Gut reactions: host microbiome interactions in the intestine in health and disease

Gut reactions: host microbiome interactions in the intestine in health and disease


>>GOOD AFTERNOON, EVERYONE. WELCOME TO THE WEDNESDAY AFTERNOON NIH DIRECTOR’S LECTURE. A FASCINATING TOPIC AND A HIGHLY QUALITIED SPEAKER TO LEAD US THROUGH INTERESTING IDEAS WITH IMMUNOLOGY, THE INTESTINE AND HEALTH AND DISEASE. FIONA POWRIE, WHO IS HERE AS OUR SPEAKER, GOT HER UNDERGRADUATE DEGREE AT THE UNIVERSITY OF BATH, Ph.D. OR D PHIL AT THE UNIVERSITY OF OXFORD IN IMMUNOLOGY AFTER POSTDOCTORAL FELLOWSHIP AT A COMPANY CALLED DNAX RESEARCH INSTITUTE SHE WENT BACK TO THE U.K. AND HAS BEEN AT OXFORD PRETTY MUCH FOR THE LAST 20 YEARS OR SO, MOVING THROUGH THE RANKS FROM FELLOW TO SENIOR RESEARCH FELLOW TO SYDNEY TRUELOVE PROFESSOR OF GASTROENTEROLOGY TO CURRENT POSITION AT DIRECTOR OF KENNEDY INSTITUTE OF RHEUMATOLOGY AT THE UNIVERSITY OF OXFORD, AND PRINCIPAL INVESTIGATOR OF THE TRANSLATIONAL GASTROENTEROLOGY UNIT, NEUFIELD DEPARTMENT OF MEDICINE, FELLOW OF ROYAL SOCIETY, REACHED THE LOUIS PRIZE FOR MEDICINE, HER INTEREST IN INTESTINE IMMUNE SYSTEM IS& SOMETHING SHE WILL SPEAK ABOUT. SHE’S INTERESTED IN EDUCATING PUBLIC AND SCIENTISTS ABOUT WHAT WE ALL DO IN THE LABORATORY, AND HAS PLAYED AN INTERESTING ROLE IN TRYING TO PROMOTE THAT TRANSLATIONAL COMPONENT TO THE PUBLIC. HER TOPIC TODAY IS ENTITLED IN A PROVOCATIVE WAY, GUT REACTIONS. WE IN THE UNITED STATES ARE FAMILIAR WITH THOSE AT THE MOMENT. I’LL SAY NOTHING MORE THAN THAT. GUT REACTIONS, HOST-MICROBIOME INTERACTIONS IN THE INTESTINE IN HEALTH AND DISEASE. PLEASE JOIN ME IN WELCOMING PROFESSOR FIONA POWRIE. [APPLAUSE] >>WELL, THANK YOU VERY MUCH, DR. COLLINS, FOR THAT VERY WARM WELCOME. IT’S ABSOLUTELY A PLEASURE TO BE BACK HERE AT THE NIH. I HAVE MANY FRIENDS AND COLLEAGUES I’VE ADMIRED THE IMMUNOLOGY THAT HAS COME OUT OF THE NIH FOR DECADES, AND THAT’S NO CHANGE, VERY EXCITING WORK AND WONDERFUL TO MEET EARLY STAGE SCIENTISTS, ALWAYS A TREAT ON THESE TYPES OF VISITS. I WANT TO TALK ABOUT OUR INTERESTS IN UNDERSTANDING HOST MICROBE INTERACTIONS IN THE INTESTINE, HOW THEY PROMOTE HEALTH AND HOW WHEN THEY BECOME DERANGED CAN LEAD TO THE DEVELOPMENT OF DISEASE. WHAT I WANT TO START WITH THOUGH IS A MESSAGE FOR EARLY STAGE SCIENTISTS IN THE AUDIENCE. I KNOW THIS IS NOT A SPECIALTY IMMUNOLOGY AUDIENCE, BUT ONE MESSAGE FOR YOU IS THAT THIS IS A GREAT TIME TO BE AN IMMUNOLOGIST BECAUSE WHAT WE’RE SEEING IS THAT IMMUNITY AND DERANGED OR DEFICIENT IMMUNE RESPONSES UNDERLIE A NUMBER OF DISEASES THAT WE HAVEN’T NECESSARILY ASSOCIATED WITH THE IMMUNE SYSTEM. SO OF COURSE WE KNOW ABOUT THE IMMUNE SYSTEM IN INFECTIOUS DISEASE, WE KNOW HOW IT CAN RESPOND ABERRANTLY IN AUTOIMMUNE AND CHRONIC INFLAMMATORY DISEASE BUT WE’RE LEARNING ALTERATIONS IN IMMUNITY AND INFLAMMATION CAN UNDERLIE DISEASES SUCH AS NEURODEGENERATION, MENTAL HEALTH, TYPE 2 DIABETES, ATHEROSCLEROSIS AND OTHERS. SO A GREAT TIME WITH THE TOOLS EMERGING IN OUR TRADE TO BE ABLE TO DECIPHER THE IMMUNE SYSTEM IN HEALTH AND DISEASE. NOW, OF COURSE, CHRONIC INFLAMMATORY DISEASES ARE COMPLEX MULTI-FACTORIAL DISEASES INVOLVING THE INTERACTION OF GENETIC AND ENVIRONMENTAL FACTORS, OFTEN PLAYING OUT THROUGH EPIGENETIC CHANGES, THAT HAVE AN IMPACT ON THE IMMUNE SYSTEM AS WELL AS TISSUE RESPONSES. AND WHEN THERE WAS A MALADAPTATION WHEN THE HOST AND ITS GENETIC LANDSCAPE AND ENVIRONMENTAL DRIVERS, THAT CAN LEAD TO CHRONIC INFLAMMATORY DISEASES. AND IN SOME INDIVIDUALS THEY WILL MANIFEST IN THE BOWEL WITH INFLAMMATORY BOWEL DISEASE, IN OTHER INDIVIDUALS INFLAMMATORY JOINT DISEASES, OR IN OTHER INDIVIDUALS INFLAMMATORY SKIN DISEASES. AND OF COURSE THE CHALLENGE NOW AS WE THINK ABOUT HOW TO TREAT THESE DISEASES IS THAT THERE ARE SHARED COMPONENTS AS WELL AS UNIQUE, AND THE UNIQUE ONES MIGHT RELATE TO PARTICULAR TISSUE SPECIFIC COMPONENTS. OF COURSE IF WE CAN UNDERSTAND AND REDEFINE DISEASE BASED ON PROCESSES THAT MAY RELATE TO IMMUNE FUNCTION IN TISSUES, THEN WE MAY BE ABLE TO REPURPOSE DRUGS ACROSS DIFFERENT DISEASES AND MOVE AWAY TO A NEW MOLECULAR TAXONOMY OF DISEASE AWAY FROM OUR SILOED UNDERSTANDING BASED SIMPLY ON MEDICAL CLINICAL SYMPTOMS TO MORE UNDERLYING PROCESS-DRIVEN TAXONOMY. NOW, OF COURSE, AN IMPORTANT FACTOR IN A NUMBER OF CHRONIC DISEASES IS RESIDENT MICROBES. WE MAY CONSIDER OURSELVES A SUPER ORGANIZATION COMPOSED OF HUMAN TISSUES AS WELL AS RESIDENT COLONISTS WHICH OF COURSE INCLUDE A VAST ARRAY OF BACTERIA, VIRUSES AND FUN GUY. OF COURSE, THEY MAKES UP A LARGE NUMBER OF CELLS IN THE INTESTINE. THERE ARE MORE BACTERIA IN THE INTESTINE THAN ANYWHERE ELSE IN THE BODY, 100 TRILLION BACTERIA INHABIT THE INTESTINE. OF COURSE THEY CONTRIBUTE THROUGH THEIR GENES, THEIR PROTEINS, AS WELL AS THEIR METABOLITES INTERACTING WITH THE HOST AND IMPACTING ON NUTRITION, IMMUNE SYSTEM DEVELOPMENT AND HOST DEFENSE. SO A HUGE AMOUNT OF INTEREST INCLUDING ECONOMIC INTEREST IN HOW TO BE ABLE TO HARNESS THE MICROBIOME, HOW IT INTERACTS WITH THE HOST, YOU TO PROMOTE HEALTH, AND OF COURSE TO UNDERSTAND MALADAPTATION OF THIS INTERFACE, HOW THIS DRIVES DISEASE. THE DISEASE WE’RE INTERESTED IN IS INFLAMMATORY BOWEL DISEASE, A POSTER CHILD, IF YOU LIKE OF THE COMPLEX MULTI-FACTORIAL DISEASES INVOLVING THE INTERACTION BETWEEN GENETIC LANDSCAPE AND ENVIRONMENTAL FACTORS THAT PLAYS OUT IN THIS INTERFACE BETWEEN THE IMMUNE SYSTEM AND THE RESIDENT MICROBIOTA. INFLAMMATORY BOWEL DISEASE IS A COMPLEX RANGE OF CLINICAL PHENOTYPES, ENCOMPASSING CROHN’S DISEASE, AND ULCERATIVE COLITIS. IT HAS AN INCREASING PREVALENCE PARTICULARLY IN EASTERN POPULATIONS, AND PROBABLY THE HETEROGENEOUS CLINICAL NATURE IS A CONSEQUENCE OF THE MULTI-FACTORIAL INTERACTIONS THAT ARE DRIVING THE DISEASE. THERE’S NO CURE FOR INFLAMMATORY BOWEL DISEASE. A LARGE PROPORTION OF PATIENTS WILL PROGRESS TO SURGERY. OF COURSE, THERE HAVE BEEN GREAT ADVANCES IN THE LAST DECADES, AND ONE OF THOSE IS ANTI-TNF PIONEERED IN PART BY MARK FELDMAN AND AT THE KENNEDY INSTITUTE FOR TREATMENT OF RHEUMATOID ARTHRITIS, ALSO APPLIED IN INFLAMMATORY BOWEL DISEASE, IN SOME INDIVIDUALS THEY WILL DO EXTREMELY WELL. BUT A LARGE PROPORTION DO NOT RESPOND. AND WE DON’T UNDERSTAND WHO THEY ARE. WE DON’T UNDERSTAND HOW TO TREAT, WHAT THE COURSE OF DISEASE IS GOING TO BE, AND OF COURSE IN THIS CHRONIC DEBILITATING DISEASE WITH INTESTINAL INFLAMMATION, THERE IS AN INCREASED RISK FOR COLON CANCER. NOW, THERE IS EVIDENCE OF DYSBIOSIS OF BACTERIAL COMMUNITIES IN THE INTESTINE AND A LARGE NUMBER OF STUDIES HAVE BEEN DONE IN THIS AREA. SO IF YOU LOOK AT THE MAKEUP OF THE DIFFERENT PHYLA PRESENT IN THE INTESTINE, IN A NORMAL INDIVIDUAL, MADE UP OF FIRMA CUTIES AND BACKTEROIDES PHYLA, THERE’S AN INCREASE IN PROTOBACTERIA FOR EXAMPLE AND OTHERS BUT THE QUESTION IS WHETHER THIS IS A COURSE OR CONSEQUENCE AND HOW WE MIGHT USE THIS INFORMATION TO BE ABLE TO DESIGN THERAPEUTIC MODALITIES. WHEN THERE’S A GENETIC SUSCEPTIBILITY GIVING IBD-LIKE SYNDROME WITH OVER 70 MODELS OF COLITIS THAT DEVELOP AS CONSEQUENCE OF GENETIC MANIPULATION OF THE HOST ALTERING VARIOUS TYPES OF PATHWAYS, BUT IN VERY FEW SITUATIONS DO THOSE GENETIC CHANGES HAVE AN IMPACT WITHOUT A REPLICATING INTESTINAL MICROBIOTA SO THIS REALLY IS AN INTERACTION BETWEEN THE HOST GENETIC INTERFACE AND THE MICROBES. OF COURSE, IN IBD SOME PATIENTS DO BENEFIT FROM PROBIOTIC THERAPIES, AND THERE ARE TRIALS LOOKING AT FECAL TRANSPLANTATION. WE HAVE EVOLVED WITH INTESTINAL MICROBES, AND WE’RE STARTING TO UNDERSTAND IN GETTING — SCRATCHING THE SURFACE, I WOULD SAY, OF THE NATURE OF THOSE MOLECULAR INTERACTIONS. SO A NUMBER OF METABOLITES THAT ARE PRODUCED BY DIVERSE ARRAY OF MICROBES HAVE A DIRECT IMPACT ON THE INTESTINAL IMMUNE SYSTEM. ONE IN PROMOTING HOST DEFENSE AT BARRIER SURFACES, TWO, METABOLITES SUCH AS SHORT CHAIN FATTY ACIDS WHICH ARE FERMENTATION PRODUCTS OF CERTAIN TYPES OF BACTERIA CAN IMPACT ON THE TYPES OF IMMUNE CELLS AND OF COURSE BACTERIA EXPRESS A NUMBER OF PATTERN RECOGNITION RECEPTOR LIGANDS THAT STIMULAE DIFFERENT INNATE IMMUNE CELLS. SO IN HEALTH, OF COURSE, THAT IS GOING TO PROMOTE PROPERLY FUNCTIONING DIALOGUE AND INTERFACE, WHEN THERE IS IT A HOPE MALADAPTATION, SOME PATHWAYS START TO DRIVE CHRONIC INTESTINAL INFLAMMATION WHERE THE SITUATION IS MALADAPTIVE. THAT’S REALLY WHAT I WANT TO TRY TO DISSECT TODAY. FRSH THERE ARE CHANGES IN THE MICROBIOME, IS THAT BECAUSE CAUSE OR CONSEQUENCE, IF WE CAN UNDERSTAND CAN WE RESET THE BALANCE? THERE’S A STRONG GENETIC CONTRIBUTION TO INFLAMMATORY BOWEL DISEASE IN EARLY ONSET, A NUMBER OF MUTATIONS, OVER 60 DIFFERENT MUTATIONS THAT HAVE BEEN SEQUENCED IN MONOGENIC DISEASES, WHERE THE PATIENTS PRESENT WITH INFLAMMATORY BOWEL DISEASE. THEY ARE DIFFERENTLY PENETRANT, SO FOR DEFICIENCIES IN IL-10, FOR EXAMPLE, AS FAR AS WE KNOW 100 PER CENT OF PATIENTS WHO HAVE A MALFUNCTIONING IL-10 RECEPTOR PATHWAY WILL DEVELOP INFLAMMATORY BOWEL DISEASE. THE VAST MAJORITY OF IBD DEVELOPS WITH PEAK ONSET OF 20 YEARS OF AGE, IN THAT SETTING GENETICS CONTRIBUTES FOR AROUND 70% OF THE RISK, THERE ARE A LARGE NUMBER OF SNPs THAT HAVE BEEN IDENTIFIED TO CONFER RISK, AND I THINK THERE WE MIGHT SUGGEST THAT THE GENETICS IS GIVING A LANDSCAPE THAT IS PERMISSIVE FOR A PARTICULAR RESPONSE TO ENVIRONMENTAL DRIVERS. BUT I THINK WHAT THE MONOGENIC DISEASES HAVE BEEN HIGHLY INFORMATIVE IN TERMS OF IDENTIFYING NON-REDUNDANT PATHWAYS, MY COLLEAGUE UHLIG MAPPED THESE, MAPPING INTO WHAT WE MIGHT CONSIDER AS CORNERSTONES OF INTESTINAL HOMEOSTASIS. THESE ARE THE ABILITY TO COMPARTMENTALIZE AND EXCLUDE ALL OF THESE RESIDENT MICROBES FROM THE UNDERLYING LIMINA P.R. O PRIA, WE ALL THE KNOW INTERACTIONS LIKE THAT. THAT MEANS HAVING APPROPRIATE EPITHELIAL CELL FUNCTION, MUCIN SECRETION WITH A STOUT EPITHELIAL BARRIERS THAT PREVENTS TRANSLOCATION OF BACTERIA. THE NEXT IS APPROPRIATE PHAGOCYTES, KILLING INVADING BACTERIA. IF THERE ARE DEFICIENCIES IN SOME OF THESE SUCHS IS IN CHRONIC GRANULOMATOUS DISEASE, WHERE A PROPORTION OF PATIENTS WILL DEVELOP INFLAMMATORY BOWEL DISEASE, THEN THAT AGAIN IS A MALADAPTATION OF A KEY ASPECT OF INTESTINAL HOMEOSTASIS. OUR OWN WORK HAS FOCUSED PARTICULARLY ON THE ROLE OF REGULATORY T CELLS, THE SO-CALLED FOX P3 POSITIVE REGULATORY T CELLS I’LL EXPLAIN IN A LITTLE MORE DETAIL IN A MINUTE. BUT A NUMBER OF MUTATIONS IN THE FOX P3 REGULATORY COR TELL AND IL-10 PRODUCTION ARE ASSOCIATED WILL GIVE RISE TO MONOGENIC INFLAMMATORY BOWEL DISEASE. SO OF COURSE UNDERSTANDING THE PATHWAYS ATTACHED TO THESE MONOGENIC DISEASES THROUGH THAT, WE MAY BE ABLE TO MAP THOSE INTO LARGER PATHWAYS, ASSOCIATED WITH THE SNPs THAT RELATE TO DIFFERENT PROCESSES THAT GO WRONG IN DIFFERENT SUBSETS OF PATIENTS. SO I’M SUGGESTING TO YOU THAT IBD IS NOT CROHN’S DISEASE AND ULCERATIVE COLITIS BUT RATHER SYNDROMIC WHERE THERE MAY BE PATIENTS THAT HAVE EPITHELIAL BARRIER DEFECTS VERSUS PHAGOCYTE DEFECTS VERSUS REDUCTION IN IMMUNE REGULATION. SO THESE ARE REGULATORY T CELLS. THIS IS A SPECIALIZED SET OF T CELLS IN THE THYMUS, TRANSCRIPTION FACTOR FOXP3. IF YOU HAVE THAT DEFICIENTS IS YOU DON’T HAVE A FUNCTIONS POPULATION OF REGULATORY T CELLS, AND DEVELOP IBD AND OTHER A– A RANGE OF OTHER INFLAMMATORY DISEASES. SO REGULATORY T CELLS DEVELOP IN THE THYMUS, EXIT INTO THE PERIPHERY, WHERE THEY CONTROL EFFECTOR AS WELL AS ADAPTIVE AS WELL AS INNATE RESPONSES, NOW IN ADDITION TO THYMIC DERIVED, NAIVE T CELLS UNDER CERTAIN CONDITIONS CAN BE DIFFERENTIATED, CAN DEVELOP INTO FOXP3 EXPRESSING CELLS, WHAT THE FUNCTIONS ARE, THE DIVISION OF LABOR IN TERMS OF CONTROLLING THE INFLAMMATORY RESPONSE. OF COURSE, THE GUT IS AN IMPORTANT LOCATION FOR THE DIFFERENTIATION OF NAIVE T CELLS INTO FOXP3 EXPRESSING CELLS. OF COURSE, THIS THOSE IMPORTANT ROLE OF TGF-BETA IN DRIVING THIS PROCESS, AND WE TOGETHER WITH YASMIN BELLCADE AND OTHERS, THE VITAMIN A METABOLITE AS A CO-FACTOR DRIVING THIS RESPONSE, DRIVING T CELLS INTO THE REGULATORY T CELL FUNCTION. WHEN YOU’RE LACKING THIS, IT LEADS TO A RANGE OF THESE DIFFERENT INFLAMMATORY DISEASES. SO, WE HAVE OVER THE LAST DECADES NOW BEEN STUDYING THE PATHWAYS THAT CONTROL INTESTINAL HOMEOSTASIS, AND THIS IS A MOUSE MODEL THAT WE DESCRIBED MANY YEARS AGO, AND INVOLVES TRANSFER OF NAIVE T CELLS INTO MICE THAT HAVE NO T CELLS OF THEIR OWN, AND THAT LEADS TO A CHRONIC INFLAMMATORY DISEASE, THAT IS DRIVEN BY THE MICROBIOTA, IF YOU TRANSFER CELLS INTO MICE THAT DO NOT HAVE A REPLICATING INTESTINAL MICROBIOME THEN THEY DON’T DEVELOP DISEASE, AND THIS DISEASE IS CHARACTERIZED BY AN INCREASE IN A NUMBER EVER T CELLS, Th1 AND Th17 RESPONSE WITH KEY CYTOKINE DRIVERS. AND IF YOU NEUTRALIZE THESE PATHWAYS, YOU INHIBIT DISEASE IN THIS MODEL. BUT YOU CAN ALSO PREVENT DISEASE, NOT ONLY PREVENT IT BUT ALSO CURE IT, IF YOU TRANSFER THE REGULATORY FOXP3 SUPPRESSING POPULATION INTO MICE THAT HAVE ESTABLISHED DISEASE, THIS IS EFFICIENT, THESE CELLS WILL HONE TO THE INTESTINE WHERE THEY PROLIFERATE AND IN IL-10 DEPENDENT FASHION WILL RESOLVE THE INTESTINAL INFLAMMATORY RESPONSE. SO A LOT OF INTEREST IN BEING ABLE TO MANIPULATE REGULATORY T CELL AXIS IN INFLAMMATORY BOWEL DISEASE, WE IDENTIFIED A NUMBER OF CYTOKINES USING THIS MODEL AND CELL SURFACE MOLECULES THAT ARE INVOLVED IN THE FUNCTION OF REGULATORY T CELLS TO BE ABLE TO BOTH PREVENT AND CURE ESTABLISHED COLITIS, AND OF COURSE THOSE PATHWAYS HAVE NOW COME OUT IN THOSE MONOGENIC FORMS OF DISEASE. SO THE MOUSE MODELS ARE HIGHLY SIMPLIFIED, AND YOU’RE LOOKING AT AN ASPECT OF INFLAMMATORY BOWEL DISEASE, BUT I THINK THAT MAPS ONTO CERTAIN MONOGENIC FORMS OF DISEASE THAT ARE ALSO VERY SEVERE. SO OFTEN YOU WILL HEAR OF THE MOUSE MODELS THEY ARE NOT PREDICTIVE. THEY DON’T REPRESENT ALL THE ASPECT AND THE CHALLENGE IS TO MAPPING DIFFERENT MOUSE MODELS TO DIFFERENT POPULATIONS, HUMAN SUBSETS OF INFLAMMATORY BOWEL DISEASE. IT IS WORTH JUST NOTING THAT CHECKPOINT BLOCKADE INDUCED COLITIS WITH ANTI-CTLA-4 IS PROBABLY HIGHLY RELEVANT TO WHAT WAS UNRAVELED IN THESE MOUSE MODELS. SO FAST FORWARDING NOW TO WHERE WE ARE, THERE’S A LOT OF INTEREST IN UNDERSTANDING HOW DIFFERENT SORTS OF BACTERIA ARE ABLE TO SELECT DIFFERENT SUBSETS OF T CELLS. SO HOW PARTICULAR TYPES OF CLOSTRIDIA, FOR EXAMPLE, FROM KENYA HONOR DA’S WORK WILL DRIVE REGULATORY T CELLS, DAN LIPPMAN DEFINING SFB, A TYPE OF BACTERIA PROMOTING Th17 AXIS AND HOW THESE CHANGES IN THE INTESTINE MAY HAVE SYSTEMIC SEQUELAE IN VARIOUS CHRONIC INFLAMMATORY DISEASES. SO THE MODEL THAT WE HAVE USED OVER SOME YEARS NOW IS INFECTION WITH THE MOUSE PATHOGEN HELICO BACTERIA, GRAM NEGATIVE THAT COLONIZES CECUM AND COLON, LIVES IN THE GEL MUSOUS NICHE ESTABLISHING LIFELONG INFECTION WHEN THERE’S A MALADAPTATION IN THE HOST AND PARTICULARLY WHEN THERE IS A DEFICIENCY IN IL-10, THIS CAN LEAD TO IMMUNE PATHOLOGY. SO WHAT WE FIND IS NORMAL MICE INFECTED WITH HELICOBACTER HEPATICUS, DAN LIPPMAN TRACKS T CELLS SHOWING HIGH FREQUENCY OF HELICOBACTER REACTIVE REGULATORY T CELLS, IN THE ADAPTED NORMAL SETTING. IF WE NOW MAKE THIS MALADAPTIVE BY TAKING OUT THE IL-10, ADMINISTERING AN ANTI-IL-10 RECEPTOR, THEN WE SEE NOW THAT WE GET CHRONIC INTESTINAL INFLAMMATION, AND THAT IS HIGHLY DEPENDENT ON THE PRODUCTION OF INTERLEUKIN 23. SO WHAT WE’RE FINDING IS THAT CERTAIN SORTS OF MALADAPTATION MAY HAVE PARTICULAR DRIVERS OF PATHOGENESIS. AND WHEN WE HAVE A RELEVANT MICROBIAL DRIVER HERE IN THE FORM OF HELICOBACTER, WITH AN IL-10 DEFICIENCY, THAT GIVES RISE TO A HIGH AMOUNT OF IL-23 AND IL-23-DRIVEN INTESTINAL INFLAMMATION. THIS MODEL IS IN FACT ANTI-TNF RETRACTORY AND DOESN’T RESPONSE WELL TO ANTI-TNF AS IT DOES TO ANTI-IL-23, THAT’S SOMETHING I’LL COME BACK TO. OVER SOME YEARS, WE HAVE PUT FORWARD A MODEL IN WHICH THERE IS A CRUCIAL BALANCE BETWEEN THE PRODUCTION OF INTERLEUKIN 10 AND INTERLEUKIN 23, THAT CONTROLS THIS RESPONSE. SO IN THE NORMAL SETTING, HELICOBACTER INDUCES A HIGH AMOUNT OF IL-10, AND THIS MIGHT BE AN IMPORTANT MUTUALISTIC RESPONSE. IT DOESN’T INDUCE IMMUNE PATHOLOGY BECAUSE IT’S DRIVING A PARTICULAR TYPE OF RESPONSE FROM THE HOST. WHEN THAT RESPONSE IS ABSENT, HELICOBACTER NOW DRIVES A HUGE AMOUNT OF INTERLEUKIN 23, AS WELL AS INTERLEUKIN 1 FROM MONOCYTES AND MACROPHAGES, AND THAT DRIVES ACCUMULATION AND INFLAMMATION, WE WANT TO UNDERSTAND HOST AND BACTERIAL FACTORS THAT CONTROL THIS BALANCE. OF COURSE, ANTI-IL-23 IS NOW COMING ALONGSIDE ANTI-TNF IN THE CLINIC FOR THE TREATMENT OF CERTAINLY PSORIASIS AND NOW THERE ARE STUDIES LOOKING AT IL-23 BLOCKADE IN INFLAMMATORY BOWEL DISEASE. SO I WANT TO TELL YOU ABOUT THREE NEW THINGS TODAY. ONE IS BACTERIAL MEDIATORS THAT PROMOTE THE REGULATORY T CELL AXIS, NEW INNATE INFLAMMATORY CHECKPOINT AND PATHWAY OF ANTI-TNF THERAPY RESIS RESISTANCE THAT MAY BE RELEVANT TO A SUBSET OF PATIENTS. HELICOBACTER INDUCES STRONG RESPONSE, HOW? A TALENTED MICROBIOLOGIST IN THE LAB ASKED THE QUESTION WHETHER HELICOBACTER MAKES ANYTHING THAT IMPACTS ON MACROPHAGES, WHICH OF COURSE ARE SENTINEL CELLS IN THE INTESTINE THAT PROMOTES THE IL-10 AXIS, HELICOBACTER SEEMS TO DRIVE IL-10 IN VIVO WITH A PHYSIOLOGICAL RELEVANCE. SO WHAT CAMILLE DID WAS TO GENERATE CULTURE SUPERNATANTS OF HELICOBACTER AND ADD THIS TO BONE MARROW DERIVED MACROPHAGES AND FOUND IT PRODUCE AND ACTIVITY THAT LED TO HIGH AMOUNT OF IL-10, COMPARED TO THE PRO-INFLAMMATORY CYTOKINE INTERLEUKIN 6. SHE WENT ON TO CHARACTERIZE THIS AS PRESENTED EVIDENTS THAT THIS IS A POLYSACCHARIDE, IT FUNCTIONS THROUGH SIGNALING THROUGH TLR 2. WHAT WAS DIFFERENT IS THE HELICOBACTER POLYSACCHARIDE FUNCTIONED DIFFERENTLY THAN AGONIST SUCH AS PAM 2 AND PAM 3, SO CAMILLE WENT ON TO LOOK AT SOME OF THE DOWNSTREAM SIGNALING COMPONENTS THAT ARE ACTIVATED FOLLOWING HELICOBACTER POLYSACCHARIDE STIMULATION VERSUS LPS OR TLF 4 AGONIST OR PAM3, A PURE AGONIST, AND FOUND THE SUPERNATANT, WELL, THIS IS A PURIFIED ENRICHED POLYSACCHARIDE ACTIVITY, PREFERENTIALLY STIMULATED A PARTICULAR SIGNALING PATHWAY DOWNSTREAM OF TLR2 INVOLVING A NUMBER OF KINASES AND ACTIVATING THE KREB TRANSCRIPTION FACTOR, PATHWAY PREVIOUSLY SHOWN TO LINK TO INDUCTION OF IL-10 AND OTHER NEGATIVE ANTI-INFLAMMATORY REGULATORY PATHWAYS. SO THIS SIGNALING THROUGH TLR2 WAS DIFFERENTIALLY INDUCING A PARTICULAR SIGNALING CASCADE PAM3 DID NOT SUGGESTING ANOTHER RECEPTOR COULD BE INVOLVED. RNA SEQUENCING OF THE DIFFERENTIAL — ACTUALLY THIS MAY BE A MICROARRAY ANALYSIS OF DIFFERENTIAL GENE EXPRESSION FROM THESE MACROPHAGES STIMULATED WITH THE POLYSACCHARIDE OR PAM3, TLR2 LIGAND, SHOWED ALSO IN ADDITION TO IL-10 VERSUS IL-6 RATIO THAT THIS POLYSACCHARIDE DIFFERENTIALLY INDUCED THE EXPRESSION OF A NUMBER OF GENES THAT WERE ASSOCIATED WITH A PROMINENT KREB BINDING SIGNATURE, SPECIAL IN TERMS OF INFLAMMATORY MOLECULES AS WELL AS TISSUE REMODELING. WE THINK THIS IS AN EXAMPLE OF MODEL PRODUCED BY HELICOBACTER THAT COUPLES INFLAMMATORY RESPONSE FOR ANTI-INFLAMMATORY AND REPAIR PATHWAY. IT DOES THIS THROUGH PROMOTING CREB PHOSPHORYLATION AND DRIVING DOWNSTREAM GENES ASSOCIATED WITH A PARTICULAR FUNCTIONAL RESPONSE THAT MAY RELATE TO MUTUALISTIC RELATIONSHIP WITH HELICOBACTER. IMPORTANT OUTSTANDING QUESTIONS, IS THERE ANOTHER RECEPTOR HERE, AND THIS MAY BE A COMMON PATHWAY OF COMMENSAL SIGNALING THROUGH TLR2 THAT PROMOTES DIFFERENTIAL GENE EXPRESSION, AND OF COURSE THERE ARE SIMILARITIES THEN TO POLYSACCHARIDE A, IDENTIFIED BY CASPER THAT SIGNALS THROUGH TRL2 SHOWN TO PROMOTE REGULATORY T CELLS, THERE IS SOME THOUGHT THERE MAY BE OTHER RECEPTORS INVOLVED, TOGETHER WITH THIS AND THAT MAY BE COMMON THEME, THIS MAY BE A MECHANISM OF MUTUALISM, AND OF COURSE UNDERSTANDING THE CREB PATHWAY IN TERMS OF INTESTINAL HOMEOSTASIS IS THE NEXT STEP. SO I WANT TO TURN NOW TO EXECUTION OF A HOST PATHWAY THAT MODULATES THE INFLAMMATORY RESPONSE TO HELICOBACTER. NOW, THIS IS A RESPONSE NOT IN A LYMPHOCYTE REPLETE MOUSE BUT PARTICULAR STRAIN OF IMMUNE DEFICIENT MICE THAT HAVE RAG DEFICIENCY, AND WHAT WE FOUND SOME YEARS AGO IS THAT THE 129 STRAIN OF RAGS BY CONTRAST WITH FOR EXAMPLE BLACK 6 RAG FOLLOWING HELICOBACTER HEPATICUS INFLECTION DEVELOPS CHRONIC INTESTINAL INFLAMMATION, INVOLVES MANY SIMILAR PATHWAYS YOU SEE IN A T CELL DEPENDENT RESPONSE, SO IT IS NEUTRAL ITSED WITH ANTI-IL-23, BUT OF COURSE THERE ARE NO T CELLS IN HERE. AND THAT LED US TO FURTHER CHARACTERIZE IL-23 RESPONSIVE CELL IN THIS SETTING WHICH IS AN INNATE LYMPHOID CELL, IT EXPRESSES TRANSCRIPTION FACTORS RELATED TO TH-17 CELLS BUT DOESN’T HAVE A T-CELL RECEPTOR BUT HAS HIGH — THESE CELLS ARE PRESENT IN THE INTESTINE, EXPRESS HIGH AMOUNTS OF IL-23 RECEPTOR, AND CAN RESPONSE TO IL-23 PROMPTLY WITH PRODUCTION OF A NUMBER OF INFLAMMATORY CYTOKINES. NOW, IN THE BLACK 6 STRAIN, THIS RESPONSE DOES NOT LEAD TO INTESTINAL INFLAMMATION. BUT IN THE 129 STRAIN, IT LEADS TO CHRONIC COLITIS, WHICH WILL ULTIMATELY PROGRESS TO COLORECTAL CANCER. NOW, IF WE DEPLETE THESE INNATE LYMPHOID CELLS, BASED ON MARKERS THAT ARE PRESENT ON THEIR SURFACE, THIS INHIBITS THE DEVELOPMENT OF THIS DISEASE. SO, THERE’S A GENETIC SUSCEPTIBILITY HERE TO HELICOBACTER-DRIVEN INNATE COLITIS, AND THIS WAS ALSO A MODEL FOR US TO IDENTIFY INNATE LYMPHOID CELLS AND NEW TYPE PRESENT AT BARRIER SURFACES THAT CAN CONTRIBUTE TO IMMUNE PATHOLOGY. OF COURSE, THERE HAVE BEEN A LARGE NUMBER OF STUDIES CHARACTERIZING THESE INNATE LYMPHOID CELLS, THERE ARE DIFFERENT SUBSETS OF INNATE LYMPHOID CELLS, I’M NOT GOING TO GET INTO ALL OF THESE. WHAT’S VERY INTRIGUING ABOUT THESE CELLS IS THAT THEY ARE PRESENT IN SMALL NUMBERS AND THEY ARE ABLE TO AMPLIFY THEIR FUNCTION. SO OUR WORK TOGETHER WITH MANY OTHER LABS IDENTIFIED THESE SUBSETS OF INNATE LYMPHOID CELLS THAT CAN SECRETE CYTOKINES, IL-3 IN RESPONSE TO IL-23 WHICH CAN BE HOST PROTECTIVE BUT WHEN THEY BECOME DERANGED CAN BE IMMUNE PATHOLOGICAL. FOR SOME REASON, IN THIS 129 STRAIN, THIS RESPONSE WAS LEADING TO HELICOBACTER-DRIVEN IMMUNE PATHOLOGY. SO WE WANTED TO MAP THE GENETIC COMPONENT THAT WAS INVOLVED, THE GENETIC SUSCEPTIBILITY TO HELICOBACTER-DRIVEN INNATE COLITIS, SO WE DID A CLASSICAL BACK-CROSSING ANALYSIS, THE 129 STRAIN IS SUSCEPTIBLE, THE BLACK 6 STRAIN IS RESISTANT, AND WE IDENTIFIED A REGION ON CHROMOSOME 3 THAT WE IDENTIFIED AS THE HICKS LOCUS, AND THIS WAS WORK OF OLIVIER BOULLARD IN THE LAB THAT COULD PROTECT THE 129 WHEN YOU HAVE THIS BIT OF B6 FROM THE DEVELOPMENT OF HELICOBACTER-DRIVEN INNATE COLITIS. SO, THE PROTECTIVE EFFECT WITH B6 ALLELE IS DOMINANT, I’M NOT SHOWING YOU ALL OF THIS. WE’VE PUBLISHED THIS. THE FUNCTIONAL RESPONSE IS CONTAINED WITHIN HEMATOPOIETIC CELLS, AND WHAT IS IN THIS 1.7 OR SO MEGABASE REGION OF CHROMOSOME 3, WELL, THERE ARE NINE GENES AND FIVE microRNAs. AND WE HAD A LOOK AT WHAT WAS IN THERE, WHAT SORTS OF CELLS WERE EXPRESSING THESE GENES, AND ONE OF THE CANDIDATE GENES THAT WE HAVE BECOME INTERESTED IN, OF COURSE I’M GOING TO TELL YOU A STORY ABOUT THIS, IS THIS ALPHA KINASE, AND THERE ARE 17 NON-SYNONYMOUS SNPs IN K 1 BETWEEN 129 AND B6 STRAIN. BUT FIRST OF ALL JUST LOOKING AT THE CHARACTERISTICS, USING THE CONGENIC STRAINS FOLLOWING HELICOBACTER INFECTION, YOU SEE THIS VERY RAPID MOBILIZATION OF INNATE IMMUNITY, LOTS OF NEUTROPHILS, MONOCYTES, EOSINOPHILS ON THE 129 BACKGROUND AND BY CONTRAST HUGELY STUNTED RESPONSE ON THE CONGENIC, WITH THAT LITTLE BIT OF CHROMOSOME 3. SO GREERA AND NATEIAN WEST PICKED THIS PROJECT UP FROM OLIVIER OBSERVATION, THE 129 HAS A VERY STRONG INTERFERON GAMMA RESPONSE GENE EFFECT EARLY ON COMPARED TO R 17, CONGENIC, AND YOU SEE THIS IN TERMS OF THE PRODUCTION OF INTERFERON GAMMA mRNA. SO WE WANTED TO LOOK AT THIS CANDIDATE GENE, ALPHA KINASE 1, AND THIS IS FROM A FAMILY THAT HAVE THIS ALPHA KINASE DOMAIN, WHICH HAVE REALLY QUITE DIVERSE FUNCTIONS THROUGHOUT THE BODY, ALPHA KINASE 1, THERE REALLY WASN’T VERY MUCH INFORMATION, SOME CELL LINE WORK SUGGESTING IT’S INVOLVED IN ACTING IN THE SIGH CO-SKELETON. A COUPLE RECENT PAPERS, IN RESPONSE TO HELICOBACTER PYLORI IN CELL LINES, OUT K-1 PLAYS AN IMPORTANT ROLE IN PROMOTING THIS PARTICULAR INNATE SIGNALING COMPLEX, LEADING TO INDUCTION OF GENES SUCH AS IL-8, SOME HOST PROTECTIVE FUNCTION OF ALPK1 IN EPITHELIAL CELL LINES FROM THE PREVIOUS LITERATURE. WE WERE THINKING IT MAY BE A SUSCEPTIBILITY GENE. IF COLLABORATION WITH SANGER INSTITUTE WE PRODUCED A KNOCKOUT OF ALPK1 ON ALSO BLACK 6 BACKGROUND,ING RESISTANT TO COLITIS, WE FOUND THAT MOUSE COMPARED TO THE HETEROZYGOTE DEVELOPS A MUCH MORE SEVERE HELICOBACTER DRIVEN INNATE INFLAMMATORY RESPONSE. WE CROSSED THAT ONTO THE LYMPHOCYTE, REPLETE SETTING, NOW TO LOOK AT THE ADAPTIVE MODEL OF COLITIS, WHERE WE HAVE HELICOBACTER INFECTION, AND NEUTRALIZATION OF INTERLEUKIN 10, AND WHAT WE FOUND IN THAT SETTING IS THAT WHEN YOU HAVE ALPK1 DEFICIENCY TOGETHER WITH HELICOBACTER INFECTION, AND IL-10 BLOCK AID, YOU HAVE A MARKEDLY WORSE COLITIS. THIS IS BASED ON COLONOSCOPY SCORE BUT ALSO HISTOLOGICALLY. IN FACT, THAT ASSOCIATES WITH THIS VERY MARKED SKEWING TOWARDS Th1 CELLS. SO WHEN YOU HAVE ALPK 1 DEFICIENCY AND HELICOBACTER INFECTION, DRIVER INTRODUCING, TAKING AWAY IL-10, ALPK1 IS AN INDEPENDENT REGULATOR BECAUSE THE DISEASE IS MARKEDLY WORSE WITH A SIGNIFICANT SKEWING OF THE RESPONSE TOWARDS A TH1 PHENOTYPE AWAY FROM IL-17 WHICH MAY IN SOME SETTINGS HAVE A HOST PROTECTIVE ROLE. NOW, ALPK1 MICE GIVEN HELICOBACTER HEPATICUS DO NOT DEVELOP OVERT COLITIS BUT CLEARLY ARE NOT NORMAL, THEY HAVE A HIGH LEVEL OF TH1 CELLS FOLLOWING HELICOBACTER INFECTION ALONE, ALTERING THE NATURE OF THE MICROBIAL SIGNALING, HOW THAT LEADS TO DIFFERENTIATION OF T CELLS FAVORING TH1 RESPONSE. SO THIS EFFECT OF ALPK1 IS TRANSFERRED WITH BONE MARROW CELLS, THE SAME AS WE FIND WITH THE CONGENIC REGION, WE THINK THIS IS THE RELEVANT GENETIC LESION. SO WE ASKED THE QUESTION, IS THERE A CELL INTRINSIC ROLE IN THE FUNCTION OF ALPK1 IN BONE MARROW IN THIS SITUATION, IN BONE MARROW DERIVED MACROPHAGES WHICH WE GROW UNDER CONDITIONS AND STIMULATED WITH HELICOBACTER HEPATICUS, RED IS ALPK1 DEEFFICIENT, BLUE IS HETEROZYGOUS, THERE’S A VERY STRONG SKEWING TOWARDS INTERLEUKIN 12. SO THE HICKS LOCUS ON CHROMOSOME 3 CONTROLS THE INNATE COLONIC INFLAMMATORY RESPONSE, SUSCEPTIBLE 129 STRAIN ACTIVATE A VERY EARLY INTERFERON GAMMA AND INTERFERON RESPONSE GENES WITH RAPID RECRUITMENT OF MYELOID CELLS, ALPK1 WE BELIEVE IS THE RELEVANT GENE THAT IS INVOLVED IN THIS LOCUS, AND ALPK1 KNOCKOUT MICE SHOW AN ENHANCED TH1 RESPONSE IN BOTH INNATE AND ADAPTIVE MODELS OF COLITIS, FUNCTION IN HEMATOPOIETIC CELLS TO CONTROL THE INFLAMMATORY RESPONSE AND HAS AN IMPORTANT CELL INTRINSIC ROLE IN MACROPHAGES. SO, WHEN THERE IS HELICOBACTER STIMULATION OF THE INNATE IMMUNE SYSTEM, THIS DOESN’T LEAD TO IMMUNE PATHOLOGY, BECAUSE THE RESPONSE IS CONTROLLED BY ALPK1. WHEN THIS IS REMOVED, THIS LEADS TO A HIGH AMOUNT PARTICULARLY OF IL-12 AND SUBSEQUENTLY THEN THE Th1 RESPONSE. AND THIS IS AN INDEPENDENT CHECKPOINT FROM THE PRODUCTION OF IL-10, SO HELICOBACTER IS úONE, IT’S ACTIVATING A RESPONSE THAT IS NORMALLY CONTROLLED BY ALPK1, BUT IT’S ALSO ACTIVATING HIGH AMOUNTS OF INTERLEUKIN 10, AND IF WE TAKE AWAY THESE TWO ADAPTATIONS, THEN NOW WE HAVE A CHRONIC INFLAMMATORY RESPONSE. SO WHAT IS THE ROLE OF ALPK1 IN PRIMING VERSUS EFFECTOR FUNCTION? AND OF COURSE HOW IS THIS RELATED TO HUMAN AUTOIMMUNE AND INFLAMMATORY DISORDERS. SO I WANT TO FINISH NOW WITH BRINGING THIS BACK AROUND TO SUBSETS OF HUMAN PATIENTS, AND HOW WE CAN ITERATE BETWEEN MOUSE MODELS AND THE HUMAN DISEASE. OF COURSE, WE HAVE AN INCREASINGLY WELL STOCKED TOOLBOX FOR BEING ABLE TO INTERVENE IN INFLAMMATORY BOWEL DISEASE ANTI-TNF, IL-23, LOTS OF OPPORTUNITIES. BUT THE MAJOR CHALLENGE IS WE DON’T KNOW WHICH ONES TO APPLY TO WHICH PATIENTS. SO DEVELOPING STRATEGIES TO PREDICT THERAPEUTIC OUTCOME WILL BE CRITICAL FOR DELIVERY OF INDIVIDUALIZED PATIENT CARE. AND OF COURSE THERE IS A BIG PROBLEM IN IBD IN TERMS OF ANTI-TNF NON-RESPONSE. THERE’S PRIMARY NON-RESPONSE. AND OF COURSE SECONDARY NON-RESPONSE, WE DON’T KNOW WHICH PATIENTS WILL DEVELOP THAT. OF COURSE, THERE ARE DIFFERENT POSSIBILITIES, IS THERE GENETIC PREDISPOSITION, IS IT JUST THE STATE OF DISEASE, JUST A KINETIC THING OF WHEN WE COME TO TREAT THEM, OR ARE THERE ACTUALLY DIFFERENT BIOLOGICAL DRIVERS, AND PATIENTS THAT ARE NONRESPONSIVE TO ANTI-TNF ACTUALLY HAVE A DIFFERENT PROCESS THAT IS DRIVING THE DISEASE. SO, NATHAN WEST AND ARMAND AGATZI IN THE LAB UNDERTOOK ANALYSIS OF TRANSCRIPTOMIC DATASETS FROM ULCERATIVE COLITIS AND CROHN’S DISEASE PATIENTS TO IDENTIFY CYTOKINES THAT WERE HIGHLY ELEVATED IN BOTH OF THOSE DISEASES. AND THEY BROUGHT THE LIST DOWN TO SIX. SOME OF THEM WE KNOW A LOT ABOUT, INTERLEUKIN 1 AND IL-6, WE DIDN’T FOLLOW THOSE BUT ACTUALLY TOP OF THE LIST WAS A CYTOKINE CALLED ONCOSTATIN M, THIS HAS BEEN SOMEWHAT OVERLOOKED, NOT CALLED INTERLEUKIN, IT’S BEEN AROUND A LONG TIME BUT THERE HASN’T BEEN VERY MUCH INVESTIGATION OF OSM IN TERMS OF GUT IMMUNITY. SO OSM IS A MEMBER OF THE IL-6 FAMILY, IT SIGNALS THROUGH HETERODIMERIC AND SIGNALING PATHWAYS DOWNSTREAM OF RECEPTOR PARTICULARLY THROUGH JAK 1 AND 2. A LOT OF INFORMATION IN TERMS OF BIOLOGY, FUNCTIONS IN THE LIVER, IN THE ACUTE PHASE RESPONSE, ASPECTS OF HEMATOPOIESIS, IT’S BEEN IMPLICATED ACROSS A RANGE OF DISEASES BUT NOT MUCH KNOWN ABOUT THE GUT. WE’VE INTERROGATED DIFFERENT PUBLICLY AVAILABLE TRANSCRIPTOMIC COHORTS. THIS IS THE PEDIATRIC RISK COHORT, LOOKING AT PRIMARY IBD, AND WHAT WE FOUND THERE IN THEIR DATASET IS THAT OSM IS ONE OF THE MOST HIGHLY DIFFERENTIALLY EXPRESSED IN IBD VERSUS CONTROL MUCOSA WITH HIGH STATISTICAL SIGNIFICANCE. SO WE GET BACK TO OUR OWN IBD COHORT, IN OXFORD, TO LOOK AT OSM AND ITS RECEPTOR. AGAIN, YOU SEE SIGNIFICANT INCREASE IN OSM GENE EXPRESSION AND ITS RECEPTOR IN SEVERELY ACTIVE IBD. SO WE WANTED TO ASK THE QUESTION, IS THERE ANY IMPACT OF OSM IN WHETHER OR NOT PATIENTS WILL RESPOND TO ANTI-TNF. THIS IS OUR DISCOVERY COHORT FROM THE ACT 1 PHASE 3 TRIAL OF ANTI-TNF ANTIBODY, THESE PATIENTS ARE REFRACTORY TO STANDARD THERAPY, COLON MUCOSAL BIOPSIES COLLECTED PRIOR TO TREATMENT WITH ANTI-TNF AND THEN THE PATIENTS ARE TREATED AND THOSE ARE HISTOLOGICAL ANALYSIS AND ENDOSCOPIC SCORING, AND WE FOUND PATIENTS THAT WERE NONRESPONSIVE TO ANTI-TNF HAD HIGH AMOUNTS OF OSM. IN FACT THIS WAS HIGHLY PREDICTIVE IN TERMS OF SPECIFICITY AND SENSITIVITY IN IDENTIFYING ANTI-TNF NON-RESPONDERS. THIS IS ANOTHER PATIENT COHORT HERE, YOU SEE PATIENTS AND HIGHER EXPRESSION OF OSM AND ITS RECEPTOR IN THE REFRACTORY PATIENTS, AND THAT RESPONSE, ALTHOUGH OSM IN THE HIGHESTLY DIFFERENTIALLY EXPRESSED, IT COMES WITH A MODULE OF INFLAMMATORY MEDIATORS, RELATED TO A NUMBER OF CHEMOKINES AND CYTOKINES. AND WHEN THERE ARE PATIENTS THAT WERE MODULE HIGH, 90% OF THOSE WERE UNRESPONSIVE TO ANTI-TNF THERAPY. BY CONTRAST THE MODULE LOW PATIENTS, THE MAJORITY OF THOSE WERE ANTI-TNF RESPONSIVE. SO WE VALIDATED THAT ACROSS A NUMBER OF DIFFERENT COHORTS. THIS JUST SHOWS YOU IN OUR OXFORD-BASED COHORT THAT THERE IS AN INCREASE IN T CELLS AND NON-T CELLS THAT ARE PRODUCING OSM IN IBD. SO SEEMED TO BE A LOT OF DIFFERENT CELL TYPES THAT CAN PRODUCE OSM, INCLUDING T CELLS AND MYELOID CELLS, BUT THE CELLS THAT RESPOND TO OSM IS HIGHLY RESTRICTED, AND THAT APPEARS TO BE PRIMARILY TO MESENCHYMAL FIBROBLAST-TYPE CELLS, AND ENDOTHELIAL CELLS. SO THIS IS JUST GATING CELLS FROM THE HUMAN INTESTINE, INTO STROMAL CELLS OR DIFFERENT COMPONENTS, AND LOOKING AT OSM RECEPTOR EXPRESSION, WHICH IS EXPRESSED ON MESENCHYMAL STROMAL CELLS AS WELL AS ENDOTHELIAL CELLS, AND INDEED ONE SEES THE STROMAL CELLS MOUNT A VERY ROBUST RESPONSE TO ONCOSTATIN M WITH EXPRESSION OF NUMBER OF DOWNSTREAM SIGNALING PATHWAYS WHICH IS DISTINCT FROM IL-1 OR IL-6 FOR EXAMPLE. IS THERE ANYTHING SPECIAL ABOUT OSM RECEPTOR HIGH FIBROBLASTS? AGAIN, LOOKING IN THE HUMAN INTESTINE WE CAN SEE THESE OSM RECEPTOR HIGH OR LOW CELLS, AND IF YOU GATE ON THE OSM RECEPTOR HIGH CELLS, THEN THEY ARE CELLS THAT EXPRESS A NUMBER OF MARKERS OF STROMAL ACTIVATION, INCLUDING CERTAIN INTEGRINS AND THE MOLECULE GP 38. SO IF WE LOOK AT OSM RECEPTOR HIGH, VERSUS LOW CELLS, THERE ARE SOME THINGS THAT SIMILARLY EXPRESS BUT THERE’S A PARTICULAR SUBSET OF CYTOKINES AND ADHESION MOLECULES HIGHLY EXPRESSED IN OSM RECEPTOR HIGH POPULATION, IN INFLAMMATORY BOWEL DISEASE. SUGGESTING THAT OSM IS SIGNALING THROUGH THESE STROMAL CELLS, AND CREATING A TYPE OF STROMAL ACTIVATION THAT MAY CONTRIBUTE TO DISEASE WITH THESE INFLAMMATORY MEDIATORS. SO FINALLY, IS THERE A ROLE FOR ONCOSTATIN M COMING BACK TO OUR MOUSE MODEL ANTI-TNF RESISTANT? INDEED IF WE INDUCE HELICOBACTER HEPATICUS WITH IL-10 BLOCKADE IN THE SETTING OF OSM DEFICIENCY COMPARED TO WILDTYPE MICE, THEN THERE IS AN ATTENUATED DISEASE. WHAT’S RELEVANT HERE IS THAT’S NOT REALLY IN THE INDUCTION AND DEVELOPMENT OF THE DISEASE, BUT MORE IN THE PERPETUATION. SO WE THINK THAT OSM IS SIGNALING, ITS FUNCTION IS COMING LATER, LATER ON. SO OSM THEN IS A PREVIOUSLY UNRECOGNIZED MEDIATOR OF INTESTINAL INFLAMMATION AND POTENTIAL BIOMARKER OF ANTI-TNF NONRESPONSIVENESS. WE BELIEVE IT ACTS THROUGH ACTIVATING STROMAL CELLS, CONSTITUTING A NOVEL SIGNALING AXIS BETWEEN LEUKOCYTES AND FIBROBLASTS, THIS MAY HAVE RELEVANCE IN A NUMBER OF DIFFERENT DISEASES, IT MAY BE A KEY HIGH LEVEL DRIVER OF IBD CHRONICITY AND SEVERITY BUT IS PARTICULARLY RELEVANT TO ANTI-TNF REFRACTORY PATIENTS. SO WHAT WE THINK IS HAPPENING WHEN THERE IS A GENETIC LANDSCAPE WITH A RELEVANT MICROBIAL DRIVER, THERE IS A DYSREGULATESSED INFLAMMATORY RESPONSE, HIGH AMOUNTS OF INTERLEUKIN 23, IL-1, TNF, BUT IN SOME PATIENTS THERE’S ALSO A REMODELING OF THE STROMA THAT INVOLVES OSM, WHICH MIGHT HAVE AN IMPORTANT ROLE IN SUSTAINING THE INFLAMMATORY RESPONSE THROUGH A NUMBER OF DIFFERENT CHEMOKINES AND ADHESION MOLECULES THAT IMPACT ON WHAT IS RECRUITED AND RETAINED IN THE INFLAMMATORY LESION. SOMETHING WE’RE LOOKING AT. SO I WANT TO FINISH HERE, AND COME BACK TO THIS CONCEPT OF A PROCESS-DRIVEN TAXONOMY OF DISEASE, WHETHER WE MAY BE ABLE TO PULL OUT PATHWAYS THAT ARE RELEVANT ACROSS DIFFERENT DISEASES, AND TO BE ABLE TO TREAT THE PROCESS AS OPPOSED TO THE CLINICAL SYMPTOMS. SO OF COURSE, ALL OF THIS WORK IS THE OUTPUT AND ATTRIBUTABLE TO THE WONDERFUL POSTDOCS AND STUDENTS WHO HAVE COME TO MY LAB OVER THE YEARS. I’VE MENTIONED THE INDIVIDUALS WHO CONTRIBUTED TO THIS WORK. WE HAVE EXCELLENT COLLABORATION WITH OUR COLLEAGUES IN GASTROENTEROLOGY, SIMON TRAVIS AND OTHERS, IN OUR STUDIES ON OSM WE HAD A VERY STRONG COLLABORATION WITH SCOTT PLEVY AND COLLEAGUES AND MADE AVAILABLE CLINICAL TRIALS NOT PUBLISHED IN DATA WE COULD USE TOWARDS UNDERSTANDING THE OSM PATHWAY, AND SPONSORS THAT SUPPORT OUR WORK. THANK YOU VERY MUCH. [APPLAUSE] >>THANKS FOR A FASCINATING PRESENTATION. WE DO HAVE TIME FOR QUESTIONS, AND AS USUAL PLEASE APPROACH THE MICROPHONES AS PEOPLE ARE ALREADY DOING SO THOSE LISTENING BY VIDEO CAN HEAR. WE’LL START OVER HERE.>>FIONA, VERY INTERESTING STORY. IN THE CASE OF THE TLR 2 LIGAND FROM HELICOBACTER HEPATICUS THAT DIFFERED FROM PAM 3, IT FORMS TWO HETERODIMERS ARE TLR 2 AND 6 THERE COULD BE TWO SIGNALING PATHWAYS FOR TWO HETERODIMERS, DID YOU GET A CHANCE TO LOOK AT WHETHER THAT ACCOUNTS FOR DIFFERENCE IN WHAT YOU’RE SEEING?>>WE WERE ABLE TO USE AN ANTIBODY ACTUALLY THAT PINNED IT DOWN TO ONE OF THOSE AND THEN THE PAM3 IS THE RELEVANT TLR2 SIX HETERODIMER, I DON’T THINK IT’S JUST THE TYPE OF TLR2 MAKEUP OF THE SUBUNIT STRUCTURE. I THINK THERE’S ANOTHER RECEPTOR HERE. I THINK IT’S HIGHLY LIKELY, WHICH IS WHY THE LIPO PEPTIDE IS NOT GETTING THE SAME SIGNALING PATHWAYS AS THE WHOLE BACTERIA.>>FIONA, THIS IS A VERY FASCINATING STORY, AND WOULD PREDICT THAT IN THOSE CASES WHERE PATIENTS ARE TNF UNRESPONSIVE THEY WOULD RESPOND TO BLOCKING THE ONCOSTATIN M PATHWAY, ARE THESE TRIALS GOING ON? HOW DO YOU BLOCK THE ONCOSTATIN M PATHWAY.>>THE ANSWER IS HAVE AN ANTI-OSM ANTIBODY, FOR EXAMPLE. THE THING AGAIN TO GET IT ON THE AGENDA, OF COURSE IBD IS A BUSY SPACE. SO, YOU KNOW, IT’S DIFFICULT TO GET NEW TRIALS IN IBD ACTUALLY BECAUSE THERE’S A HIGH FAILURE RATE AND BECAUSE WE CAN’T PREDICT THE PATIENTS. YES, I WOULD BE OPTIMISTIC EVENTUALLY WE’LL BE ABLE TO MOVE FORWARD AND TEST OUR HYPOTHESIS HERE WHICH IS ALL WE REALLY IS HAVE IN PATIENTS. I THINK IT LOOKS AN INTERESTING ONE TO GO FOR AND SOMETHING THAT NOT MANY STRATEGIES OUT THERE ARE TARGETING THE STROMAL COMPARTMENT. IT’S NOT A NOVEL CONCEPT BUT TO HAVE A CYTOKINE THAT DIRECTLY IS INVOLVED IN ACTIVATING THAT GIVES A WAY TO INTERVENE.>>REALLY INTERESTING. REGARDING ONCOSTATIN M, IF I REMEMBER PROPERLY, CAN SIGNAL THROUGH TWO DIFFERENT RECEPTORS, EITHER GP 130 WITH THE ONCOSTATIN M RECEPTOR OR GP 130 WITH LIF RECEPTOR.>>YES.>>WHAT IS THE EXPRESSION OF THE LIF RECEPTOR LIKE ON THE INTESTINAL CELLS AS WELL IN IBD?>>YEAH, THAT WAS ONE OF OUR REVIEWER QUESTIONS. [LAUGHTER] SO WE DID GO BACK TO LOOK AT LIF ON OUR STROMAL CELLS. WE DON’T SEE THE LIF RECEPTOR THERE, YEAH.>>OKAY.>>A LOT OF IMMUNOLOGICAL FACTORS HAD TO GO THROUGH THE PORTAL SYSTEM INTO THE LIVER, THERE’S NO HYPOTHESES DEVELOPING IN TERMS OF ETIOLOGY OF NON-ALCOHOLIC FATTY LIVER DISEASE, CYTOKINES AND BACTERIAL PROJECTS. DO YOU HAVE ANYTHING>>I MISSED THE FIRST PART.>>WHEN YOU HAVE BACTERIAL PRODUCTSI IMMUNOLOGIC THINGS GOING ON IN THE INTESTINES, THEY GO THROUGH THE PORTAL, THEY EVENTUALLY GO TO THE LIVER. IN TERMS OF, IN CALIFORNIA, IF THIS COULD BE ETIOLOGY OF NON-ALCOHOLIC FATTY LIVER DISEASE AMONG THE OTHER HYPOTHESES NOW OFFERED. DO YOU HAVE ANY CORRELATION OF THAT WITH WHAT YOU’RE LOOKING AT?>>NO, WE HAVEN’T DONE THAT. IF YOU THINK OF SOMETHING LIKE PRIMARY SCLEROSING COLONGITIS WITH A LIVER PHENOTYPE WITH SOME IBD MANIFESTATIONS, THERE’S A VERY STRONG GUT-LIVER AXIS, THERE’S A MICROBIOME SUPPORTIVE OF THAT AND A LOT OF INTEREST IN UNDERSTANDING THE PATHWAYS BUT WE HAVE NOT LOOKED AT THE LIVER IN THIS CONTEXT.>>OKAY. THANK YOU.>>SO THIS WAS A WONDERFUL TALK AND THIS IS VERY NICE QUESTION, PEAK OF INCIDENCE BY 20 OR MORE WHICH FOLLOWS BY A FEW YEARS, THE START OF INVOLUTION OF THYMUS SUGGESTING THERE MIGHT BE SOMETHING PRODUCED BY THE TIME THAT PREVENTS OCCURRENCE OF DISEASE (INDISCERNIBLE) ARE THOSE DISEASES MORE FREQUENT IN KIDS THAT HAVE HAD THYMUS REMOVED FOR HEART SURGERY WHEN THEY WERE VERY YOUNG?>>YOU KNOW, I DON’T — I HAVEN’T HEARD THAT. I’M NOT AWARE OF THAT. BUT I DON’T KNOW THE SPECIFIC DATA. YEAH, INTERESTING THOUGHT.>>LET ME ASK YOU ONE SORT OF FUTURISTIC QUESTION. SO YOU’RE MAPPING OUT THESE AMAZING PATHWAYS AND PLAYERS, POINTING OUT HOW IMPORTANT IT IS IN THERAPEUTICS TO TRY TO DO THIS IN A FASHION THAT RECOGNIZES DIFFERENCES BETWEEN INDIVIDUALS. SO CAN ONE IMAGINE ONCE WE HAVE THE TECHNOLOGY AND PATHWAY IS BETTER WORKED OUT THAT ONE WOULD PERSONALIZE THE INTERVENTION BASED UPON EVERY BIT OF DATA YOU COULD COLLECT ON THE INDIVIDUAL THAT MIGHT INVOLVE NOT JUST LOOKING IT’S A CIRCULATING CYTOKINES AND IMMUNE CELLS BUT ACTUALLY IN A BIOPSY HAVING TO LOOK TO SEE WHO THE ACTORS ARE AND PERHAPS NOT EVEN BY MEASURING CYTOKINES BUT DOING AS SOME ARE DOING SINGLE CELL BIOLOGY WHERE YOU LOOK AT ALL THE IMMUNE CELLS AND FIND OUT WHICH GENES ARE ON USING RNA-SEQ OF SINGLE CELLS, MAYBE THEN TRY TO FIGURE OUT FOR THAT PATIENT WHAT ARE THE ACTORS THAT YOU MOST WANT TO INTERVENE WITH, IS THAT WHERE YOU THINK THE FIELD MAY BE GOING WITH THEY THERAPEUTIC SELECTION?>>THERE’S A LOT OF EXPERIMENTAL WORK IN HEALTH AND DISEASE, AND THIS IS STARTING TO COME. HOW THIS WILL PLAY OUT INTO CLINICAL PRACTICE I THINK WE PROBABLY NEED MARKERS THAT ARE MORE ACCESSIBLE, SO IN CIRCULATING CELLS. SO A CHALLENGE WILL BE TO LOOK FOR SIGNATURES AND THEN SEE IF WE CAN DETECT THEM SYSTEMICALLY. I THINK THE THING WE’RE MISSING AT THE MOMENT IN THIS IS THE MICROBIOME COMPONENT. BECAUSE WHAT WE’RE FINDING IN THE MOUSE MODELS IS WHERE THERE’S A MALADAPTATION, IL-10 OR AUTOPHAGY DEFICIENCY, DIFFERENT MICROBES SEEM TO INTERFACE WITH THAT. SO THAT’S ANOTHER WAY OF BEING ABLE TO STRATIFY. THAT MIGHT BE MORE ACCESSIBLE AS WELL.>>ESPECIALLY THE WAY THINGS ARE GOING. IT’S BEEN A FASCINATING PRESENTATION THAT BRINGS TOGETHER A LOT OF COOL IMMUNOLOGY AND LEARNING ABOUT THE INTESTINAL TRACT. LET US THANK OUR SPEAKER AGAIN AND HEAD TO THE RECEPTION. [APPLAUSE]

4 thoughts on “Gut reactions: host microbiome interactions in the intestine in health and disease”

  1. Had laparoscopy surgery and developed IBD after. Having major health problems now that seems to be endless. Doctors don't even want to know about it.

Leave a Reply

Your email address will not be published. Required fields are marked *