>> ON BEHALF OF THE NIH DIRECTOR, IT'S MY PLEASURE TO INTRODUCE TODAY'S NIH DIRECTOR'S SEMINAR SPEAKER, CRAIG THOMPSON. DR. THOMPSON, AS Y'ALL KNOW, IS PRESIDENT AND CHIEF EXECUTIVE OFFICER OF MEMORIAL SLOAN-KETTERING CANCER CENTER WHERE HE'S BEEN THE LAST COUPLE OF YEARS. HIS BACKGROUND INCLUDES BACHELOR'S DEGREE FROM DARTMOUTH COLLEGE AND MD FROM DARTMOUTH AND UNIVERSITY OF PENNSYLVANIA. HE DID A RESIDENCY IN MEDICINE AT THE PETER BRIGHAM HOSPITAL AND UNIVERSITY HOSPITAL IN BOSTON AND HEMATOLOGY ONCOLOGY FELLOWSHIP AT FRED HUTCHINSON CANCER RESEARCH INSTITUTE, UNIVERSITY OF WASHINGTON. ONE THEME THAT YOU'LL SEE IN DR. THOMPSON'S BIO IS HE'S MOVED QUITE A LOT WHICH IS INTERESTING AND HAS BEEN VERY PRODUCTIVE FOR HIM. AFTER FRED HUTCH EXPERIENCE HE CAME TO THIS AREA AND ACTUALLY WORKED FOR SEVERAL YEARS AT THE NATIONAL NAVAL MEDICAL CENTER AS A PHYSICIAN AND SCIENTIST. AND AFTER THAT EXPERIENCE HE WENT TO THE UNIVERSITY OF MICHIGAN WHERE HE BECAME ASSOCIATE -- ASSISTANT PROFESSOR, MOVED UP THROUGH THE RAINGS, BECAME AN HHMI INVESTIGATOR, AND ALSO HAD A JOINT APPOINTMENT AT THE UNIVERSITY OF CHICAGO WHERE HE WAS DIRECTOR OF THE GWEN KNAPP SENOR FOR LUPUS AND IMMUNOLOGY RESEARCH. TURNS OUT THAT WHEN DR. THOMPSON WAS AT THE UNIVERSITY OF MICHIGAN, HE SHARED A FLOOR WITH TWO OF OUR NIH SIGNATURE TEARS, GARY NABEL AND FRANCIS COLLINS, ALL ASSISTANT PROFESSORS TOGETHER AND YOU CAN IMAGINE THE OTHER GROUP THERE, WHAT A POWERFUL INTELLECTUAL FORCE THEY MUST HAVE BEEN AT THAT UNIVERSITY. IN 1999 HE CAME TO THE UNIVERSITY OF PENNSYLVANIA, WHERE HE BECAME PROFESSOR OF MEDICINE AND INITIALLY SCIENTIFIC DIRECTOR OF THE LEONARD AND MADELINE ABRAMSON FAMILY CANCER RESEARCH INSTITUTE. CHAIRMAN OF THE DEPARTMENT OF CANCER AND ULTIMATELY DIRECTOR OF THE ABRAMSON CANCER CENTER AT THE UNIVERSITY OF PENNSYLVANIA. AND AS I MENTIONED, COUPLE OF YEARS AGO HE MOVED TO ME MEMORIAL SLOAN-KETTERING WHERE HE LEADS AN INCREDIBLE PROGRAM OF CANCER RESEARCH AND TREATMENT. SO I THINK ALL OF THESE POSITIONS REFLECT THE ENORMOUS ABILITY TO GALVANIZE EXCITING NEW DISCOVERIES IN CANCER RESEARCH. DR. THOMPSON HAS CONTRIBUTED I THINK THREE MAJOR AREAS. THE FIRST WHICH IS THE WORK HE STARTED I THINK AT THE NAVY CENTER HERE AND ALSO THE UNIVERSITY OF MICHIGAN WAS TO EXPLORE THE ROLE OF ONCOGENES IN HEMATOPOIETIC CANCER AND ALSO IN LYMPHOID DEVELOPMENT. MANY CONTRIBUTIONS THERE. AND THEN HE WORKED ON MECHANISMS OF APOPTOSIS, AND THEY WERE ROLE IN CANCER AND YOU'RE PROBABLY FAMILIAR WITH A LOT OF HIS CONTRIBUTIONS THERE. OVER ABOUT THE LAST DECADE, HE'S PIONEERED A NEW UNDERSTANDING OF THE WAY CELL METABOLISM INTERACTS WITH ISSUES RELATED TO CELL GROWTH AND DEVELOPMENT OF CANCER AND SOME OF THE WORK HE'S DONE WILL BE DISCUSSED TODAY. IN ADDITION TO HIS MANY SIGNIFICANT POSITIONS AS A SCIENTIST HIS WORK HAS BEEN RECOGNIZED BY ELECTION TO THE NATIONAL ACADEMY OF SCIENCES, THE INSTITUTE OF MEDICINE, AMERICAN ACATD MY -- ACADEMY OF ARTS AND SCIENCES AMONG OTHERS. HE'S HAD QUITE A FEW AWARDS. I WON'T GO THROUGH THEM ALL. HE'S ASSOCIATE EDITOR FOR THOSE WHO ARE INTERESTED, OF CELL, IMMUNITY AND CANCER CELL, SOME OF OUR FAVORITE JOURNALS. HE'S A BOARD MEMBER FOR MANY SIGNIFICANT ORGANIZATIONS, AND MEMBER OF THE LASKER PRIZE JURY SO WE REALLY HAVE TO BE NICE TO CRAIG. IN ADDITION HE STARTED IN HIS CAREER THREE BIOTECH COMPANIES. BUT HIS MOST IMPORTANT CONTRIBUTION IS THAT FOR MANY YEARS HE WAS CHAIRMAN OF THE BOARD OF SCIENTIFIC COUNSELORS OF THE NCI BASIC SCIENCE DIVISION. MANY OF YOU CANCER RESEARCHERS GOT TO KNOW HIM IN THAT CAPACITY. I THINK WE SHOULD WELCOME CRAIG THOMPSON WHOSE TALK TODAY IS ENTITLED IDH MUTATIONS. ONCOMETABOLITE DEREGULATION OF EPIGENETIC REMODELING. CRAIG. [APPLAUSE] >> IT'S A PRIVILEGE TO BE BACK HERE IN MASUR AUDITORIUM AND GIVE THIS TALK AND SEE SO MANY OLD FRIENDS FROM NIH DAYS AND IN SCIENCE. I HAVE SAID THIS BEFORE BUT I'M ALWAYS EMBARRASSED WHEN WE GIVE THAT KIND OF INTRODUCTION BECAUSE AS MY MOTHER TELLS ME WHEN SHE'S OCCASIONALLY COMES AN LISTENS IN AN AUDIENCE LIKE THAT IT SOUND LIKE YOU CAN'T HOLD A JOB. WHAT I AM GOING TO TALK TODAY IS GIVE AN OVERVIEW SINCE APPROPRIATE ON THESE WEDNESDAY AFTERNOON CONFERENCES TO SERVE FOR STUDENTS AND POST-DOCS TO GIVE YOU AN IDEA HOW WE GOT INTO THIS FIELD OF METABOLISM AND I PUT METABOLISM ONLY BRIEFLY IN THE TITLE AS ONCOMETABOLITE. WE WILL COVER THAT AT THE END BUT I WANTED THE STUDENTS TO SORT OF GET AN UNDERSTANDING THAT NO ONE THESE DAYS OTHER THAN KICKING AND SCREAMING GETS DRAGGED BACK INTO THE FIELD WE ALL GOT TAUGHT, WHICH IS BIOCHEMISTRY AS AN UNDERGRADUATE OR METABOLISM IN ITS LATEST FORMS GOING FORWARD. PEOPLE WHO HAVE DONE THE WORK I'M GOING TO TALK ABOUT TODAY IN MY LAB CURRENTLY ARE HERE, A NUMBER OF POST DOCS LED TO HAVE THEIR OWN LABORATORIES IN THIS FIELD OF CANCER METABOLISM BUT MOST RECENTLY NEW WORK IS A COLLABORATION WITH ROSS LE VINE AT MEMORIAL SLOAN CET REGULAR AND ERIN MELMIC AROUND THE HEMATOPOIETIC MALIGNANCIES. WHAT'S NOT SHOWN HERE MORE RECENTLY PUBLISHED WITH TIN CHAN AND (INDISCERNIBLE) ON GLIOMAS. WE HAVE HAD A LONG TERM CANCER WITH LOU CAN'TLY AND JOSH (INDISCERNIBLE) TO BRING APPROACHES TO METABOLIC STUDIES. AND A FEW YEARS AGO WITH LOU AND TAP MACK WE STARTED IGIOS PHARMACEUTICALS LOOKING INTO THE SPACE OF CANCER METABOLISM TO SEE IF H'S ANY THERAPEUTIC WAYS THE INTERVENE. THEY HAVE CONTRIBUTED TO SOME OF THE BASIC SIGN SCIENCE I'LL TALK ABOUT TODAY. THAT'S THE INTRODUCTION OF THE PEOPLE THAT DID THIS. AS I SAID, WE WERE ALL -- EVERYONE IN THIS ROOM, IT'S INTERESTING WE SPEND A SIGNIFICANT PERIOD OF TIME IN SCIENCE BUT EVERY ONE OF US SHARES A COMMON BACKGROUND, WE TOOK EXACTLY THE SAME BIOCHEMISTRY COURSE. AND IF YOU TOOK IT MULTIPLE TIMES IN HIGH SCHOOL AND COLLEGE AND THEN GRADUATE SCHOOL, YOU TOOK EXACTLY THE SAME COURSE OVER AGAIN MULTIPLE TIMES MAYBE A LITTLE MORE DEPTH. THAT'S BECAUSE THERE HASN'T BEEN A NEW METABOLIC PATHWAY INTRODUCED INTO A BIOCHEMISTRY TEXTBOOK SINCE 1983. AND THEN IT WAS ONLY AFTER TEN YEARS OF ARGUING THEY ADDED ONE STEP I WILL TALK ABOUT BRIEFLY. BEFORE THAT THE PREVIOUS NEW INTRODUCTORY STEP WAS IN THE 1960S. WHAT WE LEARNED IN FIRST INTRODUCTION TO BIOCHEMISTRY OF LIFE, IN FULLY DIFFERENTIATED TISSUES, TISSUES EXTRACTED NUTRIENTS FROM THE ENVIRONMENT, THE PREFERRED NUTRIENT WE SHARE WITH ALL CELLS IN THE BODY IS GLUCOSE THAT'S METABOLIZED TO THE HEXO SUGAR, TRIO SUGAR PYRUVATE AND FULLY BURNED AS CO-2 IN WATER IN THE FORMATION OF 36 ATP EQUIVALENTS ON AVERAGE. TO A MAXIMALLY EXTRACT THE ENERGY IN THE CARBON, CARBON BACKBONE OF THIS ORGANIC MOLECULE. THAT ALLOWS CELLS TO PRODUCE ENOUGH ENERGY TO MAINTAIN THE HOMEOSTASIS, PARTICULARLY 50% OF ALL ENERGY IN CELLS IS TO PUMP OUT THE IONS THAT ACTUALLY DIFFUSE INTO A CELL, SODIUM, SEQUESTER CALCIUM, ET CETERA, ET CETERA. ALLOWS THE REL TO SURVIVE AND OTHER DATA TO SURVIVE. WE LEARNED A MORE ADVANCED WAY OF THINKING ABOUT ORGANISMAL BIOLOGY IN THE ABSENCE OF OXYGEN CELLS CAN SURVIVE FOR BRIEF PERIODS OF TIME, PURELY THROUGH GLYCOLYTIC METABOLISM THAT ENDS IN THE PRODUCT LACTATE BUT THAT IS HIGHLY INE EFFICIENT IF THE GOAL IS TO DRAG MAXIMUM ATP PRODUCTION AND THEREFORE SURVIVAL. BECAUSE IT ONLY HAS TWO MOLLS OF ATP FOR EVERY MOLE OF GLUCOSE. THAT'S NOT A DISASTER AS THE HUSBAND AND WIFE TEAM WORKING IN THE CANCER CENTER IN 1947 WHICH THEY WON THE NOBEL PRIZE, IF YOU RECYCLE THIS LACTATE ON AN ORGANISMAL BASIS YOU ONLY GIVE UP TWO EQUIVALENTS IF YOU DO NEOGENESIS FROM THIS LACTATE AND RECOVER IT IN OXIDATIVE METABOLISM. THAT IS WHAT WE WERE TOLD THE WAY IN WHICH CELLS DEALT WITH ENERGY DEMANDS, THE WHOLE SYSTEM WAS HOMEOSTATIC SO ONCE YOU LEARNED AND PASSED THE TEST YOU CAN GO TO THE COOL STUFF. YOU CAN STUDY SUB BIOLOGY, MOLECULAR BIOLOGY, MORE RECENTLY YOU CAN STUDY STRUCTURAL BIOLOGY. BUT THERE IS A NAGGING PROBLEM ABOUT THAT, WHEN PEOPLE STUDY PROLIFERATIVE TISSUE, THE MOST RECENT EXAMPLE STUDYING BIOLOGY OF ES CELLS OR IPS CELLS, AND MAINTAIN THEM OPTIMALLY IN CULTURE THEY DON'T ADOPT THIS METABOLISM. IN FACT, WHAT THEY DO IS THEY SCAVENGE THE GLUCOSE OUT OF THE ENVIRONMENT AND EVEN IN THE PRESENCE OF MOLECULAR OXYGEN THROUGH THE CAR BEEN THEY TAKE AWAY UP AT LACTATE. THAT WAS FIRST NOTICED IN TUMOR CELLS BACK IN THE 1920s BY PIONEERING BIOCHEMIST WHO CONTRIBUTED TO MUCH OF THIS INFORMATION, WARBERG, SO THE PREFERENCIAL USE OF GLUCOSE METABOLISM THROUGH GLYCOLYSIS HAS BEEN TERMED THE WARBERG EFFECT OR AEROBIC GLYCOLYSIS WHEN OCCURS IN THE PRESENCE OF MOLECULAR OXYGEN. FOR OVER NOW 80 YEARS SCIENTISTS EVERY DECADE HAVE TRIED TO FIGURE OUT WHY IT IS TUMOR CELLS AND MORE RECENTLY PROLIFERATING NORMAL CELLS ADOPT THIS ABNORMAL METABOLISM. MY INTEREST IN THIS DID NOT STEM FROM ANY INTEREST IN METABOLISM, IT CAME FROM WHAT MIKE TALKED ABOUT. IN OUR STUDIES TO UNDERSTAND LYMPHOCYTE PROLIFERATION IN THE MID 1990s, WE CAME TO APPRECIATE THAT IN FACT EVERY ONE OF US GOES THROUGH HUGE CLONAL EXPANSIONS OF LYMPHOCYTES AS WE ENCOUNTER NEW BACTERIA OR VIRUSES IN OUR ENVIRONMENT AN FIGHT THEM OFF BY A SPECIFIC IMMUNE RESPONSE. AND THAT -- BY THIS POINT YOU HAVE HAD HUNDREDS OF THOSE ENCOUNTERS WITH FOREIGN PATHOGENS. AND IF YOU KEPT ALL THE LYMPHOCYTES THAT YOU CREATED TO PROTECT YOU DURING THE EPISODIC EVENTS OF INFECTION YOU NEED A WHEEL BARROW TO CARRY THE EXTRA CELLS WITH YOU TO MAINTAIN HOMEOSTASIS. SO OTHER INVESTIGATORS THINKING ABOUT THAT PROBLEM IN EVERY LINEAGE OF THE BODY CAME ONE THE IDEA THERE MUST BE MECHANISMS THAT DELETED CELLS FROM THE BO AND THE SIMILAR LEST MECHANISM TO DELETE IS AN ORGANIZED CELL DEATH KNOWN AS APOPTOSIS FIRST WITH GENETIC DISCOVERY IN THE WORM. A FEW YEARS AGO TO TRY TO EXPLAIN HOW IT IS WE CHEEP TRACK OF EVERY CELL IN THE HUMAN BODY, EVERYONE WALKED IN WITH 60 TRILLION CELLS AND HOW YOU COPE TRACK OF 60 TRILLION CELLS WHEN THEY ACTUALLY ANY ONE COULD DIE BY APOPTOSIS IN 15 MINUTES BECAME THE PUZZLE OF THE 1990s FOR PEOPLE INTERESTED IN THE FIELD OF CELL DEATH. THERE WAS SIMPLIFIED BY MARK RAF IN THE MID 1990s WITH A STATEMENT THAT HE WAS PRETTY SURE THAT ORGANISMAL HOMEOSTASIS IS MAINTAINED BECAUSE EVERY CELL IN YOUR BODY WAKES UP EVERY MORNING THINKING ABOUT SUICIDE AND HAS TO BE TALKED OUT BY NEAREST NEIGHBORS TO MAINTAIN HOMEOSTASIS. SO THE MODEL SAID SINGLE CELLS TAKEN FROM OUR BODY PLACED IN ISOLATION WITHOUT THE NORMAL NEAREST SIGNALS FROM THE ENVIRONMENT INITIATE OVER A DEFINED PERIOD OF TIME THIS APOPTOTIC CASCADE THAT EFFECTIVELY AND SILENTLY ELIMINATES THEM FROM THE BODY. THE ARGUMENT BECAME THAT YOU NEEDED SPECIFIC INSTRUCTION SIGNALS TO MAINTAIN THE VIABILITY OF THAT CELL OVER THE NEXT FEW YEARS. THOSE WERE NOTICE SURVIVAL SIGNALS AND AS I'LL SHOW YOU IN A MINUTE, THE LATE 1990s WE PROPOSE THE COMMONALITY OF THOSE SIGNALS IS THEY INSTRUCTED CELLS TO TAKE UP SUFFICIENT GLUCOSE TO MAINTAIN ATP PRODUCTION IN HOMEOSTASIS. SECOND CAROLLY WE NOTE TO -- SECONDARILY WE NOTE TO MAINTAIN HOMEOSTASIS FOR LYMPHOCYTES YOU NEED ADDITIONAL SIGNALS. THEY HAR MESS NOT NUTRIENTS BUT NEW TRANSCRIPTION IN TRANSLATION NECESSARY FOR A CELL TO ENTER AND PROGRESS THROUGH THE CELL CYCLE. IF THIS MODEL IS CORRECT, IT SUGGESTIONS AS I'LL SHOW YOU THIS A MINUTE TWO FUNDAMENTAL BARRIERS TO THE REGULATION OF CELL SURVIVAL AND ULTIMATELY CELL AUTONOMOUS PROLIFERATION. THE CELL NEEDS TO ACQUIRE THE INFORMATION CODE TO PROLIFERATE AND IT NEEDS THE NUTRIENTS TO ENACT THE SURVIVAL DURING THAT PROLIFERATION RESPONSE. SO STARTING WITH THE HYPOTHESIS JEFF (INDISCERNIBLE) IN 1999 NOW AT DUKE IN HIS OWN LABORATORY STARTED LOOKING FOR INDIVIDUAL CELL TYPES AN CHARACTERIZE WHAT ACTUALLY HAPPENS. SO THE SIMPLE EXPERIMENT IS YOU TAKE THE FAVORITE CELL TYPE OUT BODY, MAKE A CELL SUSPENSION AND ASKS WHAT HAPPENS TO THE CELL DURING THE NEXT TWO DAYS BEFORE IT COMMITS IRREVERSIBLY TO APOPTOSIS. WHAT HE FOUND IN EVERY CELL TYPE HE WAS ABLE TO EXAMINE IN THE ANSWER SENSE OF -- ABSENCE OF SERUM NUTRIENT TRANSPORTERS TO TAKE UP NUTRIENTS FROM THE CELL ENVIRONMENT OFF THE CELL SURFACE AND DELIVERS TO THE THE LYSESOME WHERE THEY'RE COMPLETELY DEGRADED. AT THAT TIME POINT THE CELL STARTS A CELL CLOCK. THAT CLOCK IS DEPENDENT ON THE CELL AND THE NUMBER OF INTERMEDIATE ME TABULATE METABOLITES THAT THE CELL HAS INTRACELLULARLY BUT ULTIMATELY THAT CELL OVER THE NEXT 24 HOURS HAS A DECLINING BIOENERGETIC SIGNATURE READ OUT AT MULTIPLE ORGANELLES THAT ACTIVATE SO-CALLED BH-3 PROTEINS. THAT ACT ULTIMATELY AT THE SURFACE OF THE MITOCHONDRIAL MEMBRANE TO ACTIVATE THE DEATH INDUCING PROPERTIES OF THE PRO APOPTOTIC MEMBERS OF THE BCL-2 MEMBERS BACK TO BACK. THROUGH THIS PROCESS CAUSES DISILLUSION OF THE OUTER MITOCHONDRIAL MEMBRANE AND INITIATION OF THIS ORDERLY FORM OF CELL DEATH KNOWN AS APOPTOSIS. SO WE PROPOSED AS SUGGESTED A MINUTE AGO, THAT THE KEY FACTOR THAT REGULATES THE SURVIVAL IS ACTUALLY THE LOSS OF TAKE UP NUTRIENTS AND MAINTAIN BIOENERGETICS. BUT THERE WAS AN OPPOSING VIEW, AND THAT IS THAT WHILE YOU'RE NOT BEING STIMULATEDDED BY GROWTH FACTORS IN YOUR ENVIRONMENT ANY LONGER YOU CAN TAKE A INDOLENT BIOLOGYND THAT ALLOWS YOU AT THE LEVEL YOU CAN DETECT THEM TO SEE A DECLINE IN NUTRIENT TRANSPORTERS. THEY'RE STILL THERE IN ADEQUATE QUANTITIES TO MAINTAIN VIABILITY OF THE CELL. SO WE TRY TO ADDRESS THAT QUESTION BY CREATING WITH STAN COURSEMYERS LABORATORY A MOUSE DEFICIENT OF BACK TO BACK. SO IT COULDN'T INITIATE APOPTOTIC CELL DEATH AND THEN ASKED IN CELL LINES AND THE WHOLE ORGANISM WHAT HAPPENS TO CELL SURVIVAL WHEN THE CELL STILL IS EXPERIENCED NEGLECT LOSS OF EXTRA CELLULAR SIGNALING BUT NO LONGER CAN INDUCE APOPTOSIS. WHAT WE DISCOVERED ABOUT SIX YEARS AGO WAS THAT CELL TURNS TO A ANCIENT SURVIVAL PATHWAY PREVIOUSLY WELL DESCRIBED IN SINGLE CELL EUKARYOTS AND PLANTS THE PROCESS OF MACRO AUTOPHAGY WHICH THE CELLS SEQUESTERS THE CYTOPLASM NP A VAS CL KNOWN AS AUTOPHAGOSOME AND DELIVERS THAT VESICLE CONTENTS TO THE LYSOSOME FOR PH DEPENDENT DEGRADATION OF PROTEINS AND LIPIDS BACK TO THE HE WILL MEN CONSTITUENTS TO PROVIDE SUB TRAITS FOR THE MITOCHONDRIA TO MAINTAIN BIOENERGETICS. THAT ALLOWS THE CELL TO SURVIVE 2 TO 4 WEEKS BECAUSE SELF-EATING IS NOT A LONG TERM SURVIVAL STRATEGY, YOU RUN OUT OF THINGS THAT YOU CAN EAT FROM YOUR INTERCELLULAR CONTENTS AND THE CELL DIES NECROTICLY. SO THE CRITICAL LESION THAT REGULATES CELL SURVIVAL WE HAVE BEEN WORKING ON FOR 12 YEARS IS THE LACK OF CELL AUTONOMOUS ABILITY OF MET ZONE CELLS TO TAKE UP NUTRIENTS FROM THEIR ENVIRONMENT IN THE ABSENCE OF INSTRUCTIVE SIGNALS FROM GROWTH FACTORS HORMONES OR OTHER PARAFFIN SIGNALS. SO JUST TO DEMONSTRATE THAT THIS ISEN ANIMAL THAT HAS NO APOPTOSIS. IT'S HAD DEVELOPMENTAL PROBLEMS BUT IT'S REACHED ADULTHOOD. IT'S A PERFECTLY HAPPY MOUSE. IT RUNS AROUND ITS CAGE ALL NIGHT LOOKING FOR FOOD, AND SLEEPS ALL DAY WITH BROTHERS AND SITSERS. EVERY MAJOR ORGAN IS PERTURBED BECAUSE OF ITS DEVELOPMENTAL PROBLEMS IN GETTING THERE. BUT IT'S ABLE TO BE A FULLY FUNCTIONAL MOUSE IN THE ABSENCE OF APOPTOSIS. IT DOESN'T BECOME GREATLY DISTORTED, SINCE THIS IS MEANT TO BE A CANCER SER CEREMONY THE INTERESTING THING IS WE PREDICTED BASED TON ORIGINAL MODELS OF AP POPTOSIS THIS ANIMAL IS HIGHLY TUMOR PRONE. WE HAVE KEPT ANIMALS ALIVE NOW FOR OVER TEN YEARS. WE HAVE NEVER SEEN A SPONTANEOUS TUMOR IN THESE ANIMALS. YOU BREED IN AN ONCOGENE AND YOU WILL SEE TREMENDOUS ADVANCEMENT OF THE ONCOGENICITY OF THE ONCOGENE IN ABSENCE OF APOPTOSIS. BUT WE HAVE NEVER SEEN THE ANIMALS DEVELOP SPONTANEOUS TUMORS. THAT LED FUNDAMENTALLY TO A DIFFERENT CHANGE IN AT LEAST THE WAY WE THOUGHT ABOUT THINGS. INSTEAD OF THE PARADIGM I WAS TAUGHT AS A STUDENT THAT THAT'S A FUNDAMENTAL BARRIER THAT ALL CELLS HAVE TO BECOMING SPONTANEOUSLY TRANSFORMED IN HUMAN, WHICH IS THE LOSS OF THE ABILITY TO PROLIFERATE IN A CELL AUTONOMOUS FASHION, THAT'S BECAUSE YOU NEED GROWTH FACTOR SIGNALING THAT INSTRUCTS KNEW TRANSCRIPTION IN TRANSLATION. THERE'S A SECOND BARRIER THAT EXISTS TO CELL TRANSFORMATION, BECAUSE EVEN IF ONE ACQUIRES THE CELL AUTONOMOUS ABILITY TO TAKE TRANSCRIPTION AN TRANSLATION THAT ALLOWS A CELL TO ENTER A CELL CYCLE, THIS SIGNALING AND TRANSCRIPTION TRANSLATION ARE HIGHLY ENERGY DEPENDENT AND THEY'LL DEPLETE THE CELLS INTRACELLULAR METABOLITES JUST AS QUICKLY AS IF WE ELIMINATED THE NUTRIENT TRANSPORTERS IN THE ABSENCE OF THE COMPENSATORY INCREASE IN THEIR ABILITY TO TAKE UP NUTRIENTS. SO THIS LEADS PRO APOPTOTIC IN ABSENCE OF CONCOMITANT SIGNAL, A SECOND BARRIER WHICH THE CELL MUST BE INSTRUCTEDDED TO TAKE UP ADEQUATE NUTRIENTS FROM THE ENVIRONMENT TO MAINTAIN ATP PRODUCTION AND ALSO TO MAINTAIN MACRO AN MOLECULAR SYNTHESIS. SINCE WE BOTH NEED NUTRIENTS TO SUPPORT BIOENERGETICS BUT ALSO TO SUPPORT REPLACEMENT BIOSYNTHESIS OF PROTEINS ABOUT LIPIDS. IF YOU THINK ABOUT THAT FROM A CANCER CELL POINT OF VIEW IT HAS AN INTERESTING SET OF PREDICTIONS THAT ARE ALLOW YOU TO TE THINGS. ABOUT THE TIME WE WERE THINKING ABOUT THIS, PEOPLE WERE INVESTIGATING VARIOUS ONCOGENIC FORMS OF SIGNAL TRANSDUCTION ADAPTORS, ONCOGENES TRANSCRIPTION FACTORS AN DISCOVERING THEY HAVE WHAT'S CALLED ONCOGENIC STRESS OR APOPTOSIS THAT'S EXPLAINABLE IN A MODEL WHICH THIS INCREASED BIOENERGETIC DEMAND NOT MATCHED WITH INCREASED NUTRIENT SUPPLY CREATES THE APOPTOTIC CASCADE READING OUT IN THE BKL-2 FAMILY. IT ALSO ARGUES THAT IF YOU'RE GOING HAVE INITIATING LESION OF CANCER, IF WE UNDERSTAND A CANCER IS A SERIES OF GENETIC LESIONS, THIS IS NOT A VERY EFFECTIVE LESION TO INITIATE CANCER AND THAT'S WHY SO MANY MOUSE MODELS AND INITIATE WITH THESE KINDS OF ONCOGENIC LESIONS TAKE MONTHS BEFORE YOU SEE A TUMOR TO PLAY OUT. IN CONTRAST WE ARGUED IF THERE WAS SUCH A THING, AS FUEL SIGNALING AND YOU HAD A MUTATION IN RECEPTORS AND SIGNALING THAT DIRECTED THAT PROCESS THIS PROMOATS CELL SURVIVAL BECAUSE IT TAKES UP IN RESPONSE TO THIS INCREASED SUSTAINABLE TRANSDUCTION ONCOGENIC, INCREASE GLUCOSE UPTAKE OVER DEMAND AND NEED, THAT WOULD FUEL INCREASED ATP PRODUCTION TO MAKE TRANSCRIPTION AND TRANSLATION MORE EFFECTIVE. THE CELL DEALS WITH THAT BUT ULTIMATELY CREATE BY OVER EATING THE OXYGEN ELECTRON TRANSPORT IS NOW EXCEEDED CAPACITY FOR ATP PRODUCTION AND THAT WOULD PROMOTE AS THE CELL OVEREATS THE EXTRA CARBON SECRETED AS LACTATE, THE INCREASED ATP RATIO WILL MAINTAIN BIOENERGETICS AN CELL SURVIVAL AND RESISTANCE TO APOPTOSIS AND THE REACTIVE OXYGEN SPECIES WILL TAKE THIS NOW CELL AND BEGIN TO MIEW GENERALIZE IT, AS ELECTRONS THAT'S SCAIP THE ELECTRON TRANSPORT CHAIN ARE TURNEDDED TO HYDROGEN PEROXIDE IN THE PRESENCE OF MOLECULAR IRON START TO DAMAGE BASIS WITH HYDROXYL RADICALS OR SUPEROXIDES. SO THIS IS INTERESTING BECAUSE THIS PREDICTS IF YOU TRY TO INITIATE A TUMOR, THIS IS THE KIND OF LESION YOU SHOULD INITIATE WITH. SO AS WE START TO THINK ABOUT THAT IT ASKS ARE THERE SIGNALING PARADIGMS THAT CONTROL PRIMARILY UPTAKE OF NUTRIENTS SUCH AS GLUCOSE. WE REMINDED BY OUR COLLEAGUES IN ENDOCRINOLOGY THERE WAS WELL CHARACTERIZED SIGNATURE THAT WILL TRANSDUCTION PATHWAY THAT DID EXACTLY THAT. TOLD CELLS TO TAKE UP MORE GLUCOSE THAN THEY NEED FOR THEMSELVES. AND INSTRUCTS THEM TO PACK THOSE EXTRA GLUCOSE AWAY FOR THE REST OF THE BODY . THAT'S INSULIN SIGNALING PARADIGM AND THE INSULIN RESPONSE TISSUES, MUSCLE FAT, LIVER. TAKE UP MORE GLUCOSE THAN THEY NEED IN RESPONSE TO ACTIVATION RECEPTOR TYROSINE KINASE BY ITS PROTEINATIUS LIGAND CALLED INSULIN. THAT ACTIVATION OF PROTEIN KINASE DOMAIN LEAD TO ADAPTIVE PROTEINS THAT ACTIVATE PI-3 KINASE TO CONVERT MEMBRANE PHOSPHOINTO PIP 463 THAT RECRUITS THE MEMBRANE IN SERIES TO SERINE AND KINASES PK-1 AND AKT AND INSULIN RESPONSIVE TISSUES SUFFICIENT FOR AKT IN ACTIVATED FORM TO DIRECT THE CELL TO PLACE GLUCOSE TRANSPORTERS ON THE CELL SURFACE THE TO THE CAPTURE GLUCOSE, FOS FRL LATE THAT GLUCOSE THAT IS -- PHOSPHORYLATE INTRACELLULARLY SO IT CAN'T ESCAPE AND PERMIT IT TO GLYCOLYTIC METABOLISM BY THE INCORRECT ACTIVATION OF PFK-1 BY LAST STEP DESCRIBED IN BIOCHEMISTRY OF REGULATION OF BIOENERGETICS. AKT PREFERRED SUBSTRATE IN THIS PATHWAY IS PFK-2 WHICH BRO DEUCES THE ALLOSTERIC REGULATOR, ACTIVATION OF PFK-1. THAT PATHWAY INSTRUCT IT IS CELL IN RESPONSE TO THE SUSTAINMENT AND MAGNITUDE OF SIGNAL TRANSDUCTION TO TAKE UP GLUCOSE IN EXACTLY A PRECISE PROPORTION. SO THE CELL IS NOT MAKING THE DECISION, SIGNAL TRANSDUCTION MAKES THE DECISION. WE WENT ON TO TAKE A VARIETY OF NON-INSULIN RESPONSIVE TISSUES AN SHOW IF WE TRANSFORM WITH AKT CONSTITUTIVELY ACTIVE IT DIRECTED ALL THE PROCESSES IN EVERY TISSUE WE COULD FIND AND WE FOUND THAT IN EVERY TISSUE WHETHER SURVIVAL RECEPTORS WERE IDENTIFIED, THAT THEY DIRECTED THIS PROCESS BECAUSE THEY MIMICKED THE INSULIN RECEPTOR, EXCEPT THEY WERE LINEAGE SPECIFIC TYROSINE KINASES THAT RESPONDED TO LINEAGE SPECIFIC TRANSCRIPTION FACTORS TO INITIATE THE SAME SIGNALING PARADIGM. AT THE SAME TIME HERE AT THE NIH, THEY CAME TO OUR RESCUE IN THE TCGA BECAUSE AS NEW SEQUENCING WAS DONE OF HUMAN TUMORS THE MOST COMMON ACTIVATING MUTATION IN HUMAN CANCER AFTER R ASHS IS ACTIVATING MUTATIONS OF THE CATALYTIC SUB UNIT OF PI-3 KINASE. WHEN YOU PUT THOSE MUTATIONS FOUND IN NATURALLY OCCURRING TUMORS IN HUMAN BEINGS, INTO CELL LINES THEY DIRECT EXACTLY THIS PROCESS. AND THE SECOND MOST COMMONLY LOST TUMOR SUPPRESSOR GENE IN HUMAN SPONTANEOUS TUMORS IS NEGATIVE REGULATOR OF THIS PATHWAY P-10. WHICH IS THE PHOSPHOTASE THAT INACTIVATES THE PATHWAY BY DEGRADING PIP-3. IT APPEARS THIS IS A PATHWAY KNOWN BY CANCERS, DIFFERENCE IS IT'S NOT INSULIN, IT'S LIGAND. IT'S LINEAGE SPECIFIC SURVIVAL FACTORS. YOU CAN DEMONSTRATE THIS, I'LL SHOW YOU QUICKLY THESE ARE HEMATOPOIETIC STEM CELLS GROWN OPTIMALLY IN THE PRESENCE OF IL-3 AND THEY GROW NICELY IN THOSE CONDITIONS AN UNDERGO EXPONENTIAL GROWTH IN CULTURE. IF WE PUT IN A TRANSGENE AT LOW LEVELS IN ABSENCE OF INDUCTION BUT HIGHER LEVELS WHEN INDUCED WE SEE AND DIRECT RESPONSE TO THE INTRODUCTION OF AKT OR ITS INDUCTION, THERE'S AN INCREASE CORRESPONDING INCREASE IN GLYCOLYSIS. THERE'S NO CHANGE, HOWEVER, IN THE GROWTH RATE OF THESE CELLS, THEY'RE FULLY SATURATED THEIR GROWTH NEEDS ALREADY THROUGH THE IL-3 SIGNAL TRANSDUCTION. BUT AT THE HIGHEST LEVEL GLYCOLYSIS THERE COULD BE INDUCED BY THIS LEVEL OF ACTIVATED AKT THEY REPRODUCIBLY SUPPRESS OXYGEN CONSUMPTION WHAT WARBERG DESCRIBEDDED, THIS IS MEANINGFUL REDUCTION BECAUSE THE HIGH RATE OF GLYCOLYSIS IS SATISFYING THE ATP DEMANDS. THAT'S A PROBLEM HERE. IF THE CELL TAKES UP THIS MUCH GLUCOSE IN EXCESS OF NEEDS, IT'S GOT IT SOMEPLACE TO PUT IT, THESE ARE NOT PROFESSIONAL STORAGE TISSUE, THERE'S HEMATOPOI PROGENITOR CELLS SO TO MAINTAIN CARBON HOMEOSTASIS BECAUSE THEY CAN'T PUT THE CARBON INTO THE GROWTH CURVE THEY HAVE TO GET RID OF EXTRA BY CONVERTING THE TERMINAL STEP OF THE GLYCOLYTIC PATHWAY PYRUVATE TO LACTATE AND SECRETING FROM THE CELL. THERE'S AN EXACT CORRESPONDENCE OF EXTRA CARBON TAKEN UP BY GLYCOLYSIS SECRETION BY LACTATE. THIS IS AEROBIC GLYCOLYSIS. IT'S OVEREATING GLUCOSE. AND GETTING RID OF THAT YOU CAN'T EFFECTIVELY USE TO MAINTAIN SURVIVAL OR BIOENERGETICS. THAT'S WHAT CLINICIANS HAVE BEEN USING TO DIAGNOSE CANCER FOR THE LAST DECADE. PEOPLE RECOGNIZING WARBERG DISCOVERED YOU CAN USE POSITRON EMISSION TOMOGRAPHY TO IMAGE GLUCOSE THAT OCCURS IN NATURALLY OCCURRING TUMORS BY TAKING A POSITRON EMITTER, DEOXI GLUCOSE IN THE FOREARM OF A PATIENT AND THEN SCANNING THE PATIENT OVER 15 MINUTE INTERVALS WHERE THE GLUCOSE GETS TAKEN UP IN THE BODY. IT'S A REASSURING THING FROM WHAT WE LEARNED IN ORGANISMAL PHYSIOLOGY, THE FIRST TISSUE THAT RIGHTS UP THE HEMISPHERES OF THE BRAIN. THE LIVER IS THE NEXT MOST RECOGNIZED IN TERMS OF GLUCOSE UPTAKE AND THIS IS THE HEART. BUT IN EVERY POINT IN THIS PATIENT WHICH IS INDOLENT THYROID CARCINOMA IS IMAGED, THE TISSUE THAT TAKES UP THE MOST GLUCOSE AT EVERY TIME POINT IS THIS GOIDROUS ADINOCARCINOMA OR PAPILLARY CARCINOMA OF THE THYROID. WHEN THIS TUMOR WAS SEQUENCED THIS TUMOR HAD AN ACTIVATING MUTATION IN TSH RECEPTOR, THE LINEAGE SPECIFIC TYROSINE KINASE REACCEPT TOR THAT MAKE IT IS CELL SCAVENGE THE GLUCOSE THAT FLOATS BY IN THE ENVIRONMENT. THE CELL HAS NO PROLIFERATIVE RATE BECAUSE THIS IS AN INDOLENT THYROID CANCER OVER 7 TO 10 YEAR PERIOD IN THE PATIENT'S NECK. AND IT'S NOT HYPOXIC BECAUSE IN FACT THE GLUCOSE ITSELF GOT THROUGH THE VASCULAR SYSTEM TO THIS TISSUE FIRST. THESE ARE THE NATURALLY OCCURRING TUMORS WITH ACTIVATING MUTATIONS THAT DIRECT CELLS TO TAKE UP EXTRA NUTRIENTS AND THEREFORE, A PUSH SYSTEM FOR MORE NUTRIENTS IN THE CELL NEEDS AN PUSH THE CELL TO BEING RESISTANT TO APOPTOSIS, PUSHES THE CELL TO HAVE BUILDING BLOCKS TO ENGAGE IN MACRO MOLECULAR SYNTHESIS. AT THE TIME WE PROPOSED THIS WE SAID THAT'S ACROSS A COOL INSIGHT THAT ALLOWS US TO HYPOTHESIS THE FOLLOWING THING. IT SUGGESTS TO US THAT MANY ONCOGENES THIS IS RIGHT, CELLS LACK THE AUTONOMOUS ABILITY TO TAKE UP NUTRIENTS AND MANY ONCOGENES WE PREVIOUSLY HAS NOT UNDERSTOOD WELL MIGHT PRIMARILY HAVE EVOLVED TO REGULATE CELLULAR METABOLISM IN MULTI-CELLULAR ORGANISMS. AND THAT THE NEGATIVE -- THE ANTI-APOPTOTIC GENES AND THE TUMOR SUPPRESSORS MIGHT BE NEGATIVE REGULATORS OF THAT PATHWAY SO CELLS MATCH THE BIOENERGETIC RESOURCES TO DEMANDS BEING PUT ON THEM BY CELL PROLIFERATION. SO AT THE TIME WE HYPOTHESIZE THAT, IT SEEMED LIKE A CRAZY IDEA BECAUSE BOB WINEBERG FINISHED THE SINGLE TREE TEST, THE BEAUTIFUL BOOK I URGE THE STUDENTS AND POST-DOCS TO READ WHICH HE COMPILED ALL THE KNOWN MOLECULES INVOLVED IN EXPERIMENTAL CARCINOGENOSIS OF THIS BEAUTIFUL SIGNALING DIAGRAM TO THE CELL SURFACE THROUGH RECEPTORS TO ADAPTOR PROTEINS THE TO TRANSCRIPTION. AT THE SAME TIME DON NICKELSON WAS RETIRING FROM THE YOOFORT OF BATH IN ENGLAND WHERE HE PRODUCED FOR EVERY OF US THAT'S SEEN A METABOLIC CHART ON CELL WALL, HE PRODUCED THEM. THIS IS THE 23RD OF THOSE CHARTS HE PRODUCED AND HE HAD HIS RETIREMENT PARTY, THANK GOD WE'LL NEVER NEED TO PRODUCE ANOTHER ONE BECAUSE WE KNOW EVERYTHING WE'LL EVER NEED TO KNOW ABOUT METABOLISM. AS WE WERE HYPOTHESIZING IN 2007 HOWEVER, THERE WAS NOT A SINGLE GENE ON THIS CHART THAT WAS ON THIS CHART. AND VICE VERSA. THAT SEEMED TO US IMPOSSIBLE BECAUSE THESE ARE THE TWO MOST IMPORTANT VITAL THINGS FOR VITALITY OF LIFE. THE ABILITY TO INITIATE REPLICATION THROUGH SIGNAL TRANSDUCTION AND THE BIOCHEMISTRY TO ENACT IT. SO WE WENT ON TO TRY AN ASK WHAT ARE THE ADVANTAGES OF OVEREATING GLUCOSE AND OTHER NUTRIENTS TO CELLS THAT WANT TO ENGAGE IN CELL AUTONOMOUS PROLIFERATION SUCH AS CANCER CELLS. SO WE WENT FIRST INTO THIS PROCESS OF WHY IS IT AROW BIB GLYCOLYSIS IS SUCH A SPECIAL OR SELECTED EVENT IN TUMORS. RECOGNIZING THE 1920s IT'S ABSOLUTELY RECURRENTLY DONE, IMAGED EVERY DAY, 77% OF HUMAN TUMORS HAVE THE GLYCOLYTIC PHENOTYPE AS MANIFEST BY A PET SCAN. THE FIRST THING WE DISCOVERED IS WHEN YOU ENGAGE IN GLUCOSE METABOLISM AND EXCESS OF CELL NEED OR DEMAND IT ALLOWS THE CELL TO REPROGRAM MITOCHONDRIAL PHYSIOLOGY TO THE MORE ANCIENT WAY WHICH METAZOANE OR EUKARYOTIC ORGANISMS ACQUIRE THE PATHOGEN THAT BECAME THE MITOCHONDRIA. THE INITIAL SYMBIOSIS THAT'S SHOWN EFFECTIVELY NOW BY EVOLUTIONARY SEQUENCING THE ORIGINAL SYMBIOSIS DEPENDS NOT ON ABILITY OF MITOCHONDRIA TO CARRY OUT OXIDATIVE PHOSPHORYLATION BUT PROVIDE UNIQUE SIFNTSIS EVENTS FOR THE HOST. THE TWO THAT SEEM TO OCCUR LIEU THE EUKARYOTIC KINGDOM ARE THE ABILITY TO ENGAGE IN NOVEL LIPID SYNTHESIS THROUGH THE MITOCHONDRIA AND ENGAGE IN IRON SULFUR CLUSTER FORMATION. SO WITH WITH THAT INSIGHT WE ASKED IS THERE SOMETHING SPECIAL ABOUT LIPID SYNTHESIS WHEN CELLS READ GLUCOSE AND WE DISCOVERED THE CYCLE IS ENTIRELY REPROGRAMMED IN THE MITOCHONDRIA UNDER THAT CONDITION AND CELLS AS THEY INITIATE PRODUCTION OF CITRATE IN THE FIRST STEP OF THE CYCLE, COUPLE THAT PRODUCTION TO THE EXSCREETION OF CITRATE INTO THE CYTOSOL. CITRATE ENCOUNTERS ANOTHER AKT COMPLEX, THAT ATP CITRATE LYSE IS RESPONSIBLE FOR PRODUCTION OF ALL ACETYL COA NECESSARY FOR A CELL TO ENGAGE IN NET LIPID SYNTHESIS AND IN THE FEDERATION REACTIONS. WE GOT FOOLED WE BELIEVE IN A NUMBER OF YEARS AGO THINKING CELLS HAD TONS OF ACETYL COA BECAUSE THERE'S A LOT OF ACETYL COA PRODUCED DURING TCA CYCLE BUT THAT MITOCHONDRIAL ACETYL COA IS NOT ACCESSIBLE TO THE CELLS FOR SYNTHETIC REACTIONS IN THE CYTOPLASM SO WITH SHOWED OVER THE NEXT COUPLE OF YEARS THIS ACETYL COA IS THE SOURCE OF 90% OF THE SYNTHESIS OF FATTY ACIDS USED FOR PHOSPHOLIPID SYNTHESIS AS THE CELL EXPANDS FOR LIPID BILAYERS. A LARGE PORTION OF THE CHOLESTEROL, THE CELL PRODUCES AND IS RESPONSIBLE FOR ALL THE ISOPRANOID SIGNALING MOLECULES AND OTHER THINGS THAT ALLOWS SPECIALIZED LOCALIZATION AND SIGNALING. THE JOURNALATION, PALMETTOATION THAT ALLOWS ITING TO AGGRAVATED IN MEMBRANES. MORE RECENTLY WE HAVE COME TO APPRECIAB THE DYNAMIC RANGE OF ACETYL COA PRODUCED THROUGH THE RATE GLYCOLYSIS FLUKES THROUGH THIS PATHWAY TO PRODUCE ACETYL COA IS DETERMINE OF THE LEVEL OF PROTEIN ACETYLATION PARTICULARLY THE NUCLEUS. THAT HAD A PROFOUND INFLUENCE NOT JUST ON SURVIVAL AND METABOLISM BUT ULTIMATELY HOW CELL USES REMODELING OF CHROMATIN TO INFORM LEVEL OF GENE TRANSCRIPTION IN CELLS. I'M GOING TO SHOW YOU TWO AT LEAST IDEAS, ALL PUBLISHED AND IT HAS BEEN WELL ESTABLISHED AT STARTING THE WORK DECLINING BIOENERGETICS BECAUSE OF NED RATIO WAS CRITICAL FOR THE ACTIVATION OF HISTODEACETYL AISES OF -- AS AN ANTI-CANCER GENE. WHAT THAT IS DEACETYLATE THE HISTONES AND DRIVE CHROMATIN INTO A PATTERN OF ORGANIZATION THAT MAKES IT LESS INDUESIVE FOR GENE EXPRESSION, PARTICULARLY IN THE FORM OF GENE ELONGATION. WHAT WE REALIZED IS THE P DYNAMIC REGULATION OF PRODUCTION OF ACETYL COA COULD OPPOSE THAT SIGNAL, THIS WOULD BE A LOW ENERGY SIGNAL, LOW FLUX IN THE GLYCOLYSIS PRODUCES THIS CONDITION, HIGH FLUX OF GLYCOLYSIS PRODUCES THIS CONDITION AND WE FOUND A TENFOLD DINE RICK RANGE OF ACETYL COA CYTOPALACE M BASED ON THE RATE OF GLYCOLYSIS. WE TESTED FIRST THEN FOUND HISTONE ACETYL TRANSFERASES WERE SENSITIVE IN THAT DYNAMIC RANGE OF ACETYL COA PRODUCED BY AEROBIC GLYCOLYSIS AND THE GENE GCR-5 SUB FAMILY OF HISTONE ACETYL TRANSFERASES DO THE OPPOSITE UNDER CONDITIONS OF AEROBIC GLYCOLYSIS AN DRIVE CHROMATIN INTO A PLACE WHERE GENE EXPRESSION IS ENHANCED THROUGHOUT CHROMATIN NETWORK. THAT MAKES SENSE AS CELL HAS ROBUST SOURCE OF ENERGY IT CAN MORE FAITHFULLY ENGAGE IN MORE PRODUCTION OF INITIAL -- INITIATED TRANSCRIPTS. SO I'M NOT GOING THROUGH THIS EXPERIMENT, WE DYNAMICALLY MANIPULATED MYTOE GENIC SIGNALING SO THESE ARE RESTING FIBROBLASTS SIMULATED WITH A CYTOKINE COCKTAIL AND WE HAVE DONE THAT OVER FOUR DAYS IN THE CONDITIONS OF A PHYSIOLOGIC LEVEL OF GLUCOSE OR DIABETIC LEVEL OF GLUCOSE IN THE CULTURE MEDIA. WHAT YOU CAN SEE IS THE GLOBAL ACETYLATION OF HISTONE H-3 AND 4 T SUBSTRATES OF ACETYLATION REACTION IN THE HIS DEATHTONES IS REASONABLE IN THE RESTING CELLS BUT UNDERGOES DRAMATIC REDUCTION IN MITE ENGINE -- MYTOE GENERAL STIMULATION AND IT CAN BE RETARDED IF WE DO A ATP CITRATE LIEASE. YOU HAVE INDUCTION BY MYTOE GENESIS YOU SEE 40 FOLD INDUCTION OF ACETYLATION OF HISTONE H-3 AND 4 DEPENDENT ON ATP CITRATE LIES. SO THAT SAY IT IS LEVEL OF ACETYL AIX IS CONTROLLED NEGATIVELY BY THE FAMILY OF GENES BIOENERGETICLY COMPROMISED AN POSITIVE BY FLUX THROUGH THE GLYCOLYTIC PATHWAY. SO THAT SITS A VERY BEAUTIFUL ARGUMENT IN WHICH THE GENERALIZEDDED STATE OF YOUR ABILITY TO ENGAGE IN TRANSCRIPTION VERSUS ACTIVATION VERSUS REPRESSION IS CONTROLLED BY THE LEVEL OF HISTONE ACETYLATION UNDER A VARIETY OF MARKS OTHERS HAVE WORKED ON, SO THE SUBSTRATE AVAILABILITY. SIRTUITIAN DOO DEACETYLATION BECAUSE IT NOT RECEIVING ELECTRONS BY GLYCOLYSIS AND THE GENE WHEN THERE'S A HIGH FLUX OF GLUCOSE TO THE GLYCOLYTIC PATHWAY, NAD IS LOW AND ACETYL COA IS HIGH WHICH DRIVES ACETYLATION OF HISTONES TO INCREASE TRANSCRIPTIONAL ACTIVATION AND WE HAVE BEEN ABLE TO SHOW THE PREFERENCIAL SITE IS A POSITIVE FEET FORWARD LOOP ON THE GENES TO GLYCOLYTIC METABOLISM, IT CELLS LOTS OF CELL INDEPENDENCE ON GLUCOSE. THAT'S THEN SORT OF EXPLAINS WHY IT IS IN CANCER AND MODEL ORGANISMS THAT HAVE BEEN ALL ORGANISMS STUDIED SIGNAL TRANSDUCTION THROUGH THE PI-3 KINASE AKT PATHWAY AND SECONDARILY THROUGH TOR THROUGH ARCKT DIRECTED ABILITY TO ACTIVATE TOR IS THE CONTROLLER OF CELL SIZE. BECAUSE IT ALLOWS CELLS IT DIRECTS CELLS WITHOUT TRANSCRIPTIONAL INTERVENTION TO REPROGRAM ALL CELLULAR METABOLISM. TO CAPTURE GLUCOSE EXTRA CELLULARLY COMMITTED TO THE GLYCOLYTIC METABOLISM IN SUFFICIENT QUANTITIES THAT CITRATE CAN BE PRODUCED IN EXCESS TO PRODUCE NON-ESSENTIAL AMINO ACIDS BUT LIPIDS FOR LIPID BIOSYNTHESIS AN TOR ACTIVATION REPROGRAMS AMINO ACIDS AWAY FROM CATABOLISM IN THE TCA CYCLE AND INTO TR AND HR INCREASED PROTEIN TRANSLATIONAL INITIATION AND INCREASE PROTEIN SYNTHESIS. SO UNDER ALL PHYSIOLOGIC CONDITIONS, IN ALL MODEL ORGANISMS IN THE STUDY, SPONTANEOUS ACTIVATION OF THIS SIGNALING PATHWAY I JUST TALKED ABOUT MAKES A LARGER CELL IN AUTO STAGES OF CoyoR CYCLE BECAUSE THE CELL IS SIMPLY A BAG OF FROM TEENS SURROUNDED BY A LIPID BILAYER. AND SECONDARILY THERE'S POSITIVE INFORMATION OF TWO PATHWAYS I SAID REGULATED INDEPENDENTLY IN THE MODELING THAT WE STARTED WITH, BECAUSE CITRATE INFORMS THROUGH ITS ABILITY TO MODIFY ACETYLATION INCREASED ABILITY TO TRANSCRIBE GENES AND TOR INCREASES ABILITY TO OFFICIALLY ACTIVATE TRANSLATION PARTICULARLY TRANSLATION INITIATION. SO YOU INFORM THE TWO REMAINING PROCESSES REQUIRED FOR GROWTH, PROTEIN, GENE TRANSCRIPTION AN PROTEIN TRANSLATION. SO WE WERE PRETTY HAPPY WITH THIS. ASEN EXPLANATION OF IN FACT WHAT IS THE FUNDAMENTAL BEST OF CELL SURVIVAL, ABSENCE OF THE ABILITY TO TAKE UP CARBON TO SURVIVE AND REPROGRAMMING OF THE CARBON DIRECTED ONCOGENIC STEP. WE STARTED TO THINK ABOUT THIS A FEW YEARS AGO AND THIS COULD INFORM MORE ABOUT CANCER THAN JUST GLUCOSE METABOLISM. AND WE WERE INFORMED BY THAT BECAUSE AS A MEDICAL ONCOLOGIST THOUGH I'M INTERESTED IN MUTATIONS AN CANCER IN=D THESE THINGS AND THIS AEROBIC GLYCOLYSIS IMAGED BY PET SCAN IMAGING I NEVER HAD A PATIENT DIE OF ONE BIG CELL. A CELL HAS TO DIVIDE TO PRODUCE THE LESION ASSOCIATED WITH PATHOGENESIS OF CANCER. IT'S THE EXPANDING CLONE OF CELLS AND ABILITY TO ACQUIRE NEW PROPERTIES AND ABILITY TO METASTASIS IN PARTICULARLY HUMAN PATIENTS THAT WAS IMPORTANT. YOU CANNOT BUILD ALL THE BUILDING BLOCKS NECESSARY FOR CELL DIVISION SIMPLY THROUGH THESE PATHWAYS WHAT ONE NEEDS IS ROBUST SOURCE OF REDUCED CARBON TO BUILD NUCLEIC ACIDS MISSING FROM THIS CASCADE SO WE WENT LOOKING FOR WHETHER THERE WAS A PATHWAY DEDICATED TO UPTAKE OF REDUCED NITROGEN THAT WOULD ALLOW CELLS TO INFORM NUCLEOTIDE BIOSYNTHESIS AND REMAINING SYNTHETIC REACTIONS THAT WE HADN'T YET PUT ON THIS SCAS CAID OF GROWTH. -- CASCADE OF GROWTH. LONG STORY SHORT, HEMATOPOIETIC TISSUES RECEPTOR TYROSINE KINASES INITIATE THIS WAT WAY BUT WE DISCOVERED KITE SIGNS PARTICULARLY DEDICATED TO THE JACK STAT SIGNALING PATHWAY, INFORMED THE TRANSCRIPTION OF THE ESSENTIAL FACTOR THAT SO FAR WE AND OTHERS FOUND THAT REGULATES NITROGEN UPTAKE AND THAT NITROGEN IS DIRECTED BY MIC AS TRANSCRIPTION FACTOR WHICH PUTS IN A CASCADE OF GENES THAT CONTROLS UPTAKE AND METABOLISM OF GLUTAMINE MUCH THE WAY THIS SIGNALING PATHWAY CONTROLS THE UPTAKE AND METABOLISM OF GLUCOSE. AND THE UPTAKE OF GLUTAMINE PROVIDES KNEW TROA GENERAL SOURCE FOR DE NOVO PURINE BIOSYNTHESIS, A CARBON SOURCE TO ATHROUGH GLUCOSE REDIRECTED TO RIBOSE BIOSYNTHESIS. AND IT ALLOWS A SUBSTRATE THAT CAN BE CONVERTED IF METABOLIZED IN EXCESS TO PRODUCE NAD PH TO FUEL THE SYNTHESIS OF NUCLEOTIDES AND LIPIDS. SO THAT SEEMED TO COMPLETE ALL THE MAJOR BUILDING BLOCKS THAT WE NEED FOR CELL GROWTH AND CONSISTENT WITH THAT AN INTO BURNS MADE A MOUSE IN THE IMMUNOLOGIC LINEAGE, THE B CELL LINEAGE HAS THE ACTIVATION BY ENHANCER OF AKT AND MIC AT THE EMBRYO GENESIS AND INSTEAD OF TAKING MONTHS HE IS TWO INDIVIDUAL MOUSE -- THIS MOUSE MIC B CELL MOUSE WILL PRODUCE TUMORS AROUND SIX MONTHS OF AGE AND AKT MOUSE ABOUT NINE MONTHS OF AGE IF HE BRED THEM TOGETHER AS AN INTO PRODUCED HE FOUND THE GENES COMPLIMENTED EACH OTHER THE FIRST B CELLS MADE IN THE MOUSE DAY 18 OF EMBRYO GENESIS AN EVERY ANIMAL DIES OF ITS TUMOR AT BIRTH THREE DAYS LATER. BECAUSE THE MINUTE THE CELL COMMITS TO THE B CELL LINEAGE THESE TWO PATHWAYS ARE FULLY ON AND CELLS IN CELL CYCLE TIME OF FIVE HOURS. IT DOESN'T THINK. IT SIMPLY DIVIDES. IT DIVIDES WHEREVER IT IS NO MATTER WHAT ELSE IT'S DOING. MIK CAN TELL US ABOUT NITROGEN METABOLISM AND WE DID THIS EXPERIMENT WHICH GOT US TOTALLY CONFUSED. IN COMPLETE MEDIA TISSUE CULTURE CELLS FOR THE STUDENTS IN POST DOCS THE REASON WE KNEW GLUTAMINE ONE AN IMPORTANT PART OF THIS IS THAT DIRTY LITTLE SECRET OF CELL CULTURE IS THIS WE ADD TENFOLD MORE EXCESS GLUTAMINE TO EVERY TISSUE CULTURE MEDIA IN THE WORLD. SINCE WE'RE IMMUNOLOGY LAB WE DOUBLE THAT BECAUSE WE DON'T TRUST ANYBODY ELSE. ALL THE IMMUNOLOGISTS DO IT. WE'RE NOT UNIQUE. WE THOUGHT THE GLUTAMINE WOULD BE SPECIAL SO WE DEMONSTRATED YOU TAKE MIK TRANSFORMED CELLS AND FIND IT NATURALLY OCCURRING TUMORS OR MAKE MIK TRANSFORM CELLS AND WE FOUND THERE WERE ADDICTED TO GLUTAMINE. IF WE TOOK GLUTAMINE FROM MEDIA, LEFT GLUCOSE AT 11 MILLIMOLAR AS IT WAS IN THIS TISSUE CULTURE MEDIA, KEPT THE NUTRIENTS PRESENT BUT TOOK GLUTAMINE OUT EVERY CELL DIED WITHIN 3 TO 4 DAYS IN TISSUE CULTURE MEDIA. THE SURPRISE CAME WHEN WE TRIED TO ASK WHAT WOULD RESCUE THIS DEATH, WE FOUND THE ONLY THING THAT WAS NECESSARY TO RESCUE THE CELL DEATH WAS THE CARBON BACKBONE OF THE GLUTAMINE AL KHAKI TOE GLUTERATE. WE DIDN'T NEED ANY NITROGEN TO SURVIVE THE REST. WE NEEDED SOME TO INITIATE FULL PROLIFERATION BUT NO CELL DIES IF WE JUST ADDED AL CA KETO GLUTERATE, THAT WAS A PUZZLE THAT MADE US WANT THE DIG DEEPER INTO THIS. WE LEARNED FROM THOSE STUDIES IS THAT IN FACT FOR ROBUST GROWTH TO BE ABLE TO FUEL THE CITRATE BIOSYNTHESIS THAT I TALKED ABOUT AS CITRATE IS EXPORTED YOU DON'T ONLY EXPORT THE CARBON EQUIVALENTS FROM PYRUVATE THAT I TALKED ABOUT, IT'S CONVERTED TO ACETYL COA, THEN CONDENSED WITH OXALL,O ACETATE TO PRODUCE CITRATE. AND WHEN IT'S EXPORTED AND USED FOR SYNTHESIS, AND VARIOUS SYNTHETIC REACTIONS DOWNSTREAM OF THIS YOU NEED A SOURCE OF SOMETHING TO PRODUCE EXTRA MALLAY THAT REENTERS THE TCH CYCLE AS A SUBSTRATE TO REPLENISH THE CYCLE TO PRODUCE OXALLO ACETATE. IN EFFECTIVELY GROWING CANCER CELLS TRANSFORMED WITH MIK ALL THE SUB TRAIT THAT MAINTAIN IThT IS TCA CYCLE ENTERS AS ALPHA KETO GLUTERATE FROM DEAM NATION OF GLUTERATE. SO WE WERE SEEING THE ALPHA KETO GLOWT RATE TO BUILD UP A POOL OF CELL THAT MAINTAINED INTEGRITY OF THE THE,CA CYCLE OR THAT WAS SUFFICIENT EXPLANATION. IN MIK TRANSFORMED CELLS WE FIND THE ALPHA GLUTERATE POOL E QUILL LIBRIUM WITH GLUTAMATE IS HABIT TO ONE MILLIMOLAR. THAT'S A UNIQUE POOL. WE NEVER SEE THE LEVELS COLLECTIVELY COMBINED BUILD THOSE LEVELS EXCEPT WHEN CELLS ARE INSTRUCTED TO SCAVENGE GLUTAMINE FROM THE ENVIRONMENT. SO WE ASKED WHAT'S SPECIAL ABOUT HAVING THIS LEVEL OF ALPHA GLUTERATE, THERE'S RELEVANT TO THE LAST 15 MINUTES OF MY TALK. ONE THING THAT KIM OUT, PUBLISHED BY SIX GROUPS IN SIX MONTHS INCLUDING OURSELVES IS THAT THAT GLUTAMINE METABOLISM IS SPECIAL WHEN CELLS WANT TO SURVIVE HYPOXIA. WHEN YOU WANT TO SURVIVE HYPOXIA AND GENERATE ATP AS EVERYONE KNOWS YOU ENGAGE IN GLYCOLYTIC METABOLISM AS IN THE FIRST SLIDE. THAT MEANS NO SUBSTRATES TO SUPPLY THE TCA CYCLE AND THEREFORE CARBON TO DO SYNTHETIC REACTIONS TO BUILD GROWTH. YET WHEN YOU CUT YOURSELF AN YOU HAVE A WOUND, AND YOU NEED THE GROW EITHER THE VASCULATURE BACK OR RECOVER THE INTEGRITY OF THAT TISSUE IN ABSENCE OF MOLECULAR OXYGEN YOU NEED A WAY TO BUILD THE LIPIDS AND THE NON-ESSENTIAL MAY MEE KNOW ACIDS UNDER CONDITIONS OF HYPOXIA. WITH THAT KIND OF REASONING, SIX LABORATORIES SIMULTANEOUSLY ASKEDDED I WONDER IF SIGNAL TRANSDUCTION THROUGH GLUTAMINE COULD DO THAT. WHAT WE AND TWO OTHER LABORATORIES DESCRIBED EARLIER IN THIS YEAR, WAS THAT IN FACT GLUTAMINE METABOLISM DOWNSTREAM OF MIK COULD FULLY INFORM CELL GROWTH UNDER HYPOXIA, GLUCOSE METABOLISM WOULD PROVIDE THE ATP, BUT GLUTAMINE NOW BY PROVIDING THIS LARGE POOL OF ALPHA KETO GLUTERATE IN THE TCA CYCLE THOUGH NO MOLECULAR OXYGEN SO YOU CAN MOVE T IT THIS DIRECTION, TO DO ELECTRONIC TRANSPORT, ALPHA GLUTERATE WORKS IN REVERSE AS THE TCA CYCLE IS REPROGRAMMED BE CAR BOXLATED SO THIS IS REDUCTIVE COR BOXLATION, INTO CITRATE WHICH IS THEN BY CONVERTED TO CITRATE EXPORTED TO PROVIDE THE SAME SYNTHETIC FUEL FOR ACETYLATION REACTIONS AND LIPPED BIOSYNTHESIS THROUGH GLUCOSE. AS WELLS GROW USING GLUTAMINE AS SUB TRAIT TO PROTECT THE MITOCHONDRIA PREVENTING REDOX STRESS IN THE MITE CHON KOREA AND -- MITOCHONDRIA AND PROVIDE GLUCOSE. SO IT PROVIDES A SPECIAL ADAPTIVE ABILITY TO CELLS THAT HAD LESIONS THAT SCAVENGE GLUTAMINE FROM THE ENVIRONMENT. AND THAT IS EXTENDED NOW BY SEVERAL LABORATORIES. AS WE WERE DESCRIBING THIS AND GETTING SATISFIED WITH OURSELVES THIS IS A COOL PATHWAY DOWNSTREAM OF MIK ON OWE GENE THAT HAD COMPONENTS OF -- ONCOGENE, THAT DEAM MATED -- DEAMNATE GLUE TAMNASE ENTERED IN PARTIAL TCA CYCLE WE FOUND THE NAD PH WAS PRODUCED BY ENZYME CONVERTING MALI TO PYRUVATE PRODUCING NAD PH PROVIDING ELECTRONS FOR THE SYNTHESIS REACTS REQUIRED FOR NUCLEOTIDE BIOSYNTHESIS AND LIPID BIOSYNTHESIS AND YOU COULD THROW AWAY CARBON THAT THE CELL DIDN'T NEED BY USING LDH-A A WELL MIK TARGET IN THE CYTOSOL. WHAT WAS EXCITING ABOUT THIS SHOWS HOW NERDY WE ARE, IS THAT WHEN WE ADD UP CELL GROWTH AS A PROBLEM, THE LIMITING SUBSTRATE FOR US IS LACK OF ANY KNOWN WAY TO PRODUCE ENOUGH NAD PH TO ACTUALLY FUEL CELL GROWTH. YOU CANNOT DO LIPID SYNTHESIS OR NUCLEOTIDE SYNTHESIS WITHOUT ROBUST POOL OF NAD PH. IF YOU ASK YOUR TEXTBOOKS I THINK NONE OF YOU KNOW ANY SYNTHETIC PATHWAY TO PRODUCE IT, EXCEPT A FEW OF YOU REMEMBERED BIOCHEMISTRY WELL ENOUGH BECAUSE YOU GOT THE HONORS. THAT ENZYME IS G 6 PD, THE OXIDATIVE ARM OF THE P ENTOS FOSNATE SHUNT. WE KNEW EARLY ON THAT THAT WASN'T THE POSSIBLY ANSWER FOR NAD PH FOR GROWTH BECAUSE THERE ARE 3 MILLION MEN WALKING AROUND THE MEDITERRANEAN BASIS THAT HAVE NO G-6 PD. THEY HAD NO EMBRYO GENESIS WITHOUT AND WELL STUDIED IN ALL THE MEDITERRANEAN COMPANIES THEY HAVE IDENTICAL RATES OF CANCER TO THEIR AGE MATCHED CONTROLS SO G-6 PD IS NOT IS SUB TRAIT THAT PRODUCES N ASHD PH FOR GROWTH. ONLY TWO REMAINING CANDIDATES AND WE WERE VERY EXCITED ABOUT MALIK ENZYME BECAUSE THAT CAN PRODUCE PYRUVATE IN THIS PATHWAY AND NAD PH PATRICK WARD IN THE LABORATORY SHOWEDDED THIS MIK INDUCED MALIC ENZYME PRODUCE NAD PH BUT WE DISCOVERED WE GOT ONE THING ABOUT THIS MODEL WRONG. THE ENZYME PRODUCED BY MIK IS IN THE -- HAS A MITOCHONDRIAL LOCALIZATION SEQUENCE IN HERE. SO THOUGH TRUE IT CAN PRODUCE NAD PH PRODUCTION IT WAS IN THE WRONG PLACE BECAUSE YOU PRODUCE NAD IN THE MITOCHONDRIA AND THERE ARE INDEPENDENT POOLS THAT DON'T EXCHANGE. SO NOW WE HAD A PROBLEM BECAUSE THOUGH WELL DESCRIBED WAYS TO CONVERT N DARKSH FROM THE SOL TO MITOCHONDRIA AND BACK NO KNOWN WAYS TO TRANSPORT NAD PH ACROSS THE MEMBRANE. SO PATRICK WARD IN THE LAB A COUPLE OF YEARS AGO RECOGNIZED TWO OTHER ADDITIONAL ENZYMES RELATED TO TCI CYCLE ENZYME DEHYDROGENASE THAT I TALKED ABOUT THAT CONTROLS. THEY'RE CALLED IDH-1 AND 2 AND THEY ARE NAD PH DEPENDENT ENZYMES AND THEY ARE RESPONSIBLE FOR THE REDUCTIVE RESPONSIBLE FOR REDUCTIVE CARBOXLATION, IT'S ALSO RESPONSIBLE AT LEAST IN THE DATA WE HAVE BEEN ABLE TO DERIVE SO FAR TO BE ABLE TO IN THIS REDUCTIVE CAR BOXLATION EXPORT INTO THE CYTOSOL AND REVERSE METABOLISM THROUGH HOMOLOGUE IN THE CYTOSOL TO REGENERATE ALPHA KE TO TRANSFER ONE EQUIVALENT FROM THE MITOCHONDRIA TO THE CYTOSOL. SO PATRICK BUILT UP AER CONSIDERABLE AMOUNT OF DATA IN MIK TRANSFORMED THE WAY CELLS DEAL WITH NAD PH PROBLEM IS PRODUCE IN THE MITOCHONDRIA AN EXPORT THROUGH THIS SHUTTLING PATHWAY. IT'S SUPPORT -- IN SUPPORT OF THAT ARGUMENT HE SHOWED IF YOU ELIMINATE IDH-1 OR 2 BY RNAi FROM MIK TRANSFORM TUMOR CELLS, THEY DON'T DIE BUT THEY LOSE THEIR ABILITY TO PROLIFERATE IN TISSUE CULTURE COMPARED TO THE UNTREATED RNAi CELL. SO WE WERE EXCITED TO SAY THIS IS A REALLY COOL THING, THIS IS A NEW PATHWAY OF METABOLISM IN WHICH WE EQUALIZE ELECTRONS AWAY FROM THE COMPARTMENT NORMALLY USED TO FUEL OXIDATIVE PHOSPHORYLATION. WE TRANSFER BACK TO MITOCHONDRIA TO FUEL SYNTHETIC REACTIONS WHEN THE FOLLOWING DISCOVERY WAS MADE BY BURT VOGELSTEEN AND COLLEAGUES AT HOPKINS. IT'S EXTENDED BY MANY GROUPS. THIS SLIDE IS NOT UP TO DATE. BURT WAS ABLE TO DISCOVER IN SECONDARY GLIAL BLASTOMAS THE LESION OF ISOCITRATE DEHYDROGENASE. EVERY SECONDARY GLIAL BLASTOMA IN THE BED OF INDOLENT GLIOMA, INITIATES IN EITHER THE CYTOSOL FORM OF IDH-1 OR MITOCHONDRIAL FORM OF ID H-2. T 0% OF ALL -- 90% OF ALL GRADE GLIOMAS HAVE THE GENE AND THEY APPEAR TO BE THE INITIATING GENE. SUBSEQUENTLY SHOWN THAT 60% OF SARCOMAS HAVE THIS AS INITIATING LESION, 30% ADULT AMLs AN MORE RECENTLY A LARGE PERCENTAGE OVER HALF CARCINOMAS HAVE IDH-1 OR IDH MUTATIONS. THAT DIDN'T FIT OUR DATA. WHEN BURT CHARACTERIZED THEM THESE ENZYMES LOST THEIR ABILITY TO CONVERT ISOCITRATE TO ALPHA KETO GLUTERATE. I ARGUED THAT ABILITY WAS REQUIRED FOR SHUTTLING OF NAD PH EQUIVALENTS TO EFFECTIVELY PROVIDE THE ENERGY SUBSTRATES FOR CELL GROWTH. BUT THERE WAS HOPE WHAT WE WERE THREE THINKING AND WHAT THEY WERE THINKING BECAUSE THEY NO TUSKED ALL MUTATIONS WERE MISSENSE AND THAT ALL THE MISSENSE MUTATIONS OCCURRED IN A SINGLE R GENE IDH-1 AND IT'S IDENTICAL ARGININE IN THE REACTION CENTER RGH-2. IN EVERY CASE ONE IDH-1 ALLELE OR 2 ALLELE IS ALWAYS MAINTAINED WILD TYPE. SO THERE'S WILD TYPE ACTIVITY IN THE OTHER HOMOLOGUE. SO YOU LOOK AT THAT REACTION, IT STARTS TO EXPLAIN WHAT THEY WERE FINDING BY THESE RECURRENT NATURALLY OCCURRING HUMAN MUTATIONS NOW IN FOUR MAJOR MALIGNANCIES AND SOON TO BE FIFTH. THAT IS, THIS IS THE REACTION BY THESE AMINO ACIDS THAT LACK -- THAT LINE THE SUBSTRATE BINDING SITE OF ISOCITRATE DEHYDROGENASE, THIS IS THE ISOCITRATE MODELED IN THAT SITE AND HOW IT'S COORDINATED WITH ITS -- WITH THE ENZYMES IN THE IDH-GENE. WHAT YOU CAN SEE IS THE BLUE CARBON LABELED HERE THE CO-2 THAT LEAVES IS ISOCITRATE DECAR BOXLATED INTO ALPHA KETO GLUTERATE IS THE ABILITY TO BOUND IN A SITE IS COORDINATED BY ARGININE 132 IN IDH-1 AND ARNL NEEN 172 IN IDH-2. THESE WERE MUTATIONS AND THE ONLY MUTATIONS THAT BURT VOGELSTEEN WAS ABLE TO FIND IN GLIOMAS. THAT MAKES SENSE IF YOU CAN'T COORDINATE THE THING YOU BREAK OFF THIS MOLECULE YOU CAN'T BIND ISOCITRATE EFFECTIVELY. IT'S INTERESTING THE PRODUCT OF THE REAL-QAEDA, THE NORMAL FORD REACTION BINDS WITH WILD TYPE AFFINITY THE CO-2 GROUP. WITH THAT INFORMATION (INDISCERNIBLE) PHARMACEUTICALS TRANSFORMED CELLS WITH IDH-1 MUTANT AND ASKED ARE THERE NEW METABOLITES PRODUCED. WHAT HE DISCOVERED IS A NEW METABOLITE BASED ON METABALOMICS ANALYSIS THAT READS OUT IN THE FOLLOWING WAY. IT MAKES PERFECT SENSE, THE METABOLITE IS HYDROXY GLUTERATE, IT WORKS BECAUSE ALPHA KETO GLUTERATE CAN BIND WITH WILD TYPE AFFINITY, BECAUSE IT'S NOT DEPENDENT ON ARGININE FOR THE FINDING BUT IT IS DEPENDENT ON THE ARGININE LOST FOR ABILITY TO ENGAGE IN REVERSE CAR BOXLATION TO ACCEPT A CO-2. WITHOUT THE OX GENERAL COORDINATE THE CO-2 THAT REVERSE REACTION CAN'T OCCUR BUT IT CAN STILL ACCEPT ELECTRONS FROM NAD PH EXCEPT NOW THE PRODUCT IS HYDROXYLATION OF BETA CARBON SO YOU PRODUCE 2 HYDROXY GLUTERATE INSTID OF ISOCITRATE. SO THEN YOU LOOK AT TUMORS THAT HAVE MUTATIONS IN IDH AND WHAT YOU FIND IS THAT THOSE TUMORS WHEN ISOLATEDDED, THIS IS DATA SETS THAT WE DERIVED FROM AML PATIENTS, WILD TYPE TUMORS WITH WILD TYPE IDH THAT ARE AML HAVE EITHER NO DETECTIVE IDH OR THIS IS THE LEVEL OF DETECTION OF A GC MASS SPEC. IN CONTRAST, THESE MUTATIONS ALSO SEEN IN LIEU CHEMOIAS, IDH-132, IDH-2, 172 BUILD UP THIS IS A LOG SCALE TEN MILLIMOLAR INTRACELLULAR CONCENTRATIONS OF DEOXI GLUTERATE. THAT MAKES IT THE MOST COMMON METABOLITE IN THE CELL. WE THEN SCREENED ALL LEUKEMIAS FOR 2 HG AND DISCOVERED ANOTHER GROUP HAD HIGH 2 HG WHEN SEQUENCED THEIR, ENZYMES DISCOVERED THEY HAD A MUTATION IN IDH-2 BUT NOW A DIFFERENT ARGININE 140. THAT SEALED THE DEAL FOR US BECAUSE 140 IS THE OTHER AMINO ACID AND REACTION SITE THAT COORDINATES THAT CO-2 MOVING GROUP. SO WHAT THAT SAYS, IT -- WHAT'S NOW SHOWN BY THE INVESTIGATORS GSK INVESTIGATING THIS, IS THE PREFERENCIAL OPERA OPERATIVE MOIETY INSIDE THE CELL IS HETERODIMER OF MUTANT ENZYME WILD TYPE ENZYME. THE WILD TYPE CONVERTS ISOCITRATE IN PRESENCE OF NAD AS ELECTRONIC RECEPTOR AND GLUTERATE CO-2 NAD PH. THE CONCENTRATION OF NAD PH AND ALPHA KETO GLUTERATE AT THIS REACTION SITE DRIVES THE PRODUCTION OF THE REVERSE PRODUCT 2 HYDROXY GLUTERATE ARE AT MUTANT. THIS IS A HETERODIMER NATURALLY OCCURRING AS A RESULT OF THIS MUTATION, AS A ACTIVITY THAT CONVERTS ISOCITRATE TO HG THIS IS A FACTORY TO DO THAT. SO CELLS BUILD UP A STEADY STATE THAT BUILDS UP TO A MILLIMOLAR OF 2 HG. THAT'S INTERESTING BECAUSE THIS IS A GENETIC LESION BUILDING UP, IT'S A METABOLITE SO HOW DOES THE METABOLITE INFLUENCE ANYTHING THAT HAS TO DO WITH CELL BIOLOGY. SO WE TOOK ADVANTAGE OF THE FACT THAT IN LIEU CHEMOIA, WE WERE SURPRISED TO FIND A MUTATION THAT HAD A PEN TRANSOF 30% IN OUR PATIENT, WE THOUGHT WE KNEW EVERY LEE QUMEE LOCATION THAT EXISTED BASED ON SEQUENCING. WE ASKED WERE THERE GENES MUTATED WHEN IDH WAS MUTATED OR GENES NEVER MUTATED TO SUGGEST THAT THEY EITHER DEPENDENT ON EACH OTHER FOR FUNCTIONALITY OR THEY WERE EPISTATIC TO EACH OTHER. DISCOVER AD GENE EPISTATIC WITH IDH-1 AND 2. WHEN YOU LOOK FOR PATIENT, THIS IS A CONNECTION OF PATIENT, TUMORS THAT HAD MORE THAN ONE MUTATION, THESE ARE ALL TUMORS THAT I IDH-1 OR 2 MUTATIONS IN ANY KNOWN ONCOGENE THAT CAUSED LEUKEMIA, THAT WAS TET 2. IN REVERSE PATIENTS THAT HAD TET 2 MUTATIONS COULD HAVE MUTATIONS IN ANY OTHER KNOWN ONCOGENE EXCEPT IDH-1 AND 2. HIGHLY STATISTICALLY SIGNIFICANT AND REPLICATED THROUGH MULTIPLE DATABASES. THAT GAVE AT LEAST A CLUE FOR INITIATION OF THE SECOND CONNECTION OF METABOLISM EPIENGINE TIX AND THAT WAS THE REALIZATION BY (INAUDIBLE) THAT TET 2 IS ENZYME THAT INITIATES THE DEMETHYLATION OF DNA AT CPG ISLANDS BECAUSE IT TAKES ALPHA KETO GLUTERATE, STRIPS ELECTRONS OUT AND PRODUCING SUCCINATE AND CHANCES ELECTRONS IN HYDROXYLATION REACTION THAT INITIATE THE DESTABILIZATION OF METHYL C. THAT INITIATING ULTIMATELY ACTIVELY OR PASSIVELY DNA DEMETHYLATION. TWO HYDROXY GLUTERATE BECAUSE IT IS A HOMOLOGUE OF ALPHA QUITO GLUTERATE LOOKS LIKE A COMPETITIVE SUBSTRATE TO BLOCK THE REACTION. SO WE TESTED THE IDEA, ADOPTED AN ASSAY THAT WE DESCRIBED, TRANSFECT IN TATTOO, YOU CAN SEE CELLS BECAUSE THEY HAVE A FLAG TAG THAT ARE TRANSFECTED AN THOSE ACQUIRE A HUGE HYDROXYLATION MARKSES ON DNA AS EXOGENOUS PRODUCTION OF TET 2. AND IF YOU CO-TRANSFECT IN MUTANT IDH-1 YOU BLOCK THE ABILITY OF TET-2 TO INDUCE HYDROXYLATEDDED DNA, WILD TYPE ENZYME DOES NOT DO THAT. YOU CAN SEE A ONE LOG EFFECT BY FAX ANALYSIS. SO WE WENT ON TO SAY THAT -- THIS IS SOMETHING I DONE HAVE TIME TO SHOW YOU DATA FOR, WE NOW BELIEVE THAT BURST ON STREAM OF MIC THAT PRODUCES THIS POOL OF ALPHA KETO GLUTERATE IS ESSENTIAL AS CO-FACTOR THAT ALLOWS DEMETH LACE OF DNA AND AT HISTONES TO RETURN SILENCE CHROMATIN FROM INACTIVE THE ACTIVE STATE. ON THAT BATE IF YOU CAN'T REACT ACTIVATE GENES SILENCED TO ALLOW CELLS TO ENGAGE DIFFERENTIATION PROGRAM, THEY'RE STUCK IN CURRENT OR STEM CELL LIKE STATE. 2 HYDROXY GLUTERATE AS AN ANTAGONIST PROTECTS IT FROM THAT BURST OF ENERGY TO INITIATE DEMETHYLATION OF SILENCED CHROMATIN. SO TO PROVE THAT WE DEMONSTRATED, I'M GOING OVER IN TIME, WHEN YOU EITHER TRANSFECT IN MUTANT IDH OR YOU DELETE TET-2 BY SH RNA YOU GET PERSISTENCE IN CELL CULTURE AND HEMATOPOIETIC STEM CELLS, THEY LOOSE THEIR ABILITY TO DIFFERENTIATE AND THAT FITS A MODEL OF WHAT WE ALREADY KNEW FROM THE VERY BEGINNING ABOUT AML, LOST THE ABILITY TO DIFFERENTIATE AND THAT THE LOSS OF TET-2 TO INITIATE THE OPENING OF CHROMATIN OR 2 HG TO COMPETITIVELY ALPHA KETO GLUTERATE SUBSTRATE BY TET ENZYMES BLOCKS CELLULAR DIFFERENTIATION AND THEREFORE INITIATES THE BLOCKED DIFFERENTIATED THAT CONTRIBUTES TO AML PATHOGENESIS. THE QUESTION IS WHAT ABOUT GLIOMAS BECAUSE AT THE SAME TIME WE WERE DOING THIS WORK WE LOOKED AT GLIOMAS, WE DIDN'T SEE TET MUTATIONS BUT IN DOING THOSE STUDIES WE REALIZED THAT THERE'S DIFFERENT SET OF METHYLATION MARKS THAT ALSO DEPEND ON THIS ALPHA KETO GLUTERATE BURST DOWNSTREAM OF MIK FOR DEMETHYLATION, THERE'S THE HISTONE REPRESSIVE MARKS ON HK-29 AND 27 REQUIRED TO MAINTAIN SILENCE CHROMATIN ALONG WITH METHYLATED CPG. WE WERE ABLE TO DEMONSTRATE OTHERS DEMONSTRATED THAT THE CLASS OF ENZYMES THAT DO THOSE DEMETHYLATION REACTS OF REPRESSIVE MARKS, HISTONE METHYLASES THAT DEPEND ON THE SAME ENZYME KINETICS, ALPHA KEO GLUT RATE CONVERTED TO SUCKNATE AND WE DEMONSTRATED THE HK-39 YOU HAVE 2 HG SHOWN HERE, YOU TAKE AT THISTONES -- HISTONES MODIFIED, NATURALLY OCCURRING, YOU ADD IT TO CULTURE, ADD ENZYME IN PRESENCE OF ONE MILIMOLAR YOU DEMETHYLATE THEM. ADD 2HG YOU BLOCK IT BUT IT'S COMPETITIVE AS WE DOSE UP ALPHA KETO GLUTERATE YOU REVERSE ABILITY OF 2 HG TO CONVERT. IF YOU GO INTO SPONTANEOUS HUMAN GLIOMAS WHICH YOU FIND IS H 3 K-9 IN IS OLIGOMER, THIS IS IMMUNOHISTOCHEMISTRY TO DEMONSTRATE THAT WITH HIGH K-9 METHYLATION AND THAT'S TRUE AND HIGHLY STATISTICALLY SIGNIFICANT IN MUTANT TUMORS AND THEN WHEN WE TAKE AN ASTROCYTE PRIMARY CELL LINE AND INTRODUCE MUTANT IDH WE SEE HISTONE METHYLATION AND THAT'S SUB QNTLY FOLLOWED SEVERAL GENERATIONS LATER BY DNA DEMETHYLATION. NOT AFFECTING THE TET, SEEMED TO BE COOPERATIVETY OF THE INABILITY OF DEMETHYLATE HISTONES IMPAIRS ABILITY TO DEMETHYLATE DNA. BASED ON THAT, INDEPENDENTLY OF OUR WORK, THE GROUP AT USC ON EPIGENETICS WAS ALSO LOOKING AT GLIOMAS AN ASKING FOR GLIAL BLASTOMAS ACROSS THE BOARD TO DEFINE SUBGROUPS BASED ON METH LAYTOR PHENOTYPE, THEY DEFINE A GROUP, RED IS HEAVILY METHYLATED IN THIS GROUP AND YOU CAN SEE A GROUP HERE ON THIS SIDE THAT IS HEAVILY METHYLATED BY THEIR ANALYSIS COMPARED TO ALL OTHER GLIAL BLASTOMAS THAT MAKES UP 10% OF THE GROUP. WHEN YOU ASK BY GENE EXPRESSION PHENOTYPE THOSE ARE THE PRONEURAL TUMORS EXCLIEW COLLUSIVELY. THE ONES STUCK AT TRUE STEM CELL STATE OF GLIAL BLASTOMA, THEY'RE HYPERMETHYLATED AND WHEN ONE ANALYZES FOR GENOTYPE THEY'RE ALL MUTANT FOR IDH-1S AND THEY'RE ALL IN THIS GROUP. SO INDEPENDENT TEST IN SEPARATE TUMOR THE INTERFERENCE OF METABOLISM BY THIS 2 HG BLOCKS THE ABILITY TO ACCESS NEW GENES BY DEMETHYLATION. SO WE'RE STUCK WITH A MODEL HISTONE DEMETHYLATION REQUIRES ACTIVE INTERVENTION THROUGH CATABOLITE SUBSTRATE ALPHA KETO GLUTERATE TO BE PRIOR TO SUBSTRATE FOR EXCLUSIVELY SPECIFIC ENZYMES THAT DEMETHYLATE SITES TO ACTIVATE REPRESSED CHROMATIN TO ALLOW CELLS TO DIFFERENTIATE. WHEN THAT'S BLOCKED BY 2 HYDROXY GLUTERATE AS GLOBAL INHIBITOR, THE CELL IS STUCK IN MAINTAINING THE STEM CELL STATE IT HAS. SO AT THAT POINT WE WERE THINKING THIS IS INFORMATIVE IN TERMS OF A BALANCE EPIGENETICS IS CREATING THROUGH METABOLISM TO INSTRUCT WHAT PART AND HOW MUCH GENOME SHOULD WE ACCESS BASED ON THE NUTRIENT IT IS HOST IS MAKING AVAILABLE TO THE CELL TYPE. MUTATIONS IN THAT PATHWAY ONCOMETABOLITES BY SIGNAL TRANSDUCTION GENES OR METABOLISM INFORMS THIS PROCESS. SO THAT'S ALL I HAVE TO SAY. I APOLOGIZE FOR COUPLE OF MINUTES OVER BUT HAPPY TO TAKE QUESTIONS THAT I APPRECIATE YOUR ATTENTION. I GUESS I WAS GOING TO SHOW YOU IF YOU ACTUALLY DO THIS YOU BLOCK NEURAL DIFFERENTIATION AND THAT, I'M GOING TO SHOW THAT AS A FINAL POINT F. YOU TAKE PRIMARY PUBLISHED, IF YOU TAKE PRIMARY CELLS INTRODUCE AN IDH MUTATION AND ASK NEUROSPHERES TO DIFFERENTIATE PRIMARY CELL LOSE THEIR COMPLETIBILITY TO DIFFERENTIATE WHICH IS WHAT THAT LAST SLIDE DOES. OVEREXPRESSION OF WILD TYPE IDH HAS NO EFFECT ON THAT PROCESS. THIS SEEMS TO< CONNECTION BETWEEN METABOLISM AND ABILITY OF CELLS TO ENGAGE NEW PROGRAMS BY ACCESSING NEW DNA THROUGH METHYLATION. SO WITH THAT THANK YOU VERY MUCH, AND BE HAPPY TO TAKE ANY QUESTIONS. [APPLAUSE] SOMEBODY HAS TO BE BRAVE AND BE FIRST OR WE COULD ALL GO HOME. YEAH. (OFF MIC) >> GO UP TO THE MIC. THAT'S PEOPLE OUTSIDE IN ROOM. >> THANKS FOR A GREAT TALK. EARLIER TALKING CANCER TALKING ABOUT YOU HAVE WAY TO BLOCK DIFFERENTIATION OF -- YOU CAN DO SOMETHING WITH STEM CELL THERAPY BECAUSE WE KNOW LIKE KIND OF KNOWING YOU GET THE CELLS AND DIFFERENTIATE DOWN OTHER CELL TYPES. >> SO THE SURPRISE IS, WHICH DOESN'T REALLY ANSWER THE QUESTION, COULD WE USE THIS KIND OF INFORMATION TO MAINTAIN THEM CELLS OR DO SOMETHING WITH STEM CELLS, THAT'S THE QUESTION. THE ONLY THING WE CAN DO TO CELLS TO SHOW US THE TRUE CORE PHENOTYPE AT A STATE THEY WILL NEVER DIFFERENTIATE IS IF WE STARVE FOR TWO NUTRIENTS I JUST TOLD YOU ABOUT. STAR CELLS FOR GLUCOSE AND GLUTAMINE, THEY DON'T COMMIT SUICIDE THEY RETRACT TO MAINTAINING PURE PHENOTYPE. THOUGH WE KNOW THEY'RE CAPABLE OF DIFFERENTIATION WE CAN'T ADD A GROWTH FACTOR TO MAKE THEM DO IT. IN ABSENCE OF METABOLISM, IN THE ABSENCE OF TWO INTRACELLULAR POOLS OF METABOLISM WE CANNOT FORCE A CELL TO DIFFERENTIATE. AS IF A CELL NEEDS TO BE TOLD THROUGH MEDICAL PATHWAY IT'S TIME AND YOU HAVE THE RESOURCES TO TAKE THIS LEAP OF FAITH TO GO BE SOMETHING ELSE AND IT IS NEVER DONE UNDER A LOW ENERGY CONDITION. SO THE PROBLEM IS STARVING A CELL ISN'T A GOOD WAY TO MAINTAIN THINGS OR GET MORE STEM CELLS SO IT'S AN -- WE LIKE THE IDEA, WE DID IT BUT IT DIDN'T WORK. SO WE DO THINK WHAT THIS IS SAYING, I DIDN'T HAVE TIME THE TALK ABOUT IF YOU LOOK AT ANOTHER PART OF WHAT PETER LEER PUBLISHED, THERE IS ANOTHER GROUP OF CELLS THAT ARE STUCK IN THE STEM CELL STATE. THEY HAVEN'T BEEN TALKEDDED A LOT ABOUT. THEY'RE THE ONES THAT ARE COMPLETELY HYPOMETHYLATED. AT BOTH DNA AND HISTONE LEVEL. THAT REALLY MEANS, WHAT WE'RE TRYING TO PUSH AS AN IDEA TO THINK ABOUT IS THAT IF YOU'RE STUCK WITH ALL YOUR GENES EXCEPT CORE GENES THAT MAINTAIN WHAT YOU ARE, STUCK IN A HETERO CHROMATIN STATE AND YOU CAN'T ACCESS THEM YOU'RE STUCK WITH THE DIFFERENTIATION YOU HAVE. IF ON THE OPPOSITE SIDE EVERYTHING IS OPEN, STEM CELL MAINTENANCE GENES DOMINATE P AND CELL CAN'T DIFFERENTIATE UNDER THOSE CONDITIONS WHICH MEANS LIKE EVERYTHING ELSE THERE'S BARRIERS TO REGULATING DIFFERENTIATION. YOU HAVE TO SILENCE STEM CELL MAINTENANCE, NOT SURPRISING GIVEN IPS CELL BIOLOGY, SILENCE THE GENES THAT MAINTAIN STEMNESS BUT YOU HAVE TO GAIN ACCESS TO GENES THAT ALLOW YOU TO BE THE NEXT CELL IN LINEAGE, IT'S THE DISREGULATION OF THAT, IF YOU TAKE THAT MODEL YOU CAN SEE ALMOST EVERY CANCER TYPE HAS DISREGULATION IN ONE DIRECTION OR ANOTHER BY THIS. >> BEAUTIFUL ODYSSEY THROUGH CANCER METABALOMICS. THE QUESTION I HAD IS BECAUSE THESE PATHWAYS ARE FUNDAMENTAL, DOES THAT POSE A PROBLEM FOR ONCOLOGIST THINKING THERAPEUTIC INTERVENTION? BECAUSE YOU RISK DISRUPTING THINGS THAT AFFECT CELLS IN THE BODY. >> THERE IS A LOT OF WRONG ANSWERS TO MY QUESTION, CORE METABOLISM IS CORE METABOLISM, PROKARYOTES TO SINGLE CELL EUKARYOTS TO SINGLE CELL ORGANISMS WE HAVE METABOLISMS THAT INTERVENE WITH THAT. YOU MIGHT KILL THE CANCER CELL BUT YOU KILL NORMAL CELLS AT THE SAME TIME, MORE TOXICITY THAN TRADITIONAL CHEMOTHERAPEUTIC. THE FIRST WE PUBLISHED IN MULTIPLE FRMS, IF YOU ASK HOW TO TRADITIONAL CHEMOTHERAPEUTICS THAT CURE YOU OF CANCER, OR DOCTOR USES AND CURES SIGNIFICANT PATIENT TO PATIENT, ALMOST ALL ARE IN FACT INTERVENING IN METABOLIC PATHWAYS AS FUNDAMENTAL BASIS. SO ANTI-METABOLITES, METHYL METHYL METHOTREXATE, TRADITIONAL AL COLLATING AGENTS CONSUME ALL THE NAD. SO WE ACTUALLY THINK THAT OUR SUCCESS IN CANCER THEME THERAPY HAS NOT BEEN REALIZED, MOSTLY A METABOLIC STRATEGY AND TUMOR CELLS ONCE ADDICTED THROUGH ACTIVATION OF SIGNALING PATHWAYS TO BECOME REPROGRAMMED METABOLISM TO BE ADDICTED TO THESE METABOLITES, ARE FUNDAMENTALLY DIFFERENCE THAN NORMAL CELLS THAT WHEN THEY LOSE GLUCOSE AN DON'T HAVE A DRIVING MUTATION THROUGH PI-3 KINASE PATHWAY, THERE'S NO GLUCOSE BUT I CAN METABOLIZE FATTY ACIDS. WHEN AKT IS CONSTITUTIVELY ON THEY SAY YOU DON'T GET TO DO THAT WE NEED THEM FOR GROWTH SO YOUR SURVIVAL STRATEGY DOESN'T WORK HERE AND WE WENT ON TO DESCRIBE THE TUMOR SUPPRESSOR NETWORK NEEDED FOR REACTIVATION OF BETA OXIDATION. WHEN YOHa LOSE GLUCOSE. SO YOU CAN DO REAL FOR THE STUDENTS AND POST-DOCS STUPID EXPERIMENTS WHICH IS WHAT WE DO. TAKE OUT A SINGLE METABOLITE AND HOW DOES A TUMOR CELL VERSUS NORMAL ADAPT. THAT IS HOW WE CHARACTERIZE, HALF TUMOR SUPPRESSORS FIT INTO A MODEL OF ACCOMMODATING GLUTAMINE WITHDRAWAL OR GLUCOSE WITHDRAWAL. NORMAL CELLS ACTIVATE THOSE IMMEDIATELY AND ONCOGENIC LEAGUES PREVENT THE PATH WAYS OR LOSS OF THOSE GENES AS SENSORS LOSE THAT PATHWAY. SO I THINK WE CAN EXPLOIT IT. MAYBE THIS SHOULD BE THE FINAL THING SINCE I HAVE GONE OVER. I THINK THERE'S FUNDAMENTAL DIVIDE IN ONCOLOGY RIGHT NOW. WE ARE VERY IMPRESSED. I'M VERY IMPRESSED, ALL SHOULD BE IMPRESSED WE DISCOVERED ONCOGENIC DRIVER MUTATIONS IN HUMAN TUMORS. AND WE HAVE DEVELOPED THE ABILITY TO DEVELOP INHIBITORS OF DRIVER MUTATIONS THAT WILL TURN THOSE GENES OFF. BUT THE REAL FEATURE THOUGH TREMENDOUS SUCCESS IN GLEVAK TURNING THEM OFF HASN'T HELPED PATIENTS LONG. HASN'T CURE MANY PEEP. BECAUSE CANCER HAS AN ARMAMENTARIUM OF THINGS TO GET AROUND INHIBITION OF DRIVING ONCOGENE AND IT'S HARD PHARMACO LOGICALLY TO COMPLETELY INHIBIT AN ACTIVATED ONCOGENE, EVEN WHEN WE TRY TO DO THAT. IN CONTRAST ALL THERAPIES THAT CURE PATIENTS HAVE LEFT THE ONCOGENES IN PLACE ALLOW THEM TO FORCE THE CELL INTO DOING SOMETHING THAT A NORMAL CELL EQUIVALENT MIGHT NOT DO UNDER THOSE CONDITIONS AND MAKE THAT CONDITION HARMFUL FOR THE CELL. THE ATTEMPT TO TRY AND DIVIDE WHEN THERE ARE NO NUCLEOTIDES IS A REALLY BAD IDEA. THAT IS THE BASIS OF MOST METABOLITE IS THE ANTI--- THAT'S THE BASIS OF ME THEY WILL TREK SATE. SO WE HAVE FORGOTTEN THAT APPROACH AND WE THINK METABOLISM WILL REACTIVATE THAT APPROACH. THE UNDERSTANDING THAT CANCER PATHWAYS, THOUGH THEY DO OTHER THINGS, NOT TRYING FOR STUDENTS AND POST-DOCS TO NEGATE IMPORTANCE OF THE SIGNAL TRANSDUCTION PATHWAYS, GENES NECESSARY TO PROGRESS YOU THROUGH A CELL CYCLE CHECK POINTS WE TALKED ABOUT BUT THERE HAS TO BE FUEL TO DO THAT. SO ASK A CELL TO G THROUGH CELL DIVISION AND YOU SWEEP OUT A FUEL, IT'S IN MORE TROUBLE THAN A CELL HANGING OUT SAYING I'M SUPPOSED TO OCCUPY SPACE AND KEEP ALIVE. IT HAS MORE ADAPTIVE MECHANISMS SO THERE ARE GOING TO BE WAYS TO EXPLOIT IT BUT IT'S NOT BY ELIMINATING THE ONCOGENES BUT BY LEAVING THEM IN PLACE AND MAKEING WHAT THEY FORCE THE CELL TO DO TOXIC UNDER CHANGING METABOLIC CONDITIONS. THAT'S THE APPROACH WE'RE EXPLOITING RIGHT NOW. THANK YOU VERY MUCH FOR YOUR TIME AND ATTENTION. THANKS A LOT. [APPLAUSE] >> LET ME REMIND YOU OF A RECEPTION SPONSORED BY THE FAS IN THE LIBRARY AND AN OPPORTUNITY TO ASK DR. THOMPSON REALLY BURNING QUESTIONS THAT YOU HAVE.