Institute For Medical Research Inc

WOMEN'S INTERAGENCY HIV STUDY (WIHS) IV, CHICAGO CONSORTIUM

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

MACS/WIHS COMBINED COHORT STUDY: COOK COUNTY CLINICAL RESEARCH SITE (CC_CRS)

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.

RYAN WHITE PART C OUTPATIENT EIS PROGRAM

RYAN WHITE TITLE IV WOMEN, INFANTS, CHILDREN, YOUTH AND AFFECTED FAMILY MEMBERS AIDS HEALTHCARE

REGENERATIVE WOUND HEALING VIA INFLAMMATION-MODULATING BIOMATERIALS

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

RYAN WHITE TITLE IV WOMEN, INFANTS, CHILDREN, YOUTH AND AFFECTED FAMILY MEMBERS AIDS HEALTHCARE

ISOLATION AND CHARACTERIZATION OF INTESTINAL STEM CELLS

H3AFRICA KIDNEY DISEASE RESEARCH NETWORK

RYAN WHITE PART C OUTPATIENT EIS PROGRAM

ENHANCING GLOBAL HEALTH SECURITY: EXPANDING EFFORTS AND STRATEGIES TO PROTECT AND IMPROVE PUBLIC HEALTH GLOBALLY

RYAN WHITE TITLE IV PROGRAM

NATIONAL CENTER ON DOMESTIC VIOLENCE, TRAUMA & MENTAL HEALTH

NATIONAL CENTER ON DOMESTIC VIOLENCE, TRAUMA & MENTAL HEALTH

FAMILY VIOLENCE PREVENTION AND SERVICES

BUILDING CAPACITY FOR HIV OPERATIONAL RESEARCH AND PUBLIC HEALTH EVALUATION FOR

CHICAGO ADOLESCENT TRIALS UNIT

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.

GENETIC MAPPING OF FUNCTIONAL VOMERONASAL CIRCUIT

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.

STROGER HOSPITAL OF COOK COUNTY, DEPARTMENT OF SURGERY,*

CHICAGO PREVENTION AND INTERVENTION EPICENTER (CHICAGO PIE)

SAMPLE REPOSITORY FOR HUMAN GENETIC RESEARCH

MECHANISMS OF TRANSCRIPTION REGULATION IN CHROMATIN

INVESTIGATION OF EXPERIENCE-DEPENDENT POST TRANSCRIPTIONAL REGULATION OF DROSOPHI

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

NCI COMMUNITY ONCOLOGY RESEARCH PROGRAM (NCORP) COMMUNITY SITES

BUILDING CAPACITY FOR EVIDENCE UTILIZATION AND EVALUATION WITHIN THE GOVERNMENT OF TANZANIA UNDER PEPFAR

POSSE PROJECT: A COMMUNITY-LEVEL INTERVENTION FOR BLACK YMSM

EPIDEMIOLOGY, PREVENTION AND TREATMENT OF INFLUENZA AND OTHER RESPIRATORY INFECTI

HUMAN MONOCLONAL ANTIBODIES THAT BIND BOTULINUM TOXINS

TRANSGENDER YOUTH AND PREP: PK, SAFETY, UPTAKE & ADHERENCE

ZEBRAFISH SENSORY HAIR CELL REGENERATION

THE MOLECULAR BASIS OF PLANARIAN REGENERATION

MAIN LINE HEALTH CCOP

ROLE OF THE MICROBIOTA IN DNA METHYLATION AND CRC DEVELOPMENT

DEVELOPMENT OF A WHOLE HEART MODEL OF THE J WAVE SYNDROMES AND NOVEL APPROACHES TO PHARMACOLOGIC MANAGEMENT OF ASSOCIATED LIFE-THREATENING ARRHYTHMIAS

KEEPING IT LITE: EXPLORING HIV RISK IN VULNERABLE YOUTH WITH LIMITED INTERACTION

STARVATION RESISTANCE AND RESILIENCE OF METABOLIC DYSFUNCTION IN CAVEFISH

INVESTIGATING DEVELOPMENTAL POTENTIAL BASED ON GENOME-WIDE CHROMATIN STATUS

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.

MODELS OF SELFISHNESS: MOLECULAR AND EVOLUTIONARY ANALYSES OF THE WTF MEIOTIC DRIVERS

INTRINSIC AND EXTRINSIC REGULATION OF CRANIAL MESODERM

IMPROVING ADHERENCE AMONG HIV+ RWANDAN YOUTH: A TI-CBTE INDIGENOUS LEADER MODEL

TANZANIA GLOBAL HEALTH SECURITY PRIORITIES - ADVANCING EFFORTS AND STRATEGIES TO PROTECT AND IMPROVE HEALTH SECURITY (TAGHESP)

H-STAR PROJECT: (HIV SUBSTANCE TREATMENT AND RECOVERY)

A TRANSPOSASE SYSTEM FOR INTEGRATIVE CHIP-EXO AND ATAC-SEQ ANALYSIS AT SINGLE-CELL RESOLUTION

SUSTAINING AND IMPROVING SURVEILLANCE OF HUMAN AND ANIMAL INFLUENZA AND OTHER RESPIRATORY PATHOGENS IN GHANA. (SHARPEN-FLU) - ABSTRACT INFLUENZA REMAINS A DISEASE WHICH POSES A HUGE GLOBAL PUBLIC HEALTH THREAT. MAINLY BECAUSE INFLUENZA VIRUSES CAN CAUSE DEVASTATING PANDEMICS WITH THE MOST DEVASTATING TO DATE BEING THE SPANISH `FLU? PANDEMIC, WHICH LED TO THE DEATH OF ~40 MILLION PEOPLE WORLDWIDE. THE RECENT COVID-19 PANDEMIC HAS EMPHASIZED THE NEED FOR CONSTANT SURVEILLANCE FOR RESPIRATORY PATHOGENS SUCH AS INFLUENZA AND EMERGING AND RE-EMERGING PATHOGENS LIKE SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS-2 (SARS-COV-2). EARLY DETECTION AND CHARACTERIZATION OF THESE RESPIRATORY PATHOGENS IS THE KEY TO MITIGATING THEIR EFFECTS. THE NOGUCHI MEMORIAL INSTITUTE FOR MEDICAL RESEARCH (NMIMR) WITH SUPPORT FROM THE GHANA HEALTH SERVICE (GHS) AND THE UNITED STATES NAVAL MEDICAL RESEARCH UNIT (US NAMRU#3), STARTED SURVEILLANCE FOR INFLUENZA VIRUSES IN GHANA IN 2007; WHEN OUTBREAKS OF HIGHLY PATHOGENIC AVIAN INFLUENZA A H5N1 AMONG POULTRY LED TO THE DEATH AND CULLING OF THOUSANDS OF BIRDS. THIS SENTINEL SURVEILLANCE SYSTEM PROVIDES EPIDEMIOLOGIC AND VIROLOGIC DATA ON CIRCULATING INFLUENZA VIRUS STRAINS IN THE COUNTRY. IT IS ALSO ABLE TO DETECT AND RESPOND TO INFLUENZA OUTBREAKS. THROUGH THIS ESTABLISHED INFLUENZA SURVEILLANCE SYSTEM, GHANA IS ABLE TO CONTRIBUTE TO THE WHO GLOBAL INFLUENZA AND SURVEILLANCE RESPONSE SYSTEM (GISRS) BY UPDATING DATA ON THE WHO FLUNET AS WELL AS SUBMITTING SAMPLES TO THE WHO CC IN LONDON. AGAIN, THE NMIMR COLLABORATES WITH THE VETERINARY SERVICES DIRECTORATE (VSD) TO CONDUCT SURVEILLANCE FOR AVIAN INFLUENZA AMONG POULTRY IN SELECTED AREAS IN THE COUNTRY. ALTHOUGH THE ESTABLISHED SENTINEL SYSTEM HAS BEEN EXTREMELY USEFUL, THERE ARE A FEW CHALLENGES. THIS SYSTEM IS HEAVILY BIASED TOWARDS INFLUENZA VIRUSES. THIS CURRENT COVID-19 PANDEMIC HAS DEMONSTRATED THE NEED TO INCLUDE OTHER NON-INFLUENZA RESPIRATORY VIRUSES AS WELL AS BACTERIAL PATHOGENS TO BE ABLE TO HAVE A ROBUST SYSTEM THAT CAN DETECT EARLY THREATS. AGAIN, DUE TO REGIONAL RE-ORGANIZATION, SOME N EW REGIONS CURRENTLY DO NOT HAVE INFLUENZA SENTINEL SURVEILLANCE SITES. THERE IS THE NEED THEREFORE TO EXPAND TO INCLUDE DETECTION OF OTHER PATHOGENS AS WELL AS INVOLVE NEW REGIONS AND PRIMARY HEALTHCARE FACILITIES. FURTHERMORE, THERE IS AN URGENT NEED FOR CLOSE COLLABORATIONS WITH THE VSD FOR IMPROVED CONTINUOUS SURVEILLANCE AMONG ANIMAL POPULATIONS AS MOST EMERGING AND RE-EMERGING PATHOGENS ARE ZOONOTIC IN NATURE. ULTIMATELY IN ORDER TO BE ABLE TO DETECT THREATS EARLY, A HOLISTIC ONE HEALTH APPROACH, WITH COLLABORATIONS AND DATA SHARING AMONG ALL INTEREST GROUPS IS IMPORTANT. THIS NOTICE OF FUNDING OPPORTUNITY PROVIDES GHANA WITH THE UNIQUE CHANCE TO EXPAND INFLUENZA AND OTHER RESPIRATORY PATHOGENS SURVEILLANCE IN GHANA IN ORDER TO BE ABLE TO CHARACTERIZE AND MONITOR RESPIRATORY PATHOGENS AMONG BOTH HUMANS AND ANIMALS; WHILE STRENGTHENING COLLABORATIONS AND COMMUNICATION AMONG INTEREST GROUPS FOR ADEQUATE UTILIZATION OF DATA TO PREVENT OR MITIGATE EFFECTS OF OUTBREAKS AND PANDEMICS.

STRUCTURE AND FUNCTION OF MAMMALIAN CPEB2 AGGREGATES IN NORMAL AND AD BRAIN - PROJECT SUMMARY THE AGGREGATE FAILURE OF PRE-CLINICAL AND CLINICAL TRIALS IN AD HAS DEMONSTRATED THAT AN IMPROVED FUNDAMENTAL UNDERSTANDING OF MEMORY AND HOW THE MOLECULAR COMPONENTS OF MEMORY ARE ALTERED IN THE AD DISEASE PROCESS IS NECESSARY TO DEVELOP EFFECTIVE TREATMENT. THE BROAD OBJECTIVE OF THE PROJECT IS TO IDENTIFY THE BIOCHEMICAL SUBSTRATES OF LONG-LASTING MEMORIES IN MAMMALS. THE CURRENT PROPOSAL FOCUSES ON A FAMILY OF RNA-BINDING, CYTOPLASMIC POLYADENYLATION ELEMENT BINDING PROTEIN (CPEB), THAT STABILIZES MEMORY IN INVERTEBRATES AND MICE. REMARKABLY, CPEB FAMILY PROTEIN FORMS NON-DISEASE-CAUSING AMYLOIDOGENIC AGGREGATES AND AGGREGATION OF CPEB IS NECESSARY TO STABILIZE MEMORY. AS AMYLOIDS ARE TYPICALLY LINKED TO DISEASE STATES, THE QUESTION REMAINS HOW SIMILARLY STRUCTURED ASS42 OR TAU PROTEINS CAN HAVE OPPOSING EFFECTS ON MEMORY. THEREFORE, TO DEVELOP A BETTER UNDERSTANDING OF THE RELATIONSHIP BETWEEN AMYLOIDS THAT SUPPORT MEMORY AND AMYLOIDS THAT DISRUPT MEMORY, WE WILL USE A VARIETY OF TECHNIQUES TO SOLVE THE STRUCTURE AND FUNCTION OF THE CPEB FAMILY MEMBERS, CPEB2 AND CPEB3, IN HUMAN AND MICE. IN AIM 1, WE WILL USE CRYO- ELECTRON MICROSCOPY TO SOLVE THE STRUCTURE OF CPEB AGGREGATES FROM FRESH HUMAN FRONTOTEMPORAL LOBE TISSUE COLLECTED FROM 25-50-YEAR-OLD HUMAN SUBJECTS UNDERGOING TISSUE REMOVAL UNDER THE STANDARD OF CARE FOR THEIR DISEASE. THESE TISSUES WOULD HAVE BEEN OTHERWISE DISCARDED. IN AIM 2, MICE LACKING THE ABILITY TO FORM AGGREGATES OF CPEB2 AND CPEB3 WILL BE TRAINED AND TESTED IN A ONE-TRIAL INHIBITORY AVOIDANCE TASK TO ASSAY THEIR ABILITY TO FORM, MAINTAIN, AND RECALL MEMORY. IN AIM 2 WE WILL ALSO INVESTIGATE THE CONSEQUENCE OF CPEB2 AND CPEB3 AGGREGATION IN TRANSLATION OF MRNA ENCODING SYNAPTIC PROTEINS. THE RESULTS WOULD BE THE FIRST TO PROVIDE DIRECT STRUCTURAL ANALYSIS OF A FUNCTIONAL AMYLOID LINKED TO MEMORY IN MAMMALS, THE STRUCTURAL DISTINCTIONS, IF ANY, BETWEEN FUNCTIONAL AND TOXIC AMYLOID IN THE HUMAN BRAIN, AND PRECISELY LINK CPEB2 AND CPEB3 AGGREGATION AND ACTIVITY TO ANIMALS’ ABILITY TO FORM OR STABILIZE MEMORY. THIS KNOWLEDGE WOULD PROVIDE THE FOUNDATION TO INVESTIGATE IN THE FUTURE HOW TOXIC AMYLOIDS OF ASS42 OR TAU SPECIFICALLY PERTURB MEMORY.

KEEPING IT LITE 2: EXPLORING HIV RISK IN VULNERABLE YOUTH WITH LIMITED INTERACTION AND DIGITAL HEALTH INTERVENTION (LITE-2) - ABSTRACT DESPITE ADVANCES IN HIV DIAGNOSTICS, CARE AND PREVENTION STRATEGIES, INFECTION RATES AMONG ADOLESCENT AND YOUNG ADULT SEXUAL AND GENDER MINORITIES (SGM) CONTINUE TO RISE IN THE UNITED STATES (US). THERE IS AN URGENT NEED TO DESCRIBE THE EPIDEMIOLOGY AND TRAJECTORIES OF HIV ACQUISITION IN THIS POPULATION AND TO OFFER AGE AND CULTURALLY APPROPRIATE SCALABLE PREVENTION INTERVENTIONS TO THOSE AT HIGHEST RISK OF INFECTION IN THE US. THIS PROJECT WILL ENGAGE AND RETAIN YOUNG SGM IN AN INNOVATIVE LONGITUDINAL COHORT, ENROLL THEM IN A DYNAMIC ESTABLISHED DIGITAL HEALTH RETENTION PLATFORM (HMP; HEALTHMPOWERMENT), MONITOR HIV RISK AND PREVENTION BEHAVIORS AND EXPLORE THE SOCIOECOLOGICAL FACTORS THAT INFLUENCE THE USE OF NEW HIV PREVENTION TECHNOLOGIES (UG3 PHASE), WHILE ALSO ALLOWING TARGETED TESTING OF NOVEL DIGITAL HEALTH INTERVENTIONS (UH3 PHASE). WE WILL ALSO TEST THE EFFICACY OF EXPANDING THE CORE VERSION OF HMP (HMP BASIC) BY ADDING ADHERENCE TOOLS (HMP ENHANCED) FOR THOSE WHO ARE ON PREP OR ART TO IMPROVE ADHERENCE AND PERSISTENCE. IN AIM 1, WE WILL ENROLL AND RETAIN A LARGE (N=6000; 3000/YEAR), DIVERSE COHORT OF SEXUALLY ACTIVE, SGM ADOLESCENTS AND YOUNG ADULTS, AGES 13-34, USING INNOVATIVE DIGITAL RECRUITMENT, ENGAGEMENT AND RETENTION STRATEGIES. OVER THE COURSE OF THE STUDY, WE WILL LONGITUDINALLY CHARACTERIZE THE SEXUAL BEHAVIOR, HIV TRANSMISSION RISK, AND PREP UPTAKE TRAJECTORIES OF SGM YOUTH UTILIZING EPIDEMIOLOGICAL TRAJECTORY ANALYSES TO IDENTIFY THE MOST EFFECTIVE POINTS OF INTERVENTION (AIM 2). FOR AIM 3, WE WILL LAUNCH A RANDOMIZED CLINICAL TRIAL TO EXAMINE THE EFFICACY OF HMP ENHANCED TO IMPROVE PREP ADHERENCE AMONG HIV-NEGATIVE YOUTH (N =750) AND ART ADHERENCE AMONG HIV-POSITIVE YOUTH (N =150) COMPARED TO HMP BASIC. FINALLY, WE WILL MAXIMIZE THE PRODUCTIVITY OF THE COHORT BY TESTING NEW AND INNOVATIVE DIGITAL HEALTH DEVICES, HIV/STI DIAGNOSTICS AND INTERVENTIONS, INFORMED BY THE PREVIOUS AIMS AS WELL AS EMERGING NIH PREVENTION PRIORITIES (AIM 4). OUR INVESTIGATIVE TEAM HAS DECADES OF EXPERIENCE WITH RECRUITMENT, PREVENTION AND CARE OF SGM YOUTH AND LARGE-SCALE LONGITUDINAL COHORT STUDIES. THIS STUDY WILL CAPITALIZE UPON PRODUCTIVE EXISTING PARTNERSHIPS AND DIGITAL HEALTH EXPERTISE TO ARTICULATE THE DRIVERS OF THE ONGOING HIV EPIDEMIC AMONG THE MOST VULNERABLE POPULATIONS IN THE US IN ORDER TO IDENTIFY THE MOST EFFECTIVE, EXPEDITIOUS AND SCALABLE STRATEGIES TO ADDRESS THIS ONGOING PUBLIC HEALTH CRISIS.

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.

IMPROVING EVALUATION CAPACITY WITHIN THE GOVERNMENT OF TANZANIA UNDER PEPFAR

VIOLENCE AGAINST WOMEN AND SUBSTANCE USE PREVENTION INITIATIVE - THERE IS A HIGH LEVEL OF NEED FOR COORDINATED INTIMATE PARTNER VIOLENCE (IPV) AND SUBSTANCE USE DISORDER (SUD) SERVICES IN WEST VIRGINIA, PARTICULARLY FOR PREGNANT AND PARENTING WOMEN (PPW), AS EVIDENCED BY HIGH RATES OF SUBSTANCE USE, POOR INFANT AND MATERNAL HEALTH OUTCOMES, AND RACIAL DISPARITIES IN HEALTH, HEALTHCARE ACCESS AND IPV. THE PURPOSE OF THIS PROJECT IS TO CREATE A STATEWIDE PILOT INITIATIVE TO TRAIN SUBSTANCE USE DISORDER (SUD) TREATMENT PROVIDERS ON INTIMATE PARTNER VIOLENCE (IPV) AND IPV PROVIDERS ON SUBSTANCE USE AND TO ADDRESS THE INTERSECTION OF IPV AND SUD DURING THE PREGNANCY AND POSTPARTUM PERIOD. PROJECT GOALS ARE TO ENGAGE STATE AGENCIES, IPV PROGRAMS, TREATMENT PROVIDERS, AND OTHER KEY STAKEHOLDERS FROM DIVERSE COMMUNITIES IN INCREASING THE USE OF EVIDENCE-BASED AND PROMISING PRACTICES IN SERVICE DELIVERY; TO CREATE AND DELIVER TRAINING FOR SUD, IPV, AND HEALTHCARE PROVIDERS; AND TO BUILD A COLLABORATIVE STATEWIDE NETWORK OF REGIONAL TEAMS TO PROMOTE SERVICE IMPLEMENTATION AT THE LOCAL LEVEL AND ADDRESS ACCESS AND OUTCOME DISPARITIES AND UNMET NEEDS. TO ACHIEVE THESE GOALS, THE NATIONAL CENTER ON DOMESTIC VIOLENCE, TRAUMA, AND MENTAL HEALTH IS PARTNERING WITH THE WEST VIRGINIA DEPARTMENT OF HEALTH AND HUMAN RESOURCES' BUREAU FOR BEHAVIORAL HEALTH, THE WEST VIRGINIA COALITION AGAINST DOMESTIC VIOLENCE, THE WEST VIRGINIA PERINATAL PARTNERSHIP, AND THE MARSHALL UNIVERSITY RESEARCH CORPORATION’S WEST VIRGINIA BEHAVIORAL HEALTH WORKFORCE AND HEALTH EQUITY TRAINING CENTER TO ENGAGE IN SIX KEY ACTIVITIES. FIRST, WE WILL INCENTIVIZE SUD PROVIDERS TREATING PREGNANT AND POSTPARTUM WOMEN TO RECEIVE TRAINING ON IDENTIFYING IPV IN THEIR CLIENT POPULATION. SECOND, WE WILL TRAIN SUD TREATMENT PROVIDERS TO ADDRESS IPV WITH PATIENTS AND TRAIN IPV STAFF ON SUD. THIRD, WE WILL IDENTIFY RESEARCH-BASED SERVICES FOR SCREENING, ASSESSMENT, BRIEF INTERVENTION, AND REFERRAL. FOURTH WE WILL INTEGRATE IPV AND SUD PROTOCOLS INTO MEDICAL PRAC TICE. FIFTH WE WILL INTEGRATE PERINATAL AND POSTPARTUM PROGRAMS INTO EXISTING SUBSTANCE USE PROGRAMS. SIXTH, WE WILL CONDUCT PROCESS AND OUTCOMES EVALUATIONS TO DETERMINE CHANGES IN PROVIDERS' KNOWLEDGE, SKILL, AND COMFORT IN ADDRESSING IPV/SU, DOCUMENT CHANGES IN REFERRALS AND COLLABORATION AMONG STATE PARTNERS, AND ASSESS IMPROVEMENT IN IPV-SUD HEALTH OUTCOMES AMONG PREGNANT AND POSTPARTUM WOMEN. OUR APPROACH BUILDS ON WEST VIRGINIA’S EXISTING STATEWIDE PARTNERSHIPS AND NETWORKS, INCORPORATES THE PROMISING PRACTICE OF REDUCING SUBSTANCE USE COERCION, AND RESPONDS TO THE HIGH OVERDOSE AND DRUG-RELATED DEATH RATE FOR WOMEN IN WV.

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.

SMALL TRANSLATED ORFS IN THE 3'UTR ENHANCE TRANSLATION IN VERTEBRATES - PROJECT SUMMARY THE PREVAILING DOCTRINE THAT MESSENGER RNAS (MRNAS) IN HIGHER ORGANISMS ENCODE FOR A SINGLE PROTEIN HAS UNDERGONE A DRAMATIC REVISION IN RECENT YEARS. RIBOSOME AND PROTEOMIC PROFILING HAVE REVEALED A LARGE NUMBER OF SMALL TRANSLATED OPEN READING FRAMES (ORF) WITHIN PREVIOUSLY DESCRIBED “UNTRANSLATED REGIONS” (UTRS) AND LONG NON-CODING RNAS. INDEED, SOME OF THE PEPTIDES DERIVED FROM SMALL ORFS HAVE BEEN IMPLICATED IN VARIOUS FUNDAMENTAL PROCESSES (E.G., DEVELOPMENT). TRANSLATION OF SMALL ORFS IN THE 5’UTR, KNOWN AS UPSTREAM-ORFS (UORFS), HAS BEEN SHOWN TO HAVE A PROFOUND REGULATORY EFFECT ON GENE REGULATION, INDEPENDENT OF THE ENCODED PEPTIDE. FURTHER, TRANSLATION OF UORFS VARY UNDER PATHOLOGIC CONDITIONS SUCH AS CANCER, AND MUTATIONS AFFECTING UORFS ARE ASSOCIATED WITH VARIOUS HUMAN DISEASES. WE AND OTHERS HAVE ALSO INDICATED THE EXISTENCE OF TRANSLATED SMALL ORFS IN THE 3’UTR KNOWN AS DOWNSTREAM OPEN READING FRAMES (DORFS) IN HUMAN CELLS AND ZEBRAFISH EMBRYOS. HOWEVER, CONTRARY TO UORFS, THERE HAS BEEN NO SYSTEMATIC STUDY OF DORF FUNCTIONS, AND THEIR RELATIONSHIP TO HUMAN HEALTH AND DISEASE REMAINS UNTESTED. FURTHER, GIVEN THEIR LOCATION IN THE 3’UTR, THE MOLECULAR MECHANISM BY WHICH DORFS ENGAGE THE TRANSLATIONAL MACHINERY REMAIN COMPLETELY UNKNOWN. OUR LONG-TERM GOAL IS TO UNDERSTAND HOW POST-TRANSCRIPTIONAL REGULATION (MRNA HALF-LIFE AND TRANSLATION) SHAPES GENE EXPRESSION IN VERTEBRATES, AND ITS IMPACT ON HUMAN DISEASE. THE CENTRAL HYPOTHESIS OF THIS APPLICATION IS THAT TRANSLATION OF DORFS REGULATES GENE EXPRESSION. OUR PRELIMINARY DATA STRONGLY INDICATE THAT, CONTRARY TO UORFS, DORFS STRONGLY ENHANCE TRANSLATION OF THE CANONICAL ORF AND EMERGE AS AN UNCHARACTERIZED AND POTENT REGULATORY MECHANISM ACROSS VERTEBRATES. THE OBJECTIVES ARE TO: 1) IDENTIFY FACTORS INVOLVED IN ENHANCING TRANSLATION OF THE MAIN ORF. 2) DISSECT THE REGULATORY INFORMATION DRIVING DORF TRANSLATION, AND 3) CHARACTERIZE THE BIOLOGICAL IMPACTS OF DORF-MEDIATED REGULATION. THE RATIONALE FOR THE PROPOSED RESEARCH IS TO GAIN A MECHANISTIC UNDERSTANDING OF DORF-MEDIATED REGULATION IN ORDER TO ASSESS THE POSSIBLE BIOLOGICAL IMPORTANCE OF DORF DYSREGULATION UNDER STRESS OR DISEASE CONDITIONS. THIS PROPOSAL IS CONCEPTUALLY INNOVATIVE AS IT IS BASED ON THE EXPLORATION OF A NOVEL, YET WIDESPREAD AND POTENT TRANSLATION REGULATORY MECHANISM CONSERVED ACROSS VERTEBRATES. TECHNICALLY, THIS PROPOSAL WILL COMBINE GENOMIC PROFILES (RNA-SEQ, RIBOSOME PROFILING); REPORTER (CYTOMETRY); BIOCHEMISTRY TOOLS: RNA PULLDOWNS FOLLOW BY PROTEOMICS, CRISPR-CAS-9 AND - 12A (TO EDIT) AND OUR NOVEL CAS13D TOOL (KNOCK-DOWN IN EMBRYOS); COMBINING HUMAN CELL AND ZEBRAFISH EMBRYOS. THE OUTCOMES FROM THIS PROJECT WILL HELP UNDERSTAND HOW DORFS ARE TRANSLATED, SHAPE GENE EXPRESSION AND GENERATE PHENOTYPES. THIS NOVEL FUNCTION OF THE RIBOSOME ADDS TO THE RECENTLY EMERGING REGULATORY EFFECTS OF TRANSLATION ON GENE EXPRESSION (E.G. UORF, CODON OPTIMALITY). UNDERSTANDING DORF BIOLOGY WILL PROVIDE AN ENTRY POINT AND PERHAPS EVEN A DIAGNOSTIC TOOL TO ASSOCIATE MUTATIONS WITH HUMAN DISEASES. IDENTIFYING THE MOLECULAR MACHINERY INVOLVED IN THIS PATHWAY MIGHT PROVIDE TARGETS FOR THERAPEUTIC INTERVENTIONS.

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.

HOW CELLS MONITOR THE INTEGRITY OF THEIR TRANSLATION APPARATUS

IDO2 TARGETING IN PANCREATIC CANCER

MECHANISMS OF TUMOR INITIATION UPON DISRUPTION OF THE RB/E2F INTERACTION - ABSTRACT E2F (E2F1-8) TRANSCRIPTION FACTORS ARE CRITICAL REGULATORS OF CELL CYCLE AND THEIR ACTIVITY IS PHYSICALLY REGULATED BY RB FAMILY PROTEINS (RB, P107 AND P130). DISRUPTION OF THE RB/E2F INTERACTION IS A HALLMARK OF CANCER, AS DEFINED BY HANAHAN&WEINBERG. THE ACQUIRED RESISTANCE TO ANTIGROWTH SIGNALS RESULTING FROM THIS DISRUPTION IS THOUGHT TO BE NECESSARY FOR TUMOR INITIATION. HOWEVER, HOW DOES UNRESTRICTED E2F ACTIVITY INITIATES TUMORIGENESIS IN VIVO IS POORLY UNDERSTOOD AND REMAINS A FUNDAMENTAL GAP IN OUR UNDERSTANDING OF CANCER BIOLOGY. IN PARTICULAR, WHETHER THE DIFFERENTIATION STATUS AFFECT THE CAPACITY OF A CELL TO TRANSFORM UPON DISRUPTION OF RB/E2F INTERACTION IS UNKNOWN. IN ADDITION, WHETHER UNRESTRICTED E2F ACTIVATES OTHER ONCOGENIC FEATURES BESIDES ABERRANT PROLIFERATION IS STILL OBSCURE. HEPATOCELLULAR CARCINOMA (HCC) IS THE SECOND CANCER IN TERMS OF DEATH WORLDWIDE. LIMITED UNDERSTANDING OF THE FUNCTIONAL CONSEQUENCES FOR FREQUENT GENETIC EVENTS HAMPERS THE DEVELOPMENT OF EFFICIENT THERAPEUTICS. THE RB/E2F INTERACTION IS DISRUPTED IN THE VAST MAJORITY OF HCC, AS A CONSEQUENCE OF SEVERAL EVENTS THAT TARGET UPSTREAM COMPONENTS OF THE RB/E2F PATHWAY. THEREFORE, HCC IS A RELEVANT MODEL TO INVESTIGATE THE CONSEQUENCES OF UNRESTRICTED E2F ACTIVITY FOR CANCER INITIATION. ACCORDINGLY, PAN-LIVER INACTIVATION OF RB FAMILY GENES (TRIPLE KNOCK OUT, TKO) IS SUFFICIENT TO INITIATE HCC (TKO HCC) THAT RECAPITULATE MULTIPLE FEATURES OF THE HUMAN DISEASE. STUDIES OF CARCINOGEN-INDUCED MODELS OF HCC HAVE LED TO THE CONCLUSION THAT HEPATOCYTES ARE THE SOLE SOURCE OF HCC. SPECIFIC INACTIVATION OF RB FAMILY IN HEPATOCYTES TRIGGERS A SHORT PROLIFERATIVE BURST BUT FAILS TO INITIATE HCC. THIS RESULT CHALLENGES THE CURRENT HEPATO-CENTRIC MODEL IN THE FIELD AND SUGGESTS THAT OTHER LINEAGES CAN ALSO SERVE AS A CELL OF ORIGIN FOR HCC. ACCORDINGLY, INACTIVATION OF RB FAMILY IN MULTIPLE LIVER LINEAGES REVEALS THAT TKO HCC ARISES FROM A PERIDUCTAL PROGENITOR. IN PARTICULAR, OUR PRELIMINARY DATA INDICATES THAT UNRESTRICTED E2F ACTIVITY COUPLES ABERRANT PROLIFERATION WITH CELL FATE ALTERATION IN THIS POPULATION TO INITIATE HCC. BASED ON THESE DATA, WE PROPOSE TO:1) DETERMINE THE MOLECULAR MECHANISMS THAT ALTER THE CELL FATE OF A PERIDUCTAL PROGENITOR TO SERVE AS A CELL OF ORIGIN FOR TKO HCC. 2) DETERMINE THE INDIVIDUAL AND COMPOUND ROLE OF E2F FACTORS IN TKO HCC INITIATION AND DEVELOPMENT. 3) DETERMINE THERAPEUTIC VULNERABILITIES IN TKO HCC THAT COULD SERVE AS NOVEL TREATMENT FOR PATIENTS. WE BELIEVE THAT OUR PROPOSAL WILL ADDRESS FUNDAMENTAL QUESTIONS REGARDING THE ROLE OF E2F IN CANCER INITIATION, AS IDENTIFIED ABOVE. IN ADDITION, WE EXPECT THAT OUR RESULTS WILL ESTABLISH THAT DIFFERENT CELL LINEAGES CAN SERVE AS A CELL OF ORIGIN FOR HCC, WHICH WILL HAVE IMPORTANT CLINICAL IMPLICATIONS, IN PARTICULAR REGARDING THE CLASSIFICATION OF PATIENTS AND THE DEVELOPMENT OF THERAPIES TAILORED FOR DIFFERENT CLASSES OF HCC.

NOGUCHI INSTITUTE INITIATIVE FOR NTDS ELIMINATION (NIINE)

ENHANCEMENT OF HIV/AIDS LAB TRAINING AND QA CENTER IN THE UR OF TANZANIA

POLYAMINE-STIMULATED STEM CELL RECRUITMENT IN ARSENIC-INDUCED SKIN CANCER

MAINTAINING THE INTEGRITY OF A GENOME - PROJECT SUMMARY/ABSTRACT IN ORDER TO FULLY GRASP THE MOLECULAR ORIGINS OF GENOME INSTABILITY, THE FIELD MUST UNDERSTAND AT A MOLECULAR LEVEL HOW CENTROMERES WORK TO PROMOTE THE STABLE TRANSMISSION OF CHROMOSOMES. GENOME INSTABILITY UNDERLIES A VARIETY OF HUMAN PATHOLOGIES, INCLUDING CANCER AND REPRODUCTIVE AGING. OUR LONG-TERM GOAL IS TO DETERMINE HOW THE EVOLUTIONARILY CONSERVED COHESIN COMPLEX MAINTAINS GENOME INTEGRITY THROUGH ITS ROLES IN CHROMOSOME SEGREGATION, CHROMOSOME ORGANIZATION, AND DOUBLE-STRAND BREAK REPAIR. LOSS OF SISTER CHROMATID COHESION IS SPECULATED TO BE A MAJOR CONTRIBUTOR TO CHROMOSOME INSTABILITY. THE OBJECTIVE OF THIS APPLICATION IS TO PRODUCE A MOLECULAR MODEL FOR HOW COHESIN OPERATES AT INDIVIDUAL HUMAN CENTROMERES TO ACHIEVE CENTROMERIC COHESION AND ACCURATE CHROMOSOME SEGREGATION. THE CENTRAL HYPOTHESIS IS THAT COHESIN AND DNA CATENATION TOGETHER CREATE CENTROMERE-UNIQUE LANDSCAPES OF SISTER CHROMATID COHESION TO PREVENT CHROMOSOME INSTABILITY. THE VARIATION IN HUMAN CENTROMERES AND CENTROMERIC COHESION MAY THEREFORE IMPACT THE TRANSMISSION OF EACH CHROMOSOME. WE WILL TEST THE IDEA THAT CENTROMERE-SPECIFIC COHESION MUST BE CONSIDERED AS A GENETIC DETERMINANT OF SISTER CHROMATID COHESION AND SEGREGATION IN ORDER TO HAVE A COMPLETE MODEL FOR HOW CHROMOSOMAL INSTABILITY OCCURS THROUGH TWO SPECIFIC AIMS: 1) DISCOVER THE LANDSCAPE OF CENTROMERIC COHESION AT INDIVIDUAL HUMAN CENTROMERES AND 2) EXAMINE HOW CHROMOSOME CENTROMERIC COHESION MAINTAINS EUPLOIDY. UNDER THE FIRST AIM, CALIBRATED PAIRED-END CHIP SEQ WILL BE USED TO MAP COHESIN BINDING RELATIVE TO KINETOCHORE PROTEINS AND HUMAN CENTROMERIC ARRAYS IN HUMAN TISSUE CULTURE CELLS. THIS APPROACH WILL BE COMPLEMENTED BY SUPERRESOLUTION IMAGING OF THE SAME THREE COMPONENTS (CENTROMERES, KINETOCHORES, AND COHESIN) IN CELLS, AND WILL INCLUDE IMAGING-BASED DETERMINATION OF CENTROMERE-SPECIFIC COHESION. TOGETHER THESE APPROACHES WILL PRODUCE A LINEAR AND 3D MAP OF COHESION WITHIN AND AROUND INDIVIDUAL HUMAN CENTROMERES. IN THE SECOND AIM WE WILL EXAMINE HOW CENTROMERE-SPECIFIC PATTERNS OF CENTROMERIC COHESION PREVENT CHROMOSOME MISSEGREGATION EVENTS IN CULTURED CELLS AND IN XENOGRAFT TUMOR TISSUE. THE OUTCOME WILL BE FUNDAMENTAL PRINCIPLES OF CENTROMERIC ARRAY-BASED COHESION FATIGUE AND RESULTING PATTERNS OF CHROMOSOME INSTABILITY. THE RESEARCH IS INNOVATIVE BECAUSE IT INCORPORATES THE LATEST INFORMATION ON HUMAN CENTROMERIC DNA ARRAYS, A NEW WORKING MODEL FOR THE ORGANIZATION OF CENTROMERIC DNA BY COHESION, AND NEW QUANTITATIVE MOLECULAR, GENOMIC, AND IMAGING TOOLS TO PROBE HOW CENTROMERIC COHESION ENFORCES ACCURATE SISTER CHROMATID SEGREGATION. THE PROPOSED RESEARCH IS SIGNIFICANT BECAUSE CENTROMERIC ARRAYS MAY BE UNRECOGNIZED GENETIC DETERMINANTS OF CHROMOSOME INSTABILITY. MANY TYPES OF CANCER ARE ASSOCIATED WITH SEEMINGLY RANDOM PATTERNS OF INSTABILITY THAT MAY HAVE MOLECULAR ORIGINS IN UNIQUE CENTROMERIC COHESION PROFILES. FURTHERMORE, MANY CANCERS ARE ASSOCIATED WITH MUTATIONS THAT IMPACT CHROMOSOME SEGREGATION MACHINERY, SUCH AS COHESIN. THE OUTCOME OF THIS PROJECT WILL BE A MORE COMPLETE PICTURE OF THE MECHANISMS UNDERLYING CHROMOSOMAL INSTABILITY.

THE CONTRIBUTION OF SLEEP AND CIRCADIAN DISRUPTION TO KYNURENINE PATHWAY ACTIVATION AND CARDIOMETABOLIC RISK IN WOMEN WITH HIV

PHYSIOLOGICAL ROLE OF MLL AND ELL PROTEINS IN LEUKEMIA

POWERING UP MALE PREVENTION (PUMP)

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

IDO PATHWAYS IN INFLAMMATORY PATHOGENESIS AND TREATMENT OF RHEUMATOID ARTHRITIS

FEASIBILITY OF AN INGESTIBLE SENSOR SYSTEM TO MEASURE PREP ADHERENCE IN YMSM

CHICAGO ANTIMICROBIAL RESISTANCE AND INFECTION PREVENTION EPI-CENTER (CARPE)

IDENTIFICATION OF GENES ASSOCIATED WITH MYOBLAST FUSION

DEVELOPING BEST PRACTICES OF COMMUNITY ENGAGEMENT FOR GENOMICS AND BIOBANKING IN AFRICA - CEBIOGEN

PROBING AN UNEXPLORED INTRACELLULAR PATHWAY IN DIABETES PATHOGENESIS - ABSTRACT DIABETIC NEPHROPATHY AND OTHER DIABETES COMPLICATIONS IMPOSE ENORMOUS BURDENS ON PATIENTS AND HEALTHCARE SYSTEMS, MAKING IT IMPERATIVE TO DEFINE ACTIONABLE ETIOLOGIC FACTORS AND DEVELOP EFFECTIVE, LOW-COST THERAPEUTIC INTERVENTIONS. NONENZYMATIC PROTEIN GLYCATION AND THE FORMATION OF ADVANCED GLYCATION END PRODUCTS (AGES) ARE STRONGLY IMPLICATED IN PATHOGENESIS. THE DRIVER OF AGE FORMATION IS 3-DEOXYGLUCOSONE (3DG), A HIGHLY REACTIVE DICARBONYL SPECIES THAT ALSO CAUSES ACUTE CELLULAR TOXICITIES BY DAMAGING ENZYMES AND DNA AND INFLAMING THE VASCULATURE. ACCORDINGLY, THE ABILITY TO ACCURATELY MEASURE 3DG LEVELS AND UNDERSTAND ITS ETIOLOGY ARE PARAMOUNT TO ELUCIDATING PATHOGENESIS, LIMITING ITS PATHOGENIC EFFECTS, AND IMPROVING CLINICAL MANAGEMENT OF DIABETIC COMPLICATIONS. ENDOGENOUS 3DG WAS DEEMED TO ARISE NONENZYMATICALLY FROM THE SLOW DISINTEGRATION OF GLYCATED PROTEINS IN THE BODY OR ABSORBED FROM INGESTED HEAT-PROCESSED FOODS. WE DEVELOPED NEW METHODS TO STUDY THE ENZYMATIC ACTIVITY OF FRUCTOSAMINE-3-KINASE (FN3K), AN ENZYME THOUGHT TO REPAIR GLYCATED PROTEINS AND PREVENT AGE, BUT AN END-PRODUCT OF FN3K ACTIVITY IS 3DG. WE DISCOVERED THAT 3DG LEVELS IN KIDNEY ARE HIGHER THAN PREVIOUSLY ANTICIPATED. OUR CORE HYPOTHESIS IS THAT FN3K-MEDIATED 3DG FORMATION IN CELLS IS A KEY PATHOGENIC DRIVER IN DIABETIC COMPLICATIONS. SPECIFIC AIM 1: WE WILL MEASURE 3DG ARISING IN TISSUES IN RELATIONSHIP TO PATHOGENESIS IN THE KK.CG-AY/J MURINE MODEL OF TYPE-2 DIABETES. SPECIFIC AIM 2: THE IMPACT OF A HIGH GLYCATION DIET ON 3DG LEVELS WILL BE MEASURED IN TISSUES SENSITIVE TO DIABETIC COMPLICATIONS, INCLUDING IN THE KIDNEY, HEART, AND LIVER OF THE DIABETIC MICE. SPECIFIC AIM 3: WE WILL DEFINE THE PHARMACODYNAMIC PROPERTIES AND MODES OF ACTION FOR MEGLUMINE, AN AGENT, ALREADY PROVEN SAFE, THAT WE DISCOVERED HAS UNRECOGNIZED MEDICINAL EFFECTS, HAVING PROVIDED NEPHROPROTECTION AND PREVENTED TRIGLYCERIDE ACCUMULATION IN DIABETIC MICE. SPECIFIC AIM 4: A SERIES OF FN3K ANTAGONISTS THAT WE DISCOVERED WILL BE CHARACTERIZED TO IDENTIFY A PRECLINICAL DRUG DEVELOPMENT CANDIDATE. THIS PROPOSAL OFFERS SEVERAL MAJOR INNOVATIVE ELEMENTS OF HIGH SIGNIFICANCE AND IMPACT IN DIABETES TRANSLATIONAL RESEARCH. AIM 1 WILL PROVIDE NEW DATA DEVELOPED WITH METHODOLOGY WE REFINED TO MEASURE FN3K ACTIVITY AND 3DG FORMATION MORE ACCURATELY, ADDRESSING KEY GAPS IN KNOWLEDGE. AIM 2 WILL EXPLORE THE LINKAGE BETWEEN INTRACELLULAR 3DG ELEVATION AND THE CONSUMPTION OF ‘WESTERN’ DIETS RICH IN FRUCTOSAMINES—THE SUBSTRATE FOR FN3K. DRUG SAFETY IS PARAMOUNT FOR ANY NEW DIABETES DRUG. THE DATA FROM AIM 3 WILL ACCELERATE THE DEVELOPMENT OF MEGLUMINE AS AN INNOVATIVE TREATMENT MODALITY—A COMPOUND PROVEN EXTREMELY SAFE FOR CHRONIC ADMINISTRATION—TO AMELIORATE DIABETIC NEPHROPATHY, FATTY LIVER, AND POTENTIALLY OTHER DIABETIC COMPLICATIONS. AIM 4 OFFERS OPPORTUNITY TO DELIVER FIRST-IN-CLASS ENZYME INHIBITORS AS POTENTIAL DRUG LEAD CANDIDATES. IN SUMMARY, THIS RESEARCH PROGRAM WILL ILLUMINATE AN UNEXPLORED INTRACELLULAR PATHWAY IN DIABETES PATHOGENESIS AND DELIVER UNPRECEDENTED TOOLS FOR BROADER RESEARCH INTO THE ROLE OF 3DG IN DIABETIC NEPHROPATHY AND OTHER DIABETES COMPLICATIONS.

RYAN WHITE CARE ACT TITLE IV ADOLESCENT INITIATIVE

IN VIVO ANALYSIS OF THE MECHANISMS OF NEURAL CREST MIGRATION

CONTRIBUTIONS OF PROTEIN AGGREGATION TO GENE REGULATION AND PHENOTYPIC DIVERSITY

DRUG-INDUCED REGENERATION AND RE-INNERVATION IN A MOUSE DIGIT AMPUTATION MODEL

INHIBITOR REPROGRAMMING OF A HUMAN HISTONE DEACETYLASE PROTEIN INTERACTION NETWORK

BIOCHEMISTRY OF EUKARYOTIC MESSENGER RNA SYNTHESIS

PRIMARY CARE TRAINING AND ENHANCEMENT

MOLECULAR MECHANISMS OF CHROMOSOME SEGREGATION IN YEAST

MECHANISMS OF TRANSCRIPTIONAL REGULATION IN CHROMATIN

HEALTH CARE AND OTHER FACILITIES

IDO INHIBITORS FOR COMBINATORIAL CANCER THERAPY

SPECIAL PROJECTS OF NATIONAL SIGNIFICANCE

MITOTIC ROUNDING AND PLANAR SPINDLE ALIGNMENT IN PROLIFERATING EPITHELIA

MOLECULAR MECHANISMS AND EVOLUTIONARY IMPACTS OF THE WTF MEIOTIC DRIVERS - PROJECT SUMMARY/ABSTRACT GENOMES ARE PLAGUED BY PARASITIC DNA SEQUENCES THAT DO NOT PROMOTE HEALTH OR FERTILITY. MEIOTIC DRIVE GENES ARE ONE SUCH CLASS OF GENETIC PARASITE THAT SELFISHLY MANIPULATE GAMETOGENESIS TO INCREASE THEIR OWN TRANSMISSION INTO THE NEXT GENERATION. RATHER THAN BEING PASSED TO HALF OF AN INDIVIDUAL’S OFFSPRING, LIKE REGULAR ALLELES, MEIOTIC DRIVERS MANIPULATE GAMETOGENESIS TO ENSURE THEIR TRANSMISSION TO MOST, OR EVEN ALL, THE OFFSPRING. DRIVERS ARE FOUND THROUGHOUT EUKARYOTES, INCLUDING HUMANS, BUT MANY MORE LIKELY REMAIN TO BE IDENTIFIED. CHARACTERIZING MEIOTIC DRIVERS IS IMPORTANT BECAUSE THESE PARASITES CAN HAVE MAJOR IMPACTS ON FERTILITY AND HEALTH. MEIOTIC DRIVERS CAN CONTRIBUTE TO INFERTILITY DIRECTLY BY DISRUPTING MEIOTIC CHROMOSOME SEGREGATION OR BY DESTROYING GAMETES THAT INHERIT THE COMPETING ALLELE. MEIOTIC DRIVERS CAN ALSO CAUSE INFERTILITY OR OTHER DISEASE STATES INDIRECTLY BY PROMOTING THE SPREAD OF LINKED DELETERIOUS (E.G., DISEASE-ASSOCIATED) ALLELES. DESPITE THEIR LARGE IMPACT, THERE IS RELATIVELY LITTLE MOLECULAR UNDERSTANDING OF MEIOTIC DRIVERS OR THEIR EVOLUTIONARY IMPACTS. HOWEVER, THERE ARE EMERGING THEMES THAT UNITE MANY KNOWN DRIVE SYSTEMS, INCLUDING THE USE OF A POISON/ANTIDOTE MECHANISM AND THE ASSOCIATION OF MEIOTIC DRIVE GENES WITH DISTRIBUTED DNA SEQUENCE REPEATS. THIS PROPOSAL EXPLOITS A HIGHLY TRACTABLE MODEL, THE WTF GENE FAMILY FOUND IN FISSION YEASTS, TO INVESTIGATE THE MOLECULAR MECHANISMS AND EVOLUTIONARY IMPACTS OF DRIVE SYSTEMS. THE WTF GENES ENACT DRIVE BY DESTROYING THE WTF- GAMETES PRODUCED BY WTF+/WTF- HETEROZYGOTES. EACH WTF DRIVER ENCODES BOTH A POISON PROTEIN AND A SEPARATE ANTIDOTE PROTEIN ON OVERLAPPING CODING SEQUENCES. ALL DEVELOPING GAMETES ARE POISONED, BUT THOSE THAT INHERIT THE WTF+ ALLELE ARE RESCUED BY THE ANTIDOTE. THIS PROPOSAL AIMS TO UNDERSTAND THE MECHANISMS UNDERLYING THE TOXICITY OF WTF POISON PROTEINS AND HOW THE WTF ANTIDOTE PROTEINS RESCUE THAT TOXICITY. ANALOGOUS TO OTHER DRIVE SYSTEMS, THE WTF GENES ARE FLANKED BY SHORT REPETITIVE DNA SEQUENCES (TRANSPOSON-DERIVED REPEATS OR 5S RDNA GENES). THE PROPOSED WORK WILL TEST THE IDEA THAT THE REPEATS FLANKING WTF GENES AFFECT THEIR EVOLUTION BY PROMOTING NON-ALLELIC GENE CONVERSION. FINALLY, THE PROPOSED RESEARCH PROGRAM WILL EXPLORE THE EVOLUTIONARY IMPACT OF THE WTF GENES ON THE FLANKING 5S RDNA GENES. SPECIFICALLY, WE WILL TEST THE HYPOTHESIS THAT THE WTF DRIVERS HAVE PROMOTED THE MAINTENANCE OF DELETERIOUS VERSIONS OF THE 5S RDNA GENES, WHICH ENCODE AN ESSENTIAL COMPONENT OF RIBOSOMES. THIS RESEARCH PROGRAM WILL GREATLY EXPAND OUR UNDERSTANDING OF THE MOLECULAR MECHANISMS AND MOLECULAR EVOLUTION OF MEIOTIC DRIVE SYSTEMS. THIS KNOWLEDGE WILL HELP GUIDE THE SEARCH FOR AND THE MOLECULAR CHARACTERIZATION OF MEIOTIC DRIVERS IN MORE COMPLEX SYSTEMS, INCLUDING HUMANS. MORE BROADLY, THIS WORK WILL EXPAND OUR UNDERSTANDING OF DNA PARASITES AND HOW THEY CAN DIRECTLY AND INDIRECTLY IMPACT HEALTH, PARTICULARLY INFERTILITY. THIS EXPANDED UNDERSTANDING SHOULD ULTIMATELY LEAD TO IMPROVED REPRODUCTIVE OUTCOMES IN HUMANS.

(PQA2) MAMMALIAN REGENERATION, HIGH FAT DIETS, AND BREAST CANCER: A COMMON LINK

TARGETED NANOPARTICLE DNA THERAPY FOR OVARIAN CANCER

FVPSA AMERICAN RESCUE PLAN COVID-19 TESTING, VACCINES, AND MOBILE HEALTH UNITS SUPPLEMENTAL FUNDING

PREPAREDNESS AND RESPONSE TO AVIAN AND PANDEMIC INFLUENZA IN UNITED REP OF TANZAN

ASSEMBLY AND REGULATION OF YEAST SPINDLE POLES

ELUCIDATING MECHANISMS OF AMYLOID NUCLEATION IN VIVO

MECHANISMS OF TRANSCRIPTIONAL REGULATION IN CHROMATIN

FAMILY VIOLENCE PREVENTION AND SERVICES PROGRAM

THE NETWORK BIOLOGY OF PATHOGEN-HOST INTERACTIONS DRIVING EXACERBATION IN CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD)

IMPROVING EVALUATION CAPACITY WITHIN THE GOVERNMENT OF TANZANIA UNDER PEPFAR

ANALYSIS OF METAZOAN SAGA COMPLEX FUNCTION IN GENE EXPRESSION

INVESTIGATION OF NOTCH SIGNALING IN THE REGULATION OF CILIARY BODY DEVELOPMENT AND FUNCTION

SPECIAL PROJECTS OF NATIONAL SIGNIFICANCE

TRUST IN HEALTH CARE AND RACIAL DISPARITIES IN AN AGING POPULATION

KEEPING IT LITE: EXPLORING HIV RISK IN VULNERABLE YOUTH WITH LIMITED INTERACTION

FACULTY DEVELOPMENT IN PRIMARY CARE

DEFINING THE CELLULAR AND MOLECULAR MECHANISM OF IDO2 FUNCTION DRIVING AUTOIMMUNE VS.PROTECTIVE IMMUNE RESPONSES - PROJECT SUMMARY AUTOIMMUNE DISEASES ARE CHRONIC INFLAMMATORY DISEASES THAT AFFECT MORE THAN 50 MILLION AMERICANS, LEADING TO PAIN, DISABILITY, AND INCREASED MORTALITY. WHILE DRUGS TO TREAT AUTOIMMUNE DISEASES EXIST, ALLEVIATION OF INFLAMMATORY SYMPTOMS IS OFTEN ASSOCIATED WITH SEVERE SIDE-EFFECTS, INCLUDING INCREASED SUSCEPTIBILITY TO INFECTIONS AND CANCER. OUR PROPOSAL ADDRESSES THE URGENT NEED TO IDENTIFY NEW THERAPEUTIC TARGETS TO ADDRESS THE UNDERLYING CAUSES AND ASSOCIATED SYMPTOMS OF THESE DEBILITATING DISEASES. IN THIS PROPOSAL, WE WILL INVESTIGATE INDOLEAMINE-2,3-DIOXYGENASE (IDO2) AS A NOVEL IMMUNOMODULATORY TARGET FOR AUTOIMMUNE DISEASES. IDO2 IS ONE OF TWO CLOSELY RELATED TRYPTOPHAN CATABOLIZING ENZYMES EXPRESSED PRIMARILY IN IMMUNE CELLS. USING A COMBINATION OF GENETIC AND PHARMACOLOGICAL STUDIES IN THE KRN MODEL OF INFLAMMATORY ARTHRITIS, WE IDENTIFIED IDO2 AS AN ESSENTIAL MEDIATOR OF AUTOANTIBODY-MEDIATED INFLAMMATORY RESPONSES AND DISCOVERED A NOVEL NON-ENZYMATIC FUNCTION OF IDO2 THAT DRIVES AUTOIMMUNE RESPONSES LEADING TO DISEASE. MECHANISTIC STUDIES IMPLICATE THE TRANSCRIPTION FACTOR RUNX1 AS A POTENTIAL COMPONENT OF THIS PREVIOUSLY UNKNOWN IDO2 SIGNALING PATHWAY. IDO2 APPEARS TO BE ESSENTIAL FOR ONLY SOME B CELL FUNCTIONS, AS IDO2 DEFICIENT MICE DEVELOP NORMAL IMMUNE CELL COMPARTMENTS AND MOUNT PRODUCTIVE IMMUNE RESPONSES TO CHALLENGE WITH SOME, BUT NOT ALL, MODEL ANTIGENS IN VITRO AND IN VIVO. A BETTER UNDERSTANDING OF IDO2 BIOLOGY, IN PARTICULAR THE ROLES THAT ENZYMATIC AND NON-ENZYMATIC ACTIVITY HAVE IN REGULATING IMMUNE FUNCTION IS NECESSARY TO DEVELOP IDO2 AS A POTENTIAL THERAPEUTIC TARGET FOR AUTOIMMUNE DISEASE. OUR WORKING HYPOTHESIS IS THAT IDO2 MODULATION OF IMMUNE RESPONSES INVOLVES BOTH ENZYMATIC AND NON-ENZYMATIC PATHWAYS WITH AUTOREACTIVE B CELL ACTIVATION/AUTOANTIBODY PRODUCTION DEPENDENT ON IDO2’S NON-ENZYMATIC FUNCTION. WE PROPOSE THAT THIS DIFFERENTIAL USE OF NON- ENZYMATIC VS. ENZYMATIC IDO2 PATHWAYS CAN BE SPECIFICALLY TARGETED AS A THERAPEUTIC STRATEGY TO INHIBIT IDO2 FUNCTION IN AUTOIMMUNE RESPONSES WITHOUT AFFECTING ITS ROLE IN PROTECTIVE IMMUNITY. IN THIS PROPOSAL, WE WILL USE PRECLINICAL MODELS OF AUTOIMMUNE ARTHRITIS, IDO2 AND RUNX1 CONDITIONAL KNOCKOUT MICE AND OVEREXPRESSING CELL LINES, AND WHOLE TRANSCRIPTOME RNA-SEQ ANALYSIS TO DEFINE THE CELLULAR AND MOLECULAR MECHANISM MEDIATING IDO2 NON-ENZYMATIC FUNCTION (AIM 1). WE WILL THEN USE CATALYTICALLY INACTIVE IDO2 KNOCK- IN MICE AND A NOVEL IDO2-TARGETING ANTIBODY, TOGETHER WITH IDO2-DEPENDENT MODELS OF AUTOIMMUNITY AND PROTECTIVE IMMUNITY, TO DETERMINE IF DIFFERENTIAL USE OF NON-ENZYMATIC VS. ENZYMATIC PATHWAYS DISTINGUISHES IDO2 FUNCTION IN AUTOIMMUNE AND PROTECTIVE IMMUNE RESPONSES (AIM 2). THE POTENTIAL LONG-TERM IMPACT OF THIS PROJECT WOULD MOVE TARGETING NON-ENZYMATIC IDO2 FUNCTION INTO DEVELOPMENT AS A NOVEL STRATEGY TO TREAT AUTOANTIBODY-MEDIATED AUTOIMMUNE DISEASES.

FY2021 FVPSA ARP ACT SUPPLEMENTAL

COMPUTER ASSISTED QUALITY OF LIFE AND SYMPTOM ASSESSMENT OF COMPLEX PATIENTS

ARRA - TRAINING IN PRIMARY CARE MEDICINE AND DENTISTRY: RESIDENCY TRAINING IN PRIMARY CARE

TARGETING NANOPARTICLE DNA DELIVERY TO PROSTATE TUMORS

POLYAMINES AND EPITHELIAL TUMORIGENESIS

PREVENTIVE MEDICINE RESIDENCIES

INCREASING UPTAKE OF MALE CIRCUMCISION AMONG MEN AGED 20-34 YEARS IN IRINGA & TAB

BENEFITS, HARMS, AND COST OF OSTEOPOROSIS SCREENING IN MALE VETERANS

THIS APPLICATION INVESTIGATES MICROBIAL GROWTH AND PHYSIOLOGICAL RESPONSES TO THE MULTIPLE STIMULI ENCOUNTERED IN SPACE FLIGHT ENVIRONMENTS, USING SA

ANALYSIS OF HAIR CELL REGENERATION IN ZEBRAFISH

THE PURPOSE OF THE MONITORING & SURVEILLANCE DATA FOR EFFECTIVE MALARIA CONTROL ACTIVITY IS TO IMPLEMENT TWO ACTIVITIES TO SUPPORT THE COUNTRY OF GHANA’S MALARIA CONTROL EFFORTS. THE ACTIVITIES PROPOSED IN THE COOPERATIVE AGREEMENT SUPPORT MALARIA MONITORING UNDER TWO BROAD COMPONENTS: COMPONENT 1: THERAPEUTIC EFFICACY STUDIES (TES) IN GHANA AND COMPONENT 2: ENTOMOLOGICAL MONITORING.

DIRECTING THE FATE OF CELLS TO MYOGENIC LINEAGES

MOLECULAR MECHANISMS REGULATING DROSOPHILA OVARIAN GERMLINE STEM CELLS

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

BIOCHEMISTRY OF EUKARYOTIC MESSENGER RNA SYNTHESIS

MECHANISMS OF MEIOTIC DRIVE AND THE FUNCTIONAL CONSEQUENCES OF RAPID GENOME EVOLUTION

22-22-SBBLEO2-0001 PROTECTION FROM COSMIC RADIATION..

HISTONE MODIFICATIONS REGULATING PHENOTYPE PLASTICITY: ROLE OF NON-CANONICAL WNT SIGNALING

GENE REGULATORY NETWORKS OF SYNAPTIC SPECIFICITY - PROJECT SUMMARY/ABSTRACT NEURONAL TYPE IDENTITY IS CENTRAL TO THE DEVELOPMENT AND FUNCTION OF NEURAL CIRCUITS, AS IT INSTRUCTS BOTH THE CONNECTIVITY OF NEURONS AS WELL AS THEIR SYNAPTIC AND ELECTROPHYSIOLOGICAL PROPERTIES. NEURONAL FATES ARE THOUGHT TO BE CONTROLLED BY COMBINATIONS OF TRANSCRIPTION FACTORS (TF) KNOWN AS TERMINAL SELECTORS, BUT VERY LITTLE IS KNOWN ABOUT THE GENE REGULATORY MECHANISMS THAT LINK DIFFERENTIAL TF EXPRESSION TO SPECIFIC NEURONAL FEATURES. THE DROSOPHILA VISUAL SYSTEM, A WELL-CHARACTERIZED BRAIN REGION THAT HAS AN ORGANIZATION ANALOGOUS TO THE VERTEBRATE RETINA AND CORTEX, PROVIDES THE IDEAL BALANCE OF COMPLEXITY AND ACCESSIBILITY TO INVESTIGATE THESE MECHANISMS. THE FIRST AIM OF THIS PROJECT WILL BE TO DECIPHER THE TERMINAL SELECTOR TF CODES THAT ESTABLISH AND MAINTAIN THE UNIQUE IDENTITY OF APPROXIMATELY 200 NEURONAL TYPES THAT MAKE THE DROSOPHILA OPTIC LOBES. USING A SINGLE-CELL RNA SEQUENCING (SCRNA-SEQ) DATASET I GENERATED FROM DEVELOPING OPTIC LOBES, I IDENTIFIED THE COMBINATIONS OF TFS THAT ARE STABLY MAINTAINED IN EACH NEURONAL TYPE THROUGHOUT THEIR DIFFERENTIATION. UNDER THE MENTORSHIP OF CLAUDE DESPLAN (K99 PHASE), I WILL TEST THE HYPOTHESIS THAT THESE TFS FUNCTION AS TERMINAL SELECTORS BY MODIFYING THE TF CODES OF SPECIFIC OPTIC LOBE NEURONS IN VIVO, WITH THE GOAL OF PREDICTABLY TRANSDIFFERENTIATING THEM INTO OTHER CELL TYPES. THIS WILL DEMONSTRATE THE SUFFICIENCY OF TERMINAL SELECTORS TO CONFER NEURONAL IDENTITY AND BENEFIT THE FIELD OF REGENERATIVE MEDICINE. THE CONSERVED MECHANISMS IN MAMMALIAN SYSTEMS COULD BE EXPLOITED TO INDUCE DIFFERENTIATION OF PLURIPOTENT CELLS INTO SPECIFIC NEURONS THAT COULD BE TRANSPLANTED TO TREAT BLINDNESS OR NEURODEGENERATION. THE SECOND AIM OF THIS PROJECT WILL LINK THE TERMINAL SELECTOR TFS TO THEIR DOWNSTREAM TARGETS. IN COLLABORATION WITH RICHARD BONNEAU, I WILL LEARN TO USE THE “INFERELATOR” ALGORITHM TO GENERATE COMPUTATIONAL MODELS OF GENE REGULATORY NETWORKS BY COMBINING MY EXISTING SCRNA-SEQ DATA WITH NEW CHROMATIN ACCESSIBILITY (SCATAC-SEQ) DATA I WILL ACQUIRE. DURING THE K99 PHASE, I WILL TEST THE EFFECTS OF PERTURBATING KEY PREDICTED DOWNSTREAM EFFECTORS ON THE MORPHOLOGY AND CONNECTIVITY OF A SELECT GROUP OF NEURONS TO ESTABLISH PROOF-OF- CONCEPT. I WILL THEN GENERALIZE THIS APPROACH IN THE R00 PHASE BY INFERRING GENE REGULATORY NETWORKS FOR ALL OPTIC LOBE NEURONS AT MULTIPLE DEVELOPMENTAL STAGES. THE THIRD AIM WILL BE PERFORMED IN MY INDEPENDENT LAB (R00) TO UTILIZE THE NETWORK MODELS FOR ENGINEERING PRECISE MODIFICATIONS IN VISUAL CIRCUITS. I WILL SEEK TO SELECTIVELY UNCOUPLE THE CIRCUIT THAT COMPUTES WIDE-FIELD MOTION FROM THE ONE THAT DETECTS SMALL MOVING OBJECTS. I WILL USE SYNAPTIC TRACING METHODS AS WELL AS INTRAVITAL CALCIUM IMAGING TO DEMONSTRATE THE FUNCTIONAL CONSEQUENCES OF DEVELOPMENTAL PERTURBATIONS. ALTOGETHER, THIS PROJECT WILL ESTABLISH DIRECT MECHANISTIC LINKS BETWEEN THE ENCODING OF NEURONAL IDENTITY AND THE MOLECULES THAT MEDIATE INTERCELLULAR INTERACTIONS DURING SYNAPTIC PARTNER SELECTION, WHICH ARE COMMONLY AFFECTED IN NEURODEVELOPMENTAL DISORDERS. THE MENTORSHIP I WILL RECEIVE FROM DR. DESPLAN AND DR. BONNEAU, COMBINED WITH THE IMPRESSIVE RESOURCES OF NEW YORK UNIVERSITY PROVIDE THE IDEAL ENVIRONMENT FOR PREPARING ME TO BUILD A SUCCESSFUL INDEPENDENT RESEARCH PROGRAM THAT LINK GENE REGULATION TO BRAIN WIRING.

THE PRIMARY RESEARCH PRODUCT THIS PROJECT WILL PRODUCE IS IDENTIFICATION OF THE INSERTION STRAINS OF CHLAMYDOMONAS REINHARDTII A SINGLE-CELL GREEN ALGA WHICH OPTIMIZE SURVIVAL DURING EXPOSURE TO THE COMBINED IMPACTS OF SPACE RADIATION AND MICROGRAVITY IN THE COSMIC SPACE ENVIRONMENT BEYOND THE VAN ALLEN BELTS. THE CHLAMYDOMONAS REINHARDTII STRAINS IN A MAPPED INSERTIONAL MUTANT LIBRARY ARE A TRANSFORMATIVE TOOL ALLOWING NEAR GENOME WIDE ANALYSIS OF THE GENES MEDIATING SURVIVAL ADVANTAGE DURING STIMULI SUCH AS COSMIC RADIATION AND MICROGRAVITY. THE SECONDARY SET OF RESEARCH PRODUCTS THIS PROJECT WILL PRODUCE ARE IDENTIFICATION OF THE PATHWAYS MEDIATING THE GENE CHANGES VISUALIZATION OF THE PATHWAY INTERACTIONS AND IDENTIFICATION OF ANY ENVIRONMENTS GIVING SIMILAR GENE PATHWAY SIGNATURES IN OTHER MODEL SYSTEMS. OUR GOAL IS TO IDENTIFY STRAINS OF CHLAMYDOMONAS REINHARDTII WITH GENE INSERTIONS WHICH CONVEY THE BEST SURVIVAL ADVANTAGE DURING THE EXPOSURE TO THE COMBINED IMPACTS OF SPACE RADIATION AND MICROGRAVITY FOR FUTURE USE AS PARENT STRAINS IN HYDROGEN PRODUCTION INITIATIVES. HYDROGEN IS THE LEAD CONTENDER FOR FUEL PRODUCTION FOR RETURN SPACE FLIGHT MISSIONS FROM ORBITAL BODIES BEYOND LOW EARTH ORBIT. THE METABOLISM OF SELECT MICROORGANISMS MAKES HYDROGEN FUEL. THE BEST-KNOWN AND MOST EFFICIENT HYDROGEN PRODUCER IS CHLAMYDOMONAS REINHARDTII WHICH IS POISED FOR EXPLOITATION AS AN INDUSTRIAL BIOTECHNOLOGY PLATFORM FOR BIOFUEL PRODUCTION. THE TYPE OF INVESTIGATION WE PROPOSE IS EXPERIMENTS THAT WILL BE CONDUCTED ABOARD THE EM-1 LUNAR ORBIT FLIGHT WITH ASYNCHRONOUS CONTROLS IN KENNEDY SPACE CENTER ISSES CHAMBERS (ISS ENVIRONMENTAL SIMULATORS). THE PROPOSED HARDWARE IS THE NASA BRIC-100VC. HYPOTHESIS: THE HYPOTHESIS TO BE TESTED IS THAT THE MAPPED INSERTIONAL MUTANT LIBRARY OF CHLAMYDOMONAS REINHARDTII WILL ALLOW IDENTIFICATION OF THE GENES THAT MEDIATE OPTIMAL RADIATION RESISTANCE AND IMPERVIOUSNESS TO MICROGRAVITY IN THE COSMIC SPACE ENVIRONMENT BEYOND THE VAN ALLEN BELTS. THE SPECIFIC AIMS OF THE PROPOSAL ARE: SPECIFIC AIM 1: TO EXPERIMENTALLY DETERMINE WHICH OF THE THOUSANDS OF MAPPED INSERTION MUTANT STRAINS OF CHLAMYDOMONAS REINHARDTII HAVE THE BEST LONG-TERM SURVIVAL ADVANTAGE DURING COMBINED SPACE RADIATION AND MICROGRAVITY IMPACTS IN THE COSMIC SPACE ENVIRONMENT BEYOND THE VAN ALLEN BELTS. SPECIFIC AIM 2: TO ANALYZE THE DATASET OF INSERTION STRAINS WITH THE BEST LONG-TERM SURVIVAL ADVANTAGE DURING COMBINED SPACE RADIATION AND MICROGRAVITY IMPACTS TO: PERFORM PATHWAY ANALYSIS USING GENE SET ENRICHMENT ANALYSIS (GSEA) VISUALIZE THE PATHWAY INTERACTIONS USING CYTOSCAPE/CLUEGO MAKE COMPARISON TO HOMOLOGOUS GENE SETS IN CHLAMYDOMAS SACCHAROMYCES CEREVISIAE AND OTHER MODEL ORGANISM DATABASES FOR ANY ENVIRONMENTS GIVING SIMILAR GENE PATHWAY SIGNATURES. ALTHOUGH FOCUSED ON GENE PATHWAYS FOR SURVIVAL FOR FUTURE APPLICATIONS IN HYDROGEN FUEL PRODUCTION THE DATA GENERATED WILL PROVIDE GENOME LEVEL INSIGHTS INTO THE CANCER RISKS OF PROLONGED EXPOSURE TO COSMIC RADIATION AND MICROGRAVITY. THE SPECTRUM OF CANCER RISK GENES IS KNOWN AND THIS STUDY WILL PARSE THE IMPORTANCE OF RISK GENES AND THE PATTERNS OF RADIATION RISK CHANGE WITH EXPOSURE TO THE COSMIC SPACE ENVIRONMENT BEYOND THE VAN ALLEN BELTS.

YEAST CHEMICAL GENOMICS IS A POWERFUL TOOL THAT IS BEING APPLIED TO DRUG DISCOVERY. THIS APPROACH ALLOWS THE STUDY OF COMPOUND-TARGET RELATIONSHIPS

IMPACT OF DISTINCT ECO-EPIDEMIOLOGY ON MALARIA DRUG RESISTANCE IN GHANA

TARGETING INCREASED POLYAMINE TRANSPORT OF RESISTANT MELANOMAS

EPIDEMIOLOGY AND MOLECULAR MECHANISMS OF ANTHELMINTHIC TREATMENT FAILURE IN KINTA

MENTAL HEALTH, TRAUMA AND EMPLOYMENT TA PROJECT

APPLICATION OF MAGNETIC NANOARRAY TECHNOLOGY TO THE RAPID DIAGNOSIS OF INVASIV

AN HIV INTERVENTION TAILORED FOR BLACK YMSM IN THE HOUSE BALL COMMUNITY

NOVEL MALARIA MULTI-EPITOPE VACCINE DESIGN

INVESTIGATION OF NICHE CONTROL OF GERMLINE STEM CELL LINEAGE DIFFERENTIATION

POWERING UP MALE PREVENTION (PUMP)

NEW DRUG DISCOVERY PARADIGMS FOR SYNUCLEINOPATHIES

COMPREHENSIVE FUNCTIONAL ASSESMENT OF THE HUMAN ANTIBODY RESPONSE TO ENTEROVIRUS 71

CRISPR-CAS13D: TRANSGENIC ZEBRAFISH LINES TOKNOCKDOWN MRNA - PROJECT SUMMARY DETERMINING THE FUNCTION OF GENES IS FUNDAMENTAL FOR UNDERSTANDING VERTEBRATE DEVELOPMENT, REGULATORY MECHANISMS AND HUMAN DISEASES. GENOME EDITING TECHNOLOGIES, SUCH CRISPR-CAS9, HAVE ALLOWED ASSOCIATING SPECIFIC PHENOTYPES TO PERMANENT GENE ALTERATION. HOWEVER, SOME KEY TECHNICAL AND CONCEPTUAL ISSUES REMAIN PROBLEMATIC IN VERTEBRATES, PARTICULARLY IN AQUATIC MODEL ORGANISMS SUCH ZEBRAFISH. FOR EXAMPLE, THE MATERNALLY PROVIDED MRNA CAN RESCUE THE PHENOTYPE OF HOMOZYGOUS MUTANTS; GENOTYPING STEPS ARE TEDIOUS; AND LONG NON- CODING RNA, LETHAL OR TISSUE/TEMPORAL GENES ARE DIFFICULT TO STUDY USING DNA MANIPULATION AS WELL AS COMPLEX GENOMIC LOCI. COMPLEMENTARY ‘KNOCK-DOWN’ APPROACHES ARE INVALUABLE TOOLS TO CIRCUMVENT SOME OF THESE PROBLEMS, HOWEVER, THERE WAS NO SYSTEMATIC TOOL TO KNOCKDOWN MRNAS IN ZEBRAFISH OR OTHER TELEOST FISH. OUR LONG-TERM GOAL IS TO UNRAVEL THE FUNCTION OF GENES RELATED TO REGULATORY MECHANISMS, DEVELOPMENT, AND HUMAN DISEASES. OUR RECENT PUBLICATION DEMONSTRATES THAT INJECTION OF THE CRISPR-RFXCAS13D SYSTEM INTO VERTEBRATE EMBRYOS PROVIDES A ROBUST AND COST-EFFECTIVE TECHNOLOGY TO SYSTEMATICALLY DISRUPT GENE FUNCTION. HOWEVER, THE INJECTION OF THIS SYSTEM ONLY PROVIDES TRANSIENT KNOCKDOWN FOR ~3 DAYS. THEREFORE, THE CENTRAL GOAL OF THE PROPOSAL IS TO TRANSFER THE CRISPR-RFXCAS13D TECHNOLOGY FROM BEING INJECTED INTO VERTEBRATE EMBRYOS, TO BEING ENDOGENOUSLY EXPRESSED THROUGH TRANSGENESIS. WE PROPOSE THAT THE DEVELOPMENT OF A REPERTOIRE TRANSGENIC ZEBRAFISH EXPRESSING THE RFXCAS13D ENZYME (UBIQUITOUS AND TISSUES-SPECIFIC) AND GUIDERNA WOULD FACILITATE RAPID AND VIGOROUS INVESTIGATION INTO GENE FUNCTIONS. OUR PRELIMINARY DATA INDICATE THAT TRANSGENIC EXPRESSION OF RFXCAS13D IN ZEBRAFISH IS FUNCTIONAL AND NOT TOXIC. THE OBJECTIVES ARE: 1) DEFINE THE OPTIMAL SYSTEM TO KNOCKDOWN MRNA EXPRESSION IN CRISPR-RFXCAS13D TRANSGENIC ZEBRAFISH. 2) KNOCKDOWN MRNA EXPRESSION IN A TISSUE-SPECIFIC MANNER USING SPECIFIC TRANSGENIC LINES. THIS PROPOSAL IS CONCEPTUALLY INNOVATIVE AS IT IS BASED ON THE EXPLORATION OF A NOVEL TECHNIQUE, CRISPR-RFXCAS13D, TO KNOCKDOWN MRNA IN A TISSUE-SPECIFIC MANNER IN ZEBRAFISH. THIS APPROACH HAS NEVER BEEN DONE IN VERTEBRATE MODEL SYSTEMS WHERE RNAI DOES NOT WORK. THE OUTCOMES OF THIS PROJECT WILL BE THE FIRST TRANSGENIC SYSTEM TO DISSECT AND STUDY GENE FUNCTION BY KNOCKING DOWN MRNA EXPRESSION IN ZEBRAFISH. OUR APPROACH WILL HELP THE SCIENTIFIC COMMUNITY INVESTIGATE GENE FUNCTION IN A FASTER AND TISSUE-TEMPORAL SPECIFIC MANNER, AS WELL AS ANSWERING QUESTIONS THAT ARE VERY CHALLENGING TO ADDRESS BY CURRENT METHODOLOGIES, SUCH AS THE FUNCTION OF NON-CODING RNAS OR PHENOTYPES CAUSED BY MULTIPLE GENES. MOREOVER, BY FOLLOWING THE GUIDELINES WE WILL OPTIMIZE THROUGH THIS PROJECT, RESEARCHERS MAY PRODUCE A WHOLE GUIDERNA COLLECTION, TARGETING ALL GENES, WHICH SHOULD BE AVAILABLE FOR ANYONE TO ORDER TO ADDRESS THEIR INDIVIDUAL RESEARCH QUESTIONS. FINALLY, AS WE HAVE SUCCESSFULLY IMPLEMENTED THE CRISPR-CAS13D SYSTEM BY INJECTION IN OTHER ORGANISMS SUCH AS MEDAKA, KILLIFISH, AND MOUSE EMBRYOS, OUR WORK DESCRIBED IN THIS PROPOSAL MAY SERVE AS THE FOUNDATION FOR TRANSFERRING THIS EFFICIENT KNOCKDOWN TECHNOLOGY INTO A RANGE OF OTHER SPECIES.

IN VIVO ANALYSIS OF TRKB SIGNALING DURING SYMPATHETIC NERVOUS SYSTEM DEVELOPMENT AND NEUROBLASTOMA PATHOGENESIS

THIS APPLICATION AIMS TO INVESTIGATE HOW CELLS ADAPT TO THE UNIQUE ASPECTS OF THE SPACE ENVIRONMENT, USING THE MODEL EUKARYOTIC ORGANISM, SACCHAROMYC

FUNCTION OF MATERNAL MRNA DURING ZEBRAFISHEMBRYOGENESIS - PROJECT SUMMARY OUR RESEARCH FOCUSES ON UNRAVELING THE FUNDAMENTAL PROCESSES OF EARLY VERTEBRATE EMBRYOGENESIS, PARTICULARLY THE DYNAMIC MOLECULAR MECHANISMS THAT REGULATE MATERNAL AND ZYGOTIC RNAS DURING THE MATERNAL-TO-ZYGOTIC TRANSITION (MZT). THIS CRUCIAL PERIOD INVOLVES THE DEGRADATION OF MATERNAL RNAS AND ACTIVATION OF THE ZYGOTIC GENOME, PROCESSES ESSENTIAL FOR PROPER EMBRYONIC DEVELOPMENT. OUR GOAL IS TO DISSECT THE SPATIAL AND TEMPORAL REGULATION OF MATERNAL AND ZYGOTIC TRANSCRIPTS, IDENTIFY THE FUNCTIONS OF INDIVIDUAL MATERNAL RNAS, AND UNDERSTAND THEIR ROLES IN GENE EXPRESSION, INCLUDING MRNA STABILITY AND TRANSLATIONAL REGULATION. OUR RECENT WORK HAS ESTABLISHED TRANSFORMATIVE TOOLS AND FRAMEWORKS FOR INVESTIGATING EMBRYOGENESIS, INCLUDING THE DEVELOPMENT OF THE CRISPR-CAS13D SYSTEM FOR EFFICIENT MATERNAL RNA KNOCKDOWN IN ZEBRAFISH EMBRYOS. THIS SYSTEM HAS OVERCOME LIMITATIONS IN TRADITIONAL GENETIC APPROACHES, INCLUDING LETHAL AND/OR MASKED PHENOTYPES DUE TO MATERNAL CONTRIBUTION, ENABLING US TO SYSTEMATICALLY TARGET MATERNAL RNAS AND INVESTIGATE THEIR FUNCTIONS WITH UNPRECEDENTED PRECISION. THROUGH INTEGRATIVE MULTI-OMICS APPROACHES—SUCH AS RNA-SEQ, SLAM-SEQ, RIBOSOME PROFILING, AND QUANTITATIVE PROTEOMICS—WE HAVE MADE SIGNIFICANT PROGRESS IN UNDERSTANDING CODON-DEPENDENT MRNA STABILITY, MATERNAL-ZYGOTIC GENE EXPRESSION, AND THE INTERPLAY OF TRANSCRIPTION, TRANSLATION, AND PROTEIN ACCUMULATION DURING EARLY EMBRYOGENESIS. OVER THE NEXT FIVE YEARS, WE AIM TO EXPAND THESE EFFORTS BY ADDRESSING KEY QUESTIONS SURROUNDING THE SPATIAL AND TEMPORAL REGULATION OF MATERNAL RNAS AND THEIR INFLUENCE ON EARLY DEVELOPMENT. OUR STUDIES WILL FOCUS ON: MATERNAL RNA DECAY, LOCALIZATION AND FUNCTION DURING MZT: INVESTIGATING HOW MATERNAL RNAS, SUCH AS CTH1, ARE REGULATED BY SPATIAL AND CELL-SPECIFIC DECAY MECHANISMS. THIS WILL INCLUDE IDENTIFYING CIS-REGULATORY ELEMENTS IN THEIR UNTRANSLATED REGIONS AND CHARACTERIZING THE MOLECULAR FACTORS THAT MEDIATE THEIR LOCALIZATION AND STABILITY. LEVERAGING CRISPR-CAS13D AND MULTI-OMICS TECHNIQUES TO UNCOVER THE ROLES OF MATERNAL RNAS IN ZYGOTIC GENOME ACTIVATION AND EARLY CELLULAR PROCESSES. BROADER EXPLORATION OF MATERNAL RNAS: SYSTEMATICALLY INVESTIGATING ADDITIONAL MATERNAL GENES, INCLUDING GENES ENCODING SMALL TRANSLATED OPEN READING FRAMES, GENES WITH SPECIFIC TEMPORAL EXPRESSION AND SPATIALLY LOCALIZED TRANSCRIPTS WITH CRITICAL DEVELOPMENTAL ROLES. THE OVERARCHING VISION OF OUR RESEARCH IS TO ESTABLISH A COMPREHENSIVE UNDERSTANDING OF HOW GENE REGULATION IS ORCHESTRATED AT MULTIPLE LEVELS—SPATIAL, TEMPORAL, AND MOLECULAR—DURING THE EARLIEST STAGES OF VERTEBRATE DEVELOPMENT. OUR WORK HAS BROAD IMPLICATIONS FOR ADVANCING KNOWLEDGE IN DEVELOPMENTAL BIOLOGY, REPRODUCTIVE HEALTH, AND GENE REGULATION, WITH POTENTIAL APPLICATIONS IN AREAS SUCH AS FERTILITY, REGENERATIVE MEDICINE, AND MRNA-BASED THERAPEUTICS. BY INTEGRATING CUTTING-EDGE TECHNOLOGIES WITH INNOVATIVE APPROACHES, OUR RESEARCH PROGRAM IS POISED TO UNCOVER FUNDAMENTAL PRINCIPLES OF GENE REGULATION AND MRNA STABILITY, CONTRIBUTING TO A DEEPER UNDERSTANDING OF THE MOLECULAR PROCESSES THAT DRIVE THE TRANSFORMATION OF A SINGLE CELL INTO A MULTICELLULAR ORGANISM.

IMMUNOTOXINS BASED ON THE IGG RESPONSE TO AUTOLOGOUS ANTIGENS IN CANCER PATIENTS

NOVEL APPROACHES TO PHARMACOLOGIC MANAGEMENT OF LIFE-THREATENING ARRHYTHMIAS ASSOCIATED WITH THE J WAVE SYNDROMES

MULTILINEAGE DAMFRET TO INVESTIGATE AD/ADRD PROTEIN PHASE BEHAVIOR IN NEURAL TISSUE MODELS - PROJECT SUMMARY/ABSTRACT ALZHEIMER'S DISEASE AND RELATED DEMENTIAS (AD/ADRD) ARE THE MOST COMMON FORMS OF DEMENTIA. NO TREATMENTS EXIST TO STOP OR PREVENT THEM. AD/ADRD ARE CAUSED IN PART BY THE AGGREGATION OF ONE, OR MORE OFTEN A COMBINATION OF, SPECIFIC PROTEINS. TDP-43 IS ONE OF A HANDFUL OF SUCH PROTEINS. IT CHARACTERISTICALLY TRANSITIONS FROM A DYNAMIC LIQUID PHASE OF ASSEMBLY TO A RIGID AND PATHOGENIC PHASE IN A LARGE FRACTION OF DEMENTIA CASES. THIS TRANSITION OCCURS AGAINST A BACKDROP OF PROGRESSIVE CHANGES IN THE INTERACTIONS BETWEEN DIFFERENT CELL TYPES IN THE BRAIN. AN INTERDEPENDENCE OF THESE MOLECULAR AND CELLULAR CHANGES LIKELY DETERMINES THE CLINICAL COURSES OF AD/ADRD, BUT TO UNDERSTAND THAT INTERDEPENDENCE, WE NEED NEW TOOLS WITH WHICH TO COMPARE PROTEIN PHASE TRANSITIONS AND THEIR CORRESPONDING PHENOTYPE EFFECTS IN THE CONTEXT OF INTERACTING BRAIN CELL TYPES. WE PROPOSE HERE TO DEVELOP SUCH A TOOL BY MODIFYING A CELL-BASED BIOPHYSICAL METHOD, DISTRIBUTED AMPHIFLUORIC FRET (DAMFRET) FOR USE IN HUMAN INDUCED PLURIPOTENT STEM CELL (HIPSC)-DERIVED BRAIN TISSUE MODELS (AIM 1). WE WILL SIMULTANEOUSLY USE DAMFRET TO DEVELOP TDP-43 AS A REPORTER OF CELL TYPE-SPECIFIC DIFFERENCES IN THE HOMEOSTASIS OF PROTEIN PHASE TRANSITIONS (AIM 2). BY COMPLETING THESE AIMS, WE WILL HAVE CREATED AND VALIDATED A UNIQUELY POWERFUL TOOL FOR BETTER UNDERSTANDING THE COMPLEX MOLECULAR AND CELLULAR CAUSES OF AD/ADRD.

POTENTIAL ROLE OF FETAL STEM CELLS IN LUNG TUMOR DEVELOPMENT

IDO2 TARGETING FOR PANCREATIC CANCER TREATMENT

TRAINING AND TA ON THE INTERSECTION OF VIOLENCE AGAINST WOMEN AND SUBSTANCE ABUSE: COLLABORATIVE STRATEGIES FOR IMPROVING SAFETY AND RECOVERY

NT'L TRAINING & TECH. ASSIST. CENTER ON D.V. TRAUMA AND MENTAL HEALTH

PATHOGENESIS, PHENOTYPIC VARIATION, AND PREVENTION OF CRANIOFACIAL ANOMALIES - PROJECT SUMMARY CRANIOFACIAL ANOMALIES ACCOUNT FOR ABOUT ONE-THIRD OF ALL BIRTH DEFECTS AND CONSEQUENTLY ARE A MAJOR CAUSE OF INFANT MORTALITY. TO DATE OVER 700 DISTINCT CRANIOFACIAL SYNDROMES HAVING BEEN DESCRIBED, AND ANOMALIES SUCH AS CLEFT LIP AND PALATE, MICROGNATHIA AND CRANIOSYNOSTOSIS HAVE SERIOUS LIFETIME FUNCTIONAL, AESTHETIC AND SOCIAL CONSEQUENCES THAT ARE DEVASTATING TO BOTH CHILDREN AND PARENTS ALIKE. COMPREHENSIVE SURGERY TOGETHER WITH WELL-COORDINATED AND INTEGRATED DENTAL CARE, PSYCHOLOGICAL COUNSELLING AND REHABILITATION CAN HELP AMELIORATE AND MANAGE EACH CONDITION. HOWEVER, THE RESULTS ARE OFTEN VARIABLE AND RARELY FULLY CORRECTIVE, HENCE CONSIDERABLE EFFORT NEEDS TO BE INVESTED IN DEVELOPING PREVENTATIVE THERAPIES. THIS CAN ONLY COME FROM A THOROUGH APPRECIATION OF THE ETIOLOGY AND PATHOGENESIS OF INDIVIDUAL CRANIOFACIAL MALFORMATION SYNDROMES, WHICH IS BUILT UPON A DEEP FOUNDATIONAL KNOWLEDGE AND UNDERSTANDING OF NORMAL CRANIOFACIAL DEVELOPMENT. ADVANCES IN GENOMICS CONTINUES TO ELUCIDATE THE COMPLEX ETIOLOGY UNDERLYING BIRTH DEFECTS, BUT KNOWING THE MOLECULAR GENOTYPE OF A SINGLE LOCUS IS OFTEN INSUFFICIENT FOR PREDICTING THE PHENOTYPE OF MANY MALFORMATION SYNDROMES. FURTHERMORE, BIRTH DEFECT DISORDERS ARE TYPICALLY CHARACTERIZED BY CONSIDERABLE PHENOTYPIC VARIANCE, OFTEN DUE TO GENE-ENVIRONMENT INTERACTIONS, BUT OUR UNDERSTANDING OF ENVIRONMENTAL RISK FACTORS IS POOR, AS BIOLOGICAL AND TECHNICAL CONSTRAINTS HAVE MADE DEFINING GENE-ENVIRONMENT INTERACTIONS IN BIRTH DEFECT ETIOLOGY CHALLENGING. CRANIOFACIAL MORPHOGENESIS IS ONE EXAMPLE OF A COMPLEX TRAIT IN WHICH EPISTATIC AND GENE-ENVIRONMENT INTERACTIONS LIKELY CONTRIBUTE EXTENSIVELY TO PHENOTYPIC VARIATION DURING NORMAL DEVELOPMENT AND IN THE PATHOGENESIS OF CRANIOFACIAL DYSMORPHOLOGY. HOWEVER, WHILE IT HAS BEEN POSITED THAT MOST CRANIOFACIAL VARIATION IS GENETICALLY DETERMINED, A LARGE GAP IN KNOWLEDGE EXISTS WITH RESPECT TO THE CONTRIBUTIONS OF ENVIRONMENTAL FACTORS AND GENETIC BACKGROUND IN THE PATHOGENESIS OF CRANIOFACIAL ANOMALIES. FOCUSING ON THE CRANIOFACIAL DISORDER, TREACHER COLLINS SYNDROME, THIS RESEARCH PROPOSAL: (I) UNCOVERS AND VALIDATES SPECIFIC GENE-ENVIRONMENT INTERACTIONS WHICH UNDERLIE THE ETIOLOGY AND PATHOGENESIS OF ANOMALIES OF THE HEAD AND FACE; (II) IDENTIFIES AND VALIDATES GENETIC MODIFIERS THAT CONTRIBUTE TO INTER-FAMILIAL AND INTRA-FAMILIAL VARIABILITY CHARACTERISTIC OF CRANIOFACIAL SYNDROMES; AND (III) DEFINES AND VALIDATES IN UTERO THERAPEUTIC APPROACHES TO PREVENT CRANIOFACIAL ANOMALIES.

INVESTIGATING THE ROLE OF INDOLEAMINE 2,3-DIOXYGENASE (IDO) IN BREAST CANCER METASTASIS

INCREASING ACCESS TO VICTIM SERVICES FOR VICTIMS OF DOMESTIC VIOLENCE AND SEXUAL ASSAULT WHO HAVE A SERIOUS MENTAL ILLNESS

MEASURING HEALTH-RELATED TRUST IN DIVERSE POPULATIONS

PRIMARY CARE TRAINING AND ENHANCEMENT

SPECIAL PROJECTS OF NATIONAL SIGNIFICANCE

#CHOPVIOLENCE/#CHOPHIV: A TAILORED VIOLENCE PREVENTION INTERVENTION FOR BLACK YOUNG MSM AND TRANSGENDER WOMEN FROM THE HOUSE BALL COMMUNITY TO IMPROVE HIV OUTCOMES AND DECREASE EXPOSURE TO VIOLENCE

TRANSCRIPTIONAL HIV-1 LATENCY IN ASTROCYTE AND MACROPHAGE RESERVOIRS OF THE CENTR

"VIRAL AND LATENT HIV RESERVOIR CHARACTERISTICS IN HIV PATIENTS WITH PERSISTENT LOW-LEVEL VIREMIA - PROJECT SUMMARY THE SUCCESS OF ANTIRETROVIRAL THERAPY (ART) IS USUALLY MEASURED BY THE PROPORTION OF PERSONS LIVING WITH HIV (PLWH) WHO ARE VIROLOGICALLY SUPPRESSED ON ART. HOWEVER, THE STANDARDS USED TO DETERMINE VIRAL SUPPRESSION DIFFER BETWEEN THE DEVELOPED AND THE DEVELOPING WORLD. THE DEVELOPED COUNTRIES DEFINE VIRAL SUPPRESSION AS VIRAL LOAD (VL) LESS THAN 50 COPIES PER ML, WHILE MOST OF AFRICA, ACCEPTS THE WORLD HEALTH ORGANIZATION (WHO) PRESCRIBED LESS THAN 1000 COPIES PER ML AS VIRAL SUPPRESSION IN RESOURCE-LIMITED SETTINGS (RLS). THIS HAS CREATED A CATEGORY OF PERSONS IN RLS WITH PERSISTENTLY LOW-LEVEL VIREMIA (PLLV) OF 50-999 COPIES/ML WHO ARE LUMPED TOGETHER WITH THE TRULY VIROLOGICALLY SUPPRESSED, AND THUS NOT GIVEN MUCH ATTENTION. SINCE VIRAL REPLICATION CONTINUES DURING LOW-LEVEL VIREMIA, THIS MAY FUEL THE PROLIFERATION OF RESISTANCE MUTATIONS, EXPAND THE DIVERSITY OF VIRAL STRAINS AND INCREASE THE VIRAL RESERVOIR SIZE IN THOSE WITH PLLV. HOWEVER, WHETHER THIS POPULATION HAS MORE DIVERSE VIRAL STRAINS, HAS A LARGER RESERVOIR SIZE OR SERVE AS A SOURCE OF THE CURRENT RISE IN DRUG RESISTANCE IN AFRICA HAS NOT BEEN STUDIED. THIS CREATES A CRITICAL KNOWLEDGE GAP WHICH IF NOT FILLED COULD DERAIL THE SUCCESS OF ART AND SERVE AS A BOTTLENECK IN HIV CURE EFFORTS IN AFRICA. OUR WORKING HYPOTHESIS IS THAT ART-TREATED PLWH WITH PERSISTENT LOW-LEVEL VIREMIA HAVE GREATER VIRUS DIVERSITY, LARGER RESERVOIR SIZE AND SELECT FOR DRUG RESISTANCE MUTATIONS. WE INTEND TO INVESTIGATE THIS HYPOTHESIS WITH TWO SPECIFIC AIMS: AIM 1: DETERMINE VIRAL DIVERSITY AND DRUG RESISTANCE MUTATIONS AMONG PATIENTS WITH PLLV. WE HYPOTHESIZE THAT CONTINUOUS VIRAL REPLICATION DURING LOW-LEVEL VIREMIA ON ART EXPANDS THE DIVERSITY OF VIRAL STRAINS AND FUEL THE EMERGENCE OF DRUG RESISTANCE MUTATIONS. FIRST, WE WILL PERFORM A CROSS-SECTIONAL EVALUATION TO DETERMINE THE FREQUENCY OF CLINICALLY-RELEVANT DRUG RESISTANCE MUTATIONS IN THOSE WITH PLLV. SECOND, WE WILL FOLLOW THE PATIENTS WITH PLLV FOR 18 MONTHS TO DETERMINE RESISTANCE EVOLUTION AND THE PROPORTION OF THOSE WHO BREAK THROUGH TO VL>1000. THIRD, WE WILL DETERMINE INTER- AND INTRA-PATIENT GENETIC DIVERSITY OF HIV IN THOSE WITH PLLV. AIM 2: DETERMINE THE CHARACTERISTICS OF THE HIV RESERVOIR IN HIV PATIENTS WITH PLLV. WE HYPOTHESIZE THAT PERSISTENT LOW VIREMIA FEEDS THE LATENT RESERVOIR MAKING IT LARGER AND MORE DIVERSE IN THOSE WITH PLLV COMPARED TO THE TRULY SUPPRESSED PERSONS. THE QUESTION HERE IS WHETHER PLLV ENLARGE THE SIZE OF THE RESERVOIR AND INCREASE CLONAL EXPANSION. IF THAT WERE THE CASE, IT WOULD BECOME MORE URGENT TO BRING THE VIRUS TO UNDETECTABLE IN THESE PATIENTS. THE KNOWLEDGE GAINED WILL BE USEFUL FOR OUR HIV CONTROL PROGRAMS; MAY CALL FOR CHANGES IN TREATMENT GUIDELINES IN GHANA AND AFRICA AND PROMPT MORE EXTENSIVE STUDIES AMONG PERSONS WITH PERSISTENT LOW VIREMIA IN RESOURCE- LIMITED SETTINGS.

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.

ROLE OF COMPLEMENT RECEPTOR 1 IN ERYTHROCYTE INVASION BY PLASMODIUM FALCIPARUM IN

CD4

IMPROVED DIAGNOSIS OF ACTIVE SYPHILIS AT THE POINT-OF-CARE FOR THE ELIMINATION OF CONGENITAL SYPHILIS

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.

THE ROLE OF RRNA TRANSCRIPTION AND RIBOSOME BIOGENESIS IN NEURAL CREST PROGENITORS AND STEM CELLS DURING EMBRYONIC AND POSTNATAL CRANIOFACIAL DEVELOPMENT - ABSTRACT CRANIOFACIAL ANOMALIES ARE THE MOST COMMON MALFORMATIONS PRESENT AT BIRTH AND COMPRISE UP TO 1/3 OF ALL CONGENITAL DEFECTS. MOST OF THE CRANIOFACIAL CARTILAGE, BONE AND CONNECTIVE TISSUE ARE DERIVED FROM NEURAL CREST CELLS (NCC), A MIGRATORY STEM AND PROGENITOR CELL POPULATION BORN DURING EARLY EMBRYOGENESIS. THE MAJORITY OF CRANIOFACIAL DISORDERS ARE THEREFORE THOUGHT TO BE A RESULT OF DISRUPTIONS IN NCC DEVELOPMENT3. IN ORDER TO IMPROVE THE PROGNOSIS FOR INDIVIDUALS AFFECTED WITH CRANIOFACIAL ANOMALIES, AND TO DEVELOP POTENTIAL PREVENTATIVE THERAPIES, THERE IS A CRITICAL NEED FOR A DEEPER UNDERSTANDING OF THE FUNDAMENTAL MECHANISMS THAT GOVERN THE FORMATION, MIGRATION AND DIFFERENTIATION OF NCCS DURING CRANIOFACIAL MORPHOGENESIS4. RECENT WORK IN OUR LAB DEMONSTRATED THAT RNA POLYMERASE I (POL I)-MEDIATED RIBOSOMAL RNA (RRNA) TRANSCRIPTION, WHICH IS THE RATE LIMITING STEP IN RIBOSOME BIOGENESIS, IS ELEVATED IN NCCS COMPARED TO OTHER CELL TYPES5, 6. THIS IS NECESSARY TO MEET NCC’S HIGHER PROTEIN TRANSLATIONAL REQUIREMENTS AND UNDERPINS THEIR HIGH RATE OF PROLIFERATION, EPITHELIAL TO MESENCHYMAL TRANSFORMATION, AND METABOLICALLY EXPENSIVE MIGRATION PROPERTIES5. IN SUPPORT OF THIS IDEA, NCC- SPECIFIC DISRUPTION OF POL I FUNCTION LEADS TO INCREASED NCC DEATH AND CRANIOFACIAL ANOMALIES CHARACTERISTIC OF CONGENITAL CRANIOFACIAL DISORDERS SUCH AS TREACHER COLLINS SYNDROME AND ACROFACIAL DYSOSTOSIS-CINCINNATI TYPE5, 7, 8. ADDITIONALLY, EVIDENCE FROM OTHER STUDIES SUPPORTS ELEVATED REQUIREMENTS FOR RIBOSOMES AND TRANSLATIONAL CAPACITY IN ORDER FOR STEM CELLS TO ACQUIRE NEW FATES, SUGGESTING THAT RIBOSOME BIOGENESIS ALSO PLAYS A CRITICAL ROLE DURING THE DIFFERENTIATION STAGE OF NCC DEVELOPMENT. BASED ON OUR PREVIOUS FINDINGS I HYPOTHESIZE THAT INCREASED RRNA TRANSCRIPTION AND RIBOSOME BIOGENESIS ARE ESSENTIAL FOR BOTH NCC-DERIVED PROGENITOR AND STEM CELL MAINTENANCE AND DIFFERENTIATION DURING EMBRYONIC AND ALSO JUVENILE PHASES OF CRANIOFACIAL DEVELOPMENT. TO TEST THIS HYPOTHESIS, I WILL GENETICALLY DELETE POLR1A, THE CATALYTIC CORE OF POL I, IN TAMOXIFEN-INDUCIBLE GLOBAL AND TISSUE-SPECIFIC MOUSE LINES. THEN THROUGH PHENOTYPIC AND CELLULAR ANALYSES, IN COMBINATION WITH TRANSCRIPTOMIC AND PROTEOMIC APPROACHES, I WILL DEFINE THE MECHANISMS DRIVING THE ANATOMICAL ANOMALIES THAT RESULT FROM DISRUPTIONS IN POL I FUNCTION. MY RESULTS WILL PROVIDE NEW KNOWLEDGE ON THE ROLES OF RRNA TRANSCRIPTION AND RIBOSOME BIOGENESIS IN CRANIOFACIAL PROGENITOR AND STEM CELL POPULATIONS DURING EMBRYONIC AND POSTNATAL DEVELOPMENT. THE FINDINGS CAN POTENTIALLY UNCOVER NEW THERAPIES FOR PREVENTING OR AMELIORATING RIBOSOMOPATHIES, AND IMPROVE CLINICAL OUTCOMES FOR CRANIOFACIAL PATIENTS, ESPECIALLY WHEN IT COMES TO CORRECTIVE SURGERIES, THROUGH IMPROVED POSTNATAL GROWTH PREDICTIONS.

INVESTIGATING PROTEIN SUPERSATURATION AS A DRIVER OF AGING - PROJECT SUMMARY PROTEIN AGGREGATION IS A HALLMARK OF AGING AND AGE-ASSOCIATED DISEASE, HOWEVER A CAUSAL RELATIONSHIP HAS NOT BEEN DEMONSTRATED. ELUCIDATING WHETHER THERE IS A PREDESTINED, IRREVERSIBLE DRIVING FORCE IN CELL AGING WOULD ENABLE THE DEVELOPMENT OF NOVEL THERAPIES TO DECELERATE AGING. THEREFORE, MY LONG-TERM GOAL IS TO UNDERSTAND THIS RELATIONSHIP AT A FUNDAMENTAL LEVEL. MY CENTRAL HYPOTHESIS IS THAT PROBABILISTIC, IRREVERSIBLE CONFORMATIONAL TRANSITIONS IN PHYSIOLOGICALLY SUPERSATURATED PROTEINS NOT ONLY INITIATE THE PROCESS OF CELLULAR AGING, BUT DRIVE IT!. THE ACCUMULATION OF AMYLOIDS FOLLOWING SUCH TRANSITIONS WILL COMPROMISE KINETIC PROTEOSTASIS, OR KINETIC BARRIERS FOR SUPERSATURATED PROTEINS TO REMAIN SOLUBLE, AND THIS ULTIMATELY COMPROMISES THERMODYNAMIC PROTEOSTASIS -- OR THE PROCESSES THAT MAINTAIN THE CONCENTRATIONS AND STABILITIES OF SOLUBLE PROTEINS. I WILL UTILIZE THE FOLLOWING SPECIFIC AIMS AND SYNERGISTIC APPROACHES, DISTRIBUTED AMPHIFLUORIC FRET (DAMFRET), EPIGENETIC CLOCKS, AND RNA-SEQ, TO DISTINGUISH THE KINETIC FROM THERMODYNAMIC DETERMINANTS OF PROTEIN SOLUBILITY AS A FUNCTION OF CELL AGE. IN AIM 1, I WILL COMPARE THERMODYNAMIC AND KINETIC PROTEOSTASIS AS A FUNCTION OF BIOLOGICAL AGE. TO DO SO, I WILL FIRST OBTAIN PRIMARY HUMAN FIBROBLASTS (PHFS) FROM DIFFERENTIALLY AGED DONORS, AND VALIDATE THEIR EPIGENETIC AGE USING DNA METHYLATION SIGNATURES (DNAM) REFERENCED AGAINST PREVIOUSLY DEVELOPED DNAM AGE PREDICTION ALGORITHMS, AS WELL AS RNA-SEQ. I WILL THEN PERFORM DAMFRET EXPERIMENTS IN EACH OF THE PHFS USING A PANEL OF INDUCIBLE CONSTRUCTS THAT RELIABLY AGGREGATE IN A NUCLEATION- AND/OR CONCENTRATION-LIMITED MANNER. THESE DATA WILL REVEAL THE DEGREE TO WHICH KINETIC PROTEOSTASIS AND/OR THERMODYNAMIC PROTEOSTASIS ARE IMPACTED BY BIOLOGICAL AGE. IN AIM 2, I WILL TEST IF CONFORMATIONAL NUCLEI ACCELERATE THE AGING OF PHFS. I WILL GENERATE GENERIC LIGHT-ACTIVATED OPTOSEEDS FROM OUR REPORTER LIBRARY IN AIM 1 TO ELICIT A CONFORMATIONAL TRANSITION, OR CROSS-SEEDING EVENT, IN PHFS OF YOUNG AGE. I WILL THEN USE MULTIPLE MASS SPECTROMETRY APPROACHES TO EVALUATE WHETHER THE NUCLEATION EVENT PRECIPITATED ENDOGENOUS PROTEINS, AND DETERMINE THEIR IDENTITIES. I WILL THEN DETERMINE IF THE TREATMENT ACCELERATES THE PROGRESSION OF CELL AGE VIA DNAMAGE AND RNA-SEQ. IN AIM 3, I WILL TEST IF PERTURBING KINETIC PROTEOSTASIS IN THE NUCLEUS ENHANCES THE RATE OF AGING AS COMPARED TO THE CYTOSOL. USING OUR OPTOSEEDS, I WILL ELICIT A CONFORMATIONAL TRANSITION IN THE NUCLEAR AND CYTOPLASMIC COMPARTMENTS. I WILL AGAIN USE DNAM AGE PREDICTION AND RNA-SEQ TO DETERMINE WHETHER AGE IS ACCELERATED VIA CONFORMATIONAL TRANSITIONING IN THE NUCLEUS VERSUS THE CYTOSOL. COMPLETION OF THESE AIMS WILL PROVIDE FUNDAMENTAL INSIGHTS INTO THE THERMODYNAMIC REASONS FOR WHY WE AGE. IN ADDITION, COMPLETION OF THE PROPOSED STUDIES WILL PROVIDE ME WITH A STRONG FOUNDATION TO CONTINUE MY RESEARCH AS AN INDEPENDENT INVESTIGATOR.

A GENETIC PROGRAM FOR ORGAN REGENERATION IN ZEBRAFISH

INTEGRATED GENOME-BASED STUDIES OF SHEWANELLA ECOPHYSIOLOGY

ZEBRAFISH DEVELOPMENT & GENETICS CONFERENCE

EVOLUTIONARY MECHANISMS OF GASTRULATION AND LEFT-RIGHT PATTERNING IN AMNIOTES. - PROJECT SUMMARY ALTHOUGH AS HUMANS WE APPEAR SYMMETRICAL ON THE OUTSIDE, OUR INTERNAL ORGANS ARE ASYMMETRICALLY POSITIONED ALONG THE LEFT AND RIGHT SIDES OF OUR BODY. LEFT-RIGHT (L-R) PATTERNING IS A FUNDAMENTAL BIOLOGICAL PROCESS THAT HELPS TO ENSURE THE CORRECT POSITIONING OF OUR ORGANS, AND ITS PERTURBATION IS TYPICALLY ASSOCIATED WITH CONGENITAL HEART MALFORMATIONS AND HIGH MORTALITY. IN A TYPICAL DEUTEROSTOME, INCLUDING HUMANS, PROPER L-R PATTERNING INVOLVES MOTILE CILIA IN THE L-R ORGANIZING (LRO) REGION, WHICH TRIGGER A CA2+ WAVE ON THE LEFT SIDE OF THE EMBRYO. THIS RESULTS IN ASYMMETRIC GENE EXPRESSION AND ULTIMATELY ASYMMETRIC ORGANOGENESIS. HOWEVER, MORE THAN 65% OF ALL TETRAPODS, INCLUDING REPTILES AND EVEN-TOED UNGULATES, DO NOT USE MOTILE CILIA FOR L-R PATTERNING. INSTEAD, TILTING OF THE LRO AND ASYMMETRIC CELLULAR MOVEMENTS SOMEHOW LEAD TO MOLECULAR ASYMMETRY. HOWEVER, THE MECHA- NISMS UNDERPINNING L-R ASYMMETRY IN THESE ORGANISMS ARE POORLY UNDERSTOOD. IT IS UNCLEAR HOW ASYMMETRIC CELL MOVEMENTS ORIGINATE, WHETHER THEY TRIGGER THE ASYMMETRICAL CA2+ WAVE, OR IF CILIA ARE INVOLVED IN ANY ASPECT OF L-R PATTERNING. CURRENTLY, THE CHICKEN EMBRYO IS USED TO REPRESENT THE DIVERSITY OF ALL 25,000 SPECIES OF REPTILES, AND NEW MODELS ARE REQUIRED FOR A DEEPER EVOLUTIONARY UNDERSTANDING OF FUNDAMENTAL DEVELOPMENTAL EVENTS. VEILED CHAMELEONS (CHAMAELEO CALYPTRATUS) ARE PERFECT FOR THE STUDY OF EARLY DEVELOPMENT AND EVOLUTION IN NON- AVIAN REPTILES, SINCE THEY LAY LARGE CLUTCHES OF EGGS AT PRE-GASTRULATION STAGES. THEIR LRO LACKS MOTILE CILIA, AND INSTEAD MOLECULAR ASYMMETRY IS ESTABLISHED THROUGH LARGE-SCALE MORPHOLOGICAL CHANGES. VEILED CHAMELEONS HAVE A SEQUENCED AND ANNOTATED GENOME, AND ARE AMENABLE TO CELL AND EMBRYO CULTURE, AS WELL AS LIVE IMAGING. THE OBJECTIVE OF THIS APPLICATION THEREFORE IS TO DEFINE THE MECHANISMS GOVERNING L-R PATTERNING IN CHAMELEONS AND THUS EXPAND OUR UNDERSTANDING OF AMNIOTE DEVELOPMENT AND EVOLUTION. THE CENTRAL HYPOTHESIS IS THAT CELLULAR FLOW AND LARGE-SCALE MORPHOLOGICAL CHANGES TRIGGER AN EVOLUTIONARILY CONSERVED ASYMMETRIC CA2+ WAVE, LEADING TO MOLECULAR L-R ASYMMETRY, WHICH HAS UNDERGONE EVOLUTIONARY CHANGE ACROSS AMNIOTES. THIS HYPOTHESIS WILL BE ADDRESSED IN THE FOLLOWING AIMS: (AIM 1) DETERMINE THE MECHANICS OF ESTABLISHING L-R PATTERNING IN VEILED CHAMELEON. (AIM 2) EVALUATE GENETIC CHANGES AND CONSERVATION OF THE L-R PATTERNING PATHWAY ACROSS AMNIOTES. THE PATTERNS OF CELL MIGRATION AND THE DYNAMICS OF CA2+ SIGNALING WILL BE EVALUATED THROUGH LIVE IMAGING, PROVIDING TRAINING IN ADVANCED MICROSCOPY. CRISPR/CAS9 GENE EDITING WILL BE ADAPTED FOR USE IN CHAMELEONS AND WILL INCLUDE TRAINING IN SURGERY AND VIRUS PRODUCTION. THIS STUDY WILL RESULT IN THE FIRST SCRNA-SEQ AND SCATAC-SEQ LIBRARIES FOR ASYMMETRIC GENE EXPRESSION BETWEEN THE LEFT AND RIGHT SIDES OF CHICKEN, CHAMELEON, AND MOUSE EMBRYOS, AND WILL INVOLVE COMPUTATIONAL BIOLOGY TRAINING. SUCCESSFUL COMPLETION OF THIS PROJECT WILL BE SIGNIFICANT IN THE FIELDS OF L-R PATTERNING AND EVO-DEVO, PROVIDING THE FIRST DETAILED STUDY OF THE EARLY STEPS OF L-R PATTERNING IN A NON-AVIAN, NON-MAMMALIAN AMNIOTE, WHICH MAY REVOLUTIONIZE OUR CURRENT THINKING ABOUT ROLES FOR CILIA AND CA2+ SIGNALING IN L-R PATTERNING. IT WILL ALSO LAY THE FOUNDATION FOR A SUCCESSFUL INDEPENDENT CAREER.

UNCOVERING MECHANISMS CONTROLLING CHROMOSOME-SPECIFIC BEHAVIORS DURING MEIOSIS

AN INVESTIGATION INTO THE NATURE OF NON-VOIDING CONTRACTIONS RESULTING FROM DETRUSOR HYPERREFLEXIA IN NEUROGENIC BLADDERS FOLLOWING SPINAL CORD INJUR

UNDERSTANDING RNA POLYMERASE III TRANSCRIPTION IN NEURAL CREST CELL AND CRANIOFACIAL DEVELOPMENT - PROJECT SUMMARY CRANIOFACIAL ANOMALIES ACCOUNT FOR ONE THIRD OF ALL BIRTH DEFECTS AND ARE A SIGNIFICANT CAUSE OF INFANT MORTALITY. NEURAL CREST CELLS (NCC) GIVE RISE TO THE MAJORITY OF CRANIOFACIAL BONE, CARTILAGE, AND CONNECTIVE TISSUE AND UN- DERSTANDING THEIR DEVELOPMENT IS CRUCIAL FOR ADVANCING THE PREVENTION OF CRANIOFACIAL BIRTH DEFECTS. DISRUPTIONS IN NCC DEVELOPMENT ARE KNOWN TO UNDERLIE SEVERAL CRANIOFACIAL DISORDERS INCLUDING TREACHER COLLINS SYNDROME, WHICH IS CAUSED BY MUTATIONS IN TCOF1, POLR1B, POLR1C, AND POLR1D. POLR1C AND POLR1D ARE SUBU- NITS OF BOTH RNA POLYMERASES (POL) I AND III AND ARE IMPORTANT FOR TRANSCRIPTION OF RIBOSOMAL RNA. I PREVIOUSLY DEMONSTRATED IN POLR1C AND POLR1D ZEBRAFISH MODELS THAT RIBOSOMAL RNA TRANSCRIPTION IS REDUCED LEADING TO TP53-DEPENDENT CELL DEATH OF NCC PROGENITORS WHICH RESULTS IN CRANIOFACIAL ANOMALIES. HOWEVER, HOW GLOBAL DISRUPTIONS IN POLR1C AND POLR1D SPECIFICALLY AFFECT NCC DEVELOPMENT REMAINS UNRESOLVED AND THE CONTRIBUTION OF POL III, WHICH TRANSCRIBES NON-CODING RNAS INCLUDING 5S RIBOSOMAL RNA AND TRANSFER RNAS, TO CRANIOFACIAL DEVELOPMENT IS NOT KNOWN. I HYPOTHESIZE THAT IN ADDITION TO DISRUPTION OF POL I TRANSCRIPTION IN THE PATHOGENESIS OF TREACHER COLLINS SYNDROME, POL III TRANSCRIPTION IS ALSO DISRUPTED AND CONTRIBUTES TO THE TISSUE-SPECIFIC PHE- NOTYPES OBSERVED. TRANSCRIPTS PRODUCED BY POL III, INCLUDING TRNAS, HAVE BEEN SHOWN IN MULTIPLE SYSTEMS TO BE TISSUE-SPECIFICALLY EXPRESSED. TO GENERATE A NEW UNDERSTANDING OF THE ROLE OF POL III TRANSCRIPTION SPECIFICALLY IN NCC, I WILL RECEIVE TRAINING IN PROFILING NCC FOR CHANGES IN POL III TRANSCRIPTS AND IN EVALUATING THE EFFECT OF THESE CHANGES ON TRANSLATION. IT HAS BEEN POSTULATED THAT DISTINCT PATHOGENIC VARIANTS IN POLR3A, THE LARGEST SUBUNIT OF POL III, LEAD TO DIFFERENTIAL EFFECTS ON POL III TRANSCRIPTION. IN ORDER TO TEST THIS HYPOTHESIS IN A NCC- SPECIFIC MANNER, I WILL USE HIPSCS DERIVED FROM PATIENT FIBROBLASTS WITH PATHOGENIC VARIANTS IN POLR3A AND ANALYZE THEM FOR PROLIFERATION, TRANSLATION, DIFFERENTIATION, AND POL I AND III TRANSCRIPTION. GIVEN THE PREVALENCE OF DENTAL ANOMALIES IN INDIVIDUALS WITH MUTATIONS IN POLR3A, I EXPECT TO IDENTIFY POL III-SPECIFIC EFFECTS IN A SUBSET OF NCC DERIVATIVES. IN THE INDEPENDENT PHASE OF THIS AWARD, I WILL GENERATE NEW ZEBRAFISH MODELS TO UNDER- STAND THE ROLE OF SPECIFIC VARIANTS IN POL III IN A DEVELOPMENTAL CONTEXT AND ASSESS NCC FORMATION, MIGRATION, DIFFERENTIATION, AND PROLIFERATION IN COMBINATION WITH THE EFFECT ON POL I AND III TRANSCRIPTION. THESE MODELS WILL PROVIDE NEW RESOURCES TO THE RESEARCH COMMUNITY FOR THE UNDERSTANDING OF POL III TRANSCRIPTION. ALTOGETHER, I WILL RECEIVE THE TRAINING NECESSARY TO ANALYZE POL III TRANSCRIPTION AND TRANSLATION AND MODEL PATIENT-SPECIFIC VAR- IANTS IN NCC WHICH WILL FORM THE FOUNDATION OF MY INDEPENDENT RESEARCH PROGRAM AND FURTHER MY GOAL OF UNDER- STANDING AND PREVENTING CRANIOFACIAL BIRTH DEFECTS.

NEUROSTEROIDS AS CRITICAL MODULATORS OF THE STRESS RESPONSE IN PTSD

RYAN WHITE HIV/AIDS PROGRAM PART C EIS COVID-19 RESPONSE

A NEW SYSTEM FOR MODELING SUPERNUMERARY CHROMOSOME DYNAMICS AND FORMATION DURING MEIOSIS.

EXAMINING SIGNALS THAT SCULPT CRANIAL NEURAL CREST MIGRATION

PROFILING EARLY METASTATIC MELANOMA IN VIVO USING THE CHICK EMBRYO MODEL

OPPC TARGETING TO IMPROVE PANCREATIC CANCER TREATMENT

EXTRINSIC SIGNALING IN NEURAL CREST AND CRANIOFACIAL DEVELOPMENT

FUNCTIONAL CHARACTERIZATION OF MEDIATOR COMPLEX PROTEINS IN NEURAL CREST AND CRANIOFACIAL DEVELOPMENT - PROJECT SUMMARY CRANIOFACIAL DEVELOPMENTAL DISORDERS SUCH AS CLEFT PALATE AND RETROGNATHIA (SMALLER LOWER JAW) OFTEN ARISE DUE TO DEFECTS IN NEURAL CREST CELL DEVELOPMENT AND AFFECT 1 IN 700 AND 1 IN 1,500 LIVE HUMAN BIRTHS, RESPECTIVELY. CLEFT PALATE AND RETROGNATHIA OFTEN PRESENT WITH OTHER CRANIOFACIAL ANOMALIES IN CONDITIONS SUCH AS TREACHER COLLINS SYNDROME AND DIGEORGE SYNDROME. ALTHOUGH GENETIC ANALYSES HAVE IDENTIFIED THE GENES RESPONSIBLE FOR SOME CRANIOFACIAL ANOMALIES, THE VAST MAJORITY HAVE AN UNDIAGNOSED MOLECULAR ETIOLOGY AND CELLULAR PATHOGENESIS. IN AN ENU MUTAGENESIS SCREEN, THE MED23 GENE WAS IDENTIFIED TO BE CRITICAL FOR CRANIOFACIAL DEVELOPMENT. A POINT MUTATION IN MED23 RESULTS IN CRANIOFACIAL AND VASCULAR DEFECTS, LEADING TO EMBRYONIC LETHALITY AT MID-GESTATION. MED23 BELONGS TO THE TAIL MODULE OF THE MEDIATOR COMPLEX, WHICH IS A GLOBAL TRANSCRIPTION COREGULATOR FOR GENES TRANSCRIBED BY RNA POLYMERASE II. MED23 IS UBIQUITOUSLY EXPRESSED IN MOUSE EMBRYOS, THEREBY, RAISING AN IMPORTANT QUESTION OF HOW A UBIQUITOUSLY EXPRESSED PROTEIN REGULATES TISSUE SPECIFIC DEVELOPMENT DURING EMBRYOGENESIS, PARTICULARLY IN CRANIOFACIAL DEVELOPMENT. TO UNDERSTAND THE FUNCTION OF MED23 IN CRANIOFACIAL AND NEURAL CREST CELL DEVELOPMENT, I GENERATED NEURAL CREST CELL SPECIFIC CONDITIONAL KNOCKOUTS OF MED23 THAT EXHIBIT CLEFT PALATE, RETROGNATHIA, GLOSSOPTOSIS AND CLEIDOCRANIAL DYSPLASIA. INTERESTINGLY, ENDOTHELIAL CELL SPECIFIC CONDITIONAL KNOCKOUTS OF MED23 ALSO RESULT IN CRANIOFACIAL DEFECTS TOGETHER WITH VASCULAR DEFECTS. IN THIS PROPOSAL, I WILL TEST THE OVERARCHING HYPOTHESIS THAT MED23 AND OTHER TAIL MODULE SUBUNITS OF MEDIATOR HAVE IMPORTANT AND DISTINCT TRANSCRIPTIONAL REGULATORY FUNCTIONS IN CRANIOFACIAL DEVELOPMENT VIA THE CONTROL OF NEURAL CREST CELL AND ENDOTHELIAL CELL TRANSCRIPTOMES. SPECIFICALLY, I WILL ADDRESS THE FOLLOWING AIMS. (AIM 1) IDENTIFY THE MOLECULAR MECHANISM UNDERLYING THE CRANIOFACIAL DEFECT IN NEURAL CREST CELL SPECIFIC MUTANTS OF MED23 BY ANALYZING TRANSCRIPTOMIC CHANGES IN THESE MUTANTS AS WELL AS BY IDENTIFYING MED23 DNA AND PROTEIN BINDING PARTNERS. (AIM 2) CHARACTERIZE THE PATHOGENESIS OF CRANIOFACIAL DEFECTS IN ENDOTHELIAL CELL SPECIFIC MUTANTS OF MED23 AND THEIR UNDERLYING MOLECULAR MECHANISMS BY ANALYSIS OF THE CRANIOFACIAL TRANSCRIPTOME. FURTHERMORE, I WILL TEST THE MECHANISM OF MED23-MEDIATED CONTROL OF KEY REGULATORY GENES THAT FUNCTION IN CRANIOFACIAL DEVELOPMENT AND ARE LINKED TO CRANIOFACIAL DISORDERS. SPECIFICALLY, I WILL INVESTIGATE THE MOLECULAR BASIS OF MED23 FUNCTION VIA ITS CONTROL OF RUNX2 AND SS-CATENIN IN THE MANDIBULAR MESENCHYME. (AIM 3) GENERATE AND CHARACTERIZE LOSS OF FUNCTION MUTANTS OF THE MEDIATOR TAIL SUBMODULE PROTEIN, MED24. THE BROAD IMPACT OF THIS INNOVATIVE PROPOSAL WILL ADVANCE OUR MECHANISTIC UNDERSTANDING OF THE GENE AND PROTEIN NETWORKS UNDERLYING THE FUNCTION OF GLOBAL TRANSCRIPTION CO-FACTOR MEDIATOR COMPLEX PROTEINS IN FUNDAMENTAL CELLULAR PROCESSES AND DEVELOPMENT.

DEFINING MECHANISTIC DIFFERENCES BETWEEN EMBRYONIC AND REGENERATIVE ORGANOGENESIS

RYAN WHITE TITLE III HIV CAPACITY DEVELOPMENT AND PLANNING GRANTS - THE RUTH M. ROTHSTEIN CORE CENTER (CORE CENTER) IS A CRITICAL PROVIDER OF PRIMARY MEDICAL AND SUPPORTIVE SERVICES TO PEOPLE LIVING WITH HIV/AIDS (PLWHA) THROUGHOUT THE CHICAGO ELIGIBLE METROPOLITAN AREA (EMA). CORE CENTER IS PART OF THE COOK COUNTY HIV INTEGRATED PROGRAM (CCHIP) AND COOK COUNTY HEALTH (CCH), THE LARGEST SAFETY NET HEALTHCARE PROVIDER FOR CHICAGO AND COOK COUNTY. IN 2021, THE CORE CENTER PROVIDED 14,733 AMBULATORY VISITS TO 4,683 UNDUPLICATED CLIENTS AND 11,133 VISITS FOR MEDICAL SUBSPECIALTY CARE TO MORE THAN 3,300 CLIENTS. ACCORDING TO THE ILLINOIS DEPARTMENT OF PUBLIC HEALTH (IDPH), THE BURDEN OF HIV/AIDS IN THE STATE OF ILLINOIS IS MOST CONCENTRATED IN THE NINE-COUNTY CHICAGO EMA, WHICH INCLUDES THE CITY OF CHICAGO. AS OF DECEMBER 31, 2020, 80% OF ALL PLWHA IN ILLINOIS RESIDE IN THE CHICAGO EMA, WITH 66% IN THE CITY OF CHICAGO. THIS AREA IS ALSO HOME TO THE MAJORITY OF AFRICAN AMERICAN AND HISPANIC PLWHA: 84% OF AFRICAN AMERICAN PLWHA AND 88% OF HISPANIC PLWHA. THIS PROPOSAL SEEKS $150,000 FOR A ONE-YEAR INNOVATIVE PROJECT TO ENHANCE AND EXPAND OUR PEER NAVIGATOR PROGRAM AT CORE CENTER. THE AIM IS TO DEVELOP A VIRTUAL COMMUNITY HEALTH WORKER (CHW) PROGRAM AND RECRUIT CLIENTS TO PARTICIPATE TO GAIN SKILLS TOWARD WORKFORCE DEVELOPMENT. THIS FUNDING WOULD ALLOW THE NEWLY FORMED “SEXUAL HEALTH AMBASSADOR PROGRAM EMPOWERED” INITIATIVE OR “S.H.A.P.E. PROGRAM TO EXPAND AND CONTINUE BEYOND ITS CURRENT SHORT-TERM ENDING THE HIV EPIDEMIC (EHE) FUNDING WHICH ENDS FEBRUARY 2022. S.H.A.P.E. IS A "YOUTH SEXUAL HEALTH AMBASSADOR PROGRAM" AIMED AT RECRUITING YOUNG MSM AND TRANSGENDER HIV POSITIVE PATIENTS OF COLOR WHO ARE DISPROPORTIONATELY IMPACTED BY HIV/AIDS. THE PROJECT GOALS ARE TO: TO POSITIVELY IMPACT THE OVERALL SEXUAL HEALTH OUTCOMES OF YOUNG ADULTS OF COLOR AGES 18-25 AS WE ENGAGE, EDUCATE, AND EMPOWER YOUTH TO ACCESS PRIMARY CARE, PREVENTIVE SEXUAL HEALTH SERVICES INCLUDED BY NOT LIMITED TO EDUCATION, SCREENING, AND TR EATMENT OF HIV AND OTHER STI'S. WITH THIS FUNDING, WE WILL MERGE OUR EXISTING PEER NAVIGATION PROGRAM WITH THE S.H.A.P.E. INITIATIVE TO FORM A NEW CHW PROGRAM.

RYAN WHITE TITLE III HIV CAPACITY DEVELOPMENT AND PLANNING GRANTS

DECIPHERING GENOME INTEGRITY MAINTENANCE USING CYTOGENOMICS - PROJECT SUMMARY/ABSTRACT CHROMOSOME MISSEGREGATION LEADS TO ANEUPLOIDY AND GENOMIC INSTABILITY—HALLMARKS OF CANCER AND CONTRIBUTORS TO REPRODUCTIVE AGING. UNDERSTANDING HOW CENTROMERE SEQUENCE COMPOSITION, STRUCTURE, AND EPIGENETIC BACKGROUND INFLUENCE CHROMOSOME SEGREGATION IS IMPORTANT FOR UNRAVELING THE MOLECULAR ORIGINS OF GENOME INSTABILITY THAT COULD PROMOTE CANCER EVOLUTION. CENTROMERES ARE SPECIALIZED CHROMOSOMAL REGIONS THAT SERVE AS SITES OF KINETOCHORE FORMATION AND MICROTUBULE BINDING THAT PARTITION CHROMOSOMES INTO DAUGHTER CELLS DURING CELL DIVISION. THE GERTON LAB AT THE STOWERS INSTITUTE FOR MEDICAL RESEARCH, SUPPORTED BY NCI FUNDING (R01CA266339, MAINTAINING THE INTEGRITY OF A GENOME), INVESTIGATES CENTROMERE BIOLOGY AND KINETOCHORE FUNCTION TO UNCOVER THE MECHANISMS DRIVING CHROMOSOME MISSEGREGATION AND ANEUPLOIDY. AS A RESEARCH SPECIALIST II IN THE GERTON LAB, I DESIGN AND IMPLEMENT EXPERIMENTAL STRATEGIES ADDRESSING CENTROMERE COMPOSITION, FUNCTIONAL ACTIVITY, AND CHROMOSOME DYNAMICS. OUR CURRENT RESEARCH INVESTIGATES HOW NATURAL VARIATION IN HUMAN CENTROMERIC ARRAY SIZE AND ACTIVITY IMPACTS THE ACCURACY OF CHROMOSOME SEGREGATION. MULTIPLE COMPLETE TELOMERE- TO-TELOMERE HUMAN GENOME ASSEMBLIES EXPOSED CENTROMERES AS SOME OF THE MOST VARIABLE REGIONS IN THE HUMAN GENOME. CENTROMERES OF THE SAME CHROMOSOMES CAN DIFFER BETWEEN INDIVIDUALS IN BOTH SIZE AND THE EPIGENETIC DETERMINANTS OF KINETOCHORE POSITIONING. WE HYPOTHESIZE THAT INDIVIDUAL SEQUENCE AND EPIGENETIC PATTERN VARIATIONS IN HUMAN CENTROMERES CAN ALTER THEIR INTERACTIONS WITH CELL DIVISION MACHINERY, PREDISPOSING INDIVIDUALS TO DISTINCT CHROMOSOME MISSEGREGATION EVENTS. TO TEST THIS HYPOTHESIS, WE ARE WORKING ON MAPPING THE CENTROMERIC SEQUENCE LANDSCAPE AND ASSOCIATED KINETOCHORE PROTEINS IN INDIVIDUAL HUMAN CENTROMERES, AND INVESTIGATING HOW THESE VARIATIONS IMPACT CHROMOSOME SEGREGATION IN CULTURED CELLS AND TUMOR TISSUES. I UTILIZE CUTTING-EDGE GENOMIC AND CYTOGENETIC TECHNIQUES, ALONG WITH HIGH-RESOLUTION MICROSCOPY, TO MEASURE CENTROMERE SIZES AND THE DEPOSITION OF INNER KINETOCHORE PROTEINS. THESE DATA ARE BEING INTEGRATED WITH GENETIC AND EPIGENETIC INFORMATION FROM THE TELOMERE-TO-TELOMERE (T2T) CONSORTIUM, A GLOBAL MULTI-LAB INITIATIVE FOCUSED ON GENERATING COMPLETE GENOME ASSEMBLIES. THIS COLLABORATION ALLOWS OUR GROUPS TO INTEGRATE GENOMIC DATA WITH CYTOGENETICS, A METHOD WE CALL “CYTOGENOMICS”. THIS INFORMATION WILL BE USED TO FIND THE CENTROMERE-SPECIFIC GENETIC AND EPIGENETIC SIGNATURES INFLUENCING THE ACCURACY OF CHROMOSOME SEGREGATION IN BOTH CULTURED CELLS AND XENOGRAFT TUMOR TISSUES. OUR LONG-TERM GOAL IS TO DEVELOP COMPREHENSIVE MODELS EXPLAINING HOW CENTROMERE SEQUENCES AND ACTIVITIES CONTRIBUTE TO THE RISE OF ANEUPLOIDY AND TO TRANSLATE THESE FINDINGS INTO INSIGHTS RELEVANT TO CANCER BIOLOGY. THIS RESEARCH IS INNOVATIVE BECAUSE IT LEVERAGES THE LATEST INFORMATION ON HUMAN CENTROMERIC DNA ARRAYS, INTRODUCES NEW MODELS FOR CENTROMERE ORGANIZATION, AND EMPLOYS CUTTING-EDGE MOLECULAR, GENOMIC, AND IMAGING TOOLS. THE PROPOSED WORK IS SIGNIFICANT BECAUSE IT ADDRESSES FUNDAMENTAL QUESTIONS ABOUT THE ROLE OF INNATE CENTROMERE VARIATION IN GENOMIC ALTERATIONS, PROVIDING INSIGHTS INTO CANCER BIOLOGY AND AGING.

CHARACTERIZATION OF THE SIN3A AND SIN3B HDAC COMPLEXES

ROLE OF LONG NONCODING RNAS IN PROSTATE CANCER

CAPACITY BUILDING FOR ENHANCED RESEARCH ADMINISTRATION (CABERA- II) IN AFRICA

CAPACITY BUILDING FOR ENHANCED RESEARCH ADMINISTRATION (CABERA) AT NOGUCHI MEMORIAL INSTITUTE FOR MEDICAL RESEARCH

RYAN WHITE HIV/AIDS PROGRAM PART D WICY COVID-19 RESPONSE

INTRINSIC VS EXTRINSIC DEFECTS IN NEURAL CREST CELL PATTERNING

RYAN WHITE TITLE III HIV CAPACITY DEVELOPMENT AND PLANNING GRANTS

CHARACTERIZATION OF NOD, A DROSOPHILA PROTEIN REQUIRED FOR CHROMOSOME SEGREGATION

OPENING SMALL PACKAGES: UNRAVELING ROLES FOR MICROPROTEINS DURING EARLY VERTEBRATE DEVELOPMENT - PROJECT SUMMARY HUMAN DEVELOPMENT RELIES ON HIGHLY COORDINATED CELL DIVISION, SIGNALING, MIGRATION, AND DIFFERENTIATION. ANY PERTURBATIONS OF THE PROTEIN EFFECTORS THAT ORCHESTRATE THESE CRITICAL PROCESSES CAN LEAD TO HUMAN BIRTH DEFECTS AND/OR DISEASES1-3. WHILE OUR CLASSICAL CATALOG CONTAINS AROUND 20,000 PROTEINS, EVIDENCE FROM ‘OMICS-BASED TECHNIQUES HAS GENERATED A RAPID PARADIGM SHIFT IN RNA BIOLOGY4-12. NOTABLY, RNA SEQUENCES DEFINED AS NON- CODING IN FACT PRODUCE SHORT PROTEINS (£ 100 AMINO ACIDS) CALLED MICROPROTEINS THAT MODULATE DIVERSE PROCESSES13- 40 INCLUDING HEART FUNCTION26, IMMUNITY22,27,41, AND CELL GROWTH29,30. FOR EXAMPLE, A MICROPROTEIN CALLED APELA MAINTAINS PLURIPOTENCY IN HUMAN EMBRYONIC STEM CELLS20 AND IS CRITICAL FOR ZEBRAFISH HEART DEVELOPMENT16,19. HOWEVER, ADDITIONAL MICROPROTEIN FUNCTION(S) DURING VERTEBRATE DEVELOPMENT REMAIN LARGELY UNKNOWN. ZEBRAFISH IS AN OUTSTANDING MODEL FOR INTERROGATING VERTEBRATE GENE FUNCTION. THEIR GENETIC TRACTABILITY COUPLED WITH EXTERNAL, SYNCHRONOUS DEVELOPMENT IS WELL-SUITED FOR DEVELOPMENTAL ANALYSES. FURTHER, HUNDREDS OF MICROPROTEINS HAVE BEEN IDENTIFIED ACROSS ZEBRAFISH DEVELOPMENT USING RIBOSOME PROFILING, MASS- SPECTROMETRY, AND CONSERVATION ANALYSES16,42. REMARKABLY, APELA IS THE ONLY MICROPROTEIN OUT OF THESE 400 THAT IS CURRENTLY CHARACTERIZED. A KEY BARRIER TO FURTHER MICROPROTEIN INVESTIGATION IS THAT A MAJORITY OF MESSENGER RNAS (MRNA) DURING EARLY DEVELOPMENT ARE MATERNALLY PROVIDED AND CAN MASK THE EFFECTS OF A TARGETED GENE DISRUPTION. OUR NOVEL CRISPR/CAS13D SYSTEM43,44 OVERCOMES THIS BARRIER BECAUSE IT ACTIVELY DEGRADES ITS TARGET MRNA AND THEREFORE ENABLES SELECTIVE KNOCKDOWN OF MATERNALLY PROVIDED MRNAS IN ZEBRAFISH. MY PRELIMINARY EXPERIMENTS WITH CRISPR/CAS13D HAVE REVEALED THAT KNOCKDOWN OF ONE MICROPROTEIN MRNA INHIBITS ZYGOTIC GENOME ACTIVATION AND DISRUPTS POSTERIOR PATTERNING. THIS STUDY WILL COMBINE CRISPR/CAS13D AND ‘OMICS-BASED TECHNIQUES TO INTERROGATE MICROPROTEIN FUNCTION DURING ZEBRAFISH DEVELOPMENT. AIM 1 WILL LEVERAGE CRISPR/CAS13D TO ELUCIDATE MICROPROTEINS IMPORTANT FOR EARLY DEVELOPMENT. THEN, AIM 2 WILL DETERMINE THE CELL AND MOLECULAR PROCESSES THAT RELY ON DEVELOPMENTAL MICROPROTEINS. TOGETHER, THESE AIMS WILL DEFINE AND CHARACTERIZE A POPULATION OF MICROPROTEINS INVOLVED IN VERTEBRATE DEVELOPMENT. EXPERIMENTAL APPROACHES WILL DEVELOP MY SKILLS IN BIOINFORMATICS, MOLECULAR GENETICS, DEVELOPMENTAL BIOLOGY, AND PROTEIN BIOCHEMISTRY. MICROPROTEINS CRITICAL FOR ZEBRAFISH DEVELOPMENT WILL BE INFORMATIVE FOR EXPANDING THE CATALOG OF HUMAN PROTEINS THROUGH COMPARATIVE ANALYSES. FURTHER, MICROPROTEINS WITH FUNCTIONS DURING DEVELOPMENT REPRESENT UNCHARTED THERAPEUTIC AND/OR DIAGNOSTIC OPPORTUNITIES FOR HUMAN BIRTH DEFECTS AND/OR HUMAN DISEASES WITH DEVELOPMENTAL ORIGINS.

TRITHORAX-RELATED, MLL3 AND MLL4 IN ENHANCER-MEDIATED CANCER PATHOGENESIS

RIBOSOMAL RNA TRANSCRIPTION IS SPATIOTEMPORALLY REGULATED AND FUNCTIONALLY REQUIRED DURING NEURAL CREST,BONE AND CARTILAGE DEVELOPMENT

SPECIAL PROJECTS OF NATIONAL SIGNIFICANCE

INVESTIGATING HOW TRANSCRIPTION FACTORS COOPERATE AND OVERCOME THE ENHANCER NUCLEOSOME BARRIER DURING EMBRYONIC PATTERNING - PROJECT SUMMARY THIS MISREGULATION OF GENE EXPRESSION UNDERLIES SEVERAL HUMAN DISEASES, INCLUDING MANY CANCERS, DIABETES, OBESITY, AND MULTIPLE DEVELOPMENTAL DISORDERS. GENOME-WIDE STUDIES AND NEXT-GENERATION SEQUENCING HAVE REVEALED THAT SEQUENCE VARIANTS IN ENHANCERS, CIS-REGULATORY DNA SEQUENCES THAT CONTROL SPACIOTEMPORAL GENE EXPRESSION PROGRAMS, CONTRIBUTE TO THE DEVELOPMENT OF THESE DISEASES. THESE MUTATIONS OFTEN AFFECT ENHANCER ACTIVITY, WHICH MUST BE TIGHTLY CONTROLLED SINCE ENHANCERS DRIVE TISSUE AND CELL-TYPE SPECIFIC GENE EXPRESSION PATTERNS. ONE WAY THAT ENHANCER ACTIVITY IS CONTROLLED IS THROUGH THE REGULATION OF ENHANCER ACCESSIBILITY BY THE NUCLEOSOME: THE STRUCTURAL UNIT OF CHROMATIN COMPRISED OF 147 BP OF DNA AND A HISTONE OCTAMER. ENHANCERS ARE CHARACTERIZED BY AN INTRINSICALLY STRONG NUCLEOSOME BARRIER THAT PREVENTS THE BINDING OF TRANSCRIPTION FACTORS (TFS), THE PROTEINS THAT ACTIVATE ENHANCERS, UNTIL THE PROPER CONTEXT FOR ACTIVATION IS REACHED, AT WHICH POINT TFS MUST OVERCOME THE NUCLEOSOME BARRIER AND BIND TO THE DNA. WHILE NUCLEOSOME DEPLETION IS A KEY EARLY STEP IN ENHANCER ACTIVATION, WE DO NOT YET UNDERSTAND HOW THE NUCLEOSOME BARRIER IS OVERCOME AND HOW ENHANCERS ARE MADE ACCESSIBLE FOR GENE ACTIVATION, DESPITE ACCESSIBILITY BEING A MAJOR REGULATOR OF ENHANCER ACTIVITY. CURRENT MODELS SUGGEST THAT SPECIALIZED TFS CALLED PIONEER FACTORS CAN ACCESS THEIR MOTIFS IN THE PRESENCE OF NUCLEOSOMES AND FOMENT NUCLEOSOME DEPLETION THROUGH COOPERATIVITY WITH ADDITIONAL TFS. EVEN STILL, HOW PIONEER AND NON-PIONEER TFS COOPERATE TO GENERATE CHROMATIN ACCESSIBILITY AT ENHANCERS IS NOT YET KNOWN. FURTHERMORE, HOW PIONEER FACTORS PERTURB THE NUCLEOSOMAL LANDSCAPE TO FACILITATE CHROMATIN ACCESSIBILITY AND COOPERATIVE TF BINDING IS UNCLEAR. THIS STUDY SEEKS TO IDENTIFY HOW TFS OVERCOME THE NUCLEOSOME BARRIER AT ENHANCERS USING HIGH-RESOLUTION EXPERIMENTAL AND COMPUTATIONAL GENOMICS TECHNIQUES TO MAP TF BINDING, CHROMATIN ACCESSIBILITY, AND NUCLEOSOME POSITIONING. AIM 1 WILL CHARACTERIZE HOW PIONEER AND NON-PIONEER TFS COOPERATE FOR BINDING TO THE DNA AND FOR ESTABLISHING CHROMATIN ACCESSIBILITY. THIS AIM WILL COMBINE HIGH-RESOLUTION TF BINDING (CHIP- NEXUS) AND TEMPORALLY RESOLVED CHROMATIN ACCESSIBILITY (TIME-COURSE ATAC-SEQ) INFORMATION WITH DEEP LEARNING MODELS (BPNET) THAT WILL REVEAL THE SEQUENCES AND SEQUENCE CONSTRAINTS THAT ARE IMPORTANT FOR AND PREDICTIVE OF TF COOPERATIVITY. AIM 2 WILL PROFILE GENOME-WIDE NUCLEOSOME POSITIONAL CHANGES OVER DEVELOPMENTAL TIME AT UNPRECEDENTED RESOLUTION, USING A CHEMICAL MAPPING OF NUCLEOSOME CENTERS APPROACH. THIS AIM WILL UNCOVER HOW THE NUCLEOSOME STATE AT ENHANCERS IS ALTERED OVER TIME TO GENERATE ACCESSIBILITY AND HOW NUCLEOSOMES ARE POSITIONED WITH RESPECT TO THE UNDERLYING REGULATORY DNA SEQUENCES. TAKEN TOGETHER, THESE AIMS WILL ILLUMINATE HOW TFS PIONEER THE CHROMATIN LANDSCAPE FOR ENHANCER ACTIVATION, THEREBY DEEPENING THE FIELD’S UNDERSTANDING OF THE MECHANISMS OF GENE REGULATION AND HOW MISREGULATION CONTRIBUTES TO HUMAN DISEASE.

DEFINING THE MECHANISMS OF NUCLEAR PORE COMPLEX ASSEMBLY IN FISSION YEAST

SPECIAL PROJECTS OF NATIONAL SIGNIFICANCE