Coriell Institute For Medical Research Inc

THE NIGMS HUMAN GENETIC CELL REPOSITORY COOPERATIVE AGREEMENT (U42) APPLICATION

EPIGENETIC THERAPIES - NEW APPROACHES - ABSTRACT (OVERALL) EPIGENETICS REFERS TO STABLE GENE EXPRESSION PATTERNS MEDIATED BY DNA METHYLATION AND/OR CHROMATIN REMODELING AND IS INVOLVED IN CELLULAR IDENTITY AND REPRESSION OF SPURIOUS TRANSCRIPTION, INCLUDING FROM REPETITIVE ELEMENTS. OVER THE PAST 20 YEARS, IN WORK LED IN PART BY INVESTIGATORS IN THIS APPLICATION, EPIGENETIC CHANGES WERE RECOGNIZED AS IMPORTANT DRIVERS OF CANCER FORMATION, PROGRESSION AND RESISTANCE TO THERAPY. THIS RECOGNITION, ALONG WITH THE REVERSIBLE NATURE OF THE BIOCHEMICAL MODIFICATIONS REQUIRED FOR EPIGENETICS LED TO THE FIELD OF EPIGENETIC THERAPY, WHICH AIMS TO REPROGRAM GENE EXPRESSION TO ACHIEVE A THERAPEUTIC EFFECT. THIS FIELD, WHICH STARTED WITH DNA METHYLTRANSFERASE (DNMT) INHIBITORS, HAS GROWN TO A DOZEN EPIGENETIC TARGETS AND OVER 30 DRUGS IN CLINICAL TRIALS. FOUR TARGETS HAVE MADE IT TO US-FDA APPROVAL (DNMTS, HISTONE DEACETYLASES (HDACS), EZH2 AND ISOCITRATE DEHYDROGENASES) AND TENS OF THOUSANDS OF CANCER PATIENTS BENEFIT FROM THIS EVERY YEAR. WITH THE IDENTIFICATION OF NEW TARGETS AND THE RECOGNITION THAT EPIGENETICS IS INVOLVED IN SENSITIVITY AND RESISTANCE TO CHEMOTHERAPY AND IMMUNOTHERAPY, THE CLINICAL POTENTIAL OF EPIGENETIC THERAPY HAS BEGUN TO BE EXPLORED IN EARNEST. THERE REMAIN FUNDAMENTAL CHALLENGES, FROM THE LACK OF ROBUST BIOMARKERS OF ACTIVITY, TO THE EMERGENCE OF RESISTANCE, AND TO THE UNEXPLAINED DIVIDE IN RESPONSES BETWEEN HEMATOLOGIC MALIGNANCIES AND SOLID TUMORS. THIS SPORE APPLICATION WILL ADDRESS ALL OF THESE CHALLENGES. THE SPORE TEAM CONSISTS OF INVESTIGATORS WHO ARE PIONEERS IN THE FIELDS OF CANCER EPIGENETICS AND EPIGENETIC THERAPY AND EXPLORES NEW EPIGENETIC TARGETS AND COMBINATION STRATEGIES ALONG WITH A ROBUST BIOMARKER ANALYSIS PIPELINE TO IDENTIFY PATIENTS LIKELY TO RESPOND AND PHARMACODYNAMIC MARKERS OF RESPONSE. THE TEAM LEVERAGES A STRONG CLINICAL AND TRANSLATIONAL PIPELINE BUILT THROUGH THE VAN ANDEL INSTITUTE STAND UP TO CANCER EPIGENETICS DREAM TEAM, WHICH HAS CONDUCTED 14 EPIGENETIC THERAPY CLINICAL TRIALS IN THE PAST FEW YEARS, AND IS FULLY COMMITTED TO THE CLINICAL TRIALS PROPOSED IN THIS APPLICATION. THIS EPIGENETIC THERAPY SPORE ENCOMPASSES FOUR MAJOR THEMES: (I) DEVELOP AND TEST DRUGS AGAINST NEW EPIGENETIC TARGETS (PROJECTS 1, 2), (II) MECHANISTIC AND TRANSLATIONAL STUDIES OF IMMUNOSENSITIZATION BY EPIGENETIC THERAPY (PROJECTS 1-3), (III) STUDIES OF DRUG COMBINATIONS THAT ENHANCE THE EFFICACY OF KNOWN EPIGENETIC DRUGS (PROJECTS 1-3); AND (IV) BIOMARKER STUDIES TO DEFINE SENSITIVITY AND RESISTANCE TO EPIGENETIC THERAPY IN THE CLINIC (ALL PROJECTS). THESE THEMES WILL BE ADDRESSED THROUGH 3 PROJECTS: (I) CYCLIN DEPENDENT KINASES AS EPIGENETIC THERAPY TARGETS; (II) EPIGENETIC SYNERGY BETWEEN DNMT AND EZH1/2 INHIBITORS; (III) LINKING EPIGENETIC-THERAPY INDUCTION OF INFLAMMASOME SIGNALING TO GENERATION OF A BRCANESS PHENOTYPE. THESE PROJECTS WILL BE SUPPORTED BY THREE CORES (ADMINISTRATIVE, PATHOLOGY, GENOMICS) AND A KEY GOAL WILL ALSO BE TO MENTOR THE NEXT GENERATION OF EPIGENETIC THERAPY INVESTIGATORS AND SUPPORT CUTTING-EDGE SCIENCE THROUGH THE CAREER ENHANCEMENT AND DEVELOPMENTAL RESEARCH PROGRAMS.

TAS: :97 0130: :CORIELL PERSONALIZED MEDICINE COLLABORATIVE (CPMC) CLINICAL UTILITY STUDY (CUS)".

CORIELL CELL ENGINEERING CENTER TO SUPPORT NATIONAL BIOBANKING INFRASTRUCTURE - CORIELL INSTITUTE FOR MEDICAL RESEARCH APPLICATION TO ORIP C06 FUNDING OPPORTUNITY ANNOUNCEMENT PAR- 25-061, BIOMEDICAL RESEARCH FACILITIES. PROJECT TITLE: “CORIELL CELL ENGINEERING CENTER TO SUPPORT NATIONAL BIOBANKING INFRASTRUCTURE” PROJECT SUMMARY/ABSTRACT: CORIELL INSTITUTE FOR MEDICAL RESEARCH’S MISSION IS TO CONDUCT AND SUPPORT RESEARCH TO BENEFIT HUMAN HEALTH AND HELP CURE DISEASE. A LEADER IN BIOBANKING, CORIELL HAS HOSTED NIH BIOBANKS SINCE 1972, BUILDING AND DISTRIBUTING WORLD CLASS SAMPLE COLLECTIONS, ENABLING BIOMEDICAL RESEARCH. CORIELL’S BIOBANK IS COMMITTED TO MEETING THE EVOLVING NEEDS OF RESEARCHERS BY ADOPTING NEW TECHNOLOGIES AS WELL AS DIVERSIFYING AND INCLUDING MINORITIES AND UNDERREPRESENTED POPULATIONS IN ITS BIOSPECIMEN COLLECTIONS. CORIELL SCIENTISTS ARE SUPPORTED BY THE NIH, NJ STATE, AND OTHER PARTIES, IN FIELDS INCLUDING BUT NOT LIMITED TO CANCER AND DISEASE RESEARCH, AGING BIOLOGY, BIOBANKING, AND PERSONALIZED MEDICINE. CORIELL’S PRIMARY FACILITY, 403 HADDON AVENUE, CAMDEN, NJ, IS OWNED BY, AND RESIDES ON, THE CAMPUS OF COOPER UNIVERSITY HOSPITAL. THERE IS LIMITED ROOM FOR GROWTH AND PLANS ARE UNDERWAY FOR CORIELL TO RELOCATE TO A NEW HEADQUARTERS BUILDING, ON ITS OWN SITE IN CAMDEN, IN APPROXIMATELY THREE YEARS. IF AWARDED, ORIP C06 FUNDS WOULD HELP CREATE A >7,495 SQUARE FOOT CELL ENGINEERING CENTER ON THE FIRST FLOOR OF CORIELL’S NEW FACILITY, TO ENABLE EXPANSION OF BIOBANKING INFRASTRUCTURE AND PRODUCT AND SERVICE OFFERINGS. OF THESE OFFERINGS, STEM CELLS OFFER UNIQUE INSIGHTS INTO CELL AND DEVELOPMENTAL BIOLOGY, DISEASE ETIOLOGY, AND EXPEDITION OF DRUG DISCOVERY. ADVANCES IN GENE ENGINEERING AND TOOLS LIKE 3D “MINI-ORGAN” MODELS ARE REVOLUTIONIZING RESEARCH. SUCH TOOLS ALSO REPRESENT NEW APPROACH MODELS (NAMS) THAT COMPLEMENT AND REDUCE THE NEED FOR VERTEBRATE ANIMALS IN RESEARCH. THIS CENTER WILL SERVE AS A CORE LABORATORY, OFFERING CANCER CELL LINES, NEWLY CREATED IPSCS, DIFFERENTIATED CELLS, 3D ORGANOID MODELS, CELL GENE ENGINEERING, QUALITY CONTROL, GENOMIC/EPIGENOMIC PROFILING, AND BIOINFORMATIC SERVICES, FOR NIH-FUNDED SCIENTISTS AT CORIELL, IN THE REGION, AND ACROSS THE COUNTRY. THIS PROJECT EPITOMIZES CORIELL’S MISSION TO SUPPORT ROBUST RESEARCH BY PROVIDING SCIENTISTS A NEW GENERATION OF REPRODUCIBLE, HIGHLY CHARACTERIZED, HIGH-QUALITY BIOSPECIMENS AND SERVICES TO ACCELERATE THE PACE OF BIOMEDICAL DISCOVERIES. CORIELL SAMPLES HAVE BEEN INSTRUMENTAL FOR ELUCIDATING MECHANISMS OF DISEASES TO ALLOW NEW REGENERATIVE AND PRECISION MEDICINE THERAPEUTICS. THIS CENTER WILL PROVIDE CRITICAL, AFFORDABLE NEW PRODUCTS AND SERVICES TO SUPPORT RESEARCHERS. AN EXPERIENCED AND COLLABORATIVE TEAM OF DEDICATED PROFESSIONALS, WITH AN EXPERT PRINCIPAL INVESTIGATOR, CONSTRUCTION PROJECT MANAGER, AND FACILITY MANAGER WILL LEAD THIS PROJECT. THIS PROJECT IS PART OF A LARGER NEW BUILDING INVESTMENT WHICH IS ALREADY UNDERWAY WITH AN ANTICIPATED COMPLETION DATE OF 2027-2028, AND CORIELL LEADERSHIP COMMITS TO STAFFING AND SUPPORTING THIS CENTER FOR 10+ YEARS POST-COMPLETION, USING BUSINESS TACTICS DEVELOPED AS AN INDEPENDENT RESEARCH INSTITUTE FOR OVER 70 YEARS. IN ADDITION TO INSTITUTIONAL, LOCAL AND REGIONAL BENEFITS, THIS PROJECT WILL HAVE A SUBSTANTIAL NATIONAL IMPACT. THE CENTER WILL INCREASE CURRENT CAPABILITIES >20 FOLD, SUPPORT UP TO 30 SCIENTISTS/TECHNICIANS, AND ENABLE BIOBANKING OF NEW, NEEDED SAMPLE TYPES, SUPPORTING CORIELL INTERNAL GRANTS, CONTRACTS, AND FEDERALLY FUNDED RESEARCHERS, AS WELL AS SCIENTISTS NATIONWIDE.

NHGRI SAMPLE REPOSITORY FOR HUMAN GENETIC RESEARCH - PROJECT SUMMARY SPONSORED BY THE NHGRI AND ESTABLISHED AT THE CORIELL INSTITUTE IN 2006, THE NHGRI SAMPLE REPOSITORY FOR HUMAN GENETIC RESEARCH PROVIDES A PUBLICALLY ACCESSIBLE, AND CENTRALIZED RESOURCE OF WELL-CHARACTERIZED BIOSPECIMENS FROM A WIDE RANGE OF GLOBAL HUMAN POPULATIONS, INCLUDING THE 1000 GENOMES PROJECT COLLECTION, FOR USE IN BIOMEDICAL RESEARCH. THE OBJECTIVES OF THE NHGRI REPOSITORY ARE TO STIMULATE AND FACILITATE THE STUDY OF HUMAN GENETIC AND GENOMIC VARIATION BY ESTABLISHING, MAINTAINING, AND DISTRIBUTING A REPOSITORY OF HIGH-QUALITY, RENEWABLE, REPRODUCIBLE, WELL-CHARACTERIZED, AND BROADLY CONSENTED CELL LINES AND DNA. THE NHGRI REPOSITORY IS A GLOBAL RESOURCE THAT HAS BEEN USED BY THOUSANDS OF INVESTIGATORS TO SUPPORT COUNTLESS RESEARCH STUDIES. SINCE ITS INCEPTION, OVER TWO HUNDRED THOUSAND CELL LINES, DNA AND RNA SAMPLES HAVE BEEN DISTRIBUTED TO RESEARCHERS IN 50 COUNTRIES AROUND THE WORLD, AND THOUSANDS OF SCIENTIFIC ARTICLES HAVE RESULTED FROM STUDIES USING THESE SAMPLES AND ASSOCIATED DATA. PROPOSED NHGRI REPOSITORY ACTIVITIES INCLUDE: (1) MAINTAINING INVENTORY AND DISTRIBUTING NHGRI REPOSITORY CELL LINES AND DNA SAMPLES FOR THE EXISTING REPOSITORY COLLECTION, (2) DISTRIBUTING CUSTOM PREPARATIONS AND LOTS OF DNA, HIGH MOLECULAR WEIGHT (HMW) DNA, RNA, CELL PELLETS, AND CUSTOM PLATES AND PANELS AS REQUESTED BY THE SCIENTIFIC COMMUNITY, (3) EXPANDING THE REPOSITORY TO INCLUDE BIOSPECIMEN SUBMISSIONS FROM THE HUMAN PANGENOME REFERENCE CONSORTIUM AND CREATING A NEW PANEL OF INDUCED PLURIPOTENT STEM CELLS THAT WILL ENCOMPASS A WIDE RANGE OF GENOMIC VARIATION TO MEET THE GROWING NEEDS OF THE RESEARCH COMMUNITY, AND (4) MAINTAINING A COMPREHENSIVE AND SECURE DATABASE AND PUBLIC ONLINE CATALOG FOR NHGRI REPOSITORY ACTIVITIES, AND ENGAGING THE SCIENTIFIC COMMUNITY AND PUBLIC AT LARGE THROUGH SCIENTIFIC PRESENTATIONS, CONFERENCES, EDUCATIONAL EVENTS, AND COMMUNITY REPORTS. WITH OVER 50 YEARS OF NIH-SPONSORED BIOBANKING EXPERTISE, CORIELL IS UNIQUELY QUALIFIED TO ACHIEVE THESE AIMS AND STRATEGICALLY EXPAND THE OPERATIONS AND OFFERINGS OF THE NHGRI REPOSITORY. THE GOALS OF THE NHGRI REPOSITORY ARE CONSISTENT WITH THE NHGRI’S MISSION TO SUPPORT DEVELOPMENT OF RESOURCES AND TECHNOLOGY THAT ACCELERATE GENOMIC RESEARCH. CORIELL IS A TRUSTED NIH PARTNER AND HAS THE INFRASTRUCTURE, EXPERTISE, AND TRACK-RECORD TO ENSURE THE SUCCESSFUL OPERATIONS, MAINTENANCE AND GROWTH OF THIS IMPORTANT AND UNIQUE GENOMIC RESOURCE.

SAMPLE REPOSITORY FOR HUMAN GENETIC RESEARCH

MECHANISMS AND TARGETED THERAPY OF NRF2-HIGH ESOPHAGEAL SQUAMOUS CELL CARCINOMA

ROLE OF THE MICROBIOTA IN DNA METHYLATION AND CRC DEVELOPMENT

NRF2-ACSS2 AXIS IN ALCOHOL-INDUCED METABOLIC REPROGRAMMING AND ESOPHAGEAL PATHOLOGY - PROJECT SUMMARY THIS MULTIPLE-PI R01 PROPOSAL IS DESIGNED TO INCORPORATE BOTH IN VITRO AND IN VIVO APPROACHES TO ELUCIDATE THE ROLE OF THE NRF2-ACSS2 AXIS IN ALCOHOL-INDUCED METABOLIC REPROGRAMMING AND ESOPHAGEAL PATHOLOGY. WE HYPOTHESIZE THAT THE NRF2-ACSS2 AXIS MEDIATES METABOLIC REPROGRAMMING IN ALCOHOL-ASSOCIATED ESOPHAGEAL PATHOLOGY. WE WILL TEST THIS HYPOTHESIS USING HUMAN CELLS AND GENETICALLY MODIFIED MICE THROUGH THREE INDEPENDENT AND COMPLEMENTARY SPECIFIC AIMS. IN AIM 1, WE WILL CHARACTERIZE THE ROLE OF NRF2-ACSS2 AXIS IN MEDIATING ALCOHOL-INDUCED METABOLIC REPROGRAMMING IN HUMAN ESOPHAGEAL SQUAMOUS EPITHELIAL CELLS IN VITRO. IN AIM 2, WE WILL VALIDATE THE ROLE OF THE NRF2-ACSS2 AXIS IN MEDIATING ALCOHOL-INDUCED METABOLIC REPROGRAMMING IN MOUSE ESOPHAGUS IN VIVO. IN AIM 3, WE WILL EXAMINE THE INHIBITORY EFFECTS OF NRF2 AND ACSS2 INHIBITORS ON ALCOHOL-INDUCED METABOLIC REPROGRAMMING AND ESOPHAGEAL PATHOLOGY IN VIVO. IF THE HYPOTHESIS PROVED TO BE TRUE, THESE STUDIES WILL LAY DOWN A SOLID MECHANISTIC FOUNDATION FOR NRF2/ACSS2 INHIBITION AS A NOVEL MECHANISM-BASED PREVENTIVE MEASURE AGAINST ALCOHOL-ASSOCIATED ESOPHAGEAL PATHOLOGY IN THE FUTURE.

MOLECULAR DISSECTING AND TARGETING YAP1 MEDIATED CANCER STEMNESS AND IMMUNE SUPPRESSION IN ADVANCED GASTRIC ADENOCARCINOMA - PROJECT ABSTRACT THE MAIN OBJECTIVES OF THIS PROPOSAL ARE TO ELUCIDATE THE MECHANISMS OF YAP1-MEDIATED METASTATIC NICHE AND IMMUNOSUPPRESSION IN GASTRIC ADENOCARCINOMA (GAC) WITH PERITONEAL CARCINOMATOSIS (PC) FOR NOVEL THERAPEUTIC DISCOVERIES. GAC IS A MAJOR HEALTH BURDEN IN THE US AND WORLDWIDE. PC IS COMMON AFFECTING ~45% OF GAC PATIENTS. GAC PATIENTS WITH PC HAVE SHORT SURVIVAL AND TREATMENTS ARE INEFFECTIVE. MOLECULAR UNDERSTANDING FOR PC IS LIMITED. TO ADDRESS THIS UNMET CLINICAL CHALLENGE, WE HAVE ESTABLISHED PC BANKING INFRASTRUCTURE AIMING TO UTILIZE THESE PATIENT-DERIVED SPECIMENS TO DISCOVER AND VALIDATE OUR NOVEL TARGETS. OUR PRELIMINARY RNASEQ PROFILING OF PC SPECIMENS REVEALED AN ENRICHMENT OF UNIQUE IMMUNE SUPPRESSIVE MOLECULES, INCLUDING TIM3 AND IT’S LIGAND GALECTIN-9 (GAL-9), TGF-SS AND VISTA, BUT LESS EXPRESSION OF PD-1/PDL-1 AND CTLA-4. YAP1 HAS BEEN IMPLICATED IN HUMAN DEVELOPMENT, LINEAGE PLASTICITY, AND UPREGULATED IN MANY TUMOR TYPES. HOWEVER, ITS ROLE IN MEDIATING PC METASTASES AND IMMUNE SUPPRESSION IN TUMOR MICROENVIRONMENT (TME) REMAIN UNCLEAR. OUR PRELIMINARY DATA SUGGEST THAT YAP1 IS HIGHLY EXPRESSED IN PRIMARY AND METASTATIC TUMOR CELLS OF GAC PATIENTS AND IS SIGNIFICANTLY ASSOCIATED WITH POOR SURVIVAL. GENETIC KNOCKOUT (KO) YAP1 SIGNIFICANTLY DECREASED CANCER STEMNESS TRAITS, TUMOR FORMATION AND PC IN MICE. FURTHER, DEPLETION OF YAP1 IN TUMOR CELLS CONSISTENTLY INCREASED CD3 AND CD8 T CELL RESPONSES FROM GAC. THROUGH A SINGLE CELL RNASEQ (SCRNASEQ) OF PC SAMPLES AND VALIDATION USING IMMUNOFLUORESCENT STAINING, WE NOTICED THAT YAP1, TIM3 LIGAND GAL-9 AND DKK1 ARE HIGHLY EXPRESSED IN TUMOR CELLS OF PC; WHILE TIM3 IS ENRICHED IN IMMUNE CELLS. RNASEQ FROM YAP1HIGH AND YAP1 KO PATIENT-DERIVED TUMOR CELLS REVEALED THAT GAL-9 AND DKK1 WERE SIGNIFICANTLY DECREASED UPON DEPLETION OF YAP1 AND THESE FACTORS ARE ASSOCIATED WITH POOR SURVIVAL OF PATIENTS. WE HYPOTHESIZE THAT YAP1HIGH PC CELLS ARE METASTASIS-INITIATING CELLS THAT ORCHESTRATE A NICHE BY CONFERRING CANCER STEMNESS ATTRIBUTES TO THE TUMOR CELLS AND PROMOTE TUMOR IMMUNOSUPPRESSION THROUGH ACTIVATING TIM3/GAL-9 AXIS AND INCREASING PARACRINE OF DKK1 IN PC TME. THEREFORE, SIMULTANEOUSLY TARGETING HIPPO/YAP1 AND IMMUNE CHECKPOINT (TIM3/GAL-9) COULD BE AN ENHANCED STRATEGY. TO TEST OUR HYPOTHESIS, WE PROPOSE THREE SPECIFIC AIMS: AIM 1. DETERMINE THE FUNCTIONAL RELEVANCE OF YAP1 IN PC STEMNESS AND METASTASES IN TUMOR CELLS USING NOVEL STEM CELL CLONING TECHNOLOGY AND PDX/PDO MODELS IN VIVO. AIM 2. TO INVESTIGATE THE MECHANISMS WHEREBY YAP1 MEDIATES IMMUNOSUPPRESSION IN TME OF PC. AIM 3. ELUCIDATING EFFICACY OF INHIBITION OF YAP1 ALONE OR IN COMBINATION WITH TIM3 INHIBITION USING THE PDO MODELS, KP-LUC2 SYNGENEIC MOUSE MODEL, GEMM, AND ONGOING YAP1 CLINICAL TRIAL. BY UTILIZING PATIENT- DERIVED PC CELLS, WE WILL UNCOVER FUNCTIONAL IMPORTANCE OF YAP1-MEDIATED PC AND ELUCIDATE THE MECHANISMS BY WHICH YAP1 MEDIATED IMMUNOSUPPRESSION FOR NOVEL THERAPEUTIC STRATEGIES. UPON COMPLETION OF THIS STUDY, WE WILL HAVE A STRONG RATIONALE FOR A NOVEL COMBINATION THAT COULD OVERCOME THE SHORTCOMINGS WE EXPERIENCE IN THE CLINICS TODAY.

THE ROLE OF GSK3/PPAR-/MITOPHAGY PATHWAY IN REGULATING HEMATOPOIA - THE ROLE OF GSK3/PPAR-D/FAO/MITOPHAGY PATHWAY IN REGULATING HEMATOPOIETIC STEM CELL HOMEOSTASIS AND FUNCTION ABSTRACT HEMATOPOIETIC STEM CELLS (HSCS) POSSESS THE ABILITIES TO BOTH PRODUCE STEM CELLS, A PROPERTY KNOWN AS SELF-RENEWAL, AND GIVE RISE TO ALL DIFFERENTIATED HEMATOPOIETIC LINEAGES. CLINICALLY, HSCS ARE THERAPEUTICALLY VALUABLE FOR TRANSPLANTATION IN TREATMENT OF VARIOUS HEMATOLOGIC MALIGNANCES. DESPITE REMARKABLE PROGRESS MADE IN THE RESEARCH OF HSCS DURING THE PAST THREE DECADES, THE MOLECULAR MECHANISMS REGULATING HSC HOMEOSTASIS AND FUNCTION ARE STILL NOT FULLY UNDERSTOOD. OUR PREVIOUS PUBLISHED SHOWED THAT GSK3 PLAYS AN ESSENTIAL ROLE IN REGULATING HSC HOMEOSTASIS. SPECIFICALLY, KNOCKDOWN OF GSK3 PROMOTE TRANSIENT EXPANSION AND LONG-TERM EXHAUSTION OF HSC IN VIVO. OUR NEW PRELIMINARY STUDIES DEMONSTRATED THAT GSK3 FUNCTIONS THROUGH PPAR-D/FAO/MITOPHAGY PATHWAY TO REGULATE HSC DIVISION SYMMETRY; INHIBITION OF GSK3 INDUCES MITOPHAGY AND CONVERSELY, BLOCKING PPAR-D/FAO/MITOPHAGY CAN REVERSE THE ENHANCED SELF-RENEWAL PHENOTYPE THAT IS ASSOCIATED WITH GSK3 INHIBITION. IN THIS STUDY, WE WILL FIRST EXAMINE HOW GSK3 REGULATES PPAR-D, WHICH IN TURN REGULATES MITOPHAGY AND HSC DIVISION SYMMETRY. SECONDLY, WE WILL INVESTIGATE WHETHER LOSS-OF- FUNCTION OF PPAR-D, PARK2 OR PINK1 (TWO MITOPHAGY KEY REGULATORS) REVERSE THE FUNCTIONAL DEFECT OF GSK3B-DEFICIENT HSC IN VIVO. LASTLY, WE WILL EXPLORE WHETHER GSK3 CONTROLS FAO AND LIPID METABOLISM TO REGULATE HSC FUNCTION AND HOMEOSTASIS. OUR STUDY WILL PROVIDE SIGNIFICANT NEW INSIGHTS INTO THE MOLECULAR MECHANISMS UNDERLYING GSK3-DEPENDENT REGULATION OF HSC HOMEOSTASIS AND FUNCTION. THE KNOWLEDGE LEARNED MAY FACILITATE BONE MARROW TRANSPLANTATION FOR TREATING DIVERSE HEMATOLOGICAL DISEASES.

EXPLORE THE SIGNALING MECHANISMS OF ACQUIRED RESISTANCE TO TYROSINE KINASE INHIBITORS IN AML - EXPLORE THE SIGNALING MECHANISMS OF ACQUIRED RESISTANCE TO TYROSINE KINASE INHIBITORS IN AML ABSTRACT ACUTE MYELOID LEUKEMIA (AML) IS A MALIGNANT HEMATOPOIETIC DISEASE AND THE MOST COMMON TYPE OF ACUTE LEUKEMIA IN ADULTS. ONE MAJOR OBSTACLE TO GREATER SUCCESS WITH TARGET THERAPY OF LEUKEMIA IS DRUG RESISTANCE. THE MECHANISMS UNDERLYING DRUG RESISTANCE IN AML ARE POORLY UNDERSTOOD. FLT3 IS A CYTOKINE RECEPTOR WHICH BELONGS TO THE RECEPTOR TYROSINE KINASE (RTK) CLASS III. ACTIVATING MUTATIONS IN FMS-LIKE TYROSINE KINASE 3 (FLT3) ARE NOW RECOGNIZED AS THE MOST COMMON MOLECULAR ABNORMALITY IN AML AND FLT3ITD MUTATIONS ARE FOUND IN NEARLY 30% OF AML PATIENTS. QUIZARTINIB (AC220) IS A POTENT AND SELECTIVE SECOND-GENERATION INHIBITOR OF FLT3. IT IS IN CLINICAL TRIALS FOR THE TREATMENT OF RELAPSED OR REFRACTORY FLT3ITD POSITIVE AND NEGATIVE AML PATIENTS AND AS MAINTENANCE THERAPY. REMARKABLY, THOSE CLINICAL TRIALS HAVE SHOWED VERY PROMISING RESULT. HOWEVER, DRUG RESISTANCE TO AC220 HAS ALSO BEEN REPORTED THROUGH THE EARLY CLINICAL STUDIES. TO UNDERSTAND THE UNDERLYING MECHANISMS OF DRUG RESISTANCE TO AC220, WE UNDERTOOK AN UNBIASED APPROACH WITH A NOVEL CRISPR POOLED LIBRARY TO SCREEN NEW GENES WHOSE LOSS OF FUNCTION CONFERS RESISTANCE TO AC220. IN OUR SCREEN, WE IDENTIFIED SPRY3, AN INTRACELLULAR INHIBITOR OF RTK SIGNALING, AND GSK3, A CANONICAL WNT SIGNALING ANTAGONIST, AND DEMONSTRATED THAT RE-ACTIVATION OF DOWNSTREAM RTK/RAS/ERK AND WNT SIGNALING AS MAJOR MECHANISMS OF RESISTANCE TO THE FLT3 INHIBITOR. FURTHERMORE, WE ALSO CONFIRMED OUR FINDINGS IN PRIMARY AML PATIENT SAMPLES. WE DEMONSTRATED THAT THE EXPRESSION LEVEL OF SPRY3 AND GSK3A IS DRAMATICALLY REDUCED IN AC220 RESISTANT AML SAMPLES AND SPRY3 DELETED PRIMARY AML CELLS ARE RESISTANT TO AC220. ADDITIONALLY, WE TREATED SPRY3 AND GSK3 KNOCKOUT AML CELLS WITH A POTENT MAP KINASE INHIBITOR AND SS- CATENIN INHIBITOR RESPECTIVELY, DEMONSTRATED THAT BOTH INHIBITORS RE-SENSITIZED AML CELLS TO AC220. INTRIGUINGLY, WE FOUND THAT EXPRESSION OF SPRY3 IS GREATLY REDUCED IN GSK3 KNOCKOUT AML CELLS, WHICH POSITIONED SPRY3 DOWNSTREAM OF GSK3 IN THE RESISTANCE PATHWAY. IN THIS PROPOSAL, WE HYPOTHESIZE THAT SPROUTY (SPRY) AND GSK3 PLAY CRITICAL ROLES IN THE RESPONSE TO TYROSINE KINASE INHIBITOR IN AML. THE RAS/MEK/ERK AND WNT PATHWAYS REGULATED BY SPRY3 AND GSK3 ARE IMPORTANT FOR THE ACQUIRED DRUG RESISTANCE IN AML. NEXT, WE WILL PERFORM A SERIES COMPREHENSIVE STUDY TO EXPLORE NOVEL DOWNSTREAM EFFECTORS/ INTERACTING PARTNERS OF SPRY3 AND GSK3 IN AMLS AND THE MOLECULAR MECHANISMS OF THEIR ACTION. FURTHERMORE, WE WILL EXAMINE THE POSSIBILITY TO TRANSLATE OUR FINDINGS INTO NEW CLINICAL THERAPIES. TAKEN TOGETHER, OUR STUDY IDENTIFIED NOVEL GENES WHOSE LOSS OF FUNCTION CONFERS RESISTANCE TO A SELECTIVE FLT3 INHIBITOR AND REVEALED THE UNDERLYING MECHANISM, THEREBY PROVIDING NEW INSIGHT INTO SIGNALING PATHWAYS THAT CONTRIBUTE TO THE ACQUIRED RESISTANCE IN AML. THE KNOWLEDGE LEARNED MAY LEAD TO THE DEVELOPMENT OF MORE EFFICIENT COMBINED THERAPEUTIC AVENUES FOR AML.

DISSECTING SOX9-MEDIATED CANCER STEMNESS AND IMMUNOSUPPRESSION FOR NOVEL STRATEGIES IN ADVANCED GASTRIC ADENOCARCINOMA

EXOSOMAL GALECTIN-3 MEDIATED STROMAL ACTIVATION AND IMMUNOSUPPRESSION IN ADVANCED GASTRIC ADENOCARCINOMA

IDENTIFYING FACTORS RESPONSIBLE FOR SEX DISPARITIES IN MELANOMA ETIOLOGY CORRECTION INCORPORATED TO REFLECT TRANSFER OF AWARD TO CORIELL INSTITUTE FOR MEDICAL RESEARCH INC. ON 03/12/2025

EXPAND HUMAN UMBILICAL CORD BLOOD HEMATOPOIETIC STEM CELLS WITH PPAR-A AGONISTS - EXPAND HUMAN UMBILICAL CORD BLOOD HEMATOPOIETIC STEM CELLS WITH PPAR-A AGONISTS ABSTRACT HEMATOPOIETIC STEM CELLS (HSCS) ARE DEFINED BY THEIR SELF-RENEWAL POTENTIAL AND ABILITY TO DIFFERENTIATE INTO MULTIPLE BLOOD LINEAGES. HEMATOPOIETIC STEM CELL TRANSPLANT (HSCT) IS A MAINSTAY OF LIFE-SAVING THERAPY FOR HEMATOPOIETIC MALIGNANCIES AND HYPOPROLIFERATIVE DISORDERS. THE USE OF UMBILICAL CORD BLOOD (UCB)-DERIVED HSCS PROVIDES MANY ADVANTAGES OVER ADULT HSCS, INCLUDING ENHANCED LONG-TERM IMMUNE RECOVERY, DECREASED GRAFT VERSUS HOST DISEASE, AND AVAILABILITY OF DONORS FROM A BROAD POPULATION, EXPANDING THE AVAILABILITY OF HSCT FOR GROUPS CURRENTLY UNDERREPRESENTED IN BONE MARROW REGISTRIES. THE USE OF HAPLOIDENTICAL TRANSPLANTS ADDRESSES MANY OF THESE ISSUES, BUT UCB WOULD STILL BE A HIGHLY USEFUL RESOURCE IF UCB UNITS CONTAINED SUFFICIENT NUMBERS OF HSCS. HOWEVER, THE LOW CELL DOSE IN MOST UCB UNITS LIMITS THEIR USE, AS INSUFFICIENT HSCS LEADS TO DELAYED ENGRAFTMENT, GRAFT FAILURE, AND SEVERE INFECTIOUS COMPLICATIONS. EVEN A MODEST EXPANSION OF HSCS FROM UCB CAN SOLVE MANY OF THESE PROBLEMS, AND THUS HSC EXPANSION FROM UCB HAS REMAINED AN IMPORTANT GOAL FOR THE FIELD. OUR GOAL IS TO USE CRYOPRESERVED UCB FROM THE NHLBI BIOLOGIC BIOSPECIMEN REPOSITORY TO DEVELOP NEW METHODS TO EXPAND FUNCTIONAL HSCS FOR THERAPEUTIC APPLICATIONS. RECENT STUDIES HAVE ACHIEVED EX VIVO EXPANSION OF HSCS USING CYTOKINE COCKTAILS COMBINED WITH SMALL MOLECULES, BUT THESE APPROACHES REQUIRE EXPOSURE TO HIGH CONCENTRATIONS OF CYTOKINES. CYTOKINES INDUCE DIFFERENTIATION AND IMPAIR THE SELF-RENEWAL FUNCTION OF PRIMITIVE HSCS. HSC EXPANSION WITH MINIMAL CYTOKINE EXPOSURE WOULD THEREFORE BE IDEAL FOR CLINICAL APPLICATIONS. OUR PREVIOUS STUDIES DEMONSTRATED A COMBINATION OF TWO INHIBITORS (CHIR99021 AND RAPAMYCIN) MAINTAINS HUMAN AND MOUSE LONG-TERM HSCS EX VIVO IN THE ABSENCE OF CYTOKINES OR SERUM. BASED ON THIS FINDING, WE PERFORMED A HIGH THROUGHPUT SCREEN AND IDENTIFIED SEVERAL PPAR-A AGONISTS, WHICH ARE USED CLINICALLY TO TREAT HYPERTRIGLYCERIDEMIA, THAT SIGNIFICANTLY EXPAND LONG-TERM FUNCTIONAL UCB HSC EX VIVO WHILE MINIMIZING EXPOSURE TO CYTOKINES. IN THIS PROJECT, WE WILL CARRY OUT STUDIES TO OPTIMIZE AND IMPLEMENT OUR PPAR-A AGONISTS-BASED EXPANSION METHOD USING CRYOPRESERVED UCB STORED IN THE NHLBI BIOLOGIC BIOSPECIMEN REPOSITORY.

A GENETIC BLUEPRINT FOR DIFFERENCES BETWEEN MALES AND FEMALES IN EARLY MAMMALIAN DEVELOPMENT -ALTHOUGH MALE AND FEMALE MAMMALS HAVE DIFFERENT REPRODUCTIVE ORGANS, OTHER ORGANS, SUCH AS THE HEART AND LUNGS, SEEM TO BE IDENTICAL. HOWEVER, WHEN LOOKING AT THE GENES BEING EXPRESSED IN MALE AND FEMALE CELLS IN ANY ORGAN, THERE ARE SUBSTANTIAL DIFFERENCES. SOME OF THESE ARE DUE TO THE FACT THAT FEMALE CELLS HAVE TWO X CHROMOSOMES AND MALES HAVE ONE X AND ONE Y CHROMOSOME. IN ADDITION, THE HORMONES PRODUCED BY THE GONADS CAN INFLUENCE THE GENES EXPRESSED. HOW THESE MOLECULAR DIFFERENCES ARE ESTABLISHED AND HOW THEY AFFECT FUNCTIONALITY IS NOT KNOWN. THE OBJECTIVE OF THIS PROJECT IS TO INVESTIGATE THE MOLECULAR DIFFERENCES BETWEEN MALE AND FEMALE MICE, STARTING SOON AFTER FERTILIZATION AND THROUGHOUT EMBRYONIC DEVELOPMENT BY CHARACTERIZING GENE EXPRESSION AND EPIGENETIC FEATURES AT SUCCESSIVE STAGES OF EMBRYOGENESIS. THE DATA GENERATED WILL ALLOW COMPARATIVE STUDIES WITH OTHER MODEL ORGANISMS AND LEND INSIGHT INTO HOW EVOLUTION HAS SHAPED MALE AND FEMALE GENETIC AND HORMONAL DIFFERENCES. THE PROJECT INCLUDES OUTREACH AND EDUCATIONAL PROGRAMS TO INCREASE AWARENESS OF HOW MALE AND FEMALE PHYSIOLOGIES DIFFER AMONG STUDENTS, BOTH AT THE GRADUATE AND UNDERGRADUATE LEVEL. THIS PROJECT WILL ALSO PROVIDE TRAINING OPPORTUNITIES FOR UNDERGRADUATE HANDS-ON RESEARCH EXPERIENCES. IN ADDITION, IT WILL CONTRIBUTE TOWARDS CREATING A RESOURCE TO TRAIN STUDENTS TO TEACH GENOMIC AND BIOINFORMATIC CONCEPTS TO HIGH SCHOOL STUDENTS IN THE AREA. THESE STUDENTS ARE GENERALLY FROM UNDERREPRESENTED BACKGROUNDS, ESPECIALLY IN THE STEM DISCIPLINES. BEGINNING SOON AFTER FERTILIZATION, THE X AND Y CHROMOSOMES PROGRAM AUTOSOMAL GENE EXPRESSION AND THE EPIGENOMIC LANDSCAPE, ESTABLISHING MALE- AND FEMALE-SPECIFIC GENE NETWORKS. THE MECHANISMS UNDERLYING THESE EFFECTS ARE UNKNOWN, AS WELL AS HOW MALE AND FEMALE BIASES EVOLVE ACROSS DEVELOPMENT AND IN DIFFERENT LINEAGES. THE OBJECTIVE OF THIS PROJECT IS TO FILL THIS KNOWLEDGE GAP BY INTEGRATING EXPERIMENTAL AND SYSTEMS LEVEL ANALYSES IN VITRO AND IN VIVO. IT EXAMINES THE HYPOTHESIS THAT REGULATORY FACTORS ENCODED ON THE X AND Y CHROMOSOMES DICTATE DIFFERENTIAL EXPRESSION AND EPIGENETIC PROFILES OF AUTOSOMAL GENES. HORMONES EQUALIZE SOME OF THESE DIFFERENCES, BUT OTHERS PERSIST, AFFECTING CELLULAR PHENOTYPES EVEN IN THE ADULT ORGANISM. THIS HYPOTHESIS WILL BE TESTED BY: 1) DETERMINING THE TRANSCRIPTIONAL AND EPIGENETIC EFFECTS OF DIFFERENTIALLY EXPRESSED REGULATORY FACTORS IN EARLY EMBRYOGENESIS; AND 2) IDENTIFING THE BIASES IN GENE EXPRESSION AND EPIGENETIC PATTERNS DEPENDENT ON THE X AND Y CHROMOSOMES BEFORE AND AFTER THE APPEARANCE OF GONADAL HORMONES. THESE EXPERIMENTS WILL EXPLOIT A MOUSE MODEL THAT ALLOWS SEGREGATION OF THE GENETIC AND HORMONAL COMPONENTS OF THE MALE AND FEMALE PHENOTYPES. SINCE EPIGENETIC MARKS ESTABLISHED IN EARLY DEVELOPMENT CAN BE LATENT AND RELEVANT TO GENE EXPRESSION AT LATER STAGES, THIS RESEARCH WILL ALSO SERVE AS A PARADIGM FOR UNDERSTANDING HOW EVENTS IN EMBRYOGENESIS INFLUENCE DIMORPHISMS AFTER BIRTH AND BEYOND. MOREOVER, THESE STUDIES WILL LAY THE GROUNDWORK FOR MECHANISTIC STUDIES ON THE EFFECTS OF TRANSCRIPTION AND EPIGENETIC FACTOR DOSAGE ON THE TRANSCRIPTOME. THIS AWARD REFLECTS NSF'S STATUTORY MISSION AND HAS BEEN DEEMED WORTHY OF SUPPORT THROUGH EVALUATION USING THE FOUNDATION'S INTELLECTUAL MERIT AND BROADER IMPACTS REVIEW CRITERIA.- SUBAWARDS ARE NOT PLANNED FOR THIS AWARD.

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