A BRIEF BACKGROUND ON THE STAR SERIES. WHAT WE HAVE TRIED TO DO FOR 14 YEARS, I THINK, IS TO FEATURE EXTRAORDINARY CONTRIBUTORS OR, QUOTE, STARS IN THE FIELD OF CANCER AND NUTRITION RESEARCH, AND SOMETIMES THEY LEAN MORE TOWARD THE NUTRITION, SOMETIMES TOWARD THE CANCER PART, AND TODAY WE HAVE A SUPERSTAR, SUPERNOVA, WHO WILL COVER ALL BASES ALL AT THE SAME TIME. THE SPEAKERS HIGHLIGHT THE IMPORTANCE OF THE ROLE NUTRITION PLAYS IN MODIFYING CANCER DEVELOPMENT, ALL KINDS OF PREVENTION DURING TREATMENT, ET CETERA. THE LECTURE IS AIMED TO FACILITATE INTERDISCIPLINARY INTERACTIONS AMONG BASIC SCIENTISTS, CLINICIANS, BEHAVIORAL AND SOCIAL SCIENTISTS, WHICH IS WHY WE HAVE IT HERE AT THE LIPSETT AUDITORIUM, SO WE CAN GET PEOPLE FROM NIH PROPER AND NOT JUST HOLD IT UP JUST FOR NCI IN SHADY GROVE. WE HAVE LECTURES EACH FALL AND EACH SPRING. ABOUT THE PRACTICUM, WELL, THAT'S NOT IMPORTANT. OH, OKAY. THE LAST POINT IS WORTH REPEATING. THE GOAL OF THE PRACTICUM IS PROMOTE UNDERSTANDING OF THE COMPLEX RELATIONSHIP BETWEEN DIET, CANCER PREVENTION AND ENCOURAGE INDIVIDUALS TO ACTIVELY PARTICIPATE IN RESEARCH THAT WILL HELP CLARIFY THE SPECIFIC ROLE OF FOODS AS MODIFIERS OF CANCER RISK. THAT'S PRIMARILY BECAUSE THAT'S THE HARDEST THING TO STUDY, SHOW AND EVEN THOUGH SOME PEOPLE ACT LIKE RATS, THEY REALLY AREN'T AND YOU CAN'T KEEP THEM IN A CAGE AND CONTROL EXACTLY WHAT THEY EAT. SOME MORE THAN OTHERS, OKAY. I'M INTRODUCING JOHANNA LAMPE, A FULL MEMBER AND ASSOCIATE DIVISION DIRECTOR AT PUBLIC HEALTH SCIENCES DIVISION AT FRED HUTCHINSON CANCER CENTER IN SEATTLE, ALSO A RESEARCH PROFESSOR IN THE DEPARTMENT OF EPIDEMIOLOGY AT THE UNIVERSITY OF WASHINGTON. SHE RECEIVED HER PH.D. IN NUTRITIONAL SCIENCES WITH A MINOR IN BIOCHEMISTRY AT THE UNIVERSITY OF MINNESOTA, WHERE SHE TRAINED AS A POSTDOCTORAL FELLOW IN EPIDEMIOLOGY BEFORE JOINING FRED HUTCHINSON. HER RESEARCH INTERESTS RELATE TO THE MECHANISMS BY WHICH COMPONENTS OF DIET, PARTICULARLY CONSTITUENTS OF PLANT FOOD, ALTER SUSCEPTIBILITY TO AND RISK OF CANCER. SHE FOCUSES ON CONTROLLED DIETARY INTERVENTIONS IN HUMANS, TO DETERMINE HOW INDIVIDUAL VARIATION IN BIOCHEMICAL RESPONSE, PARTLY DUE TO NATURE, PARTLY DUE TO NURTURE, TO HIGH PLANT FOOD DIETS MAY EX-MEANS THE DIFFERENCES IN DISEASE RISK. ALSO STUDY IT IS EFFECTIVE DIET ON THE GUT MICROBIAL COMMUNITY AND THE RELATIONSHIP OF THE GUT MICROBIOME TO BIOMARKERS OF CANCER RISK. NOW, WITH NO FURTHER ADO, JOHANNA LAMPE. [APPLAUSE] >> THANK YOU, MUCH, HAROLD, AND IT REALLY IS A PLEASURE TO BE HERE AND A GREAT HONOR TO HAVE AN OPPORTUNITY TO TALK ABOUT WHAT'S NOW BECOME SEVERAL DECADES WORTH OF WORK IN THE AREA OF DIET AND CANCER PREVENTION, SO I THOUGHT PARTICULARLY FOR PARTICIPANTS IN THE PRACTICUM, GIVE YOU SOME IDEA OF HOW A TRAJECTORY IN CANCER PREVENTION RESEARCH MIGHT GO. SOME ARE MORE SIR CUEDUCE OR WINDY TRAJECTORIES THAN OTHERS AND I THINK MINE IS ONE OF THOSE, BUT GIVE YOU A CHANCE TO SEE HOW THAT WORKS OUT, SO AS FAR AS THE PLAN FOR THIS AFTERNOON, A NUMBER OF OBJECTIVES AND THEREFORE AN OUTLINE WHICH I REALIZED WASN'T ALIGNED, BUT WAS A NUMBER OF INTERLOCKING CIRCLES, FOCUSING ON MECHANISMS BY WHICH DIET AND PARTICULAR THE CONSTITUENTS OF DIET AFFECT CANCER RISK, THE IMPACT -- SORRY -- IMPACT OF BOTH HOST GENETICS AS WELL AS MICROBIAL AGAIN NETS ON RESPONSE TO DIET AND HOW DOES THIS AFFECT OUR APPROACHES TO PREVENTION AND RECOMMENDATIONS FOR PREVENTION, AND THEN MOST OF MY WORK HAS BEEN CONDUCTED IN HUMANS OVER THE YEARS, AND HAVE REALLY RELIED ON TO THE EXTENT ONE CAN CONTROL HUMANS AS MUCH AS ONE CAN CONTROL RATS, WE HAVE ATTEMPTED, IN ORDER TO STUDY CANCER PREVENTION RESEARCH. JUST THINKING ABOUT THE CONTINUUM OF RESEARCH APPROACHES FOR STUDYING DIET AND CANCER, CERTAINLY RUNS ALWAYS WAY FROM THE CELLULAR LEVEL, ALL THE WAY TO LARGE-SCALE POPULATION-BASED COHORTS, WHERE YOU ARE FOLLOWING HUNDREDS OF THOUSANDS OF INDIVIDUALS OVER TIME, AND REALLY ACROSS THIS, WE HAVE A NUMBER OF DIFFERENT APPROACHES. I WOULD LIKE TO THINK ABOUT WHAT ARE THE KINDS OF THINGS WE CAN DO WITH THESE DIFFERENT TYPES OF STUDIES, AND CERTAINLY, IN RELATION TO THE CELLULAR-BASED WORK, AS WELL AS ANIMAL MODELS, TO A CERTAIN EXTENT IN HUMAN STUDIES, WHICH WE ARE CALLING SMALL-SCALE CONTROLLED INTERVENTIONS OR HUMAN MECHANISTIC STUDIES, ONE CAN FOCUS ON MECHANISMS OF ACTION, WE CAN TEST SPECIFIC NUTRIENTS OR COMPOUNDS THAT ARE PART OF THE DIET, AND WE CAN EITHER CONTROL FOR GENETIC VARIATION; I.E., IN THE CONTEXT OF AN ANIMAL STUDY, WHERE YOU EVER ALL THE SAME GENETIC BACKGROUND ON WHICH YOU ARE STUD DIRTING THIS, OR AS WE ARE SEEING MORE STUDIES COME OUT WITH USING DIFFERENT STRAINS OF THE ANIMALS, FOR EXAMPLE, IN ORDER TO CONTRAST DIFFERENT GENETIC BACKGROUND, AND SIMILARLY, AS I WILL POINT OUT WITH SOME OF OUR STUDIES, WHERE WE CAN RECRUIT PARTICIPANTS ON THEIR GENETIC BACKGROUNDS. IN CONTRAST, WHEN WE THINK ABOUT SOME OF THE MORE LARGE AND COMPLEX TYPES OF STUDIES, WE ARE REALLY LOOKING AT SYSTEMIC EFFECTS AND THIS IS THE CASE FOR ANIMAL MODELS ALSO, WHERE YOU HAVE TO TAKE INTO CONSIDERATION THE WHOLE ANIMAL, CERTAINLY AT THE LEVEL OF POPULATION, THAT'S ALSO THE CASE, AND REALLY, AS YOU MOVE INTO RANDOMIZED CLINICAL TRIALS, EITHER INDIVIDUALS OR COMMUNITIES OR IN THE POPULATION-BASED STUDIES, YOU ARE TYPICALLY DEALING SPECIFICALLY WITH FOOD OR AN OVERALL DIETARY PATTERN IN THAT IT'S VERY HARD TO CONTROL DIET IN THOSE LARGE-SCALE STUDIES. SO KEEPING THIS IN MIND, YOU CAN THINK OF THE VARIOUS WAYS THAT ONE CAN WORK BACK AND FORTH ACROSS THIS CONTINUUM, AS FAR AS STUDIES IN EPIDEMIOLOGY, IN FORMING POSSIBLE SMALL-SCALE HUMAN INTERVENTIONS, AS WELL AS TAKING THINGS TO AN ANIMAL MODEL OR WORKING FROM ANIMAL MODELS BACK AND FROTH. REALLY, I HAVE COME TO FIND WORKING IN THIS AREA HERE, THE HUMAN MECHANISTIC STUDIES, WE DRAW HEAVILY ON THE WORK IN THE PRECLINICAL ANIMAL MODELS, AS WELL AS WORKING BACK FROM THE EPIDEMIOLOGIC STUDIES, SO IT BRIDGES THAT GAP. THINKING ABOUT THE DIFFERENT COMPONENTS WE WORRY ABOUT IN RELATION TO POTENTIAL EFFECTIVE FOOD, NUTRITION, OBESITY AND PHYSICAL ACTIVITY ON CANCER RISK R THIS HAS BEEN A LONG-STANDING FIGURE THAT CAME OUT OF THE LAST WCRF ARCIR EXPERT REPORT, AND I PUT IT UP AS A REMINDER TO MYSELF, BUT ALSO THINK IT'S ALWAYS SOMETHING GOOD TO KEEP IN MIND, PARTICULARLY WHEN WE THINK ABOUT CANCER IN A WHOLE PERSON, A WHOLE ANIMAL, WE NOT ONLY HAVE THINGS THAT ARE HAPPENING AT THE CELLULAR LEVEL TO A CELL THAT COULD BECOME A CANCER CELL OR IS A DEVELOPING TUMOR, BUT WE ALSO HAVE THINGS THAT ARE SYSTEMIC THAT INFLUENCE RISK OF DISEASE; I.E., HORMONE REGULATION, OVERALL INFLAMMATION AND IMMUNE FUNCTION, THAT ALL PLAY A ROLE IN THE WHOLE ANIMAL, IN THE WHALE PERSON IN RELATION TO RISK. IF WE THINK ABOUT WHERE OUR OPPORTUNITIES FOR PREVENTION, AS FAR AS INTERVENING, CERTAINLY IT IS ALONG THE WHOLE CANCER PROCESS, EITHER FROM THE STANDPOINT OF THINKING ABOUT EXPOSURE TO POTENTIAL CAR SIN JENS, WE ARE ENCOURAGED TO AVOID EXPOSURE TO THOSE THINGS, WHETHER IT BE UV LIGHT, WHETHER IT BE WELL-COOKED MEATS, FROM THE STANDPOINT MANY OF THESE COMPOUNDS, ALTHOUGH AS THEY EXIST IN NATURE ARE NOT NECESSARILY CARSONGENIC ARE ACTIVATED THROUGH ENZYMATIC ACTIVITY TO PRODUCE COMPOUNDS THAT INTERACT WITH NORMAL CELLS TO INITIATE THE CARSON GENESIS PRO-CYST. FROM THE STANDPOINT OF THINKING ABOUT THESE OPPORTUNITIES FOR PREVENTION AND THE TIME WE ARE LOOKING AT -- THESE ARE MERE ESTIMATES, BUT THE OPPORTUNITIES OVER YEARS TO MINIMIZE THE TRANSITION FROM A CELL WITH DNA DAMAGE THROUGH TO PRODUCTION OF PRE-NEOPLASTIC CELLS AND ULTIMATELY PROGRESSION TO TUMOR, THERE ARE A LOT OF OPPORTUNITIES TO INTERACT WITH THIS AND IN THAT CONTEXT, TO THINK ABOUT MESSAGES IN RELATION TO CANCER PREVENTION AND APPROACHES THERE. SOME OF OUR FIRST WORK WE REALLY FOCUSED ON THIS COMPONENT, HOW DO YOU TRY TO MINIMIZE EXPOSURE TO COMPOUNDS THAT ARE TAKEN UP IN THE BODY AND CAN BECOME COURSEGENIC? HOW DO YOU GET RID OF THEM QUICKLY AND LOWER YOUR RISK FOR CANCER THAT WAY? SO DON'T NEED TO TELL YOU GUYS, THE HUMAN DIET IS CERTAINLY A VERY COMPLEX EXPOSURE, WE ARE NOT ONLY TALKING ABOUT THOUSANDS OF COMPOUNDS, BUT THE COMPLEXITY OF THE MIXTURE OF FOODS, AS THEY EXIST, AS WELL AS POTENTIAL SYNERGISTIC EFFECTS OF COMPONENTS OF PARTICULAR FOODS OR EVEN ACROSS DIFFERENT TYPES OF FOODS, SO REALLY THE TOTALITY OF DIET. IN THAT REGARD, WE HAVE A NUMBER OF DIFFERENT WAYS WE GO ABOUT EVALUATING EXPOSURE TO DIET, AND THAT COULD BE AT THE LEVEL OF INDIVIDUAL COMPOUNDS, INDIVIDUAL FOODS OR CLASSES OF FOODS, CERTAINLY YOU HAVE SEEN ALL THESE DIFFERENT TYPES OF STUDIES, AND REALLY MORE RECENTLY, COMING TO RECOGNIZE THAT WE DON'T ONLY EAT INDIVIDUAL FOODS, BUT WE REALLY CONSUME DIETARY PATTERNS THAT WE ARE GETTING LOT MORE RESEARCH AT THE LEVEL OF DIETARY PATTERNS TO BETTER UNDERSTAND WHAT THE IMPACT OF THE TOTALITY OF DIET IS. THAT SAID, WE HAVE IDENTIFIED A NUMBER OF DIFFERENT COMPOUNDS THAT HAVE BEEN ASSOCIATED WITH LOWER CANCER RISK. THESE JUST A FEW IN THE LAUNDRY LIST OF PHYTOCHEMICALS THAT A LOT OF THE WORK WAS DONE AT NCI LOOKING AT SOME OF THESE, IN RELATION TO THEIR POTENTIAL CHEMOPROTECTIVE EFFECT. I WILL TALK A LITTLE ABOUT THE VEGETABLES AND THE LIGHT DEMANDS IN HIGH-FIBER FOODS. ONE OF THE THINGS I THINK IS IMPORTANT TO KEEP IN MIND WHEN YOU START TO THINK ABOUT MECHANISMS BY WHICH VARIOUS PHYTOCHEMICALS ACT IS THAT EACH OF THESE, AS SING COMPOUNDS, CAN ACTUALLY INTERACT WITH A NUMBER OF DIFFERENT SIGNALING MECHANISMS, SO THEY TENT HAVE ONE TARGET RECEPTOR OR ONE TARGET PATHWAY THEY AFFECT, BUT THEY COULD BE EFFECTIVE A PANOPLY OF PATHWAYS, WHETHER IT BE INHIBITING THE NF KAPPA B MEDIATED INFLAMMATORY PATHWAY, ACTING AS AGONISTIC LIGANDS, RESULTING IN UP-REGULATION OF A NUMBER OF DETOXIFICATION OR ANTIOXIDANT ENZYMES, INHIBITING THE IGF1, IGF1 RECEPTOR PATHWAY THROUGH MTOR AND RAPTR. IN SOME CASES, THIS FIGURE IS DESIGNED TO FOCUS ON THE LIGNANDS, THEREFORE, AFFECT THE ESTI-END RECEPTOR PATHWAY AND ALSO INTERACTING AS LIGANDS FOR PPAR ALPHA AND GAMMA, INHIBITING THE LXR, SO LOOKING AT THE LAUNDRY LIST OF VARIOUS DOWNSTREAM MECHANISMS, YOU CAN SEE WHY THE OPPORTUNITY TO HAVE AN IMPACT ON A WHOLE HOST OF IMPORTANT MECHANISMS FOR CANCER POPS OUT IN THE CONTEXT OF THESE COMPOUNDS. THE OTHER THING WE NEED TO KEEP IN MIND, AND THIS WILL COME UP ACROSS SEVERAL EXAMPLES I GIVE THIS AFTERNOON, IS THE VARIOUS SOURCES VARIATION RELATED TO PHYTOCHEMICAL EXPOSURE. PART OF THIS IS OCCURRING AT THE LEVEL OF THE INTESTINE; METABOLISM TAKING PLACE IN THE EPITHELIUM IN THE SMALL INTESTINE OR FOR COMPOUNDS NOT ABSORBED, POTENTIAL FOR UNDERGOING A WHOLE HOST OF GUT MICROBIAL REACTIONS, PRODUCING COMPOUNDS WE NEVER SEE IN NATURE, BUT ONLY PRODUCED AS A RESULT OF MICROBIAL ACTIVITY, THEN FOLLOWING UP BY FURTHER METABOLISM THERE. THIS ALSO GOES ON TO AFFECT DIFFERENCES IN LIVER METABOLISM, BOTH PHASE I ACTIVATING AND PHASE 2 CONGREGATING ENZYME SYSTEMS, POTENTIAL FOR DIFFERENCES, IN THIS CASE, LOOKING AT HOST GENETIC DIFFERENCES, IN THESE METABOLIC PATHWAYS OR WITHIN THE TISSUES OF INTEREST FROM THE STANDPOINT OF PARTICULAR CANCERS, INTERACTIONS WITH DIFFERENT RECEPTORS AND DEPENDING ON GENETIC VARIATION IN THOSE, OR SIGNAL TRANSDUCTION PATHWAYS. THEN ALSO, IMPACT ON EXCRETION AND RELEASED BACK INTO THE COLON. EXCUSE ME. ALL THESE POINTS COULD AFFECT EXPOSURE TO THESE COMPOUNDS. BY WAY OF EXAMPLES TODAY, I WILL FOCUS BOTH ON HOST GENETICS IN THESE METABOLIC PATHWAYS, AS WELL AS THE GUT MICROBIAL METABOLISM. SORRY ABOUT MY VOICE. WE HAVE HAD A NUMBER OF QUESTIONS THAT HAVE BEEN PART OF OUR RESEARCH PROGRAM FOR A NUMBER OF YEARS NOW, AND REALLY, THIS STEMMED FROM MY GRADUATE WORK, WHERE I WAS IN CHARGE OF RUNNING SEVERAL CONTROLLED FEEDING STUDIES, AND WE'D ALWAYS FIND YOU PUT EVERYBODY ON THE SAME DIET, AND DESPITE THAT, YOU WOULD HAVE A WHOLE HOST OF RESPONSES OVER A WIDE RANGE, AND WE STARTED TO ASK, WHAT IS CONTRIBUTING TO THESE DIFFERENCES, AND SO FOCUSING ON BOTH THE PHYTOCHEMICALS AND THEIR MECHANISMS OF ACTION, RECOGNIZING THE MULTIPLE PATHWAYS THAT ARE AFFECTED AND THE IMPACT OF THE HOST OF GENETICS. WE ARE FORTUNATE TO HAVE A FACILITY FOR CONDUCTING THESE TYPES OF STUDIES, CALLED THE PREVENTION CENTER. IT INCLUDES A RESEARCH CLINIC, AN EXERCISE FACILITY, WHERE WE CAN DO EXERCISE INTERVENTIONS AND A HUMAN NUTRITION LAB TEAR, WHICH CONSISTS OF WHAT LOOKS LIKE A COMMERCIAL KITCHEN, LOTS OF STAINLESS STEEL SURFACES AND THE PUBLIC HEALTH DEPARTMENT COMES THROUGH ROUTINELY TO CHECK ON US, AND THEN WE HAVE A DINING AREA, WHERE THE PARTICIPANTS CAN COME TO EAT, SO WE CAN PREPARE ALL THE'S FOR OUR PARTICIPANTS, AND THEN THEY COULD COME DINE IN THE PRIVATE SIT OF OUR SPACE HERE. PEOPLE DON'T HAVE TO LOOK AT THEM AND SAY WHAT ARE YOU EATING, SO IT MAKES FOR A SAFER ENVIRONMENT FOR THEM. ONE OF THE FIRST AREAS WE FOCUSED ON, VEGETABLES. BASED ON THE LITERATURE, WE HAD GOOD EVIDENCE FROM EPIDEMIOLOGIC LITERATURE THAT INDIVIDUALLY INTAKE HAS BEEN ASSOCIATED WITH LOWER RISK OF A NUMBER OF CANCERS AND RELATIVELY CONSISTENTLY FOR A NUMBER OF YEARS. FURTHER, SOME OF THE COMPOUNDS IN CAR CIPHERROUS VEGETABLES, HAVE BEEN SHOWN TO PREVENT CANCER IN ANIMAL MODEL, DECREASE INFLAMMATION AND OX DATESSIVE STRESS AND A NUMBER OF OTHER MECHANISMS THAT ARE RELEVANT TO THE CANCER PROCESS. FIRST OF OUR STAWED DIRTS WE EMBARKED ON WAS CALLED THE ENZYME ACTIVATION STUDY, AND THAT WAS BASED ON THE PREMISE THAT THESE COMPOUNDS SEEMED TO UP-REGULATE ENZYMES INVOLVED IN CARCINOGEN METABOLISM AND THE GOAL WAS TO SEE WHETHER OR NOT WE COULD DETECT THE EVENTS IN HUMANS. WE RECRUITED OR PARTICIPANTS, BOTH MEN AND WOMEN BETWEEN THE AGES OF 20 TO 45, ON THE BASIS OF GENETIC POLYMORPHISM? THE M1 GENE, AND THIS HAD BEEN AT THE TIME IDENTIFIED AS ONE OF THE GST'S INVOLVED IN METABOLISM, AND THEREFORE, THERE WAS SOME HYPOTHESIS THERE WOULD BE A DIFFERENTIAL EXPOSURE. SO WE SCREENED A NUMBER OF INDIVIDUALS IN ORDER TO GET A BALANCE OF GSTM NULL AND 1 POSITIVE. THE M1 NULL HAVE A TRUNCATED GENE, SO THERE'S NO EXPRESSION OF THE GSTM1 ENZYME. WE PUT PEOPLE ON TO FOUR DIET TREATMENTS IN A RANDOMIZED CROSSOVER DESIGN IN THIS APPROACH. EVERYBODY GETS ALL DIETS, AND SO THEY ARE ON FOR EACH PERIOD WITH SEVEN DAYS, WITH WASHOUT PERIOD OF AT LEAST TWO WEEKS IN BETWEEN, SO THEY RECEIVED ALL OF THESE DIETS IN RANDOM ORDER, RANDOM ASSIGNED ORDER, INCLUDED A BASAL DIET, LOW IN DIETARY FIBER, LOW IN PHYTOCHEMICALS, NO HERBS AND SPICES, ANYTHING THAT MIGHT BE A SOURCE OF INCREASED ACTIVITY, THEN WE ADDED VEGGIES ON TOP OF THAT, ONION FAMILY VEGETABLES, THE CARROT FAMILY. IT INCLUDED CARROTS, DILL, PARSLEY, ONIONS, CHIVES, LEEKS AND GARLIC, AND THE BRASSICA WAS BROCCOLI, CAULIFLOWER, RADISH śWE WERE INTERESTED IN WHAT KIND OF CHANGES WE COULD SEE IN A NUMBER OF DIFFERENT ENZYMES, BOTH PHASE 2, AS WELL AS THE PHASE I. IN THIS CASE, YOU CAN MEASURE THE ENZYME ITSELF, WITH AN ALIZA. WE RELIED ON AN ACTIVITY ASSAY IN WHITE BLOOD CELLS, AND FOR NAT2, WE USED CAFFEINE DOSE THAT WE THEN LOOKED AT PRODUCTION OF CAFFEINE METABOLITES, WHICH THE CONVERSION OF CAFFEINE TO ITS METABOLITES IS CARRIED OUT BY ITS ENZYMES. FOR THOSE OF YOU WONDERING WHAT THE BASAL DIET LOOKED LIKE, THESE WERE THE FOODS ON IT. YOU PUT IT ON A WHITE PLACE, YOU CAN'T SEE IT, AND SOME OF OUR STUDY PARTICIPANTS FONDLY CALLED IT THE BASAL BORING DIET. THE THING I ALWAYS FOUND AMAZING WAS SOME PEOPLE FOUND THIS WAS THE BEST DIET AND THEY DIDN'T LIKE IT WHEN WE STARTED PUTTING VEGETABLES ON TON OF IT, BUT IT REALLY -- WHAT IT DID WAS TO STANDARDIZE THE BASE DIET, SO WE WERE REALLY COMPARING ALL ASPECTS OF THE DIET ACROSS INDIVIDUALS. THIS IS LOOKING AT BOTH IN THE GSTM 1 POSITIVE, PEOPLE WHO HAVE THIS PRODUCTION OF THIS ENZYME AND THOSE WHO DON'T, AND REALLY LOOKING AGAINST USING THE BASAL DIET AS A REFERENCE, LOOKING AT CHANGES IN -- WITH THE CRUCIFEROUS, ALIUM AND THE ONLY EFFECT WE SAW WAS INCREASE AMONG THE GSTM1 NULL INDIVIDUALS, WHERE WE SEE INCREASE IN PRESENCE OF GST ALPHA. SIMILARLY, WE SEE INCREASE IN -- WITH THE ALIUM VEGETABLES ON GSTM1, AND AS YOU CAN SEE HERE, THOSE INDIVIDUALS WHO ARE NULL FOR THE GENE ARE ALSO NULL FOR ANY ACTIVITY, COMPARED TO THOSE WHO ARE THE GSTM1 PLUS, AND BOTH OF THESE WERE SIGNIFICANTLY HIGHER. INTERESTINGLY, IT WAS FUNNY, BECAUSE WHEN WE DID THE STUDY, WE SORT OF THREW THE VEGETABLES THERE ON THE SIDE, BECAUSE PEOPLE EAT THEM, LIKE CARROTS AND CELERY AND WEREN'T NECESSARILY EXPECTING TO SEE ANYTHING, BUT BESIDES SEEING AN INCREASE IN -- WE SAW A DECREASE WITH THE AACIOUS VEGETABLES, WHICH WAS RATHER UNUSUAL, BECAUSE THERE HADN'T BEEN MANY COMPOUNDS FOUND TO INHIBIT ZIP 1A2. WE FOLLOWED UP THE WORK WITH DAVE EATEN AT THE UNIVERSITY OF WASHINGTON, AND IDENTIFIED THE IMPACT OF SORALINS AS BEING THE CONTRIBUTE ORS TO REDUCTION IN ACTIVITY, SO OF COURSE YOU FINISH ONE STUDY AND THINK I STILL HAVE MORE QUESTIONS AND WE NEED TO TRY THIS A DIFFERENT WAY. AND PART OF THE PROBLEM WITH OUR PREVIOUS STUDY WAS BOTH MEN AND WOMEN ALL GOT THE SAME AMOUNT OF VEGETABLES, SO WE HAD SEEN SEX DIFFERENCES IN RESPONSE TO THE TREATMENTS AND WE WEREN'T SURE IF IT WAS BECAUSE PER BODY WEIGHT DOUGH, PEOPLE WERE GETTING DIFFERENT AMOUNTS AND THAT WAS CONTRIBUTES, OR IF THERE WAS ACTUALLY A TRUE SEX DIFFERENCE. SO WE FOLLOWED THIS UP WITH ANOTHER STUDY, AGAIN, RECRUITING ON THE BASIS OF GSTM1, BUT ALSO ON THE BASIS OF GSTT1, IN WHICH THERE'S A SIMILAR POLYMORPHISM WHICH RESULTS IN NO PRODUCTION OF A GSTT1 ENZYME, AND CONSEQUENTLY, WE HAD A SOMEWHAT MORE COMPLICATED DESIGN, AND MY STUDY COORDINATORS WERE NOT HAPPY WITH THE IDEA WE WERE GOING TO HAVE TO SCREEN HUNDREDS OF MORE PEOPLE IN ORDER TO GET ENOUGH OF THESE COMBINATIONS OF THE LESS COMMON VARIANTS OF THE M1 NULL AND T1 NULL, SO WE ENDED UP WITH THREE GROUPS, PLUS PLUSES, M1 NULL PLUSES AND M1 NULL, GST 1 NULL. WE RAN THE STUDY FOR FOUR 14-DAY PERIODS, AND COLLECTED BLOOD SAMPLES AND URINES FOR THE CAFFEINE TEST. IN THIS CASE, WE DECIDED TO FEED AS GRAMS PER KILOGRAM BODY WEIGHT FOR VEGETABLES. YOU CAN RECOGNIZE THIS IS NOT YOUR GRANDMOTHER'S RECOMMENDATION FOR A DOSE OF VEGETABLES PER DAY, ESPECIALLY THE TWO TIMES, WHICH WAS ABOUT TWICE WHAT WE HAD FED ON WHAT IS ALSO HERE THE 1X, WHICH WAS 7 GRAMS, 14 GRAMS PER KILOGRAM BODY WEIGHT FOR THE TWO, AND A COMBINATION. OUR HYPOTHESIS WAS WELL, IF CRUCIFEROUS MAKED ZIP 1A2 GO UP, WHAT HAPPENS WHEN YOU EAT THE TWO TOGETHER? DOES IT LOOK AT THOSE YOU ARE NOT EATING ANY VEGGIES AT ALL FROM THE STANDPOINT OF IMPACT ON ZIP 1A2, SO THAT WAS THE COMBINATION. LOOKING AT THE EFFECTS ON GST ALPHA, THE GSTM1 NULL US AND COMBINATION OF THE T1 NULLS HAD THE GREATEST RESPONSE FARCE INCREASES IN GST ALPHA, MODEST EFFECT IN THIS GROUP HERE, REALLY NOT MUCH GOING ON IN THE MIDDLE ONES. IN THE CASE OF ZIP 1A2, IT DIDN'T MATTER WHAT YOUR GENOTYPE WAS. YOU SEE INCREASE AND PRETTY MUCH DOSE RESPONSE INCREASE IN ZIP 1A2 ACTIVITY, WITH BOTH DOSES OF THE CRUCIFEROUS DIETS. YOU DID SEE EQUIVALENT OF NO EFFECT WITH THE COMBINATION OF THE APYACIOUS AND THE 1X CRUCIFEROUS, SO LOOKS LIKE YOU ARE ON THE BASAL DIET. SLIGHT DECREASE HERE, BUT NOT SIGNIFICANT. THE ONE THING THAT CAME OUT OF THE STUDY, WHEN WE LOOKED AT URINARY EXCRETION AS A MEASURE FOR EXPOSURE TO THE BIOACTIVE COMPOUNDS IN CRUCIFEROUS VEGETABLES IS OVER THIS DOSE RANGE OF GRAMS PER DAY OF CRUCIFEROUS, YOU SEE FOR EACH SPECIFIC DOSE, A WIDE RANGE IN 24-HOUR URINARY EXCRETION OF ITC'S, AND THIS GETS MORE PRONOUNCED AS YOU GO INTO THE HIGHER LEVELS, SO SOME PEOPLE ARE UP TO A KILO OF VEGGIES DAY, WITH LITTLE EXCRETION OF ISOTHIGH OWE CRYATE. JUST BY WAY OF EXPLANATION FOR POSSIBLE HYPOTHESIS HERE IS THESE COMPOUNDS EXIST AS GLUCCO SINLATES IN THE PLANTS, AND WITHIN CRUCIFEROUS VEGETABLES, THERE IS ENZYMES THAT ALLOW FOR CLEAVAGE OF GLOW COURSE FROM WHEN IT CONS INTO CONTACT WITH IT. RELEASING WHAT IS CONSIDERED TO BE THE BIOACTIVE COMPONENT. WHEN YOU THINK ABOUT HOW MOST OF US EAT CRUCIFEROUS VEGETABLES, WITH TEND TOP COOK THEM, WHICH WOULD IMPEDE CONVERSION OF THE GLUCOSINLATE. FORTUNATELY, OUR GUT BACTERIA, SEVERAL OF THEM HAVE SIMILAR ENZYME, THAT ALLOWS FOR THIS CONVERSION, SO WE FOLLOWED UP OUR PREVIOUS FEEDING STUDY, THEN, WITH AN ATTEMPT TO TRY TO UNDERSTAND IF THERE WERE DIFFERENCES IN THE AMOUNT OF ACTIVITY THAT VARIOUS INDIVIDUALS HAD IN THEIR CAPACITY TO RELEASE THE ISOTHIGH OWE KINDLATE. WE RECITED A BUNCH OF PEOPLE, 200 GRAMS OF BROCCOLI, THEN WE RECOVERED THE URINE FOR 24 HOURS AFTER, AND YOU CAN SEE RANGE OF ANYWHERE FROM ABOUT 1% TO 28% OF THE DOSE BEING RECOVERED IN URINE. SO RELATIVELY MODEST RECOVERY, AS FAR AS COMPARED TO WHAT WAS ADMINISTERED, SO WE TOOK THE INDIVIDUALS WHO WERE DOWN IN THIS RANGE OF THE LOW EXCRETERS AND THOSE THAT WERE IN THE HIGHER EXCRETER RANGE, AND WITH A GLUCOCINELATE. WHAT WE FOUND WAS THE INDIVIDUALS WHO WERE PRODUCING MORE IN THEIR URINE, ALSO TENDED TO HAVE GUT BACTERIAL POPULATION THAT WAS CAPABLE OF DEGRADING IT MORE RAPIDLY, WHICH IT WOULD BE ABSORBED AND THEN ULTIMATELY SHOW UP IN THE URINE, BUT THIS REALLY ALSO REMIND ME THAT WE HAVE TO KEEP IN MIND THE VARIATION AND POTENTIAL RESPONSE OR BOTH EXPOSURE TO THESE COMPOUNDS. ANOTHER GROUP OF COMPOUNDS THAT WE HAVE BEEN WORKING ON OVER THE YEARS ARE LIGNANDS. THEY EXIST AS A WIDE VARIETY OF PLANTS AND SOME FOODS TEND TO BE HIGHER. FLAX SEED, FRUITS AND VEGETABLES. WE HAVE A WIDE RANGE OF DIETARY SOURCES OF THESE COMPOUNDS, SO EXPOSURE COULD BE RELATIVELY HIGH. THESE ARE METABOLIZED BY GUTS BACTERIA TO TWO END PRODUCTS. THIS REQUIRES A CONSORTIUM OF BACTERIA TO CARRY OUT THE THERE NEEDS TO BE CLEAVAGE OF TWO GLUCOSE, AND THEN FURTHER DEHIDE BOXLATION, WHICH IS CONSIDERED TO BE THE BIOACTIVE COMPONENT, AS FAR AS AMONG ALL OF THE VARIOUS INTERMEDIATES, APPEARS TO BE MOST BIOACTIVE, BUT THERE HAVE BEEN FEW BACTERIA FOUND THAT COULD ACTUALLY CARRY OUT THE WHOLE REACTION ALL BY THEMSELVES, AND IT REALLY IS THE IDEA THAT ONE BACTERIAL SPECIES OR STRAIN WILL CARRY OUT ONE PART OF THE REACTION, HAND OFF THE INTERMEDIATE TO SOMEONE ELSE THAT CARRIES IT ON THROUGH. FROM THE LITERATURE, WE HAVE A NUMBER OF STUDIES FROM DIFFERENT INDIVIDUALS AND DIFFERENT TYPES OF STUDIES THAT SUPPORT A REDUCED RISK OF BOTH COLON AND BREAST CANCER, AS WELL AS REDUCED RISK OF CARDIOVASCULAR DISEASE WITH EXPOSURE TO THESE COMPOUNDS, ALSO A NUMBER OF EXPERIMENTAL HUMAN STUDIES SHOWING CHANGES INEST GENERAL METABOLITE PROFILES, DECREASES IN INFLAMMATION, EFFECTS ON TUMOR GENESIS AND A NUMBER OF RECOGNIZED INTERACTIONS. THIS IS GETS TO BE A RELATIVELY OLD SLIDE, BUT I THINK IT SHOWS JUST THE VARIATION THAT YOU SEE IN INDIVIDUALS ALL GIVEN THE SAME DOSE, AND ESSENTIALLY YOU HAVE SOME PEOPLE PRODUCING LARGE AMOUNTS OF INTER-ODIOL, AND THEN YOU HAVE OTHER PEOPLE WHO ARE MAKING THE SAME AMOUNT OF BOTH AND SOME WHO DON'T MAKE ANY WHATSOEVER. SO REALLY, A HUGE RANGE IN CAPACITY TO PRODUCE THESE END PRODUCTS ON THE PART OF THE BACTERIA. IT WAS THIS WORK THAT REALLY HAD US WONDERING, IS THERE A UNIQUE GUT MICROBIAL STRUCTURE, AND ALSO, GIVEN IF IT IS HYPOTHESIZEED TO BE THE BY OWE ACTIVE COMPONENT AND THAT'S A DIFFERENTIAL COMPONENT, DO YOU SEE DIFFERENCES IN RESPONSE TO VARIOUS LIGNANDS. WE INITIALLY DID AN OBSERVATIONAL STUDY ON 107 WOMEN AND LOOKED AT THEIR GUT MICROBIAL COMMUNITY IN RELATION TO THE EXCRETION, AND IN LOOKING AT THE INDIVIDUALS ON THE BASIS, THIS IS ON THE BASIS OF 16S GENE ANALYSIS. WE FIND THAT AS IN OTHER STUDIES, YOU CAN GET THE POPULATION TO CLUSTER INTO DUMP GROUPS THAT'S DOMINATED BY CERTAIN BACTERIA. IN THIS CASE, WE HAVE THREE DIFFERENT CLUSTERS. BUT IT REALLY DIDN'T HAVE ANYTHING TO DO WITH THE INTER--TYPES. MANY OF THE BACTERIA INVOLVED IN THE PRODUCTION ARE ACTUALLY PRESENT IN VERY LOW ABUNDANCE, AND TYPICALLY, WHEN WE DO THESE TYPES OF CLUSTERING ANALYSES, IT IS REALLY DOMINANT BACTERIA THAT ARE PRESENT AT VERY HIGH PERCENTAGE THAT ARE REALLY DRIVING A LOT OF THESE ASSOCIATIONS, SO THE FACT THAT WE ARE TALKING ABOUT BACTERIA WHO ARE PRESENT MORE AT THE RANGE OF 2 TO 3%, NOT TOO SURPRISING. WHAT WE DID SEE ALSO IS THERE WAS A HIGHER GUT MICROBIAL DIVERSITY IN INDIVIDUALS WHO WERE PRODUCING AND EXCRETING MORE, COMPARED TO THOSE IN THE LOW GROUP. ALSO, LOOKING AT THE GUT MICROBIAL COMMUNITY COMPOSITION BETWEEN HIGH AND LOW EXCRETERS, THE LOW EXCRETERS ARE CLUSTERING IN THIS COMPONENT HERE, AND HIGH EXCRETERS ARE HERE, AND EVEN AFTER ADJUSTING FOR FIBER INTAKE AND ADIPOSITY, WHICH IS ALSO A CONTRIBUTOR TO GUT MICROAKIAL COMMUNITY, THIS STALE REMAINED SIGNIFICANT. SO FOLLOWING UP ON THAT AND RECOGNIZES THERE'S DIFFERENCES ACROSS INDIVIDUALS, WE CONDUCTED AN INTERVENTION STUDY TO LOOK AT THE IMPACT OF EXPOSURE TO A PLANT LIGNANDS ON A NUMBER OF PATHWAYS IMPORTANT TO CARCINOGENESIS. THIS STUDY WAS INTERESTED IN WHETHER OR NOT YOUR CAPACITY TO PRODUCE FROM PRECURSORS WOULD AFFECT CELL SIGNALING PATHWAYS DIFFERENTLY, AND WE CONDUCTED WHAT WAS CALLED THE FLAX EFFECT STUDY, BECAUSE WE RELIED ON A FLAX SEED LIGNANDS EXTRACT, AND THIS WAS A STUDY IN 42 HEALTHY MEN AND WOMEN, RANDOMIZED TO BOTH INTERVENTION AND PLACEBO AND ACROSS-OVER DESIGN. THEY RECEIVED 50-MILLIGRAMS PER DAY OF STG FOR TWO MONTHS, AND AT THE END OF THE EACH OF THOSE PERIOD, WE TOOK COLON BIOPSIES FROM THE SIGMOID COLON AND RECTUM WE USED FOR EVALUATION OF GENE EXPRESSION. WE ALSO COLLECTED STOOL SAMPLES, WHICH WE COULD THEN LOOK AT HUMAN GENE EXPRESSION IN THE STOOL SAMPLES, CALLING THE EXFOLIATED CELLS AND WE ALSO ANALYZED THE GUT MICROBIAL COMMUNITY, BUT TO GIVE YOU AN IDEA WHAT HAPPENS WHEN YOU GIVE EVERYBODY 50 MGS A DAY, THE TRIANGLES ARE ON THE BLAH SEEM BO AND THIS IS ON THE LIGNANDS SUPPLEMENT, SO YOU CAN SEE, EVEN AMONG THOSE WHO ARE RECEIVING THE SUPPLEMENT, THEY ARE DOWN IN THE WEEDS THE WHOLE WAY THROUGH. THAT REALLY MAKES NO DIFFERENCE WHAT YOU GAVE THEM. YOU WOULDN'T SEE ANY. WHERE SOME ARE PRODUCING COPIOUS AMOUNTS OVER 24 HOURS AS A RESULT, SO VERY DIFFERENT IN POTENTIAL RESPONSE. LOOKING AT THE IMPACT OF THE SDG SUPPLEMENT ON THE GUT MICROBIAL COMMUNITY, EACH ONE OF THESE LITTLE CIRCLES AND STARS IS ONE PERSON, SHOWING THE DIFFERENCE OR SIMILARITY BETWEEN THEIR GUT MICROBIAL COMMUNITY WHEN THEY ARE ON THE PLACEBO, VERSUS THE LIGNANDS TREATMENT. THERE'S NO MAJOR CLUSTERING OF ANY TREATMENT IN ANY ONE PLACE. EACH PERSON IS OFF DOING THEIR OWN THING. IN MANY CASES YOU SEE VERY LITTLE CHANGE, SHORT LITTLE LINE HERE. THIS PERSON DID HAVE A CHANGE SOMEWHAT, SO AGAIN, A LOT OF VARIATION AND RESPONSE. BUT OVERALL, THOSE INDIVIDUALS PRODUCING LARGE AMOUNTS ON THE LIGNANDS TREATMENT ARE ALL CLUSTERING OVER IN THIS AREA, SO THE LIGHTER THE COLOR, THE MORE. THE DARKER, THE LESS AMOUNT, SUGGESTING THE CAPACITY TO PRODUCE IS BEING DRIVEN BY DIFFERENCES IN THE MICROBIAL COMMUNITY STRUCTURE BETWEEN THE TWO GROUPS. IF YOU TRY TO LOOK AT SPECIFIC BACTERIAL GENERA, YOU CAN'T FIND ONE THAT'S SIGNIFICANTLY DIFFERENT, SO IT SPEAKS TO THE COMPLEXITY AND INVOLVEMENT OF THE CONSORTIUM ACROSS INDIVIDUALS. WITH REGARDS TO EFFECTS ON THE HOST RESPONSE, WE LOOKED BOTH AT BIOPSIES, AS WELL AS EXFOLIATED CELLS, SAW SUBSTANTIAL DIFFERENCES IN THE GENE EXPRESSION IN THE MUCOSA BETWEEN THE LOW AND HIGH EXCRETERS. WHEN YOU SEPARATED THEM OUT BY THOSE WHO HAD A RESPONSE TO THE TAKING THE LIGNANDS SUPPLEMENT, HIGHER ENL, YOU COULD SEE DIFFERENCES. THAT WAS BOTH IN THE BIOPSY TISSUE, AS FAR AS INCREASES IN SOME OF THE UPSTREAM REGULATORS OF INFLAMMATORY PROCESSES, TGF BETA, IL 10, SUPPRESSION IN LOW ENL EXCRETERS, THEN EVEN IN THE STOOL SAMPLES THAT WE ANALYZED FOR HUMAN GENE EXPRESSION, WE COULD SEE UP-REGULATION OF NF KAPPA B AND NOS 2, SIGNALING IN THE LOW EXCRETEERS. >> THEY WEREN'T DERIVING THE SAME BENEFIT OUT OF THE LIGNANDS SUPPLEMENT THE OTHER INDIVIDUALS WERE. SO GOING BACK TO THIS SCHEME FROM BEFORE, WE ARE SEEING EFFECTS BOTH NF KAPPA B, AS WELL AS SOME PATHWAYS I DON'T HAVE ON MY LIST HERE, SUGGESTING THINGS ARE HAPPENING THROUGH MULTI-APPROXIMATELY DIFFERENT PATH WAIVES. SO I WILL TALK ABOUT SOME OF OUR WORK EMBRACING THE COMPLEXITIES, BOTH FROM THE STANDPOINT OF THE COMPLEXITY OF DIET, AS WELL AS THE OPPORTUNITIES WE HAVE THESE DAYS WITH SOME OF THE TECHNOLOGIES TO REALLY BE ABLE TO INTERROGATE THE TOTALITY OF RESPONSES TO THESE DIETARY INTERVENTIONS. WE HAVE AN APPROACH AT FRED HUTCH THAT RELIES ON AN ANTIBODY ARRAY. THIS INCLUDES ABOUT OVER 3,000 DIFFERENT ANTIBODIES THAT COULD THEN TARGET 2100 DIFFERENT PROTEINS, SO YOU CAN RUN AND GET INFORMATION ON ALL THESE WITHIN ONE SAMPLE AND COMPARE ACROSS SAMPLES WITHIN INDIVIDUALS OR ACROSS INDIVIDUALS. WE TRIED THIS, NOT NECESSARILY A DIETARY CONSTITUENT HERE, BUT THERE'S A LOT OF INTEREST IN POTENTIAL EFFECTS OF GLUCOSEMINE, HEAVILY USED DIETARY SUPPLEMENT THAT BESIDES BEING POSSIBLY USEFUL FOR JOINT PAIN, HAS BEEN ASSOCIATED WITH LOWER RISK OF COLORECTAL CANCER AND LUNG CANCER IN A COUPLE LARGE COHORT STUDIES, SO WE WERE INTERESTED IN IF WE COULD IDENTIFY POSSIBLE MECHANISMS THAT BY WHICH THESE SUB-N'TS MIGHT BE WORKING, SO WE USED RANDOMIZEDCROSS-OVER DESIGN, PLACEBO CONTROLLED WITH A GROUP OF HEALTHY OVERWEIGHT INDIVIDUALS AND HAD TREATMENT INTERVENTION AGAINST A FASTING PLACEBO CONTROL, AND WE FOUND 508 INDIVIDUAL PROTEIN ANTIBODIES THAT WERE SIGNIFICANTLY ALTERED AFTER CORRECTION FOR FDR. I DON'T THINK I HAVE EVER SEEN P VALUES LIKE THIS IN MY LIFE BEFORE. MOSTLY I'M BARELY LUCKILY OF LESS THAN 0.05, SO THIS WAS EXCITING. WHAT WAS MORE EXCITING WAS RECOGNIZING THE PROTEINS THAT WERE SHOWING UP HERE WERE REALLY RELATED TO A VARIETY OF DIFFERENT PATHWAYS, SO RUNNING A PATHWAY ANALYSIS IN THIS CASE USING KEG, YOU SEE IMPACT ON THE CYTOKINE PATHWAYS, INTESTINAL IMMUNE NETWORK IN THE IGA PRODUCTION, TRANSENDOTHELIAL MITIGATION, A WHOLE HOST OF THINGS WE HADN'T THOUGHT ABOUT IN THE CONTEXT OF THIS SUPPLEMENT, BUT ALSO SERVED AS PROOF OF PRINCIPLE FOR POTENTIAL USE TO INTERROGATE PLASMA SAMPLES TO TRY TO UNDERSTAND THE EFFECTS OF VARIOUS DIETARY INTERVENTIONS, SO CURRENTLY, WE HAVE TAKEN THIS TO A STUD DID OF DIETARY PATTERNS. SEVERAL, AS PART OF THE ENERGETICS AND CANCER, WE CONDUCTED A FEEDING STUDY LOOKING AT EFFECTS OF LOW AND HIGH GLYCEMIC LOAD PATTERNS AND FOUR-WEEK RANDOMIZED CROSSOVER DESIGN. ONE THINGS WE HAVE COME TO LOVE ARE THE CROSSOVER DESIGNS, WHERE YOU CAN DO THIS, BECAUSE PARTICULARLY IN THE CONTEXT OF DEALING WITH THE GOOSE MOI ROW BYE OWN AND THE INDIVIDUALLY OF EACH PERSON'S MICROBIOME, IT HELPS TO HAVE EACH PERSON SERVE AS THEIR OWN CONTROL, WHEN YOU TEST A PARTICULAR INTERVENTION, SO AS LONG AS YOU DON'T HAVE TO RUN THE TREATMENT FOR TOO LONG, YOU CAN DO THAT. THESE DIETS CONTAINED THE SAME NUMBER OF CALORIES ON BOTH THE HIGH AND LOW GLYCEMIC LOAD, BUT AS YOU MIGHT IMAGINE F YOU WANT TO FEED A LOW LOAD DIET, IT WILL BE SUBSTANTIALLY HIGHER IN FIBER THAN THE HIGH LOAD DIET. THERE ARE ALSO A NUMBER OF PHYTOCHEMICALS THAT DIFFER, INCLUDING THE LIGNANDS, WHICH ARE ALMOST TWICE AS HIGH ON THE LOW GLYCEMIC LOAD, JUST BASED ON THE TYPES OF FOODS YOU WOULD CHOOSE TO FOLLOW THE LOW--LOAD DIET. THE CARB STUDY WAS INTERESTED IN CIRCULATING METABOLIC BIOMARK TERSE THAT HAVE BEEN ASSOCIATED WITH HIGHER RISK OF CANCER. THEY ARE LISTED HERE, AND AS ONE MIGHT EXPECT, WE SAW DIFFERENCES IN GLUCOSE AN INSULIN AS A RESULT OF THE TWO TREATMENTS WITH THE LOW GLYCEMIC LOAD HAVING LOWER AREA OF GLUCOSE UNDER THE CURVE, AS WELL AS INSULIN, AND DIFFERENCES IN THE IGF1 LEVELS AS A RESULT OF THE INTERVENTION. CURRENTLY, WE HAVE NOW GONE BACK TO THE FREEZER, SO TO SPEAK AND ARE USING SAMPLES FROM THIS STUDY TO TAKE AN INTEGRATED APPROACH TO UNDERSTAND THE IMPACT OF THESE TWO DIFFERENT DIET PATTERNS ON CANCER RISK PATHWAYS, SO WE GET THERE THROUGH BOTH UNDERSTANDING THE IMPACT OF THE DIET OR THE INDIVIDUAL'S BASELINE MICROBUY MICROBUY -- MYOBIOME AN PLASMA POET -- PROTEOME AND UNDERSTAND THE DIFFERENCES IN PATHWAYS THAT ARE RELEVANT FOR CANCER. WE HAVE DONE SOME INITIAL WORK ON THIS. A GRANT WAS JUST FUNDED A COUPLE YEARS AGO, SO IN FIRST PASS, SOME OF THE FIRST SAMPLES, WE SEE A GOOD DISTINCTION BETWEEN THE TWO DIETS, USING PARTIAL DISCRIMINANT ANALYSIS, WHERE YOU CAN SEE THE HIGH GLYCEMIC DIET, PEOPLE ARE CLUSTERING HERE, VERSUS THE LOW DIET CLUSTERING HERE, AND THE NUMBER OF DIFFERENT METABOLITES THAN DIFFER. ONCE YOU CONTROL FOR FALSE DISCOVERY RATE, THE ONLY THING STILL HOLDING UP AT THIS POINT, BUT AGAIN, SUGGESTING THOSE COMPONENT VERSUS A POTENTIAL -- THIS WAS JUST FIRST RUN ON THE FIRST 19 SAMPLES OUT OF 80 INDIVIDUALS, SO WE HAVE WAYS TO GO, BUT I THINK IT REALLY IS EXCITING TO THINK ABOUT COMBINATIONS OF THE BIOINFORMATICS, AS WELL AS THE APPROACHES THAT ARE AVAILABLE FOR DOING NUTRITION STUDIES THAT WILL REALLY HELP US TO PULL THESE STORIES TOGETHER IN THE FUTURE. SO JUST TO WRAP UP, ALWAYS GOOD TO KEEP IN MIND THAT, UNLIKE DRUGS OR UNLIKE THE ATTEMPT FOR THE PHYTOCHEMICALS ARE ACTING THROUGH MULTIPLE PATHWAYS THAT RELATE TO CARCINOGENESIS, SO ONLY IF YOU FOCUS ON ONE PATHWAY, YOU HAVE TO REMEMBER THERE ARE LIKELY OTHER PATHWAYS THAT ARE ALSO GOING TO BE CHANGING, SO EVERYTHING MAY NOT WITH THROUGH THE ONE PARTICULAR PATHWAY THAT YOU ARE INTERESTED IN. CERTAINLY, BOTH GUT MICROBIAL, AS WELL AS HOST GENETIC DIFFERENCES CAN RESULTS IN A RANGE OF RESPONSES TO COMPOUNDS, AND IT'S NOT ONLY BECAUSE PEOPLE WEREN'T CONSUMING THE DIET THAT YOU THOUGHT THEY WERE CONSUMING, BUT TRULY, THERE IS VARIATION ACROSS INDIVIDUALS, AND I THINK WE NEED TO DO A BETTER JOB UNDERSTANDING WHAT THAT IS AND TO WHAT EXTENT IT HAS AN IMPACT ON POTENTIAL RESPONSE TO OUR RECOMMENDATIONS FOR DIETARY CHEMOPREVENTION, RECOMMENDATIONS FOR APPROACHES TO REDUCTION IN RISK OF A NUMBER OF DIFFERENT DISEASES, AND AS WE COME TOP UNDERSTAND SOME OF THE MICROBIAL PHENOTYPES, YOU COULD USE A ME TAB HOMICS APPROACH OR TARGET ON SPECIFIC METABOLTES TO GET AN IDEA OF THE RANGE AND POTENTIAL EFFICACY OF A DIET OR SUPPLEMENT-BASED INTERVENTIONS THAT WILL BE HIGHLY DEPENDENT ON THE INDIVIDUAL AND NOT EVERYBODY GOING TO RESPOND IN THE SAME WAY. I THINK WE ALSO NEED TO WORK TOWARDS DOING A BETTER JOB AT CAPTURING AND STUDYING DIETARY PATTERNS AND THE TOTALITY OF DIET, FROM THE STANDPOINT OF THAT'S WHAT WE EAT, AND DESPITE ATTEMPTS TO -- I WAS AROUND WHEN THE FIRST ASTRONAUTS WENT INTO SPACE AND WE GOT TO EAT SPACE STICKS AND ALL THESE OTHER INTERESTING PREPARED FOODS, BUT TO THIS DAY, ASTRONAUTS WOULD BE MUCH HAPPIER WITH REAL FOOD IN SPACE, SO COMING TO WRECK NICE, WE WILL LIKELY BE EATING REAL FOOD FOR A LONG TIME YET, SO BEST TO UNDERSTAND HOW THIS IS AFFECTING US AND REALLY ACROSS THE BREADTH OF FOODS THAT HUMANS CONSUME AND APPRECIATE. JUST LIKE TO ACKNOWLEDGE MY LAB GROUP, PARTICULARLY MEREDITH HELLER, A MICROBIAL ECOLOGIST, WHO WORKED WITH US THE LAST TEN YEARS AND GRAD STUDENTS AND POST-DOCS HAVE BEEN IN THE GROUP, AS WELL AS COLLABORATORS AT FRED HUTCH AND ELSEWHERE, AND CERTAINLY, THE SUPPORT OF NCI AND NIH IN GENERAL, AND FRED HUTCH. AND THANK YOU FOR YOUR ATTENTION. [APPLAUSE] SORRY ABOUT MY CROAKY VOICE. >> OKAY, WE HAVE SET ASIDE SOME TIME FOR SOME QUESTIONS, AND AGAIN, TO REMIND THE PRACTICUM PEOPLE TO COME DOWN FRONT. AND IF YOU HAVE A QUESTION, BRING THAT WITH YOU. ANY QUESTIONS FOR JOHANNA? OKAY. FOURTH ROW, FIFTH ROW. (OFF MIC). WELL, DEFINITELY, IN THAT ONE GRAPH, IT SHOWED ABOUT HALF THE INDIVIDUALS HAD SUBSTANTIAL INCREASE IN ENL WITH THE SDG SUPPLEMENT. THE OTHER HALF, IT DIDN'T REALLY BUDGE, COMPARED TO THE PLACEBO. SO SOME PEOPLE DON'T HAVE THE GUT MICROBES NEEDED TO MAKE THAT CONVERSION. YOU WILL FIND AMONG SOME OF THOSE INDIVIDUALS WHO AREN'T MAKING -- THEY ARE MAKING INTER-O DIOL, THROUGH THE ENL, SO WHAT YOU MAY FIND, YOU WILL HAVE SOME PEOPLE WHO ARE PRODUCING SOME END, SOME WHO DON'T PRODUCE ANY OF THESE COMPOUNDS, SO THERE WILL BE A RANGE IN RESPONSES THAT YOU WILL SEE IN RESPONSE NO YOUR FLAX SEED FEEDING, AND WITH OUR STUDY, WE DIDN'T FIND ANY CHANGE -- THERE WAS NO SORT OF STATISTICALLY SIGNIFICANT RECOGNIZED CHANGE IN THE MIGHT MICROBIAL COMMUNITY AS A RESULT OF THE SUPPLEMENTATION. THIS ISN'T UNUSUAL. THERE HAVE BEEN SEVERAL REPORTS IN THE LITERATURE THAT YOU DON'T REALLY SEE A SUBSTANTIAL CHANGE AS A RESULT OF THE FLAX SEED FEEDING. I THINK SOME OF THE STUDIES WHERE THEY HAVE USED LARGER DOSES AND THEREFORE YOU HAVE AN INCREASE IN DIETARY FIBER AS A RESULT, YOU MAY SEE SOME OF THE EFFECTS THERE, BUT AT LEAST OVER SHORTER TERM FEEDINGS F YOU RAN IT OUT FOR YEAR OR MORE, YOU MAY SEE SOME EFFECTS, BUT FOR SHORT-TERM FEEDINGS OF SEVERAL MONTHS, YOU REALLY DON'T SEE MUCH, RIGHT. IT'S GOING TO BE PROBABLY THE UNDERLYING MICROBIAL COMMUNITY THAT EACH WOMAN BRINGS TO THE STUDY THAT WILL DETERMINE SOME OF YOUREST-END -- YOUR ESTI-END RESPONSES. >> USE THE MICROPHONE OR WE WILL REPEAT THE QUESTION, IF IT ISN'T TOO LONG, SO WE DON'T REMEMBER WHAT YOU ASKED. THAT'S A REALLY GOOD POINT. THE COMMENT WAS HOW DO WE KNOW WENT WE DO DIETARY PATTERN STUDIES THAT WHEN YOU ADD SOMETHING, YOU HAVE TO REMOVE SOMETHING ELSE. IS THE IT THE REMOVAL OR ADDITION THAT'S PLAYING A ROLE, AND I THINK IT'S GOD POINT, BUT IT ALSO SPEAKS TO THE IDEA THAT IF YOU ARE BELIEVING OR WANTING TO TEST EFFECT OF A PATTERN, IT'S NOT THAT ONE SINGLE FOOD THAT'S IMPORTANT. IT'S THE TOTALITY OF THAT PATTERN, SO IF YOU THINK ABOUT HOW WE CONSTRUCT DIETARY PATTERNS THAT YOU ARE CHOOSING HIGH-FIBER, LOW FAT, VERSUS HIGH FAT, LOW FIBER, IT IS REALLY WHAT FOODS ARE YOU GOING TO CHOOSE TO DO THAT, AND WHAT IS THE IMPACT ON IT. >> HAVE YOU LOOKED IF THERE IS ANY CORRELATION IN YOUR RESULTS WITH THE BODY COMPOSITION, SO SOME PEOPLE EXERCISE MORE OR LESS, SO THE AMOUNT OF FAT AND MUSCLE. HAVE YOU LOOKED IF THERE'S ANY CORRELATION LIKE THAT? >> IN RELATION TO THE -- >> THE RESULT OF THE STUDY WITH THE BODY COMPOSITION, SO THE AMOUNT OF FAT, OF THE MUSCLE, THE RESULT OF THE STUDY. >> SO IN THE CONTEXT OF THE OBSERVATIONAL STUDY, I THINK WE HAD A CUTOFF FOR BMI THAT WAS A RELATIVELY NARROW RANGE, AND THIS WAS A NUMBER OF YEARS AGO, SO WE HAVE NEVER SEEN AN ASSOCIATION BETWEEN ENL AND ADIPOSITY, PER SE. THE REASON WE CONTROLLED FOR ADIPOSITY WAS BECAUSE THERE WAS AN ASSOCIATION BETWEEN ADIPOSITY AND THE GUT MICROBIOME, BUT WE HAVEN'T SEEN THE DIRECT CORRECTION TO -- >> I UNDERSTAND THE BMI, BUT IT WAS FOR THE ADIPOSITY, IF YOU HAVE SEEN SOMETHING WITH THE ADIPOSITY ITSELF. >> WE HAD PERCENT BODY FAT, BUT WE HAVEN'T SEEN ANY DIFFERENCE THERE. >> THANK YOU. >> I'M JUST CURIOUS, NOT EVERY LAB CAN DO THAT, BECAUSE YOU HAVE 3,000, 4,000 ANTIBODIES, SO I WAS WONDERING, HAVE YOU TRIED LOOKING AT EXOSOMES INDICES? BECAUSE THEY ARE EASY TO DO IN THE PLASMA AND STABILITY IS AFFECTED BY THE PRESENCE OF -- THAT COULD BE A GOOD INDICATER. HAS ANYBODY TRIED THAT OR THINKING OF? >> WE HAVEN'T DONE THAT, BUT DEFINITELY THERE ARE GOOD CAMP IN THE LITERATURE OF APPLICATION OF THOSE ASSAYS TO INTERVENTIONS SUCH AS THESE, SO I THINK IT'S DEFINITELY ANOTHER ROUTE TO GO. >> THAT COULD BE CHEAPER THAN -- I DON'T KNOW HOW MUCH IT COSTS TO DO THAT. >> YEAH, I THINK IT WOULD GIVE YOU ANOTHER DIMENSION TO THE STORY. I ONOT SURE PIT WILL BE -- >> IT'S NOT THE SAME. >> BUT IN THE CONTEXT OF THE PROTEOME OWN, WHAT WE HAVE DONE, IF WE IDENTIIED THEM CHANGES, WE USE AN ASSAY ON A SMALLER TARGETED PLATFORM TO GO AFTER THE 12 OR 14 PROTEINS, SO THERE IS AN OPPORTUNITY FOR IT TO BE REPLICATED IN ANOTHER STUDY POPULATION, WITHOUT HAVING THAT -- >> THAT'S RIGHT T. IT WAS JUST MY IDEA, LIKE NOW WITH ALL THE INFORMATIVE TOOLS, YOU CAN ASSOCIATE -- >> YEAH, THAT IS A GREAT IDEA. >> YOU CAN DO THAT. >> YEAH, THANK YOU. GOOD POINT. OKAY, THANK YOU ALL. [APPLAUSE]