVEAFFECTED BY SYSTEMIC CHANGES IN THE ENVIRONMENT AND THE ACTIVATION OF THESE MICRO GLIAL CELLS CAN HAVE NEGATIVE EFFECT ON THE STEM CELLS IN THE BRAIN. >> IN GENERAL, I THINK THERE'S ALWAYS A BALANCE BETWEEN THE STEM CELLS AND THE NON-STEM CELL -- IN AGING, AND THAT'S WHAT I WANTED TO SAY. IT'S NOT BLACK AND WHITE. IS THAT WHAT YOU'RE ASKING? SO TO HAVE SOMATI C. DIFF REN SHAITED CELLS -- ALSO VICE VERSA. >> GEORGE MARTIN, UNIVERSITY OF WASHINGTON. IRINA, YOU EMPHASIZED A SINGLE AGENT MADE A DIFFERENCE BOTH WITH NEURAL STEM CELL REGENERATION AND SKELETAL MUSCLE REGENERATION, SO YOU DIDN'T TELL US WHAT IT WAS, BUT I WAS THINKING IT'S GENERALLY THIS TREMENDOUS SELECTIVE PRESSURE, I WOULD THINK EVEN DURING REPRODUCTIVE PHASE, TO HAVE A BACKUP SYSTEM, A REALLY GAMBLE TO JUST PUT ALL YOUR MONEY ON THE SINGLE AGENT, SO HOW DO YOU RESPOND TO THAT? >> -- I CAN'T TELL YOU -- COLLABORATIVE WORK OTHERWISE I DEFINITELY WOULD BUT FOLLOW MY PUBL KAI YOU CAN EASILY GUESS -- PATHWAYS -- CALIBRATE THEM AT YOUNG LEVEL. NOW I DON'T THINK IT WILL DO ONE SINGLE AGENT SMALL MOLECULE WHICH CALIBRATES SIGNAL AND STRENGTH PRECISELY AT YOUNG LEVELS OR CLOSE TO YOUNG LEVELS, BUT I DON'T THINK THAT WE NEED HUNDREDS OF MOLECULES -- OR DELAYING TISSUE AGING. I THINK IT'S SOMEWHERE BETWEEN 3 AND 10. >> ACTUALLY YOU HELP MY HIGH FREQUENCY HEARING LOSS, SMALL MOLECULE, BECAUSE I HAD TROUBLE UNDERSTANDING YOU. >> I APOLOGIZE FOR THAT. >> TOM, DID YOU WANT TO EXPAND ON THAT? >> I THINK THE POINT IS THAT IF YOU HAVE -- YOU MAY NOT NEED, YOU KNOW, MULTIPLE FACTORS, IF YOU HAVE ONE FACTOR THAT MAY BE SUFFICIENT TO HELP REGENERATE A TISSUE THAT DOESN'T EXCLUDE -- THAT THERE ARE MANY OTHER ONES THAT CAN DO THE SAME THING. BUT GIVEN -- ONE FACTOR MAY HAVE SIGNIFICANT IMPACT. >> PUBLISHED -- YOU CAN ACTUALLY REJUVENATE STEM CELLS WITH ONE FACTOR. I THINK IT ALWAYS DEPENDS ON WHAT KIND OF PROBLEM YOU WANT TO FIX -- COMPLEX, OR YOU HAVE -- SIMPLE MECHANISM, IT LOOKS LIKE SOMETIMES MECHANISMS CAN BE RELATIVELY SIMPLE, AND ONE FACTOR COULD BE A VERY INTERESTING SOLUTION TO THAT PROBLEM. GOES BACK TO -- >> WHAT WE ARE TARGETING AND WHAT AGING IS -- I WOULD SAY IT'S MORE DEVELOPMENTAL PART, AND IT'S PROBABLY AS SOMEBODY ELSE SAID STEM CELLS SEEM TO BE VERY PROTECTIVE OF A COUPLE OF THINGS THAT COULD LEAD TO AGING AGING -- DAMAGE, PROBABLY DIFFERENTLY HANDLED -- DIFFERENTLY HANDLED, SO IT MIGHT COME MORE INTO SIMPLE DECISION DECISION -- >> THE QUESTION IS TO GEIGER, HARTMUT GEIGER. HE MENTIONED THE CHRONIC WASTING OF CERTAIN TISSUE FOLLOWING HYPO OR HYPERACTIVITY, AND IT REMINDED ME A COUPLE OF PAPERS OF --, AND SHE WAS CLAIMING THAT INFLAMMATORY STRESS WAS ABLE TO DEGENERATE THE NICHE COMPONENTS, AND SHE IDENTIFIED WITH OTHER AUTHORS -- BETWEEN 2007 AND 2009 THAT INFLAMMATORY CYTOKINES COULD BE STOCKED INTO ADIPOSE TISSUE MUSCLE AND CONTRIBUTE TO THEIR -- INACTIVITY, WHICH I THINK IS A SUBJECT WHICH HAS BEEN -- AN ISSUE THAT HAS BEEN ADDRESSED BY PREVIOUS SESSIONS, BUT IT JUST REMINDED ME THAT THIS COULD BE A MECHANISM, SO WHAT DO YOU THINK? >> SO YOU'RE SAYING SOME OF THE CYTOKINES THAT WE SHOWED -- INFLAMMATORY -- >> FOR ADIPOSE TISSUE, THIS IS DEMONSTRATED, DOESN'T NEED TO BE DEMONSTRATED BUT MAYBE MUSCLE. >> SO I WOULD ANSWER LIKE THAT, SO THEY ARE PROBABLY PARAMETERS OF INFLAMMATION THAT MIGHT BE SEEN IN BONE MARROW WITH AGING, BUT YOU CAN ALSO INTERPRET A LITTLE DIFFERENTLY -- IT'S JUST A PURE INFLAMMATORY RESPONSE, INFLAMMATORY RESPONSE -- THIS IS MORE DIFFERENT, SOME SIMPLE BUT MORE DIFFERENT. -- DNA DAMAGE AND THIS -- LIKE SOMEBODY ELSE. I DON'T KNOW THAT CURRENTLY, IT MIGHT BE SOMETHING WE ARE CURRENTLY INVESTIGATING THAT. I'M LOOKING FORWARD TO THE -- >> I GUESS I'M NOT QUITE SURE TO UNDERSTAND THE QUESTION, BUT I WOULD SAY IF WE LOOK AT OLD STEM CELLS FROM MUSCLE, COMPARE THEM TO YOUNG STEM CELLS, THERE'S A PROMINENT SIGNATURE OF INFLAMMATION ITSELF, SO THE CELLS ARE CLEARLY RESPONDING TO SOME ENVIRONMENTAL INFLUENCE THAT IS WHATEVER WE TALKED ABOUT HERE IN TERMS OF INFLAMMATION. IS THAT WHAT YOU'RE ASKING? >> NO, IT WAS NOT EXACTLY WHAT I WAS ASKING, BUT OKAY, THE IDEA WAS TO SAY THAT INFLAMMATORY CYTOKINES MAY CONTRIBUTE TO STEM CELL AGING, BASICALLY. >> I'M SAYING THE SIGNATURE ON THE CELLS SUGGEST THAT THERE IS AN INFLAMMATORY COMPONENT TO THE -- OF AGED STEM CELLS. WHETHER WHAT COMPONENTS OF THIS CYTODINE KIE PROFILE ARE ACTUALLY -- BLOCKING THEM AND SEEING WHAT EFFECTS -- IN TERMS OF BENEFICIAL REJUVENATION OF THE STEM CELL FUNCTION. >> THERE IS NO SUCH CYTOKINE OR MOLECULE WHICH MAKES US GROW OLD. ALL OF THEM ARE -- WHICH -- YOU CANNOT TURN SOMETHING ON AND BECOME YOUNGER. YOU CAN'T ADD SOMETHING AND BECOME YOUNGER. YOUNGER. >> RESPONSE IS VERY IMPORTANT FOR TISSUE REMODELING, BUT -- >> QUESTION OVER THERE? >> I'M FROM THE HEART LUNG AND BLOOD INSTITUTE. I WANT TO CONGRATULATE THE ORGANIZE -- ENGAGING AND INTERESTING THOUGHT PROVOKING. MY FIRST QUESTION. I ALWAYS WANTED TO -- BECAUSE TODAY IS HALLOWEEN. -- BASICALLY JUST THE SCIENTIFIC BASIS FOR DRACULA'S OLD TRICK. [LAUGHTER] >> YES. >> AND SO I WAS WONDERING IF THE BLOOD TRANSPLANTATION CAN REALLY BRING SOME OF THE BENEFITS, BUT I DO HAVE ANOTHER QUESTION, AND IT'S PROBABLY SO SIMPLE -- SIMPLE THAT MAY BE ALMOST PHILOSOPHICAL AND IT'S TRIGGERED BY RICH'S TECHNOLOGY COMPARISON. SO I THINK AS MOST PEOPLE IN THE ROOM, I HAVE A PERSONAL INTEREST IN DELAYING AGING AND AGE HAD BEEN RELATED DISEASES IN SCIENTIFIC INTEREST. THE DESIGN AS ANY GOOD OILED MACHINE, AT SOME POINT WE DO HAVE TO QUIT, SO THE QUESTION IS WHETHER WE SHOULD TRY TO FIGURE OUT WHAT WOULD BE THE MOST EFFICIENT SHUTDOWN PROCEDURE, SPEAKING ABOUT SHUTDOWN, AND NOT JUST AN ABRUPT CRASH, RIGHT, LIKE WE EXPERIENCED RECENTLY, BUT SO WHAT DO YOU THINK IT'S POTENTIALLY AN ORGANIZED SHUT DOWN? I KNOW -- SAID THERE IS NO HEALTHY AGING, BUT I THINK THERE IS -- I HOPE THERE IS, I DO WANT TO SHUT DOWN AT SOME POINT, I DON'T WANT TO CRASH. SO I CAME WITH A FEW SPARE PARTS, THE STEM CELLS, AND STIEMENTS I WILL RUN OUT OF THEM AND WHAT'S NEXT? >> IN TERMS OF THIS CONCEPT, THERE ARE SYSTEMIC THINGS THAT CAN PERHAPS REJUVENATE OR SYSTEMIC -- THAT ACTUALLY APPEARED IN THE FIRST SCIENTIFIC JOURNAL IN 1966 IN PHILOSOPHICAL TRANSACTIONS, THE VERY FIRST JOURNAL. PARDON? [INAUDIBLE] >> YEAH, YEAH, BUT DRACULA DIDN'T PUBLISH IT. SO SINCE THE BEGINNING OF THE SCIENTIFIC LITERATURE, THIS CONCEPT THAT THERE WERE HORMONAL FACTORS THAT COULD CONTROL YOUTH AND AGING HAS APPEARED, A AND SO I THINK NOW WE'RE STARTING TO GET TO THE POINT WHERE WE CAN REALLY FIGURE OUT WHAT SOME OF THOSE ARE, BOTH FROM PROTEOMIX TECHNOLOGY, DEEP SEQUENCING, MOLECULAR FINGERPRINT. IT'S HARD TO THINK ABOUT THESE THINGS AND WORK ON THEM WITHOUT THINKING ABOUT HEALTHY AGING AND HOW LONG DO WE WANT TO LIVE. >> I WAS JUST GOING TO SAY, TONY IS GOING TO ADDRESS THE QUESTION, THE FIRST PART OF YOUR QUESTION WHICH WAS THE BLOOD TRANSFUSION. >> WE HAVE EXPERIMENTS THAT SHOW THAT WE CAN TAKE PLASMA FROM YOUNG ANIMALS AND REJUVENATE BRAIN ACTIVITY AND COGNITION IN OLD ANIMALS, AND MY SON IS PLAYING DRACULA FOR HALLOWEEN. [ LAUGHTER ] >> CAN I ADD SOMETHING? SO I ASK YOU -- FOUNDATION -- EXTENSION FOUNDATION AND YOU -- FUNDING DK PROJECT IN OUR LAB WHICH IS BLOOD ANASTOMOSIS, SEE HOW LONG -- WE CAN CONNECT AND DISCONNECT -- PRECISE MANNER, YOU CAN REMOVE AND ADD COMPONENTS TO PLASMA, AND SO WE SEE THE NEXT GENERATION EXPERIMENT TO LOOK AT -- ORGAN REJUVENATION BUT I WOULD LIKE TO CAUTION EVERYBODY THAT THE THING THAT -- YOUNG COMPONENTS IN BLOOD, YOUNG COMPONENTS, PARABIOSIS IS THE CONNECTION OF DIFFERENT ORGAN -- AND IN CULTURE, WHEN WE MIX YOUNG CELLS WITH OLD BLOOD, OLD BLOOD -- [INAUDIBLE] >> STEWART KIM FROM STANFORD. DO STEM CELLS AGE WITH THE SAME MECHANISM AND DO THEY AGE WITH THE SAME RATE? DIFFERENT STEM CELLS? >> WELL, IF YOU LOOK AT, FOR EXAMPLE, MARKERS OF DNA DAMAGE, YOU CAN SEE THIS IN -- FROM DIFFERENT DEPARTMENTS. AGAIN, IT IT WOULD DEPEND A LITTLE BIT HOW YOU WOULD DEFINE WHAT AN AGING STEM CELL IS, AND AND -- ACROSS DIFFERENT STEM CELL POPULATIONS, BUT USING THE COMPARISON WE TALKED ABOUT AT THIS MEETING, IF WE LOOKED AT CELLULAR FUNCTIONS OR CELLULAR CHARACTERISTICS, IT'S ALWAYS HARD TO TALK ABOUT RATES OF AGING IN DIFFERENT TISSUES BECAUSE THEIR MANIFESTATIONS ARE WHAT WE MEASURE AS THE AGING PROCESS. >> IF YOU LOOK AT YOUR SATELLITE STEM CELL FUNCTION COMPARED TO THE DERRICK ROSSI MYELOID SWITCH, ARE THEY AGING AT THE SAME RATE? >> SO HAIR FOLLICLE STEM CELLS AND OTHERS WILL DECLINE MUCH MORE ROUGHLY, NEURAL STEM CELLS DECLINE MUCH MORE RAPIDLY THAN WHAT WE SEE IN THE MUSCLE OR CERTAINLY IN THE INTESTINE, IN THE BLOOD. SO THERE WILL BE DIFFERENT KINETICS CAN DEPENDING ON WHAT YOU MEASURE. SO I WOULD SAY FROM WHAT WE HAVE MEASURES FOR, THERE ARE DIFFERENT RATES OF AGING AND DIFFERENT STEM CELL POPULATION FOR DIFFERENT REASONS. SO THAT'S ONE THING. THEN -- BUT YOU ASKED ABOUT WHETHER THEY AGE WITH THE SAME MECHANISM, AGAIN, I THINK THIS GETS DOWN TO THE BASIC TOPIC OF THE SUMMIT OF WHAT ARE THE MECHANISMS OF AGING AND THEN CAN WE APPLY THAT TO DIFFERENT STEM CELL COMPARTMENTS. >> I CAN TELL YOU FOR MUSCLE AND BRAIN, STEM CELLS AGE -- ONE FUNDAMENTAL -- AND THIS IS OUR OUR -- [INAUDIBLE] >> AND TO ADD TO THAT, I THINK YOU'RE ASKING A VERY LOADED QUESTION IN THE SENSE OF STEM CELL BIOLOGY, BECAUSE IT'S RELATED WHERE HOW YOU DEFINE STEM CELL. GLUSH TRANSPLANT REGENERATES THE BLOOD AND REGENERATES -- SO THIS FUNCTIONAL DEFINITION DOESN'T HOLD IN AN AGE CONTEXT BECAUSE THE PREDICTION IS UNFAIR. SO ARE YOU STILL LOOKING AT THE SYMPTOMS OR ARE YOU JUST -- MARKER AND IT'S NOT A SYMPTOM ANYMORE, SO IS THAT STILL A VERY DIFFICULT QUESTION, WHICH HAS NOT BEEN ANSWERED. >> I WOULD LIKE TO ADD, BECAUSE STEM CELL AGING IS NOT THE LEAST EXPENSIVE ONE, A LOT OF -- TO LOOK AT THE BEGINNING AT THE END OF THE GAME -- STEM CELL, KINETICS -- TO LOOK AT, THAT'S MY -- SOMETIMES WE DO NOT DO THE KINETICS TOO OFTEN. CAN GIVE YOU -- EXAMPLE OF STEM CELL AGING IN MICE AND YOU CAN COMPARE -- THERE'S SOME DATA AVAILABLE AND EVEN IN IDENTICAL -- KINETICS BEHAVIOR UPON STEM CELL AGING, DIFFERENT FROM -- EACH THERE WE DON'T HAVE PRECISE KE NE TICS ON THERE, SO IT WOULD BE INTERESTING TO LOOK AT MORE PRECISELY, THIS IS A A LOT OF WORK, BECAUSE THE STEM CELLS FUNCTION ASSAYS ARE NOT EASY TO RUN, TAKING BUT IT'S DEFINITELY A QUESTION ONE SHOULD LOOK AT IN MORE DETAIL, >> I CAN JUST ADD A LITTLE BIT TO THAT, AND I THINK IF THERE IS A GREAT DEAL OF INTEREST IN STEM CELL RESEARCHERS TO COMPARE THE MECHANISM THAT CONTROL DIFFERENT STEM CELLS IN DIFFERENT TISSUES, AND THIS IS SO FAR, THERE ARE MECHANISMS THAT HAVE BEEN TESTED IN DIFFERENT TISSUES, AND THEY DO SHARE SOME MECHANISM -- AND SOME MECHANISMS ARE -- THE FIELD NEEDS TO BE TESTED, SO IT HAS BEEN TESTED IN ONE TISSUE AND THEN TESTED THE OTHERS. FOR EXAMPLE, OXIDATIVE STRESS HAS BEEN TESTED IN A NUMBER OF TISSUES, SO THAT'S ONE EXAMPLE THAT THEY DO SHARE A COMMON MECHANISM. >> NEXT QUESTION? >> SO -- SO WITH RESPECT TO THE NATION, HAS ANYBODY LOOKED AT -- AND THIS GOES BACK TO THE QUESTION A LITTLE BIT DIFFERENT ON THE INFLAMMATORY CYTOKINES. HAS ANYBODY LOOKED AT -- I DON'T KNOW WHETHER YOU CAN DO IT IN VITRO, WHERE YOU WOULD LOOK AT THE RESPONSE OF STEM CELLS TO AN ENVIRONMENT WHERE YOU WOULD HAVE SIN HE SENT CELLS OR CELLS PRODUCING THE SECRETORY PHENOTYPE? IN OTHER WORDS, WOULD A TISSUE -- IT SEEMS TO ME FOR WHAT WE'VE SAID, WHICH IS VERY INTERESTING, IS THAT THE MILLEAU FOR OLD TISSUE WOULD BE DIFFERENT THAN FROM A YOUNG ANIMAL. I LIKE THE POINT WHERE THE FACT THAT MOST OF THE EXPERIMENTERS IN THIS AREA ARE KIND OF FOR GETTING THAT, WORKING WITH YOUNG MICE. I WAS WONDERING, FROM AN OLD ANIMAL PERSPECTIVE, WOULD THE BILTD BILDUP OF HE? SEN -- >> THIS IS A GREAT IDEA, THE TYPE OF IDEA I WOULD EXPECT -- JUDY AND I HAVE DISCUSSED VERY COOL EXPERIMENTS HOW TO ANSWER THIS QUESTION. I'M PRETTY SURE THAT THAT'S -- CONNECTION BETWEEN SENESCENCE AND AGING OF CIRCULATORY MILLEAU HAS NOT BEEN WELL ANSWERED BUT DUE TO -- LABORATORIES ARE GOING TO COLLABORATE AND YOU HAVE -- ANSWER THE QUESTION. >> EXPERIMENTS -- IF YOU ASK THIS QUESTION -- LEUKEMIA TYPE OF EXPERIMENT, CORRECT? YOU CAN EASILY SEE THAT IN VIVO, OR EX-VIVO, AGED STROMA CELLS, WE DO PRIMARY EXPERIMENTS, WE TAKE IT OUT, KEEP IT -- OR WE DO IN VIVO TRANSPLANT, THAT -- STEM CELL BEHAVIOR. WHETHER THIS IS THE AGED SECRETORY PHENOTYPE -- BUT THERE'S MUCH MORE GOING ON. WHETHER THIS IS AGE -- CONTEXT, I DON'T KNOW EITHER. -- SOMETIMES DON'T WORK VERY WELL BECAUSE STEM CELLS LIKE TO HAVE SOME -- SO YOU CANNOT REALLY SEPARATE IT FROM -- EVERYTHING GOES DOWN -- SO PEOPLE ARE GETTING AT THAT, IT'S NOT THAT SIMPLE, BECAUSE WE WOULD LIKE TO -- IN MODIFYING THE NICHES FOR YOUR PROTEINS AS TOM SAID IS NOT THE MOST FUN EXPERIMENT TO DO, AND IT'S NOT A QUICK ONE, BECAUSE YOU MIGHT HAVE TO GO THE GENETIC ROUTE AND THIS IS NOT SIMPLE. WE ARE SLOWLY MOVING TOWARDS THAT, I'M TRYING TO FIGURE OUT -- >> [INAUDIBLE] UNDERSTANDING THE NICHE WHERE THE STEM CELL RESIDES AND THEN -- BETWEEN STEM CELLS -- IS EVEN MORE ON THE FRONTIER. SO NOT RELATED TO AGING BUT RELATED TO MALIGNANCY -- SHOW FOR EXAMPLE THAT -- PREDICTION OF -- FACTOR, WHICH ACCUMULATE FIBROTIC TISSUE WHICH IS VERY REFRACTORY TO STEM CELL MAINTENANCE -- NICHE, SO THERE'S THIS KIND OF FEEDBACK MECHANISM WHICH WILL COME MORE AND MORE INTO PLAY BETWEEN -- ACTING ON THE STEM CELL AND THE STEM CELLS THEMSELVES -- AND WHAT'S -- IS THAT ADIPOSE SITE -- THERE IS A LOT OF -- WHICH WOULD COME TO PLAY AND -- >> I THINK THERE ARE -- FROM PEOPLE IN A COMPLETELY DIFFERENT DIFFERENT -- [INAUDIBLE] YES, THE NICHE STEM CELL INTERACTION FRONTIER FOR -- SYSTEM A LOT OF UNKNOWN BUT IT'S LESS OF A FRONTIER BECAUSE NICHES MUCH MORE EASIER TO ACCESS AND IS MUCH BETTER DEFINED AND I JUST WANT TO EMPHASIZE AGAIN THAT YOU DON'T ALWAYS KNOW IF THE YOUNG BLOOD TURNS INTO ALL BLOOD MOLECULARLY AND FUNCTIONALLY BECAUSE OF THE CONTRIBUTION OF SENESCENT CELLS. >> I JUST WANTED TO SAY THAT OLDER MUSCLE, I DON'T KNOW WHAT OLDER MUSCLE LOOKS LIKE IN MICE, BUT IN OLDER INDIVIDUALS, FREQUENTLY THE MUSCLE IS VERY HEAVILY INFILTRATED WITH FAT, AND I BELIEVE THAT IT CHANGES THE ASSOCIATION WITH THE VASCULATURE AND WITH THE NERVOUS SYSTEM AS WELL, AND I DON'T THINK PEOPLE HAVE REALLY BEEN TALKING VERY MUCH ABOUT THE HETEROGENEITY IN THE OLDER POPULATION. WE SEE THIS IT PRIMARILY IN PEOPLE WHO HAVE A GREAT DEAL OF DISUSE OF THE MUSCLE. AND I THINK THAT IT'S AN IMPORTANT ISSUE ESPECIALLY WHEN YOU TALK ABOUT -- AFFECTING THE NICHE. >> I THINK THERE HAS BEEN SOME STUDY -- WHAT THEY'RE SUGGESTING IS THAT IT -- CAN CAN ACTUALLY ACT AS PART OF THE NICHE AND -- IMPACT STEM CELL FUNCTION -- AT LEAST -- EVIDENCE. DIFFERENT PARTS OF THE ANIMAL AND THERE'S A CORRELATION WHERE -- STEM CELL FUNCTION IS DECREASED. -- ANIMAL MODEL WHERE -- MANIPULATED AND THEN THE -- ARE DIFFERENT IN STEM CELL FUNCTION. I THINK THAT'S THE BEST EVIDENCE. >> IT WOULD BE ALSO -- YOU ENGINEERED -- IF YOU TAKE OUT -- STROMA, AND YOU SEE THE DIFFERENCES -- SO I DIDN'T TALK ABOUT CELL ALREADY CHANGES YET, FOR SURE YOU CAN QUANTIFY THEM -- WHAT IS THE CONTRIBUTING FACTOR -- I THINK IS WHAT YOU'RE SAYING CORRECT? >> YES, THERE WAS AN INTERESTING PAPER SHOWING A TREMENDOUS AMOUNT OF ADD POE NECKTIVE PRODUCED BY BONE MARROW FAT SUGGESTING IN POPULATION STUDIES WE SEE AN INCREASE IN ADIPONECTIN, BUT IT'S ALSO AN ENDOCRINE FUNCTION IN OLDER ANIMALS AND ALSO A PARABIOSIS EXPERIMENT SHOWING REJUVENATION OF CO SITES AS WELL. >> THE QUESTION IS STILL CORE LATIVE OR IT'S CAUSATIVE. THIS IS WHAT I WANT TO GET A. CORRELATIVE IT'S STRAIGHTFORWARD, CAUSATIVE, NOT THAT SIMPLE BUT AS RICHARD SAID, WE NEED TO KNOW WHAT WE'RE DOING BEFORE WE MOVE FORWARD. SO WE LOOK FORWARD TO IDENTIFY CAUSATIVE CHANGES >> I AGREE, IT WOULD BE NICE IF WE HAD WAYS TO GET AT HUMAN MUSCLE. >> NEXT QUESTION? OR LAST QUESTION? >> JUST WANTED TO GO BACK TO THE ISSUE OF TELOMERASE THE PANEL SEEMED TO BE QUITE UNIFORM IN THAT IT'S NOT SO IMPORTANT TO MAINTENANCE OF STEM CELLS, BUT SO MARIA BRASKO DID PUBLISH A FEW YEARS AGO THAT I THINK HIGHER LEVELS OF TELL MER ACE IN -- MICE DOES IMPROVE THE HEALTH OF RENEW AABLE -- SOME RENEWABLE TISSUES, I THINK THE SKIN AND INTESTINE, AND ALSO EXPAND THE LIFESPAN OF THE MICE. SO WOULD YOU PROPOSE THEN THAT THAT'S A STEM CELL INDEPENDENT PHENOTYPE? >> I THINK THE -- IN STEM CELL SPECIFIC COMPARTMENTS, I THINK UNTIL WE DO THAT, WE JUST DON'T KNOW THE ANSWER. I THINK THAT'S A PROVOCATIVE EXPERIMENT, BUT I THINK IT'S, AGAIN, NOT DIRECTLY LOOKING AT -- SO I THINK THE JURY IS OUT. >> YEAH. I DON'T THINK SHE DID -- I DON'T RECALL BUT I DON'T THINK THAT SHE DID SHOW THAT THERE WAS ANY SPECIFIC -- ANY IMPROVED MAINTENANCE OF STEM CELLS SPECIFICALLY. SO I THINK YOU'RE RIGHT ABOUT THAT. >> STEM CELLS THEMSELVES -- ACTIVITY AND THAT'S WHY THIS -- BETWEEN CANCER CELLS AND -- CANCER CELLS AND CANCER STEM CELLS HAVE BEEN MADE AS WELL. >> RIGHT. THIS RAISES AN INTERESTING QUESTION, STEM CELL INDEPENDENT AFFECT -- AND WHAT IS THAT. >> WELL, STEM CELLS DO SHOW -- WHEN YOU REALLY -- TO UNDERGO A LOT OF -- SO IT'S NOT THAT THEY ARE IMMORTAL -- OR EQUIVALENT OF CANCER -- WHAT I VIEW AS PROTECTIVE STRATEGY, YOU WANT TO MAINTAIN -- BECAUSE YOU WANT TO MAINTAIN YOUR TISSUE FUNCTION -- WELL THE NICHE IS FACTOR -- ALL OF THAT CONTRIBUTES TO MAINTAINING BALANCE -- STEM CELLS AND WHEN SOMETHING GOES WRONG, THERE IS CONSEQUENCE FOR WHAT -- >> BUT I THINK YOUR POINT WELL TAKEN THAT WE DON'T KNOW WHETHER TELOMERASE EXPRESSION IS IN ANY WAY RATE LIMITING. WE DON'T KNOW THAT. SO I THINK THAT, WE CAN DO -- >> AGAIN THE -- SYSTEM -- 2007 PUBLISHED -- LAB -- I THINK THERE WAS NO BENEFICIAL EFFECT LONG TERM -- TRANSPLANT, AND FOR SURE WE KNOW THAT TELOMERASE HAS -- INDEPENDENT FUNCTION -- SLOWLY TO LIGHT, IT COULD BE WELL THAT THERE IS DIFFERENTIATED CELLS FUNCTIONS OF -- THAT DO THE NOT HAVE TO -- THE FIRST ONE AND THAT COULD CONTRIBUTE. AND -- IT WOULD ALSO CONTRIBUTE TO STEM CELL MAINTENANCE BUT I THINK -- TELL MERE LENGTH FOR NORMAL LIFETIME, IT'S PROBABLY NOT THE MAJOR FACTOR. >> THAT'S TRUE, BUT I THINK WE ALSO NEED TO RECOGNIZE -- I'M NOT IN THE TEE LOW MUIRFIELD BUT I THINK THERE IS A FEELING THAT IT'S NOT JUST TELOMERE LENGTH WHICH IS IMPORTANT. YOU CAN HAVE A DYSFUNCTIONAL TELOMERE, WHICH IS SEEMINGLY A LONG TELOMERE. >> AGREE ON THAT. BUT AGAIN, IT'S NOT THAT EASY TO CONCLUDE THAT THEM RAISE IS THE REJUVENATION FACTOR. >> RIGHT. OKAY. THANKS VERY MUCH. >> ONE MORE QUESTION. >> THANKS. ROBERT BROOK. JUST WANTED TO KNOW, DO YOU FIND ANY MERITS WITH ANY OF THE OFF-SHORE STEM CELL CLINICS OR ANY STEM CELL CLINICS IN THE U.S.? I KNOW IT'S PRETTY CONTROVERSIAL, BUT THEN IF YOU DON'T, DO YOU SEE ANY WAY OR EXPERIMENTS YOU WOULD WANT TO DO WITH THEM TO TAKE MORE ADVANTAGE OF THE PEOPLE THAT DO GO TO THEM THEM? >> THIS IS OBVIOUSLY A VERY -- TOPIC. THE -- STEM CELL RESEARCH IS VERY INVOLVED IN TRYING TO LOOK AT THESE STEM CELL CLINICS AND GET A SENSE OF WHICH CLINICS ARE BASED ON SCIENCE AND WHICH ONES ARE NOT. CLEARLY THERE ARE MANY THAT ARE NOT SO I'M NOT GOING TO SPEAK FOR THE GROUP OR THE SOCIETY IN TERMS OF WHAT THAT IS, BUT THE -- SO IN CALIFORNIA, THE CALIFORNIA INSTITUTE FOR REGENERATIVE MEDICINE IS SETTING UP HOPEFULLY WITHIN THE NEXT COUPLE OF YEARS CLINICS WITHIN CALIFORNIA THAT WILL BE REALLY SET UP TO IDENTIFY DISEASES THAT CAN BE APPROACHED WITH -- THERAPIES BASED ON THE SCIENCE THAT'S BEEN SUPPORTED BY RESEARCH IN THE U.S. AND AROUND THE WORLD, AND OTHER THAN THAT, WE ALL KNOW THE PROBLEMS OF SUCH CLINICS AROUND THE WORLD AND PEOPLE WILL SPEND A LOT OF MONEY TO GO THERE. I THINK IT'S MORE OF A POLITICAL ISSUE THAN A SCIENTIFIC ISSUE IN TERMS OF WHAT TO DO ABOUT IT. >> I THINK WE HAVE TO WRAP UP. THANK YOU TO THE PANEL AGAIN. [APPLAUSE] >> IF NIH RULES PERMIT ME, I'D LIKE TO MAKE A STATEMENT AND THEN A FOLLOW-UP. I'D LIKE TO THANK THE CO-CHAIR, PANELISTS AND THE AUDIENCE, AND PEOPLE WHO HAVE TRAVELED FROM AROUND THE WORLD TO BE HERE. FOR AN EXCITING FINAL SESSION AND FOR ALL THE THOUGHTFUL PRESENTATIONS AND DISCUSSIONS OVER THE LAST TWO DAYS. THANK YOU. [APPLAUSE] NOW IT'S MY PLEASURE TO INTRODUCE DR. RICHARD -- TO GIVE THE CLOSING REMARKS. DR. HODES RECEIVED HIS M.D. FROM HARVARD MEDICAL SCHOOL. ET CETERA. HERE'S DR. HODES. [APPLAUSE] >> THANK YOU. AT THE END OF THESE TWO DAYS, I APPRECIATE THE OPPORTUNITY TO RECAP A LITTLE WHAT IT'S BEEN ABOUT AND THEN SPEND THE MOST OF THE FEW LAST MINUTES WITH YOU HERE THANKING PEOPLE WHO DESERVE THAT GRAT TEUT. IT'S BEEN INTERESTING OVER THE LAST YEARS TO SEE A CONVICTION GROWING AMONG MANY OF US. I HOPE EVIDENCE BASED THAT QUESTIONS OF AGING AND THE MECHANISMS THAT UNDERLIE AGING ARE BOTH INTERESTING AND TRACTABLE, AND WITH THAT NOTION HAS COME THE CONVICTION THAT THE RELEVANCE OF THE BIOLOGY OF AGING TO PROCESSES STUDIED BY MANY WHO WOULD SELF-IDENTIFY THEIR FOCUS WITH PARTICULAR ORGAN, DISEASES OR IN THE CASE OF NIH, INSTITUTES, REALLY DEMANDS A BREADTH OF COMMITMENT FOR THE SAKE OF THE SCIENCE AND BRINGING TOGETHER THE PERSPECTIVES THAT WILL BEST SERVE OUR UNDERSTANDING OF AGING AND SO THAT THE UNDERSTANDING OF AGING CAN IN TURN BEST SERVE ITS APPLICATION TO THE MANY EXOAPTS OF THE NIH MISSION TO HEALTH. AND I THINK HAD MEETING HAS BEEN A PARTICULARLY IMPORTANT JUNCTURE IN THAT REGARD, IN THE SENSE WE'VE BEEN TALKING OUT OF CONVICTION ABOUT THE IMPORTANCE AND RELEVANCE OF INVOLVEMENT OF MANY. WE'VE TAKEN THAT CONVICTION, I THINK IT AT NIH AND THE ACTIONS OF ALL HUMAN SCIENTIFIC COMMUNITY WITH THE CREATION OF GEROSCIENCE, I GUESS THE WORD BEGAN IN 2007 AT THE BUCK INSTITUTE, IT'S BEEN PICKED UP CONGRESSIONALLY, BY NIH, AND NOW WITH THIS GEROSCIENCE INTEREST GROUP, WHICH BEGAN THROUGH EFFORTS AM THOSE I'LL THANK, CERTAINLY PHILIPPE -- HAS BEEN PARAMOUNT WITH A GET-TOGETHER, IN FACT, IN THIS BUILDING IN A ROOM ASSOCIATED WITH ANOTHER MEETING WE'RE HAVING ABOUT FIVE OR SIX INSTITUTE DIRECTORS THAT TALK ABOUT THEIR WILLINGNESS TO COMMIT THEMSELVES TO SUCH AN EFFORT, IT WAS SUCCESSFUL, NOW EXPANDED TO 20 AT CURRENT TALLY OF THE INSTITUTES WHO ARE COMMITTED AGAIN NOT JUST IN NAME BUT IN EFFORT TO MAKING A COMMITMENT TO PURSUING THE SCIENCE HERE. SO WE MAY WELL ASK MOST IMPORTANTLY WHAT IS THIS GOING TO LEAD TO. I THINK IN FACT, WHAT HAS ALREADY HAPPENED IN THIS MEETING IS AN IMPORTANT PART OF WHAT IT HAS AND WILL LEAD TO, THE VERY EVIDENT VIBRANT CONVERSATIONS THAT HAVE HAPPENED AMONG PEOPLE FROM VARIOUS BACKGROUNDS HAS ALREADY, I HOPE, SERVED TO STIMULATE US AND YOU TO PURSUE SCIENCE AND ACCELERATE THE DIRECTION OF SCIENCE THAT WE'RE TAKING FROM OUR OWN SIDE AT NIH IN THESE NEXT DAYS BEGINNING WITH A SESSION TOMORROW, WE'LL BE PUTTING TOGETHER TRYING TO ASSIMILATE THE VERY COMPLEX AND VIBRANT INPUTS THAT HAVE COME FROM MANY OF YOU. I TAKE IT THERE WILL BE STIMULATING YOUR OWN SCIENTIFIC INTERESTS, IT WAS STIMULATING OURS AS WELL AND I ENCOURAGE YOU TO MAINTAIN CONTACT WITH THOSE OF US AT NIA AND ALL THE INSTITUTES THAT HAVE BEEN COLLABORATING HERE TO MAKE SURE WE MAKE A GO OF THIS. WHETHER THERE WILL BE SPECIFIC AREAS TO IDENTIFY THAT ARE WORTHY OF INITIATIVES AT A TIME WHEN YOU ARE STRUGGLING TO SUPPORT ALL OF YOU AS BEST WE CAN WITH BUDGET CONSTRAINTS AS THEY ARE, WE WILL SEE, BUT WE'RE CERTAINLY NOT GOING TO BE BASHFUL ABOUT FOLLOWING SCIENTIFIC IMPERATIVES. SO LET ME TURN TO THAT FROM GIVING THANKS CONGRATULATIONS FROM THOSE WHO HAVE MADE THIS POSSIBLE. THE ORGANIZERS, A ROUND OF APPLAUSE FOR THEM, I'D SAY. [APPLAUSE] >> IN ADDITION TO THE REPRESENTATIVES OF THE MANY INSTITUTES YOU'VE SEEN HELPING TO ORGANIZE SESSIONS, TO THE CHAIRS WHO DID A MARVELOUS JOB OVERALL, IN WATCHING FOR THE RED LIGHTS, FORCING CONSTRAINTS LED TO A GOOD BALANCE OF PRESENTATIONS AS WELL AS VERY, VERY ACTIVE INTERACTION FROM MANY OF YOU WHO HAVE BEEN HERE THROUGH THESE TWO DAYS. I'D ALSO LIKE TO THANK ALL OF YOU, ABOVE ALL, FOR TAKE ITING THE TIME TO COME HERE, BE WITH US, AND IF THERE'S ANYBODY I'LL ASK TO STAND UP, IT'S OUR PARTNERS WHO HAVEN'T BEEN RECOGNIZED MAYBE AS PUBLICLY DAN PERRY, SUE PESHIN. FOLKS FROM THE ALLIANCE FOR AGING RESEARCH AND THE JAIR LOGICAL SOCIETY OF AMERICA WHO WERE THE PARTNERS THAT MADE THIS POSSIBLE IN MANY WAYS AND NOT JUST THIS MEETING, BUT THESE ARE THE FOLKS WHO HAVE BEEN TALKING ACROSS INSTITUTES FOR THE NIH WHO HAVE BEEN TALKING ON THE HILL AND RESPONSIBLE FOR A NUMBER OF US BEING THERE TO SPEAK TO A SENATE SUBCOMMITTEE YESTERDAY, ALL PART OF AN EFFORT THAT HAS MADE GEROSCIENCE NOT ONLY A NAME OF AN INTEREST GROUP HERE AT NIH BUT A LANDMARK, A HALLMARK FOR WHAT'S HAPPENING IN SCIENCE, IT'S IN CONGRESSIONAL LANGUAGE, IT'S IN THE MINDS OF ALL OF US, AND MOST IMPORTANTLY, I THINK WE NEED TO MAKE IT A REALITY OF A RIGOROUS AND LEGITIMATE SCIENCE AS WE ARE SEEING ITS EVOLUTION HAPPEN NOW, SO THANK YOU ALL, THANK YOU FOR BEING HERE, AND SAFE TRAVELS HOME AND WE LOOK FORWARD TO INTERACTIONS IN THE FUTURE WITH YOU, ENRICHED AND INFORMED BY THESE LAST TWO DAYS. THANK YOU. [APPLAUSE]