Sanford

CANCER CENTER SUPPORT GRANT

BURNHAM CENTER FOR CHEMICAL GENOMICS

PROTEIN GLYCOSYLATION IN THE COAGULOPATHY AND INFLAMMATION OF SEPSIS

CENTER FOR PEDIATRIC RESEARCH

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CENTER FOR CANCER BIOLOGY RESEARCH

GLOBAL INNATE IMMUNE RESPONSES TO HIV-1 INFECTION

TARGETING PTEN - AN UPSTREAM DOWNSTREAM AND OFFSTREAM APPROACH

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COLLABORATIVE RESEARCH CENTER FOR AMERICAN INDIAN HEALTH

PRENATAL ALCOHOL IN SUDDEN INFANT DEATH SYNDROME AND STILLBIRTH (PASS) NETWORK

AGING AS A RISK FACTOR AND TARGET FOR PREVENTION OF LIVER CANCER - PROJECT SUMMARY – OVERALL THE INCIDENCES OF LIVER CANCER (PRIMARILY HEPATOCELLULAR CARCINOMA (HCC)) ARE INCREASING AND DISEASE OUTCOME IS POOR. CONSEQUENTLY, THERE IS AN URGENT NEED FOR NEW THERAPIES AND BETTER PREVENTIVE STRATEGIES. AGE IS A MAJOR RISK FACTOR FOR HCC. IN LINE WITH THE GEROSCIENCE HYPOTHESIS, WE HYPOTHESIZE THAT AGING DRIVES A DYSFUNCTIONAL MITOCHONDRIAL, EPIGENETIC AND METABOLIC NETWORK THAT PROMOTES AND EXACERBATES AGE-ASSOCIATED DYSREGULATION OF IMMUNE FUNCTION AND INFLAMMATION IN LIVER. LOSS OF HOMEOSTASIS ACROSS MULTIPLE SYSTEMS IS PERMISSIVE FOR NEOPLASTIC LIVER DISEASE. WE FURTHER HYPOTHESIZE THAT DYSREGULATED CHRONIC INTERFERON SIGNALING IS CENTRAL TO THIS PATHOGENIC NETWORK. WE WILL DISSECT THIS NETWORK AND TEST THE CONSEQUENCE OF CHRONIC INTERFERON SIGNALING, TO UNDERSTAND WHY THE INCIDENCE OF LIVER CANCER INCREASES WITH AGE. WE WILL ALSO INVESTIGATE APPROACHES THAT TARGET THIS NETWORK FOR THEIR ABILITY TO PREVENT AND COMBAT LIVER CANCER. OUR OVERALL SPECIFIC OBJECTIVES ARE: OBJECTIVE 1. INVESTIGATE AGE-ASSOCIATED CHANGES TO MITOCHONDRIA, CHROMATIN, METABOLISM (SPECIFICALLY, BILE ACIDS) AND INNATE AND ADAPTIVE IMMUNITY, THEIR CAUSAL ROLE IN HCC AND UNDERLYING MECHANISMS. OBJECTIVE 2. INVESTIGATE HOW INTERACTIONS BETWEEN THESE DIFFERENT SYSTEMS AND AGE-DEPENDENT DYSREGULATION OF THESE INTERACTIONS CONTRIBUTES TO HCC. OBJECTIVE 3. TEST THE HYPOTHESIS THAT AT LEAST SOME OF THESE AGE-ASSOCIATED ALTERATIONS AND CONSEQUENT PREDISPOSITION TO HCC ARE DEPENDENT ON CHRONIC INTERFERON SIGNALING IN AGED TISSUE. OBJECTIVE 4. INVESTIGATE APPROACHES THAT TARGET AGE DYSREGULATION, FOR EXAMPLE SUPPRESSORS OF CHRONIC INTERFERON ACTIVATION, MITOHORMETIC INTERVENTIONS, RAPAMYCIN, SENOLYTICS, BILE ACID MODULATORS AND IMMUNE-MODULATORS, FOR THEIR ABILITY TO SUPPRESS THE ONSET OF LIVER CANCER AND BETTER COUNTER ESTABLISHED CANCER. SINCE AGE IS THE BIGGEST SINGLE RISK FACTOR FOR HCC, IT FOLLOWS THAT A MOLECULAR UNDERSTANDING OF THE AGE- DEPENDENCE OF HCC CAN LEAD TO IMPROVED DISEASE MANAGEMENT THROUGH RISK ASSESSMENT, EARLY DETECTION, PROGNOSTICATION AND THERAPY. MOREOVER, AN UNDERSTANDING OF HOW HCC DEVELOPS DURING AGING CAN ALSO LEAD TO PREVENTATIVE INTERVENTIONS. THIS PPG WILL DEFINE CRITICAL MOLECULAR MECHANISMS UNDERPINNING AGE-DEPENDENCE OF HCC. WE WILL ALSO PROMOTE APPROACHES FOR IMPROVED RISK ASSESSMENT THROUGH APPLICATION, TESTING AND REFINEMENT OF A TRANSCRIPTOME-BASED “TUMORIGENIC INDEX” TO QUANTITATE THE RISK OF HCC. FINALLY, BASED ON OUR DISCOVERIES, WE WILL TEST A PANEL OF CANDIDATE INTERVENTIONS FOR THOSE THAT CAN PREVENT AND COMBAT HCC.

SANFORD COMMUNITY CANCER PROGRAM OF THE NORTH CENTRAL PLAINS

PURPOSE: AMERICAN RESCUE PLAN ACT AWARDED AS ECONOMIC ASSISTANCE TO ELIGIBLE U.S. AIRPORTS AND ELIGIBLE CONCESSIONS AT THOSE AIRPORTS TO PREVENT, PREPARE FOR, AND RESPOND TO THE COVID-19 PANDEMIC. ACTIVITIES TO BE PERFORMED/EXPECTED OUTCOMES: THIS GRANT PROVIDES ECONOMIC RELIEF FUNDS FOR COSTS RELATED TO OPERATIONS, PERSONNEL, CLEANING, SANITIZATION, JANITORIAL SERVICES, DEBT SERVICE PAYMENTS, AND COMBATING THE SPREAD OF PATHOGENS AT THE AIRPORT. INTENDED BENEFICIARY: THIS GRANT WILL PROVIDE FEDERAL FUNDING FOR AIRPORTS ASSOCIATED WITH SANFORD, FLORIDA.

TRANSDISCIPLINARY APPROACHES TO AMERICAN INDIAN AND RURAL POPULATION HEALTH RESEARCH

PURPOSE: REHABILITATE RUNWAY LIGHTING; RECONSTRUCT RUNWAY VISUAL GUIDANCE SYSTEM; REHABILITATE RUNWAY; AIRFIELD SAFETY MITIGATION MEASURES. ACTIVITIES TO BE PERFORMED/EXPECTED OUTCOMES: THIS PROJECT REHABILITATES 9,700 FEET OF EXISTING PAVED RUNWAY 9L/27R TO MAINTAIN THE STRUCTURAL INTEGRITY AND MINIMIZE FOREIGN OBJECT DEBRIS TO EXTEND ITS USEFUL LIFE. . THIS PROJECT RECONSTRUCTS THE PRECISION APPROACH PATH INDICATOR SYSTEM FOR EXISTING RUNWAY 9L/27R, AT BOTH ENDS, THAT HAS REACHED THE END OF ITS USEFUL LIFE. THIS PROJECT REHABILITATES THE EXISTING LIGHTING ON RUNWAY 9L/27R TO EXTEND ITS USEFUL LIFE. THIS PROJECT REMOVES TAXIWAY K1, CURRENTLY ALIGNED WITH RUNWAY 9C, IDENTIFIED AS A HOT SPOT BY THE FEDERAL AVIATION ADMINISTRATION TO ENHANCE SAFETY. THIS GRANT FUNDS PHASE 2, WHICH CONSISTS OF CONSTRUCTION. INTENDED BENEFICIARY: THIS GRANT WILL PROVIDE FEDERAL FUNDING FOR AIRPORTS ASSOCIATED WITH SANFORD, FLORIDA.

CENTER FOR BIOBEHAVIORAL MECHANISMS OF EATING BEHAVIOR - OVERALL CENTER FOR BIO-BEHAVIORAL MECHANISMS OF EATING BEHAVIOR PROJECT SUMMARY/ABSTRACT WE PROPOSE TO ESTABLISH A CENTER FOR BIO-BEHAVIORAL MECHANISMS OF EATING BEHAVIOR (CBM-EB) AT SANFORD RESEARCH, FARGO ALONG WITH THE UNIVERSITY OF NORTH DAKOTA SCHOOL OF MEDICINE AND NORTH DAKOTA STATE UNIVERSITY (NDSU). THE PROPOSED CBM-EB WILL INITIALLY CONSIST OF FIVE THEMATICALLY RELATED RESEARCH PROJECTS FOCUSING ON MECHANISMS RELATED TO EATING DISORDERS AND MALADAPTIVE EATING BEHAVIOR. TO SUPPORT THE PROJECT DIRECTORS OF EACH THESE FIVE RESEARCH PROJECTS, WE WILL PROPOSE A RESEARCH ASSESSMENT SERVICES CORE COMPRISED OF A WIDE ARRAY OF ASSESSMENT METHODOLOGIES THAT COULD BE INNOVATIVELY INTEGRATED IN EATING RELATED RESEARCH, INCLUDING ASSESSMENT MODALITIES IN THE AREA OF PSYCHOPATHOLOGY ASSESSMENT, BIOLOGICAL ASSAYS, NEUROIMAGING AND NEUROBIOLOGICAL ASSESSMENT, EATING BEHAVIOR ASSESSMENT, AND COGNITIVE ASSESSMENT. IN ADDITION TO THESE CORE RESEARCH ASSESSMENT METHODOLOGIES, WE PROPOSE A BIOMEDICAL STATISTICS CORE TO PROMOTE SCIENTIFICALLY VALID AND APPROPRIATE DATA COLLECTION AND ANALYSES FOR THE STUDY OF MECHANISTIC PROCESSES, AS WELL AS A HUMAN SUBJECTS CORE TO SUPPORT PROJECT DIRECTORS MANAGEMENT OF REGULATORY ASPECTS OF HUMAN SUBJECTS RESEARCH. ADDITIONALLY, WE PROPOSE A CLINICAL RESEARCH TRAINING PROGRAM WHICH INTEGRATES SEMINARS, COLLOQUIA, AND CLOSE MENTORSHIP TO ENHANCE THE DEVELOPMENT OF OUR ENTRY LEVEL INVESTIGATORS. WE BELIEVE THAT THESE RESOURCES WILL FACILITATE TRANSITION OF OUR PROJECT DIRECTORS TO INDEPENDENT INVESTIGATORS WHO CONDUCT CLINICALLY RELEVANT RESEARCH.

CLINICAL DEVELOPMENT OF AN MGLU2 POSITIVE ALLOSTERIC MODULATOR TO TREAT NICOTINE ADDICTION

SPATIAL MAPPING SENESCENT CELLS ACROSS THE MOUSE LIFESPAN BY MULTIPLEX TRANSCRIPTOMICS AND EPIGENOMICS - PROJECT SUMMARY CELLULAR SENESCENCE, CHARACTERIZED BY STABLE PROLIFERATION ARREST AND SECRETION OF PRO-INFLAMMATORY FACTORS, IS NOT ONLY A HALLMARK OF AGING, BUT ALSO A KEY CONTRIBUTOR TO AGE-ASSOCIATED DISEASES IN HUMANS. AS THE US POPULATION IS AGING, THERE IS AN ADDED URGENCY TO GAIN A BETTER UNDERSTANDING OF CELLULAR SENESCENCE IN DIFFERENT TISSUES OVER THE LIFESPAN. UNFORTUNATELY, WE STILL LACK THE KNOWLEDGE TO UNAMBIGUOUSLY DEFINE SENESCENCE AT THE MOLECULAR AND CELLULAR LEVELS, DUE TO ITS HETEROGENEOUS PHENOTYPES. TO ADDRESS THIS MAJOR GAP IN KNOWLEDGE, WE PROPOSE TO ESTABLISH A TISSUE MAPPING CENTER THAT FOCUSES ON THE IDENTIFICATION AND CHARACTERIZATION OF SENESCENT CELLS IN HEALTHY MOUSE BRAIN, BONE MARROW, BREAST, COLON AND LIVER. OUR RESEARCH STRATEGY BUILDS ON RECENT ADVANCES IN SINGLE CELL EPIGENOMICS TECHNOLOGIES THAT OUR TEAM DEVELOPED AND THE KNOWLEDGE THAT SENESCENT CELLS EXHIBIT CHARACTERISTIC CHANGES IN THE CHROMATIN LANDSCAPES AND HISTONE MODIFICATIONS ALONG WITH GENE EXPRESSION LEVELS AT MARKER GENES OF CELLULAR SENESCENCE. WE WILL DEPLOY CUTTING-EDGE SINGLE CELL IN SITU AND TISSUE DISSOCIATIVE MULTI-OMIC TOOLS THAT HAVE BEEN WELL ESTABLISHED IN OUR CENTER TO PRODUCE COMPREHENSIVE SINGLE CELL RESOLUTION MAPS OF THE TRANSCRIPTOME AND EPIGENOME IN MALE AND FEMALE MOUSE BRAIN, BONE MARROW, BREAST, COLON AND LIVER, AND TO PROVIDE QUALITATIVE AND QUANTITATIVE SPATIAL MAPS OF THE NORMAL BURDEN OF SENESCENT CELLS IN THESE VITAL ORGANS, ACROSS THE LIFESPAN OF TWO MOUSE STRAINS. WE WILL RIGOROUSLY VALIDATE THE NEWLY DEFINED SENESCENT CELL POPULATIONS USING PHARMACOLOGIC AND GENETIC APPROACHES TO ERADICATE SENESCENT CELLS OR SUPPRESS THEIR INFLAMMATORY PHENOTYPE, AND ORTHOGONAL STATE-OF-THE-ART AND CONVENTIONAL ASSAYS FOR CELLULAR SENESCENCE. WE WILL GENERATE WHOLE GENOME SINGLE CELL DNA METHYLATION DATA TO LINK OUR SPATIAL ATLAS TO MEASUREMENT OF EPIGENETIC AGE, A CANDIDATE PREDICTOR OF BENEFICIAL VERSUS DETRIMENTAL EFFECTS OF SENESCENT CELLS. WE EXPECT THAT COMPREHENSIVE SINGLE CELL ATLASES OF EPIGENOME AND TRANSCRIPTOME WILL ENABLE US TO IDENTIFY AND CHARACTERIZE CELLULAR SENESCENCE IN DIFFERENT TISSUE CONTEXTS AND DURING AGING. WE EXPECT THAT THE PLANNED RESEARCH WILL PROVIDE A REFERENCE FOR FUTURE STUDIES THAT SEEK TO CHARACTERIZE AND TARGET SENESCENT CELLS ASSOCIATED WITH OR PRECEDING DISEASE IN BRAIN, BONE MARROW, BREAST, COLON AND LIVER.

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PRECLINICAL STUDIES FOR THE DEVELOPMENT OF SELECTIVE MGLU2 POSITIVE ALLOSTERIC MODULATORS TO TREAT SUBSTANCE USE DISORDERS

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SANFORD ONE CARE: TRANSFORMING PRIMARY CARE FOR THE 21ST CENTURY

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SAFETY/TOXICOLOGY, ADME AND CMC ACTIVITIES TO SUPPORT THE ASSESSMENT OF THE MGLU2 PAM SBP-9330 IN A PHASE 2 CLINICAL STUDY IN SMOKERS - PROJECT SUMMARY THIS RESUBMISSION APPLICATION, “SAFETY/TOXICOLOGY, ADME AND CMC ACTIVITIES TO SUPPORT THE ASSESSMENT OF THE MGLU2 PAM SBP-9330 IN A PHASE 2 CLINICAL STUDY IN SMOKERS”, IS IN RESPONSE TO PAR-22-202 AND REPRESENTS THE CONTINUATION OF OUR CURRENT WORK FUNDED THROUGH 07/31/2023 BY U01 DA051077 “CLINICAL DEVELOPMENT OF AN MGLU2 POSITIVE ALLOSTERIC MODULATOR TO TREAT NICOTINE ADDICTION”. CIGARETTE SMOKING REMAINS ONE OF THE LEADING CAUSES OF DEATH AND DISEASE WORLDWIDE. ONLY THREE TYPES OF MEDICATIONS FOR SMOKING CESSATION HAVE BEEN APPROVED BY THE UNITED STATES FOOD AND DRUG ADMINISTRATION (FDA) (BUPROPION, VARENICLINE, AND NICOTINE REPLACEMENT THERAPY), ALL OF WHICH HAVE POOR EFFICACY AND TOLERABILITY. POSITIVE ALLOSTERIC MODULATORS (PAMS) OF THE METABOTROPIC GLUTAMATE RECEPTOR SUBTYPE 2 (MGLU2) DECREASE NICOTINE SELF-ADMINISTRATION AND CUE-INDUCED REINSTATEMENT OF NICOTINE SEEKING IN ANIMAL MODELS, PROVIDING SUPPORT FOR MGLU2 AS A VALID TARGET FOR THE TREATMENT OF NICOTINE ADDICTION. OUR INVESTIGATIONAL DRUG, THE SMALL MOLECULE MGLU2 PAM SBP-9330 (CHARACTERIZED DURING PRIOR GRANT U01 DA041731), HAS RECENTLY BEEN EVALUATED IN HEALTHY NONSMOKERS AND SMOKERS IN A PLACEBO- CONTROLLED, RANDOMIZED, AND DOUBLE-BLIND PHASE 1 CLINICAL STUDY (NCT04948827). TO DATE, THE DATA FOR THE HEALTHY NONSMOKERS HAVE BEEN UNBLINDED AND ANALYZED, REVEALING THAT SBP-9330 WAS WELL TOLERATED, WITH NO SERIOUS ADVERSE EVENTS OR SAFETY CONCERNS. HUMAN PHARMACOKINETIC DATA REVEALED THAT PLASMA EXPOSURES WERE NEARLY DOSE-PROPORTIONAL, ACCUMULATED APPROXIMATELY 2-FOLD OVER 14 DAYS OF DOSING, AND WERE SUFFICIENTLY HIGH FOR POTENTIAL EFFICACY. THESE PROMISING DATA SUPPORT CONTINUATION OF THE CLINICAL DEVELOPMENT OF SBP-9330.THE STUDIES PROPOSED IN THIS GRANT APPLICATION ARE REQUIRED BY THE FDA BEFORE WE CAN DOSE SBP-9330 FOR LONGER DURATION IN TOBACCO SMOKERS IN A PHASE 2 STUDY AND BEYOND. TO THIS END, OUR SPECIFIC AIMS ARE: (1) PERFORM THE PRECLINICAL ABSORPTION, DISTRIBUTION, METABOLISM, AND EXCRETION (ADME) STUDIES NEEDED TO SUPPORT THE PHASE 2 CLINICAL DEVELOPMENT OF SBP-9330 IN SMOKERS; (2) PERFORM THE PRECLINICAL TOXICOLOGY STUDIES NEEDED TO SUPPORT PHASE 2 CLINICAL DEVELOPMENT OF SBP-9330 IN SMOKERS; AND (3) PERFORM THE CHEMISTRY, MANUFACTURING, AND CONTROLS (CMC) ACTIVITIES NEEDED TO SUPPORT PHASE 2 CLINICAL DEVELOPMENT IN SMOKERS. TO ACHIEVE THESE SPECIFIC AIMS, WE HAVE RETAINED THE SAME HIGHLY EXPERIENCED AND QUALIFIED MULTIDISCIPLINARY TEAM OF INVESTIGATORS THAT HAS COLLABORATED PRODUCTIVELY DURING THE PREVIOUS GRANT (U01 DA041731) AND THE CURRENTLY ACTIVE GRANT (U01 DA051077). WE HAVE ALSO COMPILED AN EXTENSIVE DATA PACKAGE TO SUPPORT THE FURTHER CLINICAL DEVELOPMENT OF SBP-9330. ACHIEVEMENT OF THE MILESTONES IN THIS PROPOSAL WILL PROVIDE THE ADDITIONAL DATA AND DRUG SUBSTANCE NEEDED TO ADVANCE SBP-9330 INTO A PHASE 2 PROOF-OF-CONCEPT CLINICAL STUDY IN SMOKERS. OUR TEAM HAS THE DEPTH AND BREADTH OF EXPERTISE AND EXPERIENCE TO EXECUTE THE PROPOSED RESEARCH PLAN, AS EVIDENCED BY THE COMPLETE AND TIMELY ACHIEVEMENT OF THE MILESTONES FOR BOTH U01 DA041731 AND U01 DA051077 IN THE PAST SIX YEARS.

MECHANISMS OF CARDIOVASCULAR REMODELING

GENETIC CONTROL OF THE HEART BY TINMAN AND NEUROMANCER

LA JOLLA-PARKINSON'S DISEASE CENTER GRANT

ULTRASTRUCTURAL BASIS OF MECHANOTRANSDUCTION IN MATRIX ADHESIONS

REWIRED SIGNALING AT THE NEXUS OF MELANOMA METASTASIS AND RESISTANCE

GLYCOSYLTRANSFERASE FUNCTION IN DEVELOPMENT AND DISEASE

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CARBOHYDRATE-DEPENDENT ADHESION IN NORMAL AND TUMOR CELLS

LA JOLLA INTERDISCIPLINARY NEUROSCIENCE CENTER CORES

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TARGETING EPH RECEPTORS IN CANCER

PUBLIC HOUSING CAPITAL FUND

ENHANCEMENT OF A PREDICTIVE TOXICOLOGY TOOLBOX VIA MULTI-PARAMETRIC DATA INFORMED QSAR MODELING OF BENCHMARK TOXINS

MOLECULAR PATHOBIOLOGY OF SOLUBLE TREM2 IN ALZHEIMER'S DISEASE.

EPIGENETICS & SIGNALING IN HESC COMMITMENT AND DIFFERENTIATION INTO MUSCLE

PURPOSE: RECONSTRUCT TAXIWAY. ACTIVITIES TO BE PERFORMED/EXPECTED OUTCOMES: THIS PROJECT RECONSTRUCTS 2,000 FEET OF THE EXISTING PAVED TAXIWAY A PAVEMENT THAT HAS REACHED THE END OF ITS USEFUL LIFE. THIS GRANT FUNDS THE FINAL PHASE, WHICH CONSISTS OF CONSTRUCTION. INTENDED BENEFICIARY: THIS GRANT WILL PROVIDE FEDERAL FUNDING FOR AIRPORTS ASSOCIATED WITH SANFORD, FLORIDA.

CHARACTERIZATION, OPTIMIZATION, AND DEVELOPMENT OF DUAL MGLU2/3 POSITIVE ALLOSTERIC MODULATORS FOR OPIOID USE DISORDER - PROJECT SUMMARY OPIOID USE DISORDER (OUD) IS A SIGNIFICANT PROBLEM WORLDWIDE AND A SURGE IN METHAMPHETAMINE (MA) USE HAS EMERGED IN CHRONIC OPIOID USERS. GIVEN RECENT INCREASES IN CO-ABUSE, THERE IS A DIRE NEED FOR NOVEL TREATMENT STRATEGIES THAT PREVENT RELAPSE TO DRUG USE IN BOTH OUD AND MA USE DISORDER (MUD). LONG-TERM DRUG EXPOSURE INDUCES ENHANCED GLUTAMATE (GLU)-MEDIATED SYNAPTIC PLASTICITY, WHICH UNDERLIES EXCESSIVE PHYSIOLOGICAL AND BEHAVIORAL RESPONSES TO DRUG-RELATED CUES. OPIOID AND MA EXPOSURE ALSO ACTIVATE MICROGLIA AND ASTROCYTES, PROMOTING RELEASE OF PRO-INFLAMMATORY CYTOKINES. ALL THESE FACTORS INCREASE THE RISK OF RELAPSE TO DRUG USE. NORMALIZATION OF ABERRANT GLU ACTIVITY CAUSED BY CHRONIC DRUG USE REPRESENTS A NOVEL THERAPEUTIC STRATEGY TO PREVENT RELAPSE IN OUD/MUD. THE ACTIVATION OF METABOTROPIC GLU RECEPTOR SUBTYPES 2 AND 3 (MGLU2/3) USING AGONISTS OR POSITIVE ALLOSTERIC MODULATORS (PAMS) DECREASES PSYCHOSTIMULANT SELF-ADMINISTRATION (SA) AS WELL AS CUE-INDUCED REINSTATEMENT (RI) IN ANIMALS. MOREOVER, PRECLINICAL STUDIES INDICATE THAT MGLU2/3 ACTIVATION HAS PROMISE FOR TREATING STRESS- AND ANXIETY-RELATED DISORDERS IN HUMANS AND CAN SYSTEMATICALLY AUGMENT SLEEP. HOWEVER, THE RELATIVE CONTRIBUTION OF MGLU2 VERSUS MGLU3 ACTIVATION FOR TREATING OUD/MUD IS NOT YET KNOWN. ACTIVATION OF PRESYNAPTIC MGLU2 AUTORECEPTORS RESULTS IN DOWNSTREAM INHIBITION OF GLU RELEASE IN THE NUCLEUS ACCUMBENS, WHICH IN TURN ATTENUATES DRUG RI. IN ADDITION, ACTIVATION OF MGLU3, WHICH IS HIGHLY EXPRESSED IN ASTROCYTES, LEADS TO THE RELEASE OF THE ANTI-INFLAMMATORY CYTOKINE TRANSFORMING GROWTH FACTOR BETA (TGF-SS). THE SPECIFIC LOCALIZATION AND SIGNAL TRANSDUCTION OF MGLU2 AND MGLU3 RECEPTORS LEAD TO OUR OVERARCHING HYPOTHESIS THAT DUAL ACTIVATION OF BOTH MGLU2 AND MGLU3 WILL PROVIDE A CLEAR ADVANTAGE FOR THE TREATMENT FOR OUD AND MUD OVER MGLU2 ACTIVATION ALONE. WE HAVE RECENTLY SYNTHESIZED AND CHARACTERIZED SBI-0799220, A PAM WITH EQUAL POTENCY FOR MGLU2 AND MGLU3 AND SBI-0801315, A PAM WITH >50-FOLD SELECTIVITY FOR MGLU2 VS MGLU3. PRELIMINARY DATA INDICATE THAT SBI-0801315 ATTENUATES OXY CUE-INDUCED RI AND SA, AND THAT SBI-0799220 ATTENUATES MA SA. HOWEVER, A DIRECT COMPARISON OF MGLU2/3 WITH MGLU2 PAMS IN MODELS OF OUD AND MUD HAS NOT YET BEEN CONDUCTED. THE GOAL OF THIS APPLICATION IS TO ADVANCE MGLU2/3 PAMS AS A NOVEL TREATMENT FOR PREVENTING RELAPSE TO OUD, EXAMINE THEIR POTENTIAL FOR TREATING MUD, AND SIMULTANEOUSLY OPTIMIZE MGLU2/3 PAMS. WE WILL DETERMINE THE IN VIVO EFFICACY OF MGLU2/3 AND MGLU2-PREFERRING PAMS TO ATTENUATE OXY/MA SA AND RI, OXY/MA-INDUCED WITHDRAWAL, MA-INDUCED NEUROINFLAMMATION, AND OPIOID-INDUCED ANTINOCICEPTION IN RATS. SIMULTANEOUSLY, WE WILL OPTIMIZE THE PHARMACOLOGICAL AND PHARMACEUTICAL PROPERTIES OF OUR MGLU2/3 PAM SERIES INCLUDING ABSORPTION, DISTRIBUTION, METABOLISM, EXCRETION, AND PHARMACOKINETICS. WE HAVE ASSEMBLED A MULTIDISCIPLINARY TEAM OF INVESTIGATORS THAT HAS THE KNOWLEDGE AND EXPERIENCE TO ACHIEVE THESE OUTCOMES. SUCCESSFUL COMPLETION OF THESE STUDIES WILL EXPEDITE DEVELOPMENT OF A NOVEL MGLU2/3 PAM TOWARDS INVESTIGATIONAL NEW DRUG (IND)-ENABLING STUDIES AND ULTIMATELY, A NOVEL TREATMENT FOR PREVENTING RELAPSE IN OUD.

ASSEMBLY, DYNAMICS AND EVOLUTION OF CELL-CELL AND CELL-MATRIX ADHESIONS

DEVELOPMENT OF SBI-553, AN ALLOSTERIC MODULATOR OF NTR1, FOR THE TREATMENT OF SUBSTANCE USE DISORDERS - PROJECT SUMMARY OPIOID USE DISORDER (OUD) IS A MAJOR PUBLIC HEALTH CRISIS IN THE UNITED STATES, WITH >50,000 OVERDOSE DEATHS IN 2017 ALONE. ADDICTION TO OPIOIDS IS INTIMATELY RELATED TO THE PHYSIOLOGY OF THE BRAIN’S DOPAMINE-BASED REWARDING SYSTEM. THE NEUROTENSIN 1 RECEPTOR (NTR1) IS FOUND AMONG DOPAMINE PRE AND POST-SYNAPTIC NEURONS IN THE CENTRAL NERVOUS SYSTEM, AND ACTS AS A MODULATOR OF DOPAMINERGIC SYSTEMS. EVEN THOUGH SUBSTANTIAL EVIDENCE POINTS TOWARDS NTR1 AS A MOLECULAR TARGET FOR TREATING ADDICTION DISORDERS, FEW NON- PEPTIDE BRAIN PENETRANT NEUROTENSIN MODULATORS HAVE BEEN IDENTIFIED, AND ORTHOSTERIC NTR1 LIGANDS DISPLAY SIDE EFFECTS INCLUDING HYPOTENSION AND HYPOTHERMIA THAT HAVE LIMITED THEIR CLINICAL DEVELOPMENT. RECENTLY, WE HAVE DISCOVERED A SERIES OF BRAIN-PENETRANT NTR1 MODULATORS, INCLUDING A LEAD COMPOUND SBI-553, WITH A UNIQUE MECHANISM OF ACTION AT NTR1. SBI-553 IS AN ORALLY AVAILABLE AND BRAIN PENETRANT B-ARRESTIN BIASED ALLOSTERIC MODULATOR OF NTR1, WHICH SHOWS EFFICACY IN A RANGE OF ADDICTION MODELS. THE LITERATURE CONTAINS NO DOCUMENTED REPORTS OF SUCH MOLECULES AMONG POSITIVE OR NEGATIVE ALLOSTERIC MODULATORS OF GPCRS. IN ADDITION, THE UNIQUE PROFILE OF SBI-553 APPEARS TO CIRCUMVENT THE CLINICALLY LIMITING SIDE EFFECTS OF HYPOTHERMIA AND HYPOTENSION DISPLAYED BY ORTHOSTERIC NTR1 LIGANDS. WHILE POTENTIALLY HIGH RISK, THE ACTIVITY OF SBI-553 HAS BEEN VALIDATED IN VITRO AND IN VIVO, AND THE INITIAL SAFETY PROFILING INDICATES NO ISSUES THAT WOULD PRECLUDE FURTHER DEVELOPMENT. THE GOAL OF THE APPLICATION IS TO DEVELOP SBI-553 AS A TREATMENT FOR OPIOID USE DISORDERS. A 2-YEAR UG3 PHASE IS PROPOSED WITH THE PRIMARY OBJECTIVE OF DETERMINING THE SUITABILITY OF SBI-553 (OR A BACKUP COMPOUND) ENTERING IND-ENABLING STUDIES FOLLOWED BY A 2-YEAR UH3 PHASE WITH THE PRIMARY OBJECTIVE OF COMPLETING A PHASE 1 SINGLE ASCENDING DOSE STUDY. DURING THE UG3 PHASE, THE TEAM WILL COMPLETE PHARMACOKINETIC (PK) STUDIES AND THE PRECLINICAL SAFETY PROFILE OF SBI-553 IN A RELEVANT SPECIES; IDENTIFY A BACKUP COMPOUND TO SBI-553; AND CONDUCT DEFINITIVE CELLULAR AND IN VIVO PHARMACOLOGY STUDIES WITH SBI-553 (AND/OR POTENTIAL BACKUP) IN MODELS OF ADDICTION TO INFORM BEST CLINICAL INDICATION AND CLINICAL ENDPOINTS (PK/PHARMACODYNAMIC). DURING THE UH3 PHASE, THE TEAM WILL ENSURE COMPLETION OF GMP SYNTHESIS, STABILITY TESTING AND FORMULATION; GLP TOXICOLOGY STUDIES; AND THE PHASE 1 SINGLE ASCENDING DOSE STUDY.

GENETIC ANALYSIS OF DROSOPHILA FUNCTIONAL AGING

MOLECULAR PATHOGENESIS AND TREATMENT OF HYPOPHOSPHATASIA

NEW CONGENITAL DISORDERS OF GLYCOSYLATION: THERAPY AND MODELS

NANOTHERAPY FOR VULNERABLE PLAQUE

NEURON-GLIA COMMUNICATION IN DEVELOPMENT

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FUNCTION OF CD19 IN B-CELL DEVELOPMENT & DIFFERENTIATION

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PURPOSE: REHABILITATE TAXIWAY LIGHTING; REHABILITATE TAXIWAY; AIRPORT WILDLIFE HAZARD ASSESSMENT/MANAGEMENT PLAN. THIS GRANT INCLUDES FUNDING BY THE AMERICAN RESCUE PLAN ACT OF 2021 TO INCREASE THE FEDERAL SHARE TO 100 PERCENT FOR THE AIRPORT IMPROVEMENT PROGRAM (AIP). ACTIVITIES TO BE PERFORMED/EXPECTED OUTCOMES: THIS PROJECT CONDUCTS AN AIRPORT WILDLIFE HAZARD ASSESSMENT. THIS PROJECT REHABILITATES 12,100 FEET OF THE EXISTING TAXIWAY B AND CONNECTORS TO MAINTAIN THE STRUCTURAL INTEGRITY OF THE PAVEMENT, MINIMIZE FOREIGN OBJECT DEBRIS, AND MAINTAIN SAFE AIRCRAFT OPERATIONS. THIS PROJECT REHABILITATES 12,100 FEET OF THE EXISTING TAXIWAY B LIGHTING SYSTEM TO EXTEND ITS USEFUL LIFE AND ENHANCE SAFE AIRFIELD OPERATIONS DURING LOW VISIBILITY CONDITIONS. THIS GRANT FUNDS THE FINAL PHASE, WHICH CONSISTS OF CONSTRUCTION TO REHABILITATE 3,280 FEET. THIS GRANT FUNDS THE FINAL PHASE, WHICH CONSISTS OF THE CONSTRUCTION TO REHABILITATE 3,280 FEET. INTENDED BENEFICIARY: THIS GRANT WILL PROVIDE FEDERAL FUNDING FOR AIRPORTS ASSOCIATED WITH SANFORD, FLORIDA.

THE CITY OF SANFORD IS SEEKING EDA FUNDING TO RECONSTRUCT THE EXISTING SEAWALL AROUND MARINA ISLE TO PROVIDE A STATE-OF-THE-ART FACILITY THAT WILL INCREASE THE SEAWALL HEIGHT TO PROTECT FROM FUTURE FLOODING AND INCREASED INTENSITY OF STORMS. THIS PROJECT AT MARINA ISLAND (AKA MARINA ISLE) IS PART OF A LARGER ONGOING EFFORT BY THE CITY OF SANFORD TO CONTINUALLY IMPROVE FLOOD PROTECTION FOR THE AREA. APPROXIMATELY 2,024 FEET OF SEAWALL WILL BE FORTIFIED AND RAISED IN ELEVATION BY TWO FEET. THE SCOPE OF WORK FOR THE PROPOSED INVESTMENT INVOLVES THE RECONSTRUCTION OF A PORTION OF SANFORD'S MARINA ISLE SEAWALL THAT HAS BEEN IN PLACE FOR OVER 50 YEARS AND HAS BEEN HEAVILY IMPACTED BY 2022 STORMS (HURRICANE IAN AND TROPICAL STORM NICOLE). THE SEAWALL PROTECTS A PRIME DOWNTOWN DEVELOPMENT AREA KNOWN AS MARINA ISLE, A PENINSULA CONSTRUCTED IN THE 1960'S IN THE LAKE MONROE PORTION OF THE ST. JOHNS RIVER. MARINA ISLE IS THE FOCAL POINT OF THE DOWNTOWN SANFORD AREA BUT IS UNDERUTILIZED DUE TO PREVIOUS FLOODING, AND A BLIGHTED MARINA FACILITY. RECONSTRUCTION OF THE SEAWALL WILL ALLOW REDEVELOPMENT TO MOVE FORWARD WITH A BETTER PROTECTED FACILITY. PROJECT TASKS WILL INCLUDE SEAWALL DESIGN, PERMITTING AND CONSTRUCTION.

DESCRIPTION:BROWNFIELDS ARE REAL PROPERTY, THE EXPANSION, DEVELOPMENT OR REUSE OF WHICH MAY BE COMPLICATED BY THE PRESENCE OR POTENTIAL PRESENCE OF A HAZARDOUS SUBSTANCE, POLLUTANT, OR CONTAMINANT. THIS AGREEMENT WILL PROVIDE FUNDING UNDER THE INFRASTRUCTURE INVESTMENT AND JOBS ACT TO THE CITY OF SANFORD TO CONDUCT REMEDIATION ACTIVITIES AS AUTHORIZED BY CERCLA 104(K)(3) IN SANFORD, MAINE. ACTIVITIES:SPECIFICALLY, THIS AGREEMENT WILL PROVIDE FUNDING TO THE RECIPIENT TO CLEAN UP A BROWNFIELD SITE. ADDITIONALLY, THE RECIPIENT WILL COMPETITIVELY PROCURE AND DIRECT A QUALIFIED ENVIRONMENTAL PROFESSIONAL TO CONDUCT ENVIRONMENTAL SITE ACTIVITIES, WILL CREATE A COMMUNITY INVOLVEMENT PLAN AND ADMINISTRATIVE RECORD FOR THE SITE, AND WILL REPORT ON INTERIM PROGRESS AND FINAL ACCOMPLISHMENTS BY COMPLETING AND SUBMITTING RELEVANT PORTIONS OF THE PROPERTY PROFILE FORM USING EPA'S ASSESSMENT, CLEANUP AND REDEVELOPMENT EXCHANGE SYSTEM (ACRES).SUBRECIPIENT:NO SUBAWARDS ARE INCLUDED IN THIS ASSISTANCE AGREEMENT.OUTCOMES:FURTHER, THE RECIPIENT WILL REMEDIATE 1 BROWNFIELD SITE AND ANTICIPATES HOLDING UP TO 3 COMMUNITY MEETINGS, FINALIZING 1 ANALYSIS OF BROWNFIELD CLEANUP ALTERNATIVES, AND SUBMITTING 16 QUARTERLY REPORTS. WORK CONDUCTED UNDER THIS AGREEMENT WILL BENEFIT THE RESIDENTS, BUSINESS OWNERS, AND STAKEHOLDERS IN AND NEAR SANFORD, MAINE.

CENTER FOR CANCER BIOLOGY RESEARCH - THIS COBRE PHASE III APPLICATION AIMS TO CONTINUE SUPPORT FOR THE CENTER FOR CANCER BIOLOGY RESEARCH (CCBR) WHICH HAS ESTABLISHED A STRONG RESEARCH PROGRAM WITHIN SANFORD RESEARCH, SIOUX FALLS, SD. THE ORIGINAL MISSION OF THE CCBR WAS TO ESTABLISH AND GROW CANCER RESEARCH WITH A FOCUS ON BASIC AND TRANSLATIONAL ACTIVITIES. DURING PHASE I COBRE FUNDING, THE CCBR WAS ESTABLISHED WITH FIVE PROJECT LEADERS. IMPORTANTLY, CRITICAL INFRASTRUCTURE, IN THE FORM OF SCIENTIFIC CORES, WAS DEVELOPED AND A PILOT GRANT PROGRAM WAS INITIATED. THE SUCCESS OF PHASE I WAS MARKED BY ATTAINMENT OF R01 LEVEL FUNDING BY PROJECT LEADERS WHO GRADUATED FROM COBRE FUNDING AND REMAIN SUCCESSFUL TO THIS DAY. IN ADDITION, PROJECT LEADERS PUBLISHED IMPORTANT SCIENTIFIC PAPERS IN PEER-REVIEWED JOURNALS AND THE SCIENTIFIC CORE FUNCTIONS BEGAN EXPANDING AS DID THEIR EXPERTISE. THE POWER OF THE PILOT GRANT PROGRAM WAS DEMONSTRATED BY SCIENTIFIC PUBLICATIONS AND OBTAINING OF FUNDING BY AWARDEES. THESE MILESTONES ENABLED THE AWARDING OF COBRE PHASE II FUNDING FOR THE CCBR. WHILE THE MISSIONS OF THE CCBR REMAINED UNCHANGED, THE INFLUX OF PHYSICIAN/SCIENTISTS TO THE CCBR PROMPTED AN EXTENSIVE FOCUS ON TRANSLATIONAL CANCER RESEARCH; A THEME THAT REMAINS STRONGLY IN PLACE. PHASE II PROJECT LEADERS WERE ALSO SUCCESSFUL IN OBTAINING EXTRAMURAL FUNDING AND GRADUATED FROM COBRE SUPPORT. MOREOVER, THE SCIENTIFIC CORES FURTHER EXPANDED THEIR SERVICES AND BEGAN A MORE SYSTEMATIC APPROACH (SMARTSHEETS) TO ORDER, TRACK AND REPORT BACK SERVICES TO THEIR USERS. IN ADDITION, FEE-FOR-SERVICE SCHEDULES WERE DEVELOPED FOR THE SCIENTIFIC CORES AS THEY PREPARED FOR ATTAINING SELF-SUFFICIENCY. THE PILOT GRANT PROGRAM CONTINUED TO PROVIDE FUNDING TO INVESTIGATORS WHO UTILIZED THESE RESOURCES TO SUCCESSFULLY PUBLISH AND OBTAIN FUNDING. PHASE III COBRE FUNDING WILL ENABLE THE CCBR'S SCIENTIFIC CORES TO REACH A STATE OF SELF-SUSTAINABILITY WHILE CONTINUING TO OFFER CRITICAL SERVICES TO CCBR MEMBERS AS WELL AS NON-CCBR MEMBERS OF SANFORD RESEARCH AND INVESTIGATORS FROM REGIONAL AND NATIONAL INSTITUTIONS. A CRITICAL COMPONENT OF THIS PHASE III APPLICATION WILL FUND THE CCBR'S PILOT GRANT PROGRAM. THESE PILOT GRANTS HAVE PROVEN TO BE ESSENTIAL SOURCES OF FUNDING FOR NEW INVESTIGATORS AND MORE SEASONED FACULTY THAT WANT TO TEST INNOVATIVE HYPOTHESES OR REQUIRE RESOURCES TO GENERATE DATA NECESSARY TO SECURE EXTRAMURAL FUNDING. HISTORICALLY, THE CCBR'S PILOT GRANT AWARDEES HAVE DEMONSTRATED THAT THIS FUNDING RESULTS IN PEER-REVIEWED SCIENTIFIC PUBLICATIONS AND ENABLES THEM TO OBTAIN FUNDING FROM EXTERNAL SOURCES. MOREOVER, A LARGE PERCENT OF PILOT GRANT PROJECTS UTILIZES THE CCBR'S SCIENTIFIC CORES THEREBY CONTRIBUTING TO THEIR SELF-SUSTAINABILITY. THE CCBR'S SUCCESSES OVER THE PAST DECADE OF COBRE FUNDING ARE A TESTAMENT TO THE TRANSFORMATIVE POWER OF THIS FUNDING MECHANISM. OBTAINING THE FINAL PHASE OF COBRE FUNDING WILL FURTHER STRENGTHEN THE SCIENTIFIC CORES, TRANSLATIONAL RESEARCH, SCIENTIFIC CULTURE OF INNOVATIVE SCIENCE AND THE RESEARCH INFRASTRUCTURE OF THE CCBR THAT BEGAN WITH PHASE I COBRE FUNDING.

HEPARAN SULFATE IN SKELETAL DEVELOPMENT AND DISEASES

ALTERED REVERSE TRANSCRIPTASE-DEPENDENT GENE DIVERSIFICATION MECHANISMS IN ALZHEIMER'S DISEASE BRAINS

A NOVEL ROLE FOR THE UPR COMPONENT, ATF6 IN AD-ASSOCIATED NEUROPROTECTIVE PATHWAYS

SIOUX COMMUNITY CANCER CONSORTIUM

SREBPS IN REGULATION OF LIPID METABOLISM

REGULATION OF NUTRIENT STRESS-INDUCED MACROPINOCYTOSIS IN PANCREATIC DUCTAL ADENOCARCINOMA

ROLE OF HYPERNUTRITION AND METABOLIC STRESS IN NON-ALCOHOLIC STEATOHEPATITIS (NASH) DRIVEN HEPATOCELLULAR CARCINOMA (HCC) - PROJECT SUMMARY – OVERALL HEPATOCELLULAR CARCINOMA (HCC) IS THE MOST COMMON PRIMARY LIVER CANCER AND A RISING CAUSE OF CANCER MORTALITY. HCC RELATED TO METABOLIC STRESS IS ON THE RISE, WITH NONALCOHOLIC STEATOHEPATITIS (NASH) RAPIDLY BECOMING THE DOMINANT ETIOLOGY. WE HYPOTHESIZE THAT STRESS-INDUCED METABOLIC REPROGRAMMING VIA NRF2 AND ATF6 ALTERS, GLUCOSE, LIPID, AND CHOLESTEROL METABOLISM TO AUGMENT GLYCOLYSIS AND INDUCE MITOCHONDRIAL DYSFUNCTION THROUGH STARD1 AND SAB, WHICH COLLABORATES WITH GENETIC ALTERATIONS TO ACCELERATE NASH PROGRESSION TO HCC THROUGH A SELF-SUSTAINED DOWNWARD SPIRAL. INTERFERENCE WITH KEY COMPONENTS OF THIS NETWORK MAY SUPPRESS HCC PROGRESSION AND CAN YIELD NOVEL INTERCEPTIVE STRATEGIES. OUR PRELIMINARY DATA INDICATE THAT INTERACTIONS BETWEEN THE CYTOSOL AND NUCLEUS (AKT;NRF2), THE ENDOPLASMIC RETICULUM (ER; ATF6) AND MITOCHONDRIA (STARD1+SAB) CONTRIBUTE SYNERGISTICALLY TO NASH-DRIVEN HCC. ELIMINATION OF ANY OF THESE COMPONENTS ATTENUATES HCC AND THEIR ACTIVATION EXACERBATES HCC PROGRESSION IN MURINE MODELS. TOGETHER, OUR OUTSTANDING TEAM OF INVESTIGATORS WILL ADDRESS THE PATHOLOGICAL SIGNIFICANCE AND MECHANISMS UNDERLYING THE CROSSTALK BETWEEN THESE PATHWAYS VIA 3 PROJECTS: PROJECT 1 WILL INTERROGATE HOW THE NEWLY DISCOVERED NRF2- FBP1 TUG-OF-WAR IS REGULATED TO SUPPORT HCC PROGRESSION. IT WILL IDENTIFY THE FIRST CELLS IN WHICH NRF2 AND AKT ARE ACTIVATED TO DRIVE FBP1 DEGRADATION AND THE CUES THAT TRIGGER THIS SWITCH. LINEAGE TRACING WILL DETERMINE WHETHER NRF2-HIGH HCC DIRECTLY ORIGINATES FROM FBP1-HIGH SENESCENT NASH HEPATOCYTES AND THE ROLE OF DIET- INDUCED DNA DAMAGE IN HEPATOCYTE SENESCENCE AND MUTAGENESIS WILL BE ELUCIDATED. PROJECT 2 WILL DEFINE HOW ATF6 DRIVES HCC IN RESPONSE TO METABOLIC STRESS. IT WILL UNCOVER THE MECHANISM BY WHICH ER STRESS AND THE UNFOLDED PROTEIN RESPONSE TRANSDUCER ATF6 PROMOTES NASH PROGRESSION TO HCC, LIKELY THROUGH NRF2 AND FBP1 DEGRADATION, THE REPROGRAMMING OF LIPID AND CHOLESTEROL METABOLISM, AND WILL TEST WHETHER AND HOW ATF6 ABLATION OR INHIBITION ABROGATE NASH-DRIVEN HCC. PROJECT 3 WILL EXPLORE HOW METABOLIC STRESS AND CHOLESTEROL TRIGGER MITOCHONDRIAL DYSFUNCTION THROUGH STARD1 AND SAB TO DRIVE HCC. MITOCHONDRIAL OUTER MEMBRANE PROTEINS STARD1 AND SAB MAY CONTRIBUTE TO NASH AND HCC DEVELOPMENT THROUGH ALTERED CHOLESTEROL TRAFFICKING AND BILE ACID SYNTHESIS, MITOCHONDRIAL DYSFUNCTION THAT TRIGGERS ROS PRODUCTION, RESULTING IN NRF2 AND ATF6 ACTIVATING OXIDATIVE AND/OR ER STRESS. THIS PPG OFFERS AN UNPRECEDENTED OPPORTUNITY FOR ADVANCED UNDERSTANDING OF NASH-DRIVEN HCC VIA PROXIMAL METABOLIC REGULATORS AND DNA DAMAGE CONTROLLED BY NRF2, ATF6, AND STARD1/SAB. EXTENSIVE COLLABORATIVE INTERACTIONS AND COORDINATED ADMINISTRATIVE RESOURCES (CORE A), SHARED USE OF DIETARY AND TRANSGENIC MOUSE MODELS, METABOLIC PROFILING/FLUX STUDIES (CORE 1) AND EXTENSIVE “OMIC” AND BIOINFORMATIC ANALYSES (CORE 2) WILL FURTHER ENHANCE COLLABORATIONS AND ENSURE UNIFORM SUCCESS THROUGHOUT ALL PROJECTS. THIS P01 OFFERS A UNIQUE TRANSLATIONAL OPPORTUNITY TO DEFINE HOW ENERGY RICH DIETS IMPACT REGULATORS THAT PROMOTE NASH PROGRESSION TO HCC, GENERATING NEW PREVENTIVE AND THERAPEUTIC STRATEGIES.

TRANSFORMATIVE RESEARCH ON SOMATIC GENE RECOMBINATION IN THE NORMAL AND ALZHEIMER'S DISEASE-RELATED DEMENTIA BRAIN

ROLE OF THE UNFOLDED PROTEIN RESPONSE IN BETA CELL

IMPROVE EXISTING AIRPORT

REGULATION OF FACTOR VIII SECRETION

IMPROVE EXISTING AIRPORT

A PHARMACOCHAPERONE-BASED STRATEGY FOR IDENTIFYING CHEMICAL PROBES OF BRAIN-DERIVED ORPHAN GPCRS

PRESENILINS IN PROTEIN TRAFFICKING

CENTER ON PROTEOLYTIC PATHWAYS(RMI)

MODIFIERS OF PROINSULIN INFLUENCE T2D SUSCEPTIBILITY

CONTROL OF LIPOGENESIS AND HEPATIC STEATOSIS BY CASPASE-2

COUNTERMEASURES FOR BIOTERROISM TARGETING CELLULAR HOST FACTORS

ROLES OF AN APP-BINDING PRO-APOPTOTIC PROTEIN IN MEDIATING NEURONAL CELL DEATH

SANFORD HEALTH CERTIFIED COMMUNITY BEHAVIORAL HEALTH CLINIC - THIS PROJECT WILL ESTABLISH THE SANFORD HEALTH CERTIFIED COMMUNITY BEHAVIORAL HEALTH CLINIC WHICH WILL PRIMARILY SERVE THE POPULATIONS OF REGION II IN NORTH CENTRAL MINNESOTA, HOME TO NEARLY 81,000 PEOPLE. THE PRIMARY GOALS OF THIS PROJECT ARE TO ESTABLISH A CERTIFIED COMMUNITY BEHAVIORAL HEALTH CLINIC IN NORTH CENTRAL MINNESOTA, IMPROVE OUR CARE COORDINATION, DECREASE THE NUMBER OF INPATIENT PLACEMENTS OUTSIDE OF OUR REGION, AND FULLY INTEGRATE BEHAVIORAL HEALTH INTO OUR HEALTH SYSTEM. WE EXPECT TO REACH 2,300 UNDUPLICATED INDIVIDUALS IN YEAR ONE AND 2,500 IN YEAR TWO FOR A TOTAL OF 4,800 THROUGHOUT THE ENTIRE GRANT PERIOD. THROUGH THIS PROJECT WE WILL MEET THE FOLLOWING OBJECTIVES: INCREASE GROWTH AND STABILITY OF OUR MOBILE CRISIS TEAM; COMPLETE AN AMBULATORY WITHDRAWAL MANAGEMENT UNIT; DEVELOP A PERSON/FAMILY CENTERED CARE COORDINATION TEAM; ESTABLISH A MANDATORY TREATMENT/DISCHARGE PLANNING PROCESS THROUGHOUT OUR CONTINUUM OF CARE; TRANSITION ALL CCBHC SERVICES LINES TO OUR HEALTH SYSTEM’S EHR, EPIC; AND REDUCE THE NUMBER OF PATIENTS REFERRED TO INPATIENT PSYCHIATRIC HOSPITALS. THE TARGET POPULATION RESIDES IN THE COUNTIES OF BELTRAMI, CLEARWATER, HUBBARD AND LAKE OF THE WOODS IN NORTH CENTRAL MINNESOTA. BELTRAMI COUNTY HAS A SIGNIFICANT AMERICAN INDIAN POPULATION, LOCATED IN CENTRAL VICINITY TO THREE INDIAN RESERVATIONS: RED LAKE, WHITE EARTH AND LEECH LAKE WHICH ACCOUNTS FOR NEARLY 22% OF AMERICAN INDIANS RESIDING IN THE COUNTY, COMPARED TO THE NATIONAL AVERAGE OF 1.3%. BELTRAMI COUNTY HAS THE HIGHEST OUT OF HOME PLACEMENT RATE AMONG ALL MN COUNTIES, WITH 11% OF ALL YOUTH IN THE COUNTY PLACED OUT OF THE HOME IN 2017. IN 2018, THERE WERE 884 ADMISSION IN BELTRAMI COUNTY WITH THE HIGHEST POPULATION OF THOSE ADMISSION BEING AMERICAN INDIAN (70%) AND SECOND BEING WHITE (25.5%). THE NEEDS ASSESSMENT ALSO INDICATES THE NUMBER ONE CONDITION SURROUNDING ADMISSION TO TREATMENT IS A CONDITION OF PROBATION/PAROLE AT 35.1% BELTRAMI COUNTY, AND THE SURROUNDING AREA, IS FACING THE CHALLENGE OF CREATING A FINANCIALLY STABLE CONTINUUM OF MENTAL HEALTH SERVICES AT A TIME WHEN COSTLY RESIDENTIAL INTERVENTIONS ARE INCREASING. THROUGH FULLY INTEGRATING BEHAVIORAL HEALTH INTO OUR LARGER HEALTH SYSTEM, USING THE CCBHC MODEL, WE WILL ALLOW FOR THE SEAMLESS PROVISION OF HEALTHCARE AND SERVE THE WHOLE PERSON. INTEGRATING SERVICE PROGRAMS AND QUALITY CARE COORDINATION AS WELL AS IMPROVING ACCESS TO ALL CCBHC REQUIRED SERVICES WILL ALLOW FOR IMPROVED ABILITY TO PROVIDE LOCAL SERVICE SUPPORTS AND FAMILY SUPPORT AND TO HELP PATIENTS MOVE TO EACH LEVEL OF CARE WITH THE SUPPORT THEY NEED. SERVICES THAT WILL BE PROVIDED THROUGH THIS GRANT INCLUDE: CRISIS MENTAL HEALTH SERVICES, INCLUDING 24-HOUR MOBILE CRISIS TEAMS, EMERGENCY CRISIS INTERVENTION SERVICES, AND CRISIS STABILIZATION; SCREENING, ASSESSMENT, AND DIAGNOSIS, INCLUDING RISK ASSESSMENT; PATIENT-CENTERED TREATMENT PLANNING; COMPREHENSIVE OUTPATIENT MENTAL HEALTH AND SUBSTANCE USE SERVICES; CASE MANAGEMENT; PSYCHIATRIC REHABILITATION SERVICES; SOCIAL SUPPORT OPPORTUNITIES; AND CLINICAL MONITORING.

DEFINING CHROMOSTASIS - A CANDIDATE REGULATOR OF HEALTHY AGING AND LONGEVITY

NEUROTENSIN 1 ALLOSTERIC MODULATOR FOR THE TREATMENT OF PAIN - PROJECT SUMMARY THE NIH HEAL INITIATIVE AIMS TO TACKLE TWO ONGOING HEALTH CARE CRISES: OPIOID USE DISORDER AND UNCONTROLLED PAIN. PHARMACOLOGICAL APPROACHES TARGETING BOTH CRISES ARE URGENTLY NEEDED. THE NEUROTENSIN RECEPTOR 1 (NTR1) HAS BEEN A SOUGHT-AFTER TARGET FOR THE TREATMENT OF BOTH PAIN AND ADDICTION, BUT DEVELOPMENT OF BALANCED NTR1 AGONISTS IS PRECLUDED BY THEIR SEVERE SIDE EFFECTS. AS A G PROTEIN-COUPLED RECEPTOR (GPCR) NTR1 SIGNALS THROUGH THE ACTIVATION OF G PROTEINS AND Β-ARRESTINS (E.G., ARRB2). ARRB2 ATTENUATES DRUG REWARD AND SUPPRESSES PAIN VIA REGULATION OF BOTH GPCRS AND NON-GPCRS, INCLUDING THE NMDA RECEPTOR. WE HAVE DEMONSTRATED IN RODENTS THAT SBI-553, A NOVEL ARRB2-BIASED ALLOSTERIC MODULATOR (BAM) OF NTR1, ATTENUATES OPIOID REWARD AND THE REINFORCING EFFECTS OF PSYCHOSTIMULANTS WITHOUT THE SIDE EFFECTS CHARACTERISTIC OF BALANCED NTR1 SIGNALING. CRYO- EM STUDIES DEMONSTRATE THAT SBI-553 BINDS AT AN INTRACELLULAR ALLOSTERIC SITE AND ACTS LIKE A MOLECULAR GLUE, DIRECTING SIGNALING TO ARRB2 IN THE PRESENCE OR ABSENCE OF AGONISTS. RECENTLY WE HAVE SHOWN THAT NTR1 BAMS PRODUCE POTENT ANTINOCICEPTION IN RODENT MODELS, A FINDING THAT BUILDS OFF PREVIOUS REPORTS THAT NEUROTENSIN, NTR1’S ENDOGENOUS LIGAND PROVIDED MORE POTENT PAIN RELIEF THAN MORPHINE THROUGH A MECHANISM THAT IS INDEPENDENT OF OPIOID RECEPTORS. NTR1 BAMS RAISE NOCICEPTIVE THRESHOLDS IN WILD-TYPE MICE, BUT NOT IN NTSR1−/−OR ARRB2−/− (KNOCKOUT) MICE, CONFIRMING THE MECHANISTIC REQUIREMENT FOR NTR1 AND ARRB2. LOCAL OR SYSTEMIC DELIVERY OF NTR1 BAMS REDUCED HYPERSENSITIVITY IN MOUSE MODELS OF POSTOPERATIVE AND NEUROPATHIC PAIN AND SUPPRESSED EXCITATORY SYNAPTIC TRANSMISSION AND NMDAR/ERK SIGNALING IN SPINAL CORD NOCICEPTIVE NEURONS. ADDITIONALLY, NTR1 BAMS SUPPRESSED C-FIBER-INDUCED RESPONSES IN VIVO AND ACTION POTENTIAL FIRING IN MOUSE AND HUMAN NOCICEPTIVE SENSORY NEURONS IN VITRO. COLLECTIVELY, OUR FINDINGS INDICATE THAT NTR1 BAMS ARE AN EXCITING AND NOVEL APPROACH TO DEVELOP A FIRST-IN-CLASS NON-OPIOID DRUG FOR THE TREATMENT OF PAIN. IN THIS APPLICATION WE OUTLINE OUR PLAN TO OPTIMIZE AND DEVELOP SUCH A DRUG FOR POSTOPERATIVE PAIN. IN THE UG3 STAGE, WE WILL LEVERAGE THE STRUCTURE ACTIVITY RELATIONSHIPS DEVELOPED AROUND SBI-553 AND, THROUGH A STRUCTURE-BASED DESIGN LEAD OPTIMIZATION PROCESS, REFINE AND IMPROVE THE PROPERTIES OF THE LEADS. CO-STRUCTURES OF LEADS WITH NTR1 WILL BE DETERMINED THROUGH CRYOEM, SUCH THAT THE ITERATIVE MEDICINAL CHEMISTRY, DESIGN-MAKE-TEST CYCLE IS INFORMED BY COMPUTATIONAL PREDICTIONS USING BOTH ARTIFICIAL INTELLIGENCE AND STRUCTURE-BASED DRUG DESIGN. IN THE UH3 PHASE THE OBJECTIVE IS TO DERIVE ADVANCED LEADS WITH EXCELLENT IN VIVO POTENCY IN ACUTE INFLAMMATORY PAIN, SURGICAL PAIN, AND CHRONIC NEUROPATHIC PAIN MODELS, OUTSTANDING DRUG-LIKE PROPERTIES, AND NO LIMITING CV OR CNS SAFETY RISK. IN THE UH3 PHASE, THE SELECTED CLINICAL CANDIDATE WILL BE SUBJECTED TO A STANDARD BATTERY OF REQUIRED IND-ENABLING STUDIES, LEADING TO AN INVESTIGATIONAL NEW DRUG APPLICATION (IND). UPON ACCEPTANCE OF THE IND, WE WILL CONDUCT A PHASE I, SINGLE ASCENDING DOSE STUDY IN HEALTHY VOLUNTEERS TO ASSESS THE SAFETY, TOLERABILITY AND PK OF OUR CLINICAL CANDIDATE.

SURVIVAL MECHANISMS FOR APOPTOTIC CASPASE

TARGETED NANOPARTICLES FOR BREAST CANCER DIAGNOSIS AND THERAPY

HVEM-BTLA SYSTEM IN INFLAMMATION

ANTI-HERPESVIRUS SIGNALING BY LTALPHABETA/LIGHT CYTOKINES

IMPROVE EXISTING AIRPORT

COMPARATIVE RESISTOMICS OF GRAM-NEGATIVE BACTERIAL PATHOGENS - ABSTRACT INCREASING ANTIBIOTIC RESISTANCE NECESSITATES EXPANDING RESEARCH INTO THE MECHANISMS BY WHICH BACTERIAL PATHOGENS ACQUIRE AND PERPETUATE DRUG RESISTANCE. DESPITE RAPIDLY EXPANDING GENOMIC MAPPING OF RESISTANCE-CONFERRING MUTATIONS IN CLINICAL ISOLATES AND LABORATORY STUDIES, OUR KNOWLEDGE OF DYNAMICS AND MECHANISMS UNDERLYING EVOLUTION OF ANTIMICROBIAL RESISTANCE IS STILL INSUFFICIENT. TO FILL-IN THIS GAP, THE AUTHORS OF THIS PROPOSAL COMBINE EXPERIMENTAL EVOLUTION IN A CONTINUOUS CULTURING DEVICE, MORBIDOSTAT, WITH TIME- RESOLVED ULTRADEEP GENOMIC SEQUENCING OF EVOLVING BACTERIAL CULTURES. THE UTILITY OF THE DEVELOPED MORBIDOSTAT-BASED WORKFLOW IS SUPPORTED BY PUBLISHED AND ONGOING STUDIES WITH ESTABLISHED ANTIMICROBIALS AND EXPERIMENTAL DRUG CANDIDATES. THE PRELIMINARY RESULTS OF COMPARATIVE RESISTOMICS STUDIES OVER A RANGE OF GRAM-NEGATIVE BACTERIAL SPECIES PROVIDED INITIAL SUPPORT TO A PREMISE THAT EVOLUTION OF DRUG RESISTANCE IN MORBIDOSTAT PROCEEDS VIA A LIMITED SET OF TRAJECTORIES DEFINED BY A COMBINATION OF RESISTANCE AND FITNESS CONSTRAINS APPROXIMATING CLINICAL EVOLUTION, WHICH FAVORS SELECTION OF LOW-FREQUENCY/HIGH-FITNESS OVER HIGH- FREQUENCY/LOW-FITNESS MUTANTS. A COMPARATIVE RESISTOMICS APPROACH ENABLES MAPPING OF BOTH UNIVERSAL AND STRAIN-SPECIFIC MECHANISMS AS DEMONSTRATED IN A RECENT PROOF-OF-CONCEPT STUDY ON EXPERIMENTAL EVOLUTION OF CIPROFLOXACIN RESISTANCE IN THREE GRAM-NEGATIVE BACTERIA. THE PROPOSED 5-YEAR PROJECT WILL TEST THE CENTRAL HYPOTHESIS AND EXTEND EXPLORATION OF ANTIMICROBIAL RESISTOME BY PURSUING THE FOLLOWING SPECIFIC AIMS: (I) IN AIM 1, THE ESTABLISHED MORBIDOSTAT-BASED WORKFLOW WILL BE USED TO DETERMINE MAJOR MECHANISMS DRIVING RESISTANCE TO BROAD-SPECTRUM CLINICAL ANTIBIOTICS, CIPROFLOXACIN, COLISTIN, TIGECYCLINE AND MEROPENEM, IN FOUR DIFFICULT-TO-TREAT GRAM-NEGATIVE BACTERIAL PATHOGENS, ACINETOBACTER BAUMANNII ATCC17978, P. AERUGINOSA ATCC27853, E. COLI ATCC25922, AND K. PNEUMONIAE ATCC13883; (II) IN AIM 2, A REPRESENTATIVE PANEL OF SELECTED CLONES WILL BE SYSTEMATICALLY CHARACTERIZED TO ASSESS THE EFFECTS OF INDIVIDUAL MUTATIONS AND COMBINATIONS THEREOF ON ACQUIRED RESISTANCE AND FITNESS; (III) AIM 3 WILL LEVERAGE A MORIBIDOSTAT-BASED WORKFLOW TO MAKE FIRST STEPS TOWARD EXPERIMENTAL EVOLUTION OF MULTIDRUG RESISTANCE FOCUSING ON A. BAUMANNII AND STARTING FROM CLONES SELECTED IN SINGLE-DRUG EVOLUTION STUDIES. THE RESULTS THAT WILL BE OBTAINED IN ALL PLANNED STUDIES WILL BE A SUBJECT OF IN-DEPTH BIOINFORMATICS ANALYSIS (INCLUDING COMPARISON WITH PUBLIC DATA FOR CLINICAL ISOLATES), PREDICTIVE MODELING, INTEGRATION AND SHARING WITH BROAD RESEARCH COMMUNITY VIA A SPECIALIZED WEB-SITE ON INTEGRATIVE GENOMICS OF EVOLUTION OF ANTIMICROBIAL RESISTANCE (IGEAR). THE PROPOSED STUDY IS EXPECTED TO HAVE TRANSLATIONAL IMPACTS IN ADVANCING METHODOLOGY TO SUPPORT RATIONAL OPTIMIZATION OF ANTIBIOTIC TREATMENT REGIMENS AND DEVELOPMENT OF NEW DRUGS WITH MINIMIZED RESISTIBILITY.

SPECIAL DIABETES PROGRAM FOR INDIANS

MOLECULAR MECHANISM OF BCL2-DEPENDENT APOPTOSIS

POST-SURGICAL PREDICTORS OF DEPRESSION AND WEIGHT REGAIN AFTER BARIATRIC SURGERY

DEVELOPMENT OF SBI-553, AN ALLOSTERIC MODULATOR OF NTR1, FOR THE TREATMENT OF SUBSTANCE USE DISORDERS

RURAL MATERNITY AND OBSTETRICS MANAGEMENT STRATEGIES PROGRAM

PRECLINICAL DISCOVERY OF NOVEL FARNESYLTRANSFERASE INHIBITORS FOR THE TREATMENT OF ALZHEIMER'S DISEASE AND RELATED TAUOPATHIES - PROJECT SUMMARY ALZHEIMER’S DISEASE (AD) IS A FATAL DISEASE THAT CURRENTLY AFFLICTS ALMOST SIX MILLION AMERICANS. WITH AN AGING POPULATION, WE RISK A PUBLIC HEALTH CRISIS BY 2050, UNLESS EFFECTIVE TREATMENTS ARE IDENTIFIED. DESPITE EXTENSIVE RESEARCH, THERE ARE CURRENTLY NO DRUGS THAT SLOW OR ALTER THE COURSE OF DISEASE. AD IS DEFINED BY THE PRESENCE OF SS-AMYLOID (ASS) PLAQUES AND INTRANEURONAL TAU INCLUSIONS CALLED NEUROFIBRILLARY TANGLES (NFTS) IN THE BRAIN. DRUG CANDIDATES THAT REDUCE ASS PLAQUES HAVE NOT, YET, BEEN SHOWN TO HAVE CLINICAL BENEFIT, AND GROWING DATA SUGGESTS THAT IT MAY BE MORE IMPORTANT TO TARGET NFTS OVER ASS PLAQUES TO PREVENT COGNITIVE DECLINE. RECENTLY, THE MACROAUTOPHAGY-LYSOSOMAL PATHWAY OF PROTEIN DEGRADATION HAS EMERGED AS A COMPELLING TARGET FOR REDUCING PATHOGENIC TAU IN THE BRAIN. OUR HYPOTHESIS IS THAT INCREASING THE RATE OF TAU DEGRADATION WILL REDUCE TAU LEVELS AND STOP, OR GREATLY SLOW, THE RATE OF TAU AGGREGATION. WE RECENTLY DISCOVERED A NOVEL PATHWAY TO ACCOMPLISH THIS OBJECTIVE. INHIBITING THE FARNESYLATION OF RHES, A GTPASE PROTEIN IN THE RAS FAMILY, ACTIVATES THE LYSOSOME AND RESULTS IN THE SELECTIVE DEGRADATION OF PATHOLOGICAL TAU. CONFIRMATION OF THE THERAPEUTIC HYPOTHESIS WAS ACHIEVED BY ADMINISTERING A FARNESYLTRANSFERASE INHIBITOR (FTI) IN A MOUSE MODEL OF TAUOPATHY, WHICH REDUCED TAU PATHOLOGY AND ATTENUATED BEHAVIORAL ABNORMALITIES IN THE MICE. KNOWN FTIS ARE NOT SUITABLE FOR HUMAN DEVELOPMENT AS CNS DRUGS. OPTIMIZED FOR CANCER INDICATIONS, THEY ARE EFFICIENTLY PUMPED OUT OF THE BRAIN BY EFFLUX PROTEINS. WE PROPOSE A THREE-PRONGED APPROACH TO IDENTIFY CHEMICAL MATTER THAT CAN REACH PHARMACOLOGICALLY SIGNIFICANT AND DOSE-PROPORTIONAL BRAIN LEVELS. FOR TWO OF THE KNOWN INHIBITORS, L-778,123 AND LONAFARNIB, WE WILL MAKE STRATEGIC CHANGES TO THE STRUCTURES, ELIMINATING FUNCTIONAL GROUPS THAT SERVE AS RECOGNITION SUBSTRATES FOR THE EFFLUX PUMPS. CONCURRENTLY, WE WILL INITIATE A HIGH- THROUGHPUT SCREEN OF A CHEMICAL LIBRARY WITH CHEMICAL PROPERTIES CONSISTENT WITH KNOWN CNS DRUGS. AS A THIRD STEP, WE WILL ENGAGE IN A MULTI-MILLION COMPOUND ARTIFICIAL INTELLIGENCE-BASED VIRTUAL SCREEN WITH ATOMWISE TO IDENTIFY NOVEL FTIS. BY GENERATING X-RAY CO-CRYSTAL STRUCTURES OF THE MOST PROMISING HITS AND USING COMPUTER- AIDED DRUG DESIGN, WE PLAN TO ACCELERATE THE PROCESS OF HIT VALIDATION, LEAD DISCOVERY, AND OPTIMIZATION TO IDENTIFY SMALL MOLECULE DRUG CANDIDATES. WE WILL ADVANCE INHIBITORS TO AN IN VIVO PHARMACODYNAMIC MODEL AND SELECT COMPOUNDS WITH LINEAR PHARMACOKINETIC/PHARMACODYNAMIC (PK/PD) RELATIONSHIPS THAT CAN BE ADVANCED INTO THE CLINIC. THREE OF THE TOP COMPOUNDS WILL BE TESTED FOR EFFICACY IN A TAUOPATHY ANIMAL MODEL USING DOSES DERIVED FROM THE PK/PD RELATIONSHIP. SHORT-TERM STUDIES WILL IDENTIFY COMPOUNDS THAT REDUCE ALL PATHOGENIC TAU SPECIES. THE MOST EFFICACIOUS COMPOUND WILL BE MOVED INTO LONG-TERM DOSING STUDIES TO EVALUATE LIFE-SPAN EXTENSION AND REDUCTION IN NFT FORMATION.

SIGNAL TRANSDUCTION AND CELL DEATH REGULATION

CYCLIC EPHA4 PEPTIDE ANTAGONISTS FOR NEUROPROTECTION IN ALS

ER STRESS AND UPR IN NON-ALCOHOLIC STEATOHEPATITIS AND HEPATOCELLULAR CARCINOMA

NOVEL ROLES FOR THE ALZHEIMER'S DISEASE (AD) RISK GENE, SORLA IN NEUROPROTECTION IN AD

CONTROL OF MUSCLE GENE EXPRESSION BY SIGNALING PATHWAYS

TARGETING NEUROINFLAMMATION IN AD WITH NOVEL CX3CR1 AGONISTS

LIGAND FUNCTIONAL SELECTIVITY IN EPHA2 RECEPTOR SIGNALING

DEVELOPMENT OF SMAC MIMETICS AS LATENCY-REVERSING AGENTS

BRAIN PATHOLOGY AND FUNCTION IN A CHRONIC MOUSE MODEL OF ZIKV TRANSMISSION

CHARACTERIZING THE NUCLEAR PORE COMPLEX-T CELL RECEPTOR CONNECTION

MERITCARE HOSPITAL COMMUNITY CLINICAL ONCOLOGY PROGRAM

PLEKHA7 AND BETA-CATENIN INTERACT TO REGULATE MUTANT KRAS

ROLE OF FBXW7-MEDIATED PROTEASOMAL DEGRADATION IN MYOFIBERS IN DETERMINING MUSCLE STEM CELL POOL SIZE

MECHANISMS OF INITATION OF SKELETAL MINERALIZATION

SOMATIC GENE RECOMBINATION OF MAPT IN SINGLE CELLS FROM FRONTOTEMPORAL LOBAR DEMENTIA (FTLD) BRAINS WITH TAU PATHOLOGY

PURPOSE: RECONSTRUCT AIRCRAFT RESCUE AND FIRE FIGHTING VEHICLE; IMPLEMENT WILDLIFE MITIGATION MEASURES. ACTIVITIES TO BE PERFORMED/EXPECTED OUTCOMES: THIS PROJECT IMPLEMENTS WILDLIFE HAZARD ATTRACTANT IMPROVEMENTS UNDER THE AIRPORT'S WILDLIFE HAZARD MITIGATION PLAN TO ENHANCE SAFETY. . THIS PROJECT REPLACES 1 EXISTING 1,500 GALLON AIRCRAFT RESCUE AND FIRE FIGHTING VEHICLE TO MEET 14 CFR PART 139 SAFETY REQUIREMENTS THAT HAS REACHED THE END OF ITS USEFUL LIFE. THIS GRANT FUNDS PHASE 2, WHICH CONSISTS OF CONSTRUCTION. INTENDED BENEFICIARY: THIS GRANT WILL PROVIDE FEDERAL FUNDING FOR AIRPORTS ASSOCIATED WITH SANFORD, FLORIDA.

MECHANISMS THAT COUPLE IRREGULAR DEVELOPMENT OF FETAL MELANOBLASTS TO PREMATURE EXHAUSTION OF ADULT MELANOCYTE STEM CELLS - PROJECT SUMMARY MAINTENANCE OF TISSUE FUNCTION DURING ADULTHOOD, AND HENCE SUPPRESSION OF TISSUE DEGENERATION AND DISEASE, DEPENDS ON MAINTENANCE OF STEM CELL POPULATIONS. ADULT STEM CELLS ARE EPIGENETICALLY PROGRAMMED AND A CULMINATION OF A SERIES OF DEVELOPMENTAL DECISIONS INITIATED IN THE EMBRYO. IN UTERO ENVIRONMENTAL EXPOSURES ON THE EMBRYO CAN INFLUENCE ADULT AND LATE-LIFE DISEASE, LIKELY IN PART VIA EFFECTS ON STEM CELL DEVELOPMENT THAT ARE TRANSMITTED TO MAINTENANCE AND FUNCTION OF STEM CELLS IN THE ADULT. HOWEVER, THE MOLECULAR LINKS BETWEEN EMBRYONIC DEVELOPMENT AND LONG-TERM MAINTENANCE OF STEM CELL FUNCTION AND PHENOTYPE IN ADULTS ARE POORLY DEFINED. WE WILL EMPLOY LINEAGE-SPECIFIC GENETIC INACTIVATION OF A HISTONE CHAPERONE TO UNDERSTAND HOW EMBRYONIC DEVELOPMENTAL INTEGRITY OF MELANOBLASTS (MB) IMPACTS ON MAINTENANCE ADULT MELANOCYTE (MC) STEM CELLS (MCSC). THE HISTONE CHAPERONE HIRA DEPOSITS HISTONE VARIANT H3.3 INTO ACTIVE GENES, PROMOTERS AND ENHANCERS. THROUGH IN VITRO AND IN VIVO STUDIES AND SINGLE CELL RNA-SEQ OF MOUSE EMBRYO MELANOBLASTS (MBS) FROM WILD TYPE MICE AND MICE LACKING EXPRESSION OF HIRA IN EMBRYONIC MBS, WE HAVE UNCOVERED A ROLE FOR HIRA IN SUSTAINING THE PAX3/SOX10-MITF MB SPECIFICATION PATHWAY. INACTIVATION OF HIRA IN MBS DEPLETES THE NUMBER OF MBS IN EARLY/MID STAGE EMBRYOS. HOWEVER, THIS EMBRYONIC DEFECT IS RESCUED BY BIRTH AND YOUNG MICE EXHIBIT NORMAL NUMBERS OF MELANOCYTIC CELLS, AND ONLY A VERY SUBTLE PIGMENTATION DEFECT. NEVERTHELESS, IN NEW-BORN MICE, HIRA KNOCK OUT (KO) MELANOCYTIC CELLS EXHIBIT A HIGHER FREQUENCY OF TELOMERE-ASSOCIATED DNA DAMAGE FOCI, INDICATING THAT HIRA KNOCK OUT MCSC AND/OR MELANOCYTES HARBOR MOLECULAR DAMAGE, EVEN IN NEW-BORN MICE. INDEED, MELANOBLASTS AND MELANOCYTES FROM NEW-BORN HIRA KO MICE RESPOND POORLY TO PRO-PROLIFERATIVE CHALLENGE IN VITRO AND IN VIVO, AND THESE MICE SHOW MARKED ACCELERATED MCSC AND MELANOCYTE DEPLETION AND DRAMATICALLY ACCELERATED HAIR GREYING DURING ADULTHOOD. BUILDING ON THESE EXTENSIVE PRELIMINARY DATA, WE WILL INVESTIGATE THE ROLE OF HIRA IN DIFFERENTIATION AND DEVELOPMENT OF THE MELANOCYTIC LINEAGE, AND INVESTIGATE THE LINKS BETWEEN ABNORMAL EMBRYONIC DEVELOPMENT AND ADULT STEM CELL DEPLETION DURING ADULTHOOD AND AGING. DYSREGULATION OF THE PAX3/SOX10-MITF SIGNALING PATHWAY CONTRIBUTES TO DEVELOPMENTAL DISORDERS AND MELANOMA. THESE STUDIES TO DEFINE HIRA'S ROLE IN THE PAX3/SOX10-MITF AXIS CAN PROMOTE THERAPEUTIC INTERVENTIONS TO COMBAT THESE DEVELOPMENTAL AND NEOPLASTIC DISORDERS. MOREOVER, COMPLETION OF THESE SPECIFIC AIMS WILL ADDRESS HOW THE INTEGRITY OF EMBRYONIC DEVELOPMENT OF TISSUE SPECIFIC STEM CELLS IMPACTS MAINTENANCE OF THOSE STEM CELLS DURING ADULTHOOD.

REGULATION OF ER STRESS-INDUCED CELL DEATH

IMPROVING PROINSULIN FOLDING TO AMELIORATE TYPE II DIABETES - PROJECT SUMMARY DIABETES IS AMONG THE FASTEST GROWING HEALTH CHALLENGES OF THE 21ST CENTURY, AFFECTING >30 MILLION PEOPLE, WITH =80,000 DEATHS ANNUALLY, AND INVOLVING ~15% OF U.S. NATIONAL HEALTH EXPENDITURES. TYPE 2 DIABETES (T2D) IS THE MOST COMMON FORM OF DIABETES, WHICH IS LINKED TO AN INSUFFICIENT AMOUNT OF CIRCULATING INSULIN BECAUSE OF THE BODY’S INSENSITIVITY TO THE HORMONE. MAINTENANCE OF THE INSULIN STORAGE POOL REQUIRES SYNTHESIS OF ~6000 PROINSULIN (PI) MOLECULES/SS-CELL/SECOND, EACH DELIVERED TO THE ENDOPLASMIC RETICULUM (ER) FOR FOLDING. EVEN MORE MOLECULES ARE NEEDED IN STATES OF INSULIN RESISTANCE. SIGNIFICANTLY, WE DISCOVERED THAT PI ENTERS ABERRANT DISULFIDE-LINKED INTERMOLECULAR COMPLEXES, EVEN IN HEALTHY (HUMAN AND MURINE) ISLETS. UNDER CONDITIONS THAT DEMAND INCREASED INSULIN PRODUCTION (EVEN PREDIABETES), THESE COMPLEXES DRAMATICALLY INCREASE, THUS LIMITING INSULIN PRODUCTION. WE SHOW NEW KEY EVIDENCE THAT THESE ABERRANTLY FOLDED PI COMPLEXES CAN BE RESOLVED TO MONOMERIC PI WITHIN THE ER. WE RECENTLY ELUCIDATED THE FIRST MAP OF THE HUMAN PI INTERACTOME IDENTIFYING PI FOLDING MODIFIERS. THE MOST SIGNIFICANT PI INTERACTOR IN HUMAN ISLETS IS THE ER CHAPERONE BIP AND WE PRESENT NEW EVIDENCE (BOTH GAIN OF FUNCTION, AND LOSS OF FUNCTION) THAT THIS INTERACTION, SUPPORTED BY BIP CO- CHAPERONES, IS ABSOLUTELY REQUIRED FOR PRODUCTIVE PROINSULIN FOLDING (AND LIMITING MISFOLDING), LEADING TO SUCCESSFUL ANTEROGRADE TRANSPORT. FOR OUR STUDIES WE GENERATED A NOVEL BIP-TAGGED MOUSE THAT CAN FOR THE FIRST TIME IDENTIFY FUNDAMENTAL STEPS IN PI FOLDING ESSENTIAL FOR INSULIN PRODUCTION. MOREOVER, WE SHOW THAT INCREASED EXPRESSION OF BIP AND ITS CO-CHAPERONE P58IPK DRAMATICALLY REDUCES ACCUMULATION OF THE HIGH MOLECULAR WEIGHT PI COMPLEXES. THUS, OUR DISCOVERIES OPEN THE POSSIBILITY THAT PHARMACOLOGIC INTERVENTION MAY IMPROVE CHAPERONE-DEPENDENT PI FOLDING, AND THIS MAY ATTENUATE T2D. AS WE BEGIN TO ELUCIDATE THE HUMAN PI FOLDING PATHWAY, WE ARE DEVELOPING PARALLEL ANIMAL MODELS TO DETERMINE HOW PI FOLDS/MISFOLDS. HERE WE PROPOSE TO: 1) MECHANISTICALLY DISSECT HOW BIP AND ADDITIONAL PI INTERACTORS IN THE ER ORCHESTRATE SUCCESSFUL PI FOLDING AND DETERMINE WHICH STEP(S) OF PI FOLDING GO AWRY IN T2D; 2) IDENTIFY HOW THE PI INTERACTOME CHANGES IN HUMAN T2D; DETERMINE THE FUNCTION OF ALTERED PI INTERACTIONS IN ISLETS FROM PATIENTS WITH T2D; AND UTILIZE NOVEL ASSAYS TO MEASURE PRODUCTIVE PI FOLDING/TRAFFICKING IN SS-CELLS. WE WILL INTEGRATE PHYSIOLOGIC STUDIES OF HUMAN ISLETS WITH NOVEL GENETIC AND BIOCHEMICAL APPROACHES TO GENERATE A COMPREHENSIVE UNDERSTANDING OF HOW PI FOLDING HOMEOSTASIS IMPACTS SS-CELL FUNCTION IN HEALTH AND DISEASE. WE BELIEVE THAT THIS HYPOTHESIS IS A HIGH-IMPACT IDEA ESSENTIAL TO THE MISSION OF THE NIDDK, AND WE NOW BRING TOOLS, ASSAYS, AND APPROACHES THAT ARE NOT CURRENTLY AVAILABLE ANYWHERE ELSE.

NG2-PG IN TUMOR VASCULARIZATION AND PROGRESSION

OVERRIDING THE IMMUNE EVASION TACTICS OF CORONAVIRUS

DISCOVERY OF SELECTIVE INHIBITORS FOR THE EPHA4 KINASE

VIRULENCE FACTORS AND CELL DEATH

ECONOMIC DEVELOPMENT INITIATIVE, COMMUNITY PROJECT FUNDING, AND MISCELLANEOUS GRANTS

LINKING THE MICROBIOME AND IMMUNE-CHECKPOINT IN MELANOMA BY RNF5

SAFE SCHOOLS/HEALTHY STUDENTS GRANTS

LEAD APPLICANT ENTITY ORGANIZATION TYPE: NONPROFIT HEALTHCARE ORGANIZATIONLEAD APPLICANT LOCATION: SIOUX FALLS, SOUTH DAKOTAREQUIRED PARTNERSEMPLOYER PARTNERS: ALL EMPLOYER PARTNERS FALL UNDER THE PARENT COMPANY OFSANFORD HEALTH BUT HAVE SEPARATE EINS.SANFORD MEDICAL CENTERSANFORD MEDICAL CENTER FARGOSANFORD BISMARCK MEDICAL CENTERSANFORD BEMIDJI MEDICAL CENTERSANFORD HEALTH NETWORKSANFORD HEALTH NETWORK NORTHTHE EVANGELICAL LUTHERAN GOOD SAMARITAN SOCIETYEDUCATION OR TRAINING PARTNERS:NORTH DAKOTA STATE UNIVERSITYSOUTH DAKOTA STATE UNIVERSITYWORKFORCE DEVELOPMENT ENTITIES:NORTH DAKOTA WORKFORCE DEVELOPMENT DIVISIONWORKER ORGANIZATION LABOR UNIONS:SOUTH DAKOTA ASSOCIATION OF HEALTHCARE ORGANIZATIONSNORTH DAKOTA NURSES ASSOCIATIONUFCW LOCALOPTIONAL PARTNERS:SANFORD HEALTH MILITARY AND VETERANS AFFAIRSGEOGRAPHIC SCOPE: NATIONAL PROJECT SERVING NORTH DAKOTA, SOUTH DAKOTA, MINNESOTA, AND IOWATOTAL FEDERAL FUNDING REQUESTED: 2,954,400TOTAL MATCH PROPOSED: 679,947PROJECT TITLE: STUDENT NURSE TO NURSE PRECEPTOR CAREER PATHWAYPROPOSED NUMBER OF PARTICIPANTS WHO START TRAINING: 825SUMMARY OF PROJECT PURPOSE AND EMPLOYMENT AND TRAINING ACTIVITIES OFFERED: SANFORD HEALTH IS APPLYING FOR FUNDING TO SUPPORT THE DEVELOPMENT OF THE HEALTHCARE WORKFORCE IN ND, SD, IA AND MN SPECIFICALLY, THIS PROJECT WILL INCREASE THE NUMBER OF WELL-PREPARED NURSES IN THE REGISTERED NURSE OCCUPATION. IN THE PROPOSED CAREER PATHWAY, INTERNS AND ACCELERATED BSN STUDENTS WILL RECEIVE THEIR RN DEGREE, SUCCESSFULLY TRANSITION TO PRACTICE, AND BECOME CERTIFIED NURSE PRECEPTORS THROUGH OUR ADVANCED PRECEPTOR CURRICULUM. THIS PATHWAY WILL IMPROVE READINESS FOR PRACTICE, CLINICAL JUDGMENT, AND CRITICAL THINKING SKILLS. THE PROPOSED TRAINING ACTIVITIES INCLUDE PAID INTERNSHIPS, PRECEPTORSHIPS, AN ACCELERATED BSN DEGREE PROGRAM, AND PRECEPTOR CERTIFICATION.TARGET POPULATION(S) TO BE SERVED: BSN STUDENTS, NEW NURSES, AND NURSE PRECEPTOR CANDIDATES IN ND, SD, MN, AND IA. PRIORITY POPULATIONS INCLUDE INDIVIDUALS IN RURAL OR FRONTIER COUNTIES, AMERICAN INDIAN ALASKAN NATIVE INDIVIDUALS, AND VETERANS.TRAINING TRACK: NURSING CAREER PATHWAYSTARGETED OCCUPATIONS: REGISTERED NURSERECOGNIZED POSTSECONDARY CREDENTIALS OFFERED: BSNSUBRECIPIENT ROLES: NORTH DAKOTA STATE UNIVERSITY (NSDU) IS THE ONLY SUBRECIPIENT ON THIS APPLICATION. NDSU WILLWORK WITH SANFORD HEALTH TO JUMP START AN ACCELERATED BSN DEGREE PROGRAM IN ALIGNMENT WITH GOAL 1 OF THE PROPOSED WORK PLAN.

FRAGMENT-BASED DISCOVERY OF STEP MODULATORS IN ALZHEIMER'S DISEASE ADMINISTRATIVE SUPPLEMENT - PROJECT SUMMARY STRIATAL-ENRICHED TYROSINE PHOSPHATASE (STEP) IS A NEURON-SPECIFIC PROTEIN TYROSINE PHOSPHATASE (PTP) AND A NOVEL THERAPEUTIC TARGET FOR ALZHEIMER’S DISEASE (AD), A DEBILITATING NEURODEGENERATIVE DISORDER FOR WHICH NO CURE CURRENTLY EXISTS. MULTIPLE STUDIES INDICATE THAT LEVELS OF STEP ARE ELEVATED IN AD AND OTHER NEURODEGENERATIVE AND NEUROPSYCHIATRIC DISORDERS, INCLUDING PARKINSON’S DISEASE, SCHIZOPHRENIA, AND FRAGILE X SYNDROME. THE DATA SUGGEST THAT INCREASED STEP ACTIVITY INTERFERES WITH SYNAPTIC FUNCTION AND CONTRIBUTES TO THE CHARACTERISTIC COGNITIVE AND BEHAVIORAL DEFICITS IN THESE DEVASTATING DISEASES. CROSSING STEP KO MICE WITH MOUSE MODELS OF AD, SCHIZOPHRENIA, OR FRAGILE X SYNDROME COMPLETELY REVERSED THE COGNITIVE AND BEHAVIORAL DEFICITS, GENERATING PROGENY WITH LOWER STEP LEVELS INDISTINGUISHABLE FROM WT MICE. SIMILAR EFFECTS WERE OBSERVED WHEN STEP WAS PHARMACOLOGICALLY INHIBITED BY OUR TOOL COMPOUND TC-2153. THESE STUDIES VALIDATE STEP AS A NOVEL DRUG TARGET FOR THE TREATMENT OF AD AND OTHER NEURODEGENERATIVE AND NEUROPSYCHIATRIC DISORDERS. TC-2153 IS THE ONLY REPORTED STEP INHIBITOR WITH CELLULAR AND IN VIVO ACTIVITY. HOWEVER, TC-2153 AND SIMILAR COMPOUNDS ARE KNOWN TO REACT WITH CELLULAR THIOLS AND MODIFY DNA, PRECLUDING THIS INHIBITOR FROM FURTHER PRECLINICAL STUDIES. OTHER REPORTED STEP INHIBITORS SUFFER FROM POOR SELECTIVITY FOR STEP AND LACK OF EFFICACY UNDER PHYSIOLOGICAL CONDITIONS. TO OVERCOME THESE TYPICAL CHALLENGES FOR INHIBITORS TARGETING PTPS - ENZYMES THAT HAVE A HIGHLY CONSERVED AND HIGHLY CHARGED ACTIVE SITE - WE SET OUT TO DISCOVER SMALL MOLECULES THAT BIND TO LESS CONSERVED, ALLOSTERIC SITES IN STEP. USING INNOVATIVE FRAGMENT-BASED SCREENING TECHNOLOGIES, WE IDENTIFIED 19 FRAGMENTS, LOW MOLECULAR WEIGHT COMPOUNDS ADHERING TO THE “RULE OF 3”, THAT SELECTIVELY BIND TO NOVEL, LESS CONSERVED SITES IN STEP WITH UP TO SINGLE DIGIT MICROMOLAR AFFINITY. THESE FRAGMENTS ARE IDEAL STARTING POINTS FOR GENERATING STEP-TARGETED DEGRADERS SUCH AS PROTEOLYSIS TARGETING CHIMERAS (PROTACS) WITH PHYSICOCHEMICAL PROPERTIES SUITABLE FOR CROSSING THE BLOOD-BRAIN BARRIER (BBB). IN FACT, IN PRELIMINARY EXPERIMENTS WE HAVE DEMONSTRATED THAT A PROTAC BASED ON OUR BEST FRAGMENT CAN EFFECTIVELY REDUCE STEP LEVELS IN VIVO. BASED ON THESE EXCITING RESULTS, WE PROPOSE A STEP-TARGETED PROTEIN DEGRADATION APPROACH AS A THERAPEUTIC STRATEGY IN AD. IN AIM 1, WE WILL OPTIMIZE OUR FRAGMENT BINDERS FOR LIGAND EFFICIENCY AND STEP SELECTIVITY. IN AIM 2, WE WILL GENERATE STEP PROTACS FROM THE MOST PROMISING BINDERS, ASSESS THEM IN OUR ESTABLISHED TESTING FUNNEL, AND OPTIMIZE THEM FOR DRUG METABOLISM AND PHARMACOKINETICS (DMPK), PHARMACODYNAMICS (PD), AND TOXICITY (TOX) PROPERTIES. THE GOAL IS TO GENERATE NON-TOXIC STEP DEGRADERS WITH EXPOSURE AND EFFICACY IN THE BRAIN. IN AIM 3, WE WILL TEST WHETHER CANDIDATE PROTACS ARE ABLE TO IMPROVE COGNITION IN MOUSE AD MODELS. OUR STUDIES ARE DESIGNED TO PROVIDE PROOF-OF-CONCEPT (POC) FOR STEP DEGRADERS AS NOVEL THERAPEUTICS IN AD. WE EXPECT TO DEVELOP AT LEAST ONE LEAD SERIES AND ONE BACKUP SERIES OF EFFECTIVE PROTACS THAT ARE READY FOR PRECLINICAL DEVELOPMENT TOWARDS A FIRST-IN-CLASS EARLY TREATMENT OPTION FOR AD.

A GENOMIC/METABOLOMIC STRATEGY TO CHARACTERIZE CARDIAC MITOCHONDRIAL DYSFUNCTION

HOMEOSTATIC ROLE OF IRE1A-XBP1-PDI1 IN HEPATIC LIPID METABOLISM

IMPROVE EXISTING AIRPORT

GENERATION OF RETINOID SIGNALS DURING DEVELOPMENT

MICROGLIAL HEPARAN SULFATE IN THE MODULATION OF APOE FUNCTION AND NEURODEGENERATION - HEPARAN SULFATE (HS), A SULFATED GLYCAN EXPRESSED AT THE CELL SURFACE AND IN EXTRACELLULAR MATRIX, HAS LONG ATTRACTED ATTENTION AS A PUTATIVE FACTOR INVOLVED IN ALZHEIMER'S DISEASE (AD), BASED ON CIRCUMSTANTIAL EVIDENCE FROM IN VITRO AND CLINICOPATHOLOGICAL STUDIES. NEVERTHELESS, THE FUNCTIONAL SIGNIFICANCE OF HS IN AD, ESPECIALLY IN THE LATE-ONSET TYPE (LOAD) THAT COMPRISES MORE THAN 90% OF AD CASES, IS ELUSIVE. RECENT HUMAN GENOMIC STUDIES IMPLICATE APOE AND MICROGLIA AS DOMINANT CONTRIBUTORS TO NEURODEGENERATION IN LOAD. THE APOE GENE IS THE STRONGEST KNOWN GENETIC RISK FACTOR FOR LOAD. ACCUMULATING DATA INDICATE THAT APOE PROTEINS EXERT IMMUNOMODULATORY EFFECTS ON MICROGLIA, AND THAT THIS FUNCTION OF APOE IS AT LEAST PARTLY MEDIATED BY THE TREM2 RECEPTOR, ANOTHER AD RISK FACTOR THAT IS EXPRESSED BY MICROGLIA. AMONG THE THREE HUMAN APOE ALLELES, THE APOE4 ALLELE CONFERS AN INCREASED RISK FOR LOAD, WHILE THE APOE2 ALLELE CONFERS A DECREASED RISK RELATIVE TO THE MORE COMMON APOE3 ALLELE. THE MOLECULAR BASIS OF THESE ALLELE- SPECIFIC RISK VARIATIONS IS ONE OF THE KEY UNANSWERED ISSUES IN AD RESEARCH. IN THIS CONTEXT, IT IS INTERESTING TO NOTE THAT APOE PROTEINS BIND HS IN AN ISOFORM-SPECIFIC MANNER — APOE4 EXHIBITS 2- TO 3-FOLD GREATER AFFINITY FOR HS THAN APOE2 AND APOE3, THUS APPARENTLY CORRELATING WITH THEIR RELATIVE AD RISKS. FURTHERMORE, A RECENT REPORT REGARDING THE UNIQUE CASE OF A COLUMBIAN WOMAN, WHO CARRIES A HIGHLY DETRIMENTAL PSEN1E280A MUTATION, SUGGESTS THAT HER APOE3 "CHRISTCHURCH" MUTATION, WHICH ABOLISHES THE AFFINITY OF APOE3 FOR HS, CONFERS STRONG PROTECTION AGAINST NEURODEGENERATION AND COGNITIVE IMPAIRMENT. ALSO SUGGESTING THE FUNCTIONAL INVOLVEMENT OF HS IN LOAD ARE RECURRENT REPORTS OF GENETIC ASSOCIATION OF HS SULFOTRANSFERASE GENES WITH LOAD. IN A SERIES OF PRELIMINARY STUDIES, WE HAVE OBTAINED MULTIPLE PIECES OF EVIDENCE SUGGESTING THAT THE STRONG INTERACTION OF HS WITH APOE4, RELATIVE TO ITS INTERACTIONS WITH APOE2 AND APOE3, IS THE KEY UNDERLYING MECHANISM BY WHICH APOE4 EXERTS A DETRIMENTAL EFFECT ON THE BRAIN. SIGNIFICANTLY, WE HAVE FOUND THAT HAPLOINSUFFICIENT REDUCTION IN HS EXPRESSION LEADS TO THE MITIGATION OF SYNAPSE LOSS AND MICROGLIAL ACTIVATION IN THE PS19 TAUP310S MOUSE MODEL. WE HYPOTHESIZE THAT HS PLAYS A CRITICAL ROLE IN MEDIATING THE APOE4 EFFECT ON MICROGLIAL RESPONSE AND FUNCTION, AND THAT THE DETRIMENTAL EFFECT OF THE APOE4 ALLELE IS MAINLY DUE TO ITS STRONGER INTERACTION WITH THE 3-O SULFATE-RICH HS SPECIES EXPRESSED IN MICROGLIA. WE WILL: (1) DETERMINE THE ROLE OF HS IN APOE-TREM2 INTERACTION AND SIGNALING IN MICROGLIA; (2) DETERMINE THE ROLE OF MICROGLIAL HS IN APOE/ABCA1-MEDIATED CHOLESTEROL EFFLUX; AND (3) EXAMINE IN VIVO EFFECTS OF MICROGLIAL HS ON BRAIN PATHOLOGY AND FUNCTION IN AD MOUSE MODELS. THIS RESEARCH WILL PROVIDE ENTIRELY NOVEL INSIGHTS INTO THE ROLE OF HS IN MICROGLIA, AND MOREOVER, INTO LONG-TERM QUESTIONS REGARDING THE ISOFORM-SPECIFIC RISK OF APOE VARIANTS. SUCCESSFUL COMPLETION OF THIS PROJECT WILL REVEAL THE APOE4-HS INTERACTION AS A PROMISING THERAPEUTIC TARGET FOR REDUCING AD RISK IN APOE4 CARRIERS.

UNLOCKING REGENERATIVE POTENTIAL THROUGH IN VIVO GENETIC REPROGRAMMING

REGULATION AND FUNCTION OF STROMAL MACROPINOCYTOSIS IN PANCREATIC DUCTAL ADENOCARCINOMA - PROJECT SUMMARY RECENT YEARS HAVE WITNESSED AN APPRECIATION OF THE ROLE THAT METABOLIC ADAPTATION PLAYS IN CONFERRING SURVIVAL ADVANTAGES TO CELLS THAT ENCOUNTER THE HARSH NUTRIENT-POOR CONDITIONS OF THE TUMOR MICROENVIRONMENT. OF PARTICULAR RELEVANCE TO THIS PROPOSAL IS THE NOW WIDELY ACCEPTED NOTION THAT MACROPINOCYTOSIS, AN ENDOCYTIC MECHANISM OF FLUID-PHASE UPTAKE, FUNCTIONS IN TUMORS AS AN AMINO ACID SUPPLY ROUTE. BY STIMULATING THE UPTAKE OF EXTRACELLULAR PROTEIN AND TARGETING IT FOR LYSOSOMAL DEGRADATION, MACROPINOCYTOSIS PROVIDES CELLS WITH A SOURCE OF PROTEIN-DERIVED AMINO ACIDS, ALLOWING TUMORS TO CIRCUMVENT AMINO ACID DEPLETION AND SURVIVE NUTRIENT STRESS. PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) TUMORS ARE DEFICIENT IN GLUTAMINE, A VITAL NUTRIENT THAT SUPPORTS TUMOR GROWTH. OUR PUBLISHED WORK IN PDAC CELLS HAS ESTABLISHED THAT GLUTAMINE DEPLETION HAS THE CAPACITY TO MODULATE MACROPINOCYTOSIS – DIALING THE PROCESS UP OR DOWN AS REQUIRED. INTERESTINGLY, OUR PRELIMINARY DATA PRESENTED IN THIS PROPOSAL DEMONSTRATE FOR THE FIRST TIME THAT GLUTAMINE SCARCITY CAN ALSO STIMULATE MACROPINOCYTOSIS IN CANCER-ASSOCIATED FIBROBLASTS (CAFS). MECHANISTICALLY, WE HAVE ATTRIBUTED GLUTAMINE STRESS-INDUCED MACROPINOCYTIC UPTAKE IN CAFS TO A CAMKK2-AMPK SIGNAL THAT LEADS TO THE RAC1- DEPENDENT ACTIN CYTOSKELETON DYNAMICS THAT ARE REQUIRED FOR MACROPINOCYTOSIS. AMPK IS A BIOENERGETIC STRESS SENSOR THAT IS MOST OFTEN STUDIED IN THE CONTEXT OF GLUCOSE STARVATION, WHICH UNLIKE GLUTAMINE DEPLETION, DOES NOT BOOST CAF MACROPINOCYTOSIS. NOTABLY, NOT MUCH IS KNOWN ABOUT AMPK ACTIVATION AND FUNCTION DURING GLUTAMINE DEPLETION. OUR PRELIMINARY STUDIES SUGGEST THAT MACROPINOCYTOSIS HAS A DUAL PURPOSE IN CAFS – IT CAN SERVE TO SUSTAIN CAF VIABILITY AND FUNCTION, AND IT CAN PROVIDE SECRETED AMINO ACIDS TO NOURISH THE TUMOR CELLS. IMPORTANTLY, OUR IN VIVO AND IN VITRO EXAMINATIONS OF MACROPINOCYTOSIS IN NORMAL FIBROBLASTS, AS WELL AS IN CAFS ORIGINATING FROM OTHER TUMOR TYPES, SUGGEST THAT GLUTAMINE DEPLETION-INDUCED STROMAL UPTAKE IS UNIQUE TO PANCREATIC CAFS. BASED ON THESE DATA, OUR CENTRAL HYPOTHESIS IS THAT GLUTAMINE SCARCITY SELECTIVELY DRIVES CAMKK2-AMPK-DEPENDENT MACROPINOCYTOSIS IN CAFS, AND THAT STROMAL MACROPINOCYTOSIS IS A PROCESS THAT CAN BE HARNESSED IN PDAC THERAPY. IN THIS PROPOSAL, WE WILL 1) EXAMINE THE MOLECULAR MECHANISMS DRIVING THE SELECTIVE ROLE OF GLUTAMINE IN CAF MACROPINOCYTOSIS, 2) FUNCTIONALLY CHARACTERIZE STROMAL MACROPINOCYTOSIS IN PDAC, AND 3) DETERMINE WHETHER THE STROMAL REORGANIZATION THAT OCCURS WITH MACROPINOCYTOSIS INHIBITION CAN BE LEVERAGED FOR PDAC THERAPY. THIS PROJECT WILL BE OF GREAT SIGNIFICANCE, NOVELTY AND IMPACT, AS IT WILL CONSTITUTE THE FIRST EVALUATION OF THE ROLE OF MACROPINOCYTOSIS IN THE PDAC TUMOR STROMA AND THE FIRST ANALYSIS TO SELECTIVELY LINK GLUTAMINE STARVATION TO CAMKK2-AMPK SIGNALING. MOREOVER, BECAUSE MACROPINOCYTOSIS IS IMPORTANT IN BOTH THE TUMOR CELLS AND THE STROMA, OUR WORK COULD HAVE TREMENDOUS IMPACT ON THE DEVELOPMENT OF NOVEL THERAPEUTIC MODALITIES FOR PDAC.

DEVELOPING ER STRESS INHIBITORS FOR TREATING ALS

HYBRID NANOTECHNOLOGIES FOR DETECTION AND SYNERGISTIC THERAPIES FOR BREAST CANCER

ELUCIDATING A MICROGLIAASSOCIATED ROLE FOR SORLA IN MODULATING AD PATHOGENESIS - PROJECT SUMMARY MUTATIONS IN SORLA (ENCODED BY SORL1) IDENTIFIED THROUGH GWAS AND WHOLE EXOME SEQUENCING ANALYSIS HAVE BEEN LINKED TO INCREASED ALZHEIMER’S DISEASE (AD) RISK. ALTHOUGH A NEURONAL ROLE FOR SORLA IN SUPPRESSING AMYLOIDOGENIC APP PROCESSING AND CONSEQUENT ASS GENERATION HAS BEEN ESTABLISHED, SORLA EXPRESSION IS ~8-FOLD HIGHER IN HUMAN MICROGLIA COMPARED TO NEURONS, THUS IMPLICATING A MICROGLIAL ROLE FOR SORLA IN AD PATHOGENESIS. SO FAR, FUNCTIONAL ROLES FOR SORLA IN MICROGLIA HAVE NOT YET BEEN DESCRIBED. HERE, WE USED CRISPR/CAS9 EDITING METHODS TO INTEGRATE AD-ASSOCIATED A528T AND R744X MUTATIONS INTO THE SORL1 (ENCODING SORLA) LOCUS IN HUMAN H9 EMBRYONIC STEM CELLS, WHICH WERE SUBSEQUENTLY DIFFERENTIATED INTO HUMAN MICROGLIA-LIKE (HMGL) CELLS. COMPARING TRANSCRIPTOMIC PROFILES BETWEEN WILDTYPE AND AD-ASSOCIATED SORL1 (A528T, R744X) AND TREM2 (R47H) MUTANT HMGLS REVEALS PATHOGENIC MICROGLIA GENE SIGNATURES SUCH AS INDUCED APOE EXPRESSION PREVIOUSLY DESCRIBED IN AD MOUSE MODELS AND HUMAN AD BRAIN. OUR RESULTS ALSO SHOW THAT CULTURED SORL1R744X AND TREM2R47H HMGLS FEATURE DEFECTS IN ASS UPTAKE IN AN APOE-DEPENDENT MANNER, ALONG WITH IMPAIRED ASS CLEARANCE AND PLAQUE ASSOCIATION IN MOUSE BRAIN XENOTRANSPLANTS BY MICRODIALYSIS/HISTOLOGY. THESE RESULTS PROVIDE PIONEERING EVIDENCE THAT SORLA DYSFUNCTION CAN CONFER PATHOGENIC EXPRESSION SIGNATURES AND IMPAIR MICROGLIAL FUNCTION. WE HYPOTHESIZE THAT MICROGLIAL DYSFUNCTION WILL VARY ACCORDING TO DOMAIN-SPECIFIC MUTATIONS IN THE SORLA EXTRACELLULAR REGION, AND THAT EARLY AND LATE ONSET SORLA MUTATIONS MAY SHOW DIFFERENTIAL EFFECTS ON MICROGLIA DYSREGULATION AND MICROGLIA/NEURON INTERACTION. USING OUR GENE EDITING/HUMAN MICROGLIAL MODELING AND ANALYSIS PLATFORM, WE WILL EXPAND OUR SORLA MUTANT EMBRYONIC STEM CELL (ESC) PANEL TO INCLUDE REPRESENTATIVE EARLY AND LATE AD ONSET MUTATIONS WITHIN EACH OF THE FUNCTIONAL DOMAINS IN THE SORLA EXTRACELLULAR REGION. WE WILL CHARACTERIZE GENE EXPRESSION PROFILES OF WILDTYPE (WT) AND SORLA MUTANT ESC-DERIVED MICROGLIA (“XHMGS”) IN VIVO BY RNASEQ/PROTEOMIC ANALYSIS, AS WELL AS FUNCTIONAL ASPECTS OF MICROGLIAL FUNCTION (ASS UPTAKE, CYTOKINE PROFILES) IN XHMGS XENOTRANSPLANTED IN AD MOUSE BRAIN. AS OUR RESULTS INDICATE THAT SORLA MUTATIONS SUCH AS R744X AND A528T UPREGULATE APOE WHICH MAY TRIGGER CERTAIN ASPECTS OF MICROGLIA FUNCTION, WE WILL ALSO TEST WHETHER APOE AND OTHER POTENTIAL DRIVERS OF MICROGLIA DYSFUNCTION EPISTATICALLY MEDIATE DOWNSTREAM PATHOGENIC BEHAVIOR IN SORLA MUTANT MICROGLIA XENOTRANSPLANTS IN AD MOUSE BRAIN. WE WILL ALSO EXPLORE CELLULAR MECHANISMS ASSOCIATED WITH VARIOUS SORLA MUTATIONS THAT DRIVE CELLULAR DYSFUNCTION IN ESC-DERIVED MICROGLIA AND NEURONS; TO THIS END, WE WILL INVESTIGATE HOW SORLA MUTATIONS IN HUMAN MICROGLIA (XHMGS) INTERACT WITH NEURONS IN MOUSE AD BRAIN XENOTRANSPLANTS, AS WELL AS WT AND SORLA MUTANT HMGLS/NEURONS IN CO-CULTURE. THESE RESULTS WILL PROVIDE US WITH MECHANISTIC INSIGHT INTO HOW VARIOUS MUTATIONS CAN TRIGGER MICROGLIA DYSFUNCTION, AND POTENTIALLY DESCRIBE HOW ASPECTS OF MICROGLIAL AND NEURONAL-RELATED SORLA PATHWAYS ARE AFFECTED TO ALTER AGE-RELATED ONSET IN AD PATHOGENESIS.

MECHANISMS OF NUP210 REGULATION OF MUSCLE DEVELOPMENT AND REGENERATION

IMPROVE EXISTING AIRPORT A

IMPROVE EXISTING AIRPORT

PSYCHOSOCIAL ISSUES AND BARIATRIC SURGERY

BRAIN-PENETRANT GPR88 AGONISTS AS NOVEL THERAPEUTICS FOR OPIOID ABUSE - SUMMARY SUBMITTED IN RESPONSE TO RFA-DA-22-031, THIS APPLICATION DESCRIBES A MULTIDISCIPLINARY EFFORT TO DEVELOP NOVEL, BRAIN-PENETRANT, SMALL-MOLECULE GPR88 AGONISTS TO ATTENUATE ADDICTION-RELEVANT BEHAVIORAL RESPONSES TO OPIOID DRUGS AND AMELIORATE WITHDRAWAL SYNDROME IN OPIOID-DEPENDENT INDIVIDUALS. GPR88 IS AN ORPHAN G-PROTEIN COUPLED RECEPTOR (GPCR) WITH CONCENTRATED EXPRESSION IN STRIATOPALLIDAL (INDIRECT PATHWAY) MEDIUM SPINY NEURONS (MSNS) IN THE STRIATUM. RECENT OBSERVATIONS IN TRANSGENIC MICE DEMONSTRATE THAT GPR88 EXERTS AN INHIBITORY INFLUENCE OVER OPIOID RECEPTOR SIGNALING IN THE STRIATUM. BUILDING ON THIS AND OTHER RECENT PROMISING PRECLINICAL OBSERVATIONS, WE PROPOSE TO LEVERAGE OUR EXPERTISE IN NEUROBIOLOGICAL MECHANISMS OF OPIOID USE DISORDERS AND SMALL-MOLECULE DRUG DISCOVERY TO FORMALLY VALIDATE GPR88 AS A DRUG TARGET FOR OPIOID USE DISORDERS (OUD) AND ESTABLISH A DRUG DEVELOPMENT PATH FORWARD BY DISCOVERING GPR88 AGONIST LEAD COMPOUNDS. WE WILL LEVERAGE AN IN VIVO MURINE MODEL OF OPIOID DEPENDENCE WE HAVE DEVELOPED AND GPR88-/- MICE ALONG WITH AVAILABLE GPR88 PHARMACOLOGICAL TOOLS. UNFORTUNATELY, PUBLISHED GPR88 AGONISTS HAVE LIMITATIONS THAT RESTRICT THEIR UTILITY IN VIVO AND PREVENT THEIR DEVELOPMENT INTO THERAPEUTICS. TO ADDRESS THIS SHORT COMING WE WILL DEVELOP NOVEL, POTENT GPR88 AGONISTS WITH PROPERTIES OPTIMIZED FOR THE TREATMENT OF OPIOID DEPENDENCE. WE ARE WELL POSITIONED TO ACCOMPLISH THIS TASK. WE RECENTLY CONDUCTED A GPR88 HIGH THROUGHPUT SCREEN (HTS), USING AN INNOVATIVE PHARMACOCHAPERONE ASSAY FORMAT, AND IDENTIFIED A NOVEL CHEMICAL SCAFFOLD EXEMPLIFIED BY SBI-‘2037 AS A VALIDATED GPR88 AGONIST. IN ADDITION, WE HAVE ALREADY DEVELOPED ROBUST CELL-BASED ASSAYS (AND APPROPRIATE COUNTER-SCREENS) TO RELIABLY MONITOR GPR88 FUNCTION AND ITS IMPACT ON OPIOID RECEPTORS. LEVERAGING THESE ASSETS, WE WILL CONDUCT A MEDICINAL CHEMISTRY CAMPAIGN TO INCREASE POTENCY AND SELECTIVITY OF THE SBI-‘2037 SCAFFOLD WITH A CRITICAL PATH CONSISTING OF CELLULAR ASSAYS FOCUSED ON THEIR UTILITY IN OUD THAT INCLUDES EARLY ASSESSMENT OF ABSORPTION, DISTRIBUTION, METABOLISM AND EXCRETION (ADME) AND BRAIN PENETRATION PROPERTIES. TO INCREASE THE OVERALL PROBABILITY OF SUCCESS, WE INTEND TO CONDUCT AN ADDITIONAL GPR88 HTS SCREENING CAMPAIGN IN SEARCH OF ONE OR MORE BACK UP SERIES. WE HAVE ESTABLISHED A STRINGENT SET OF CRITERIA FOR GPR88 AGONIST LEADS, AND ONLY COMPOUNDS THAT MATCH THIS PROFILE WILL BE ADVANCED INTO EFFICACY TESTING IN THE INTRAVENOUS OXYCODONE SELF- ADMINISTRATION PROCEDURE IN WILD-TYPE AND GPR88 KNOCKOUT MICE. THIS MULTIDISCIPLINARY RESEARCH PLAN CAPITALIZES ON THE UNIQUELY RELEVANT SCIENTIFIC AND DRUG DISCOVERY EXPERTISE OF OUR TEAM OF COMMITTED INVESTIGATORS AND IS AN INITIAL STEP TOWARDS OUR ULTIMATE GOAL OF DEVELOPING GPR88 AGONISTS AS NOVEL THERAPEUTICS TO FACILITATE ABSTINENCE IN OPIOID-DEPENDENT INDIVIDUALS.

DEVELOPMENT OF AN EFFECTIVE THERAPY AGAINST POST-EXPOSURE ANTHRAX

EPIGENETIC CONTROL OF HEMATOPOIETIC STEM CELLS

BIOBEHAVIORAL FACTORS AND THEIR IMPACT ON STRESS-RELATED EATING IN OBESITY - PROJECT SUMMARY/ABSTRACT ELUCIDATING THE BIOBEHAVIORAL FACTORS THAT LEAD TO PROBLEMATIC EATING, AND ULTIMATELY INCREASE THE RISK FOR WEIGHT GAIN, IS CRITICALLY NEEDED GIVEN THE EPIDEMIC OF OBESITY IN THE UNITED STATES. WHILE THE RELATIONSHIP BETWEEN STRESS AND PROBLEMATIC EATING HAS BEEN WELL-ESTABLISHED, OTHER FACTORS THAT IMPACT THIS ASSOCIATION REMAIN POORLY UNDERSTOOD. ONE IMPORTANT VARIABLE THAT APPEARS TO IMPACT THE STRESS-EATING RELATIONSHIP IS THE MANNER IN WHICH ONE RESPONDS TO STRESSFUL SITUATIONS. STRESS REACTIVITY IS COMMONLY ASSESSED BY THE MANNER IN WHICH CORTISOL CHANGES IN RESPONSE TO A STRESSOR. FOR THOSE WHO TEND TO BE HIGHLY REACTIVE TO STRESS, IN MOMENTS OF HIGH STRESS, THEY APPEAR TO BE MORE LIKELY TO OVEREAT. EVIDENCE SUGGESTS THAT THE GUT MICROBIOTA INFLUENCES STRESS REACTIVITY, AND THIS APPEARS TO BE AN IMPORTANT BIOLOGICAL FACTOR THAT INFLUENCES THE STRESS-EATING RELATIONSHIP. ADDITIONALLY, INDIVIDUALS WHO PLACE A HIGH REWARD VALUE ON FOOD MAY BE MORE IMPACTED IN MOMENTS OF STRESS AND MAY ULTIMATELY OVEREAT OR MAKE PROBLEMATIC FOOD CHOICES IN THOSE INSTANCES. IN THIS STUDY, WE PROPOSE TO PAIR BIOLOGICAL AND BEHAVIORAL ASSESSMENT TO EXAMINE THE NOVEL BIOBEHAVIORAL MODEL OF STRESS-RELATED EATING. THROUGH A COMBINED LABORATORY AND NATURALISTIC APPROACH, WE WILL INVESTIGATE THE EXTENT TO WHICH SPECIFIC TAXA AND GENES OF THE GUT MICROBIOTA, STRESS REACTIVITY, AND THE REWARD VALUE OF FOOD MODERATE THE STRESS-EATING RELATIONSHIP. THROUGH THIS MULTI-METHOD APPROACH, WE WILL ALSO USE A COMBINATION OF MOMENTARY AND DAILY ASSESSMENT TO COLLECT REAL-TIME DATA ON STRESS AND ITS ASSOCIATION WITH DIETARY INTAKE. THIS STUDY WILL OVERCOME LIMITATIONS OF PRIOR RESEARCH THROUGH ITS USE OF MOMENTARY DATA COLLECTION AND NOVEL PAIRING OF BIOLOGICAL AND BEHAVIORAL VARIABLES. ELUCIDATING KEY FACTORS THAT LEAD TO PROBLEMATIC EATING IN MOMENTS OF HIGH STRESS HAS THE POTENTIAL TO LEAD TO PERSONALIZED TREATMENT APPROACHES FOR WEIGHT REDUCTION AND THE PREVENTION OF OBESITY.

LEAD OPTIMIZATION OF NOVEL ML-IAP ANTAGONISTS TO TREAT LUNG CANCER

MAGNETIC RESONANCE OF CARDIAC C13 FLUX & METABOLISM RATE

NEW DOWN SYNDROME BRAIN ORGANIZATION REVEALED BY SINGLE-CELL GENOMICS - PROJECT SUMMARY/ABSTRACT NEARLY 100% OF DOWN SYNDROME (DS; TRISOMY OF HUMAN CHROMOSOME 21) INDIVIDUALS THAT LIVE INTO THEIR 5TH DECADE OF LIFE AND BEYOND, SHOW ALZHEIMER’S DISEASE (AD)-LIKE DEMENTIA AND NEUROPATHOLOGY (DS-AD), REPRESENTING A PROMINENT DS COMORBIDITY THAT HAS RECENTLY BEEN REPORTED AS THE LEADING CAUSE OF DEATH FOR DS ADULTS. BEYOND TRISOMY OF CHROMOSOME 21, WHICH INCLUDES THE KEY AD GENE, AMYLOID PRECURSOR PROTEIN (APP), THE MOLECULAR MECHANISMS UNDERLYING DS-AD HAVE RESISTED IDENTIFICATION. THERE ARE NO DISEASE-MODIFYING THERAPIES (DMTS) TO PREVENT OR TREAT DS-AD, WHICH COULD IMPROVE DS QUALITY OF LIFE AND LIFESPAN. THIS INCLUDE PROPOSAL WILL TRANSFORM OUR UNDERSTANDING OF DS BY REVEALING THE TRANSCRIPTOMIC (RNA) AND GENOMIC (DNA) SINGLE-CELL LANDSCAPE OF THE AGING DS INDIVIDUAL, AND ESPECIALLY THE DS-AD BRAIN, BY CENTERING UPON A NOVEL HUMAN BRAIN MOLECULAR MECHANISM THAT MIGHT UNDERLIE DS-AD: SOMATIC GENE RECOMBINATION (SGR) AND RESULTANT GENOMIC AND TRANSCRIPTOMIC HETEROGENEITY. SGR HAS THE POTENTIAL TO CHANGE THE DNA BLUEPRINT OF DS BRAIN CELLS RESULTING IN FUNCTIONAL CONSEQUENCES FOR BRAIN CELLS THAT COULD EXPLAIN DS-AD ONSET AS WELL AS OTHER DS BRAIN COMORBIDITIES SUCH AS AUTISM AND EPILEPSY. SGR HAS NOT BEEN EXAMINED IN DS BRAINS, WHICH IF OPERATIONAL, WOULD PROVIDE A FUNDAMENTALLY NEW VIEW ON HOW GENES AND GENE DOSAGE ACT TO PROMOTE DS-AD OVER TIME. SGR IS KNOWN TO ACT ON APP IN NORMAL AND SPORADIC AD NEURONS, RESULTING IN THOUSANDS OF NEW APP VARIANTS WITHIN INDIVIDUAL HUMAN BRAINS AND HAS BEEN INDEPENDENTLY CONFIRMED IN THE SCIENTIFIC LITERATURE. THE PROVEN INCREASES IN DS BRAIN GENE EXPRESSION, COMBINED WITH THE IDENTIFIED LINKAGE OF GENE EXPRESSION TO SGR, IMPLICATES GENES TRANSCRIPTIONALLY INCREASED BY TRISOMY 21 AS NEW TARGETS FOR SGR IN DS-AD. APP IS LIKELY THE “TIP OF THE ICEBERG” FOR DS-AFFECTED GENES IN NEURONS AND NON-NEURONAL CELLS, WITH IMPLICATIONS FOR BOTH DS AS WELL AS OTHER STATES OF THE NORMAL AND DISEASED HUMAN BRAIN. THREE INDEPENDENT, BUT DEEPLY-CONNECTED, RESEARCH ELEMENTS (RES) WILL BE COMPLETED BY A PROVEN, COLLABORATIVE TEAM OF MOLECULAR BIOLOGISTS, NEUROSCIENTISTS, NEUROLOGISTS, BIOENGINEERS, AND BIOINFORMATICIANS TO TEST THE HYPOTHESIS THAT SGR CONTRIBUTES TO DS BRAIN DEFICITS AND DS-AD BY ALTERING KNOWN AND UNKNOWN DISEASE GENES AT THE SINGLE-CELL TRANSCRIPTOMIC AND GENOMIC LEVEL WITHIN THE DS BRAIN. RE1 AND RE2 WILL USE CUTTING-EDGE SEQUENCING TECHNOLOGIES TO INTERROGATE THE TRANSCRIPTOMIC AND GENOMIC HETEROGENEITY OF SINGLE NUCLEI FROM DS AND DS-AD BRAINS COMPARED TO CONTROLS, AND WILL INFORMATICALLY INTEGRATE TRANSCRIPTOMIC EXPRESSION, CHROMATIN ACCESSIBILITY, NOVEL ISOFORM DETECTION AND GENOMIC MOSAICISM FORMS, INCLUDING GENCDNAS, WITHIN SINGLE CELLS ACROSS CELL TYPES AND WITH AGE. RE3 WILL EXPLORE THE FUNCTIONAL CONSEQUENCES SGR IN PRIMARY NEURONAL AND INDUCED PLURIPOTENT STEM CELL (IPSC) MODELS TOWARDS UNDERSTANDING THE FUNCTIONAL IMPLICATIONS OF DISEASE ENHANCED SGR, AND THE THERAPEUTIC OPPORTUNITIES THAT SGR UNVEILS.

DEVELOPMENT OF STEP ALLOSTERIC INHIBITORS AS NOVEL THERAPEUTICS FOR ALZHEIMER'S DISEASE

MOLECULAR DETERMINANTS OF TLR TRAFFICKING

IMPROVE EXISTING AIRPORT

NOVEL NEUROPROTECTIVE ROLES FOR THE ALZHEIMER'S DISEASE RISK GENE SORLA IN TAU PATHOLOGY AND PATHOGENESIS - NUMEROUS MUTATIONAL VARIANTS IN THE SORLA (SORL1) GENE ARE ASSOCIATED WITH INCREASED ALZHEIMER’S DISEASE (AD) RISK, AND ALTHOUGH GROWING EVIDENCE FROM OUR GROUP AND OTHERS SUPPORT A NEUROPROTECTIVE ROLE FOR SORLA IN SUPPRESSING EFFECTS ASSOCIATED WITH AΒ, VIRTUALLY NOTHING IS KNOWN WITH RESPECT TO WHETHER SORLA CAN MODULATE PROTEOTOXIC EFFECTS ASSOCIATED WITH TAU. HERE, WE PRESENT EVIDENCE THAT TRANSGENIC UPREGULATION OF SORLA (“SORLA TG”) CAN REDUCE TAU PHOSPHORYLATION, AS WELL AS PATHOLOGICAL VENTRICULAR DILATION IN 9 MONTH- OLD (9MO) PS19 (P301S TAU) MOUSE BRAIN. PROTEOMIC ANALYSIS REVEALS CHANGES IN PROTEOMIC PROFILES RELATED TO SYNAPTIC DYSFUNCTION AND MICROGLIAL ACTIVATION, INCLUDING UPREGULATION OF APOE AND THE COMPLEMENT COMPONENT, C1Q IN PS19 MOUSE HIPPOCAMPUS; THESE CHANGES WERE LARGELY SUPPRESSED IN SORLA TG/PS19 HIPPOCAMPUS, INDICATING THAT SORLA UPREGULATION CAN POTENTIALLY REVERSE PATHOLOGICAL CHANGES ASSOCIATED WITH TAU PROTEOTOXICITY. HISTOLOGICAL ANALYSIS INDICATES THAT C1Q AND ASTROGLIOSIS IS HIGHLY UPREGULATED IN PS19 HIPPOCAMPUS, AND THESE EFFECTS WERE REDUCED IN SORLA TG/PS19 ANIMALS. INTERESTINGLY, C1Q HAS BEEN PREVIOUSLY SHOWN TO LABEL SYNAPSES FOR MICROGLIAL UPTAKE AND ELIMINATION IN BOTH J20 (HAPP) AND PS19 MOUSE BRAIN, WHICH TOGETHER, SUGGESTS THAT SORLA POTENTIALLY MEDIATES NEUROPROTECTIVE EFFECTS THROUGH SUPPRESSION OF DELETERIOUS CHANGES ASSOCIATED WITH SYNAPTIC FACTORS AND GLIOSIS IN PS19 HIPPOCAMPUS. OUR PRIOR WORK ALSO DEMONSTRATES THAT UPREGULATION OF A CLEAVED SOLUBLE SORLA (SSORLA) FORM IN SORLA TG ANIMALS IS ALSO PROTECTIVE IN NEURONS, AND ENHANCES NEURITE REGENERATION WITH NEURONAL INJURY. WE THEREFORE HYPOTHESIZE THAT SORLA CAN MEDIATE NEUROPROTECTIVE EFFECTS TO LIMIT TAU PATHOLOGY/PROTEOTOXICITY AND DETERMINE HERE, WHETHER AND HOW SORLA OVEREXPRESSION, DELETION OR SSORLA CAN AFFECT TAU PATHOLOGY AND PATHOGENESIS. AS OUR RESULTS DEMONSTRATE THAT SORLA TG/PS19 ANIMALS CAN REVERSE PATHOLOGICAL AND PROTEOMIC FEATURES ASSOCIATED WITH LATE-STAGE (9MO) TAU PATHOLOGY, WE WILL ALSO CHARACTERIZE CHANGES AT EARLY AND MID- STAGES OF TAU PATHOLOGY, AND CHARACTERIZE CELL-TYPE SPECIFIC TRANSCRIPTOMIC CHANGES IN 9MO HIPPOCAMPUS IN PS19 MICE THAT MAY BE POTENTIALLY SUPPRESSED IN SORLA TG/PS19 ANIMALS BY SNRNASEQ ANALYSIS. WE WILL ALSO CHARACTERIZE EFFECTS OF SORLA UPREGULATION ON GLIAL-ASSOCIATED SYNAPTIC UPTAKE AND PRUNING IN PS19 AND SORLA TG/PS19 ANIMALS, AND COMPARE EFFECTS OF INTRANEURONAL TAU DISPERSION FROM THE MEC TO DG REGION IN WILDTYPE (WT) VS SORLA TG BRAIN USING AN AAV-TAU PROPAGATION MODEL USED IN OUR LAB (AIM 1) (ZHU ET AL., 2022). WE WILL ALSO DETERMINE EFFECTS OF SORLA DELETION ON TAU PATHOLOGY, GLIOSIS AND SYNPATOTOXICITY (AIM 2), AND CHARACTERIZE EFFECTS OF AD-ASSOCIATED SORLA MUTATIONAL VARIANTS (T2134M, E2055X) THAT POTENTIALLY UPREGULATE SSORLA ON MICROGLIAL FUNCTION AND SORLA/SSORLA EXPRESSION IN AN ESC-BASED HUMAN MICROGLIA CELLS IN VITRO, AND IN HUMAN MICROGLIAL XENOTRANSPLANT MODELS IN PS19/HMCSF MOUSE BRAIN IN VIVO (AIM 3). TOGETHER, RESULTS FROM THIS STUDY WILL PROVIDE INSIGHT INTO A PREVIOUSLY UNDESCRIBED ROLE FOR SORLA AND SSORLA IN TAU PATHOLOGY.

PRENATAL BLOOD-BORNE LIPIDS IN POST-HEMORRHAGIC HYDROCEPHALUS

DISCOVERY OF NOVEL INHIBITORS TARGETING TRANS-GOLGI NETWORK ACIDIFICATION IN PANCREATIC CANCER - PROJECT SUMMARY PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) IS PROJECTED TO BE THE SECOND LEADING CAUSE OF CANCER-RELATED DEATHS IN THE UNITED STATES BY 2030. WITH A 5-YEAR RELATIVE SURVIVAL RATE OF JUST 10%, PDAC HAS THE HIGHEST DEATH RATE AMONG THE MOST COMMON CANCERS, UNDERSCORING THE URGENT NEED FOR NEW THERAPEUTIC STRATEGIES THAT IMPROVE CLINICAL OUTCOMES. RECENT YEARS HAVE WITNESSED A GROWING APPRECIATION OF THE ROLE THAT METABOLIC REPROGRAMMING PLAYS IN CONFERRING SURVIVAL ADVANTAGES TO PDAC CELLS, AND THE TARGETING OF METABOLIC PROCESSES IS CONSIDERED A PROMISING AREA FOR THE DEVELOPMENT OF NOVEL THERAPIES. IN CONTRAST TO NORMAL CELLS, CANCER CELLS FAVOR GLYCOLYSIS FOR ENERGY PRODUCTION, A PHENOMENON CALLED THE WARBURG EFFECT. AS A GLYCOLYTIC BYPRODUCT, TRANSFORMED CELLS PRODUCE AN EXCESSIVE AMOUNT OF H+ IONS. THIS CYTOSOLIC ACCUMULATION OF H+ WOULD BE DETRIMENTAL TO CELL FITNESS; THEREFORE, TUMOR CELLS HAVE EVOLVED MECHANISMS TO ACHIEVE PH HOMEOSTASIS. USING PDAC AS A MODEL SYSTEM, WE DISCOVERED THAT CYTOSOLIC PH IN CANCER CELLS IS REGULATED BY ORGANELLAR SEQUESTRATION OF H+ IONS VIA COMPARTMENTAL ION TRANSPORT AND THAT THE TRANS-GOLGI NETWORK (TGN) CAN ACT AS A “SINK” FOR CYTOSOLIC H+. IMPORTANTLY, NORMAL CELLS DO NOT EMPLOY THIS HOMEOSTATIC MECHANISM. USING THE NHE7 NA+/H+ ANTIPORTER, WHICH IS PRIMARILY LOCALIZED TO THE TGN, WE HAVE SHOWN THAT TARGETING NHE7 CAUSES ACIDIFICATION OF THE CYTOSOL AND SUBSEQUENT CELL DEATH IN PDAC CELLS BUT NOT NORMAL CELLS. WE HAVE FURTHER VALIDATED THESE FINDINGS IN MOUSE MODELS OF PDAC. PATIENT-CENTRIC DATA ANALYSES SHOW THAT TGN LOCALIZED TRANSPORTERS ARE FREQUENTLY OVEREXPRESSED IN PDAC AND THEIR ELEVATED EXPRESSION IS CORRELATED WITH WORSE OVERALL SURVIVAL. TO ROBUSTLY QUANTIFY PH IN THE LUMEN OF THE TGN, WE HAVE DEVELOPED AN INNOVATIVE TGN-TARGETED PH BIOSENSOR ASSAY, WHICH HAS BEEN OPTIMIZED FOR HIGH-THROUGHPUT (HT) SCREENING OF SMALL MOLECULE COMPOUND LIBRARIES TO IDENTIFY NOVEL TGN-SELECTIVE ACIDIFICATION INHIBITORS. WE HAVE VALIDATED OUR TGN PH BIOSENSOR HT ASSAY IN A SET OF CLINICAL TRIAL AND APPROVED DRUGS, AS WELL AS KINASE INHIBITORS. TO COMPLEMENT OUR HT SCREEN, WE HAVE DESIGNED A CUTTING- EDGE SECONDARY AND TERTIARY ASSAY PIPELINE TO FURTHER VALIDATE TGN-SELECTIVE COMPOUNDS. WE HYPOTHESIZE THAT NOVEL GOLGI–SPECIFIC SMALL MOLECULE INHIBITORS OF ORGANELLE ACIDIFICATION WILL SELECTIVELY PREVENT CANCER CELLS FROM ACHIEVING PROPER PH HOMEOSTASIS, RESULTING IN ACIDIC CYTOSOLIC PH AND CELL DEATH. IN THIS GRANT WE WILL: 1) PERFORM A LARGE-SCALE HT SCREEN TO IDENTIFY INHIBITORS OF TGN ACIDIFICATION AND CONFIRM THE HITS, 2) VALIDATE THE CONFIRMED HITS USING SECONDARY ASSAYS AND PRIORITIZE HIT SCAFFOLDS, AND 3) MAP COMPOUND EFFECTS TO TGN COMPONENTS AND SELECT AND CHARACTERIZE FINAL PROBE(S). SUCCESSFUL COMPLETION OF THIS WORK WILL PROVIDE NOVEL SMALL MOLECULE CHEMICAL PROBES VALIDATED TO INTERFERE WITH TGN ACIDIFICATION AND CAUSE CANCER-SELECTIVE CELL DEATH, SERVING AS STARTING POINTS TO BE FURTHER DEVELOPED INTO SAFE AND EFFECTIVE THERAPEUTIC AGENTS FOR PDAC.

IMPROVE EXISTING AIRPORT

EATING AND QUALITY OF LIFE POST BARIATRIC SURGERY

DRUG DISCOVERY FOR FATTY ACID SYNTHASE IN ONCOLOGY

CENTER FOR PEDIATRIC RESEARCH - OVERALL PROJECT SUMMARY THE PRIMARY GOAL OF THIS APPLICATION IS TO CONTINUE TO HARNESS THE CAPABILITIES OF THE CENTER FOR PEDIATRIC RESEARCH TO SUPPORT BASIC AND TRANSLATIONAL SCIENTISTS STUDYING MECHANISMS UNDERLYING CHILDREN’S DISEASE AND ASSOCIATED TREATMENT OPTIONS. THE ORIGIN OF MANY PEDIATRIC DISEASES IS FROM ALTERED DEVELOPMENTAL PROGRAMMING RELATED TO THE PROCESSES OF CELL PROLIFERATION, MORPHOGENESIS, MIGRATION, DIFFERENTIATION, AND PROGRAMMED DEATH. THESE DEVELOPMENTAL PROCESSES ARE AT THE ROOT OF PEDIATRIC DISEASE AND ARE DISRUPTED THROUGH GENETIC DISORDERS, ABERRANT FETAL PROGRAMMING, ALTERED GROWTH & DEVELOPMENT, AND ENVIRONMENTAL PRESSURES. OUR MULTIDISCIPLINARY CENTER APPLIES GENETIC, BIOCHEMICAL, CELL, AND MOLECULAR APPROACHES ACROSS SEVERAL MODEL ORGANISMS TO CHARACTERIZE ALTERATIONS DURING DEVELOPMENT AS THEY PERTAIN TO PEDIATRIC DISEASES AND DISORDERS. FOR PHASE III WE WILL CONTINUE TO BUILD UPON THE SUCCESS OF PHASE I AND II EFFORTS IN CREATING A CRITICAL MASS OF INDEPENDENTLY FUNDED INVESTIGATORS WITHIN THE CENTER FOR PEDIATRIC RESEARCH BY REFINING OUR FOCUS TO SUPPORT AND MENTOR BOTH NEW AND ESTABLISHED INVESTIGATORS. CENTER SUCCESS WILL BE ACHIEVED THROUGH THE FOLLOWING AIMS: 1) MAINTAIN A SUPPORTIVE ENVIRONMENT FOR THE TRAINING AND MENTORSHIP OF SCIENTISTS STUDYING PEDIATRIC DISEASE TO STRENGTHEN AND GROW THE CENTER MEMBERSHIP; 2) UTILIZE AND ENHANCE EXISTING CORE RESOURCES THAT SUPPORT PEDIATRIC RESEARCH WITH A FOCUS ON LONG-TERM SUSTAINABILITY; 3) EXPAND TRAINING AND LEARNING OPPORTUNITIES RELEVANT TO PEDIATRIC DISEASE; AND 4) EVALUATE SUCCESS OF THE CENTER. THROUGH THESE AIMS, THE CENTER FOR PEDIATRIC RESEARCH WILL DEVELOP A STRONG FOUNDATION IN BASIC AND TRANSLATIONAL RESEARCH BY FOSTERING A COLLABORATIVE ENVIRONMENT FOR SCIENTISTS AND PHYSICIANS. STRENGTHENED BY OUR INSTITUTION’S STRONG COMMITMENT TO CHILDREN’S MEDICINE, PHASE III FUNDING WILL BE INSTRUMENTAL IN ENHANCING PEDIATRIC RESEARCH EFFORTS IN THE CENTER, THE REGION, AND BEYOND.

TAS::97 0130::TAS CELLULAR SENTINELS TOXICITY PLATFORM

FXR SIGNALING PATHWAY IS A VALID TARGET FOR CHEMOPREVENTION IN COLORECTAL CANCER

LEAD OPTIMIZATION OF NOVEL MGLU2 NEGATIVE ALLOSTERIC MODULATORS

MECHANISM OF ER PROTEIN MISFOLDING-INDUCED MITOCHONDRIAL DYSFUNCTION

OPTIMIZATION OF DRUG-LIKE PROPERTIES OF CRFBP-CRF2 NEGATIVE ALLOSTERIC MODULATORS FOR ALCOHOL USE DISORDER - PROJECT SUMMARY THIS APPLICATION, “OPTIMIZATION OF DRUG-LIKE PROPERTIES OF CRFBP-CRF2 NEGATIVE ALLOSTERIC MODULATORS FOR ALCOHOL USE DISORDER”, IS IN RESPONSE TO PAR-22-031 “DRUG DISCOVERY FOR NERVOUS SYSTEM DISORDERS (R01 CLINICAL TRIALS NOT ALLOWED)”. ALCOHOL USE DISORDER (AUD) REMAINS A HUGE CLINICAL AND PUBLIC HEALTH PROBLEM WITH NO EFFECTIVE PHARMACOLOGICAL RECOURSE, AFFECTING 28.3 MILLION ADULTS IN THE UNITED STATES, AND IS THE 3RD LEADING PREVENTABLE CAUSE OF DEATH. THE ONLY FDA-APPROVED MEDICATIONS FOR AUD ARE DISULFIRAM, NALTREXONE, AND ACAMPROSATE, ALL OF WHICH EXHIBIT LIMITED EFFICACY AND HAVE LIMITING CONTRAINDICATIONS. HENCE, THERE IS A CRITICAL NEED TO DEVELOP MORE EFFECTIVE THERAPEUTICS TO TREAT AUD. WHILE MANY FACTORS CONTRIBUTE TO THE DEVELOPMENT AND MAINTENANCE OF AUD, INCREASING ATTENTION IS BEING PAID TO POTENTIALLY DRUGGABLE TARGETS WITHIN THE STRESS SYSTEM. THE PRIMARY REGULATOR OF THE STRESS RESPONSE, CORTICOTROPHIN-RELEASING FACTOR (CRF), EXERTS ITS EFFECTS BY BINDING TO CRF1 AND CRF2 RECEPTORS AND, ALSO, A SECRETED 37-KD CRF-BINDING PROTEIN (CRFBP). IN ADDITION, CRFBP UNDERGOES SPONTANEOUS CLEAVAGE INTO A 27-KD N-TERMINAL FRAGMENT, CRFBP(27KD), THAT BINDS CRF AND A 10- KD C-TERMINAL FRAGMENT, CRFBP(10KD), THAT DOES NOT. WE HYPOTHESIZE THAT CRFBP HAS DUAL EXCITATORY AND INHIBITORY EFFECTS ON CRF FUNCTION AND, THUS, ETHANOL CONSUMPTION, AND THAT THE CRFBP-CRF2 INTERACTION REPRESENTS A NOVEL PHARMACOLOGICAL TARGET FOR THE TREATMENT OF AUD. TO TEST THIS HYPOTHESIS, WE DEVELOPED CHEMICAL PROBES SPECIFIC FOR THE CRFBP(10KD)-CRF2 COMPLEX IN TWO LEAD SERIES THAT ACT AS NEGATIVE ALLOSTERIC MODULATORS (NAMS) OF THE CRFBP-CRF2 COMPLEX ONLY IN THE PRESENCE OF CRFBP(10KD). OUR RECENT STRUCTURE- ACTIVITY RELATIONSHIP (SAR) STUDIES HAVE PROVIDED CRFBP-CRF2 NAMS WITH GOOD ON-TARGET POTENCY AND SELECTIVITY PROFILES IN VITRO, BUT ADDITIONAL CHEMICAL OPTIMIZATION IS REQUIRED TO PRODUCE CHEMICAL PROBES THAT ARE READY FOR COMPREHENSIVE IN VIVO EVALUATION. THESE IN VIVO PROBES WOULD ALLOW US TO ESTABLISH THE ROLE OF CRFBP IN ALCOHOL CONSUMPTION AND FACILITATE THE DEVELOPMENT OF EFFECTIVE TREATMENTS TARGETING CRFBP FOR AUD. THUS, OUR OVERALL OBJECTIVE IS TO DEVELOP SYSTEMICALLY ACTIVE CRFBP-CRF2 NAMS SUITABLE FOR ADVANCED IN VIVO PROOF- OF-CONCEPT STUDIES FOR THE TREATMENT OF AUD. ACCORDINGLY, OUR SPECIFIC AIMS ARE: (1) DESIGN AND SYNTHESIZE NOVEL CRFBP-CRF2 NAMS WITH OPTIMAL DRUG-LIKE PROPERTIES; (2) CHARACTERIZE NOVEL CRFBP-CRF2 NAMS IN ASSAYS MEASURING POTENCY, SELECTIVITY AND DRUG-LIKE PROPERTIES; AND (3) DEMONSTRATE IN VIVO PROOF-OF-CONCEPT FOR SELECT CRFBP-CRF2 NAMS IN RODENT MODELS OF AUD. THE CRFBP-CRF2 NAMS GENERATED WILL PROVIDE POWERFUL IN VIVO PROBES FOR TESTING THE ROLE OF THE CRFBP-CRF2 INTERACTION IN VIVO. WE ARE WELL-POSITIONED TO DEVELOP POTENT AND SELECTIVE SMALL MOLECULE CRFBP-CRF2 NAMS WITH EXCELLENT PHARMACOKINETIC PROPERTIES FOR IN VIVO PROOF-OF- CONCEPT STUDIES IN RODENT MODELS OF AUD. THIS MULTIDISCIPLINARY RESEARCH PROGRAM HAS THE POTENTIAL FOR SIGNIFICANT SCIENTIFIC AND MEDICAL IMPACT BY CONTRIBUTING TO THE DISCOVERY OF NEW MEDICATIONS FOR AUD.

HYALURONAN IN BRAIN EXTRACELLULAR SPACE AS A REGULATOR OF GLYMPHATIC CLEARANCE OF ABETA

CHARACTERIZING NEUROPROTECTIVE ROLES FOR SNX27 SIGNALING PATHWAYS IN AD AND DS

MECHANISMS OF HEMATOPOIETIC ACUTE RADIATION SYNDROME INDUCTION AND RADIOPROTECTION THROUGH SPHINGOSINE 1-PHOSPHATE RECEPTOR 1 SIGNAL MODULATION

LEAD OPTIMIZATION OF NOVEL CRFBP-CRFR2 COMPLEX MODULATORS FOR ALCOHOL USE DISORDER

POLYGENIC BASIS OF CARDIAC FUNCTION IN DROSOPHILA MELANOGASTER

PROTEIN SIALYLATION AND DE-SIALYATION IN CELL SURFACE GLYCOPROTEIN HOMEOSTASIS AND DISEASE - SUMMARY PROTEIN SIALYLATION IS A POST-TRANSLATIONAL MODIFICATION PRODUCED BY MEMBERS OF A FAMILY OF SIALYLTRANSFERASES. WE RECENTLY DISCOVERED THAT THE RESULTING SIALIC ACID LINKAGES ARE INVOLVED IN THE REGULATION OF GLYCOPROTEIN ABUNDANCE AND FUNCTION IN AN INTRINSIC HOMEOSTATIC MECHANISM THAT IS TARGETED BY MULTIPLE PATHOGENS. ENVIRONMENTAL FACTORS ARE DOMINANT IN THE ORIGINS OF THE HUMAN INFLAMMATORY BOWEL DISEASES (IBDS) AND SEASONAL BACTERIAL INFECTIONS HAVE BEEN IMPLICATED. WE THEREFORE DEVELOPED A MOUSE MODEL OF REPEATED HUMAN FOOD POISONING COMPRISED OF RECURRENT LOW-TITER NON-LETHAL GASTRIC INFECTIONS OF THE GRAM-NEGATIVE BACTERIAL PATHOGEN SALMONELLA ENTERICA TYPHIMURIUM (ST), A LEADING CAUSE OF HUMAN FOODBORNE ILLNESS WORLDWIDE. IN THIS UNIQUE MODEL, THE ST PATHOGEN WAS RAPIDLY CLEARED BY THE HOST, HOWEVER A PROGRESSIVELY SEVERE AND PERSISTENT COLITIS DEVELOPED SIMILAR TO ULCERATIVE COLITIS (UC). WE DEMONSTRATED THAT PATHOGENESIS WAS LINKED TO THE DISABLING OF A PROTECTIVE MECHANISM IN THE HOST INVOLVING THE ANTI-INFLAMMATORY INTESTINAL ALKALINE PHOSPHATASE (IAP) GLYCOPROTEIN ENZYME PRODUCED BY ENTEROCYTES. RECURRENT ST INFECTIONS RESULTED IN TOLL-LIKE RECEPTOR-4 (TLR4) INDUCTION OF NEURAMINIDASE (NEU) ACTIVITY AND HOST NEU3 ABUNDANCE WITH NASCENT IAP DE-SIALYLATION AND ENDOCYTIC DEGRADATION, THEREBY REDUCING IAP HALF-LIFE, ABUNDANCE, AND FUNCTION. IAP DEFICIENCY WAS SIMILARLY ACQUIRED IN MICE LACKING THE ST3GAL6 SIALYLTRANSFERASE RESULTING IN SPONTANEOUS COLITIS. THE DISEASE MECHANISM IN BOTH CASES WAS TLR4-DEPENDENT AND LINKED TO REDUCED DEPHOSPHORYLATION AND DETOXIFICATION OF THE LIPOPOLYSACCHARIDE-PHOSPHATE PRODUCED BY COMMENSAL BACTERIA OF THE COLON. IN HUMANS, SIMILAR MODULATION OF IAP AND NEU3 HAVE BEEN REPORTED IN COLITIS, AND GENETIC DEFICIENCY OF IAP CAUSES COLITIS, SUPPORTING THE RATIONALE FOR ONGOING CLINICAL TRIALS OF IAP AUGMENTATION AND NEURAMINIDASE INHIBITION. HOWEVER, THE KEY INVOLVEMENT OF NEU3 REMAINS TO BE ESTABLISHED. RESEARCH PROPOSED HEREIN WILL DIRECTLY TEST THE ROLE OF HOST NEU3 IN THE ONSET AND PROGRESSION OF COLITIS AMONG NEU3-NULL MICE. IN ADDITION, THE POSSIBILITY THAT OTHER ENTERIC GRAM-NEGATIVE PATHOGENS SIMILARLY PROVOKE IAP DEFICIENCY WILL BE INVESTIGATED WITH THE DEVELOPMENT OF OTHER RECURRENT NON-LETHAL GASTRIC INFECTION MODELS USING RELATED GRAM-NEGATIVE ENTERIC PATHOGENS INCLUDING THE HYPERVIRULENT SALMONELLA ENTERICA CHOLERAESUIS SEROVAR. WE HAVE RECENTLY MEASURED ELEVATED NEU ACTIVITY AND NEU3 PROTEIN ABUNDANCE IN THE COLON DURING RECURRENT ST INFECTION ASSOCIATED WITH THE DE-SIALYLATION OF MUCIN– 2 (MUC2), THE MAJOR GLYCOPROTEIN COMPONENT OF THE PROTECTIVE MUCIN BARRIER. THE MECHANISM OF EROSION OF THE MUCIN BARRIER IN COLITIS REMAINS UNKNOWN BUT PLAYS A LARGE ROLE IN PATHOGENESIS. WE HAVE RECENTLY LINKED EROSION OF THE MUCIN BARRIER TO REDUCTIONS OF MUC2 PROTEIN SIALYLATION, LIKELY BY ST3GAL6. REMARKABLY, NEU TREATMENT INCREASES MUC2 PROTEOLYSIS WITH REDUCED MUC2 ABUNDANCE. RESEARCH PROPOSED HEREIN WILL DETERMINE THE ROLE OF MUC2 SIALYLATION BY ST3GAL6 IN GENERATING AND MAINTAINING THE PROTECTIVE MUCIN BARRIER. TOGETHER THESE STUDIES WILL PROVIDE IMPORTANT ADVANCES PERTAINING TO THE PREVENTION AND TREATMENT OF COLITIS.

TRANSFORMATIVE RESEARCH ON THE NORMAL AND ALZHEIMER'S DISEASE BRAIN THROUGH STUDIES OF NEURONAL GENE RECOMBINATION

REACH ND: REDUCING BARRIERS, EXPANDING ACCESS, AND ADVANCING TRAUMA CARE FOR CHILDREN TO HEAL IN NORTH DAKOTA - REACH ND IS LED BY TREATMENT COLLABORATIVE FOR TRAUMATIZED YOUTH (TCTY) HOUSED WITHIN SANFORD RESEARCH NORTH AND REPRESENTS A CONSORTIUM OF ORGANIZATIONS WITH THE SHARED MISSION OF REDUCING BARRIERS, EXPANDING ACCESS, AND ADVANCING TRAUMA CARE FOR CHILDREN TO HEAL IN NORTH DAKOTA. REACH ND AIMS TO INCREASE ACCESS TO TRAUMA-FOCUSED, EVIDENCE-BASED TREATMENTS (EBTS) FOR UNDERSERVED/UNDER-RESOURCED CHILDREN (3-18) AND FAMILIES IMPACTED BY TRAUMA IN ND. TO INCREASE ACCESS, REACH ND WILL ADDRESS BEHAVIORAL HEALTH DISPARITIES, REDUCE BARRIERS TO ACCESS AND ENGAGEMENT IN TRAUMA EBTS, AND IMPROVE FRAMEWORKS FOR SUSTAINABILITY. THE PROJECT WILL COLLABORATE WITH PARTNERS INCLUDING CHILDREN’S ADVOCACY CENTERS AND NCTSN CATEGORY II TSAS TO ACHIEVE GOALS AND OBJECTIVES THAT INCLUDE (1) PROVIDING CARE COORDINATION USING INTENSIVE ENGAGEMENT STRATEGIES AND EFFECTIVE EBTS (TF-CBT, CFTSI, SPARCS) VIA TELEHEALTH; (2) CONDUCTING TRAINING ON TELEHEALTH AND COMPLEX TRAUMA FOR PROVIDERS; (3) ADAPTING TREATMENT RESOURCES FOR RURAL POPULATIONS, AMERICAN INDIAN/ALASKAN NATIVE CHILDREN, AND LINGUISTIC MINORITIES; AND (4) INCREASING PROFESSIONAL AND COMMUNITY TRAUMA AWARENESS AND ENGAGEMENT USING OUTREACH EDUCATION AND MEDIA STRATEGIES. GOALS & OBJECTIVES GOAL 1: REDUCE BARRIERS AND INCREASE ACCESS TO TRAUMA SERVICES THROUGHOUT THE RURAL AND FRONTIER REGIONS OF NORTH DAKOTA WITH EQUITABLE CARE DELIVERY AS CORE CONSIDERATION. 1. PROCEDURES FOR INTENSIVE ENGAGEMENT & TELEHEALTH. 2. CARE COORDINATION WITH ENHANCED ENGAGEMENT STRATEGIES. 3. ASSESSMENTS & TRAUMA EBTS VIA TELEHEALTH. 4. SIGNIFICANTLY REDUCED TRAUMA SYMPTOMS AT DISCHARGE. 5. DECREASE BARRIERS TO TELEHEALTH TREATMENT BY ENABLING AT-HOME ACCESS. 6. TRANSLATE OR ADAPT TREATMENT RESOURCES. GOAL 2: ADVANCE TRAUMA CARE PRACTICES AND NETWORK OF TREATMENT PROVIDERS IN NORTH DAKOTA THROUGH TRAINING, CONSULTATION, AND STATEWIDE AND NATIONAL COLLABORATION. 1. TRAIN TREATMENT PROVIDERS ON TELEHEALTH DELIVERY OF TF-CBT. 2. HOST QUARTERLY EDUCATION SERIES FOR CLINICAL TEAMS ON TOPICS IN COMPLEX TRAUMA. 3. HOST STATEWIDE TELEHEALTH BIANNUAL CONSULTATIONS TO DEVELOP RESOURCES FOR TELEHEALTH. 4. PROTOCOL FOR NEW PROVIDER TELEHEALTH ONBOARDING. 5. COLLABORATE WITH NCTSN CAT II TSA ON TRAIN-THE-TRAINER MODELS TO SUSTAIN TRAINING. GOAL 3: INCREASE PROFESSIONAL AND COMMUNITY TRAUMA AWARENESS AND ENGAGEMENT USING OUTREACH EDUCATION AND MEDIA STRATEGIES. 1. APPEAR ON NORTH DAKOTA TODAY SHOW TO DISCUSS TRAUMA AND ACCESSING SERVICES. 2. DISSEMINATE INFORMATION ABOUT TRAUMA AND ACCESSING CARE AT COMMUNITY OUTREACH EVENTS. 3. CREATE 2 VIDEOS DISCUSSING TRAUMA TREATMENT, TELEHEALTH, AND ACCESSING SERVICES. 4. HOST EDUCATION EVENTS FOR CHILD-SERVING PROFESSIONALS IN ND. WE ANTICIPATE SERVING 120-150 CLIENTS AND 101-110 PROFESSIONALS ANNUALLY AND 1205 TOTAL CLIENTS AND PROFESSIONALS THROUGHOUT THE FIVE-YEAR PERIOD OF PERFORMANCE.

IMPROVE EXISTING AIRPORT

VIRULENCE MECHANISMS OF VIRAL BCL-2 HOMOLOGS

MATURATION OF NORMAL AND DISEASE-SPECIFIC HUMAN STEM CELL-DERIVED CARDIOMYOCYTES

ANTIGEN-SPECIFIC B CELL TOLERANCE IN THE GUT-ASSOCIATED LYMPHOID TISSUE (GALT)

NLR FAMILY PROTEINS: MECHANISMS AND REGULATION

SIGNALING INTERACTIONS IN CELL SURVIVAL AND INVASION

NEW HYDROCEPHALUS THERAPIES THROUGH INTERRUPTION OF LIPID SIGNALING AND INFLAMMATORY PATHWAYS USING NOVEL DRUG-LIKE COMPOUNDS

MSX2-WNT SIGNALING IN CARDIOVASCULAR CALCIFICATION

EPHA2 RECEPTOR SIGNALING IN BREAST CANCER MECHANOTRANSDUCTION - SUMMARY MECHANICAL FORCES GENERATED BY A RIGID EXTRACELLULAR MATRIX (ECM) IN THE TUMOR MICROENVIRONMENT PLAY A KEY ROLE IN TUMOR PROGRESSION AND METASTASIS. WE RECENTLY DISCOVERED A CRITICAL ROLE OF EPHA2 NON-CANONICAL SIGNALING IN PROMOTING EPITHELIAL-MESENCHYMAL TRANSITION (EMT) AND TUMOR INVASION AND METASTASIS IN RESPONSE TO INCREASING ECM STIFFNESS IN THE TUMOR MICROENVIRONMENT. FURTHERMORE, WE FOUND THAT ACTIVATION OF EPHA2 CANONICAL SIGNALING BY THE EPHRINA1 LIGAND POTENTLY INHIBITS STIFFNESS-INDUCED BREAST CANCER CELL INVASIVENESS. BASED ON THESE RESULTS, WE HYPOTHESIZE THAT EPHA2 FUNCTIONS AS A KEY RHEOSTAT THAT INTEGRATES BOTH MECHANICAL AND BIOCHEMICAL CUES FROM THE TUMOR MICROENVIRONMENT TO REGULATE MECHANOSIGNALING IN BREAST CANCER PROGRESSION AND METASTASIS. WE PROPOSE TO COMBINE BIOCHEMICAL, 2D AND 3D CELL CULTURE ASSAYS, AND IN VIVO XENOGRAFT MODELS TO ELUCIDATE EPHA2 SIGNALING MECHANISMS IN BREAST CANCER MALIGNANCY INDUCED BY ECM RIGIDITY THROUGH THREE SPECIFIC AIMS. (1) DETERMINE HOW ECM RIGIDITY ACTIVATES EPHA2 NON-CANONICAL SIGNALING TO PROMOTE EMT AND INVASION. (2) DETERMINE HOW EPHA2 CANONICAL SIGNALING BLOCKS EMT AND INVASION INDUCED BY ECM RIGIDITY. (3) EXAMINE THE ROLE OF EPHA2 NON-CANONICAL AND CANONICAL SIGNALING IN BREAST CANCER INVASION AND METASTASIS IN VIVO. TOGETHER, THESE STUDIES WILL INFORM ON THE POTENTIAL USEFULNESS OF ACTIVATING EPHA2 CANONICAL SIGNALING FOR INHIBITION OF BREAST CANCER INVASIVENESS AND METASTASIS.

DISCOVERY OF GLUTARYL-COA DEHYDROGENASE INHIBITORS FOR MELANOMA AND PANCREATIC CANCER - PROJECT SUMMARY MELANOMA IS AN EXTREMELY AGGRESSIVE CANCER WITH HIGH MORTALITY. ITS PHENOTYPIC PLASTICITY AND HETEROGENEITY ENABLE IT TO ADAPT TO DIVERSE PHYSIOLOGICAL SETTINGS AND DEFEAT TREATMENT APPROACHES. TARGETED THERAPIES AND CHECK POINT INHIBITORS BECAME AVAILABLE IN THE PAST DECADE; HOWEVER, THESE DRUGS WORK IN ONLY A SUBSET OF PATIENTS AND DRUG RESISTANCE EVENTUALLY EMERGES, EVEN IN INITIAL RESPONDERS. THEREFORE, NEW TARGETS AND CLINICAL APPROACHES FOR MELANOMA ARE AN UNMET MEDICAL NEED. WE IDENTIFIED GLUTARYL-COA DEHYDROGENASE (GCDH) AS ONE SUCH TARGET. GCDH EXPRESSION CORRELATES WITH AGGRESSIVE CANCERS AND LOW SURVIVAL IN MELANOMA PATIENTS. OUR DATA DEMONSTRATES THAT GCDH KNOCKDOWN RESULTS IN APOPTOTIC CELL DEATH IN MELANOMA CELLS. MELANOMAS SEEM UNIQUELY SENSITIVE TO TOXIC GLUTARATE METABOLITES RESULTING FROM GCDH DEFICIENCIES, SINCE SUPPRESSION OF THE DEHYDROGENASE E1 AND TRANSKETOLASE DOMAIN CONTAINING 1 (DHTKD1) ENZYME, CATALYZING THE PRECEDING REACTION IN THE CATABOLIC PATHWAY AND CONVERTING 2-OXOADIPATE TO GLUTARYL-COA, RESCUES MELANOMA CELLS FROM APOPTOSIS. PANCREATIC CANCER CELLS, BUT NOT OTHER CANCERS NOR NORMAL CELLS, SHARE THIS OVERRELIANCE ON GCDH AND UNDERGO APOPTOSIS UPON GCDH KNOCKDOWN. WE HYPOTHESIZE THAT SMALL-MOLECULES INTERFERRING WITH GCDH WILL RESULT IN THE OBLITERATION OF MELANOMA CELLS THROUGH APOPTOSIS. WE PROPOSE TO IDENTIFY CHEMICAL PROBES OF GCDH TO FURTHER VALIDATE THE ENZYME AS A MOLECULAR TARGET FOR MELANOMA. IN A PILOT SCREEN, WE ESTABLISHED AND VALIDATED ALL OF THE ASSAYS PROPOSED HEREIN. WE WILL PERFORM LARGE-SCALE HTS, HIT CONFIRMATION AND OPTIMIZATION, AND VALIDATE THE IDENTIFIED HITS IN A PANEL OF DIVERSE CELL LINES FOR MELANOMA AND OTHER CANCERS. COMPOUNDS IDENTIFIED WILL PROVIDE DESIRABLE PHARMACOLOGICAL TOOLS TO STUDY THE PATHOPHYSIOLOGY OF GCDH IN MELANOMA AND OTHER CANCERS, AND THE MOLECULAR MECHANISMS OF LYSINE METABOLISM LIABILITY IN MELANOMA, AS WELL AS PROVIDE POTENTIAL STARTING POINTS FOR FUTURE THERAPEUTIC TREATMENTS. THIS 4-YEAR PROJECT WILL PURSUE THE FOLLOWING SPECIFIC AIMS, CONSISTENT WITH THE EXPECTATIONS IN PAR-20-271: AIM 1 GENERATE GCDH PROTEIN, OPTIMIZE CONDITIONS AND PERFORM SCREENING TO IDENTIFY COMPOUNDS TARGETING GCDH. OPTIMIZE TERTIARY ASSAYS FOR HIT VALIDATION. PRIMARY ASSAYS WILL TARGET BINDING ACTIVITY AND INHIBITION, AND TERTIARY ASSAYS WILL MONITOR CELLULAR TARGET ENGAGEMENT AND PROTEIN LEVEL. AIM 2 PERFORM HIT SELECTION, CONFIRMATION AND PROFILING USING A PANEL OF SECONDARY ASSAYS. FUNCTIONAL HIT PROFILES WILL BE ESTABLISHED USING INHIBITION ASSAYS FOR A REPRESENTATIVE OF ACYL-COA DEHYDROGENASES AND MECHANISM OF ACTION STUDIES BY USING PROTEIN THERMAL SHIFT WITH SUBSTRATE ANALOGS AND ENZYME KINETIC STUDIES TO ESTABLISH COMPETITION PROFILES WITH THE SUBSTRATES. AIM 3 PERFORM HIT VALIDATION AND SCAFFOLD PRIORITIZATION USING BIOCHEMICAL AND CELLULAR ASSAYS. GCDH TARGET ENGAGEMENT WILL BE CONFIRMED IN AN IN-CELL PROTEIN STABILITY ASSAY. EFFECTS ON CELLULAR PROTEIN LEVELS WILL BE EVALUATED USING ENDOGENOUS-TAGGED GCDH IN MELANOMA CELLS. BEST SCAFFOLDS WILL BE VALIDATED IN CELLULAR ASSAYS TO MONITOR APOPTOSIS, CELL VIABILITY, AND SPECIFICITY TOWARDS MELANOMA AND PANCREATIC CANCER.

LYMPHOTOXIN ALPHA BETA AND LIGHT CYTOKINE SYSTEMS

(PQD1) TREATMENT-EMERGENT RESISTANCE TO NEDD8-ACTIVATING ENZYME INHIBITION

IDENTIFYING GENES IN BETA-AMYLOID PRODUCTION

CONTROL OF PROTEIN SYNTHESIS BY THE UPS UNDER STRESS

LEAD OPTIMIZATION OF NOVEL INHIBITORS OF THE THIOESTERASE DOMAIN OF FASN

CYTOPLASMIC CHROMATIN FRAGMENTS IN CELL SENESCENCE - NOVEL MECHANISMS AND INTERVENTIONS - PROJECT SUMMARY CELLULAR SENESCENCE IS A CAUSE OF CELL AND TISSUE AGING. SENESCENCE IS CAUSED BY A RANGE OF CELLULAR STRESSES AND CHARACTERIZED BY AN IRREVERSIBLE PROLIFERATION ARREST AND A POTENT PRO-INFLAMMATORY PHENOTYPE, THE SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP). SENESCENCE-ASSOCIATED PROLIFERATION ARREST AND SASP COOPERATE IN TUMOR SUPPRESSION, BY ARRESTING PROLIFERATION OF DAMAGED PRE-MALIGNANT CELLS AND PROMOTING IMMUNE CLEARANCE OF THE DAMAGED CELLS. HOWEVER, OVER THE LONGER TERM, AS A SOURCE OF CHRONIC INFLAMMATION, SASP ALSO PROMOTES TISSUE AGING AND DISEASE. CONSEQUENTLY, THERE IS CURRENTLY MUCH EFFORT DEVOTED TO DEVELOPMENT OF PHARMACOLOGIC APPROACHES TO ELIMINATE SENESCENT CELLS TO PROMOTE HEALTHY AGING. HOWEVER, THESE SO-CALLED SENOLYTIC DRUGS TEND TO SHOW UNWANTED TOXICITIES. AN ALTERNATIVE, PERHAPS LESS TOXIC APPROACH, IS TO USE SENOMORPHIC DRUGS TO SPECIFICALLY INHIBIT THE PRO-AGING SASP. IMPORTANTLY, INHIBITION OF SASP DOES NOT NECESSARILY IMPAIR THE TUMOR SUPPRESSIVE ROLE OF SENESCENCE. ON THE CONTRARY, ELIMINATION OF SASP CAN PREVENT CANCER. HENCE, IT IS IMPORTANT TO DEFINE THE MECHANISM OF SASP ACTIVATION, BECAUSE ITS INHIBITION MAY BE AN APPROACH TO COMBAT THE PRO-AGING EFFECTS OF SENESCENT CELLS. RECENTLY, WE SHOWED THAT SENESCENT CELLS SHED FRAGMENTS OF NUCLEAR CHROMATIN INTO THE CYTOPLASM, CYTOPLASMIC CHROMATIN FRAGMENTS (CCF), VIA A NUCLEUS-TO-CYTOPLASMIC BLEBBING PROCESS. CCF ARE VERY STRONGLY POSITIVE FOR A DNA DAMAGE MARKER, PHOSPHORYLATED HISTONE H2AX. FORMATION OF CCF DEPENDS UPON A NOVEL MANIFESTATION OF THE CELLULAR RECYCLING PROCESS AUTOPHAGY, SPECIFICALLY NUCLEAR AUTOPHAGY. CCF SIGNAL THROUGH CYTOPLASMIC DNA SENSORS TO ACTIVATE NFB, THE MAJOR TRANSCRIPTIONAL ACTIVATOR OF SASP. MOST RECENTLY, WE HAVE DEFINED AN UNANTICIPATED UPSTREAM TRIGGER OF CCF AND SASP, NAMELY DYSFUNCTIONAL MITOCHONDRIA IN SENESCENT CELLS. DYSFUNCTIONAL MITOCHONDRIA ARE THEMSELVES ALREADY LINKED TO CHRONIC INFLAMMATION AND AGING. WE HYPOTHESIZE THAT NUCLEAR EXPULSION OF CCF HARBORING DNA DOUBLE STRAND BREAKS IS A MECHANISM FOR SENESCENT CELLS WITH IMPAIRED DNA REPAIR PATHWAYS TO DECREASE THE INTRANUCLEAR LOAD OF TOXIC DNA DOUBLE STRAND BREAKS. WE ALSO HYPOTHESIZE THAT DYSFUNCTIONAL MITOCHONDRIA TRIGGER AUTOPHAGY-DEPENDENT FORMATION OF CCF IN SENESCENT CELLS BY RETROGRADE MITOCHONDRIA-TO-NUCLEUS SIGNALING, INVOLVING JNK KINASE AND THE TRANSCRIPTION FACTORS HYPOXIA INDUCIBLE FACTOR 1A (HIF1A), THE “JNK-CCF” AND “HIF-CCF” PATHWAYS, RESPECTIVELY. DELINEATION OF THE JNK-CCF AND HIF-CCF PATHWAYS AND THEIR INTERACTIONS WILL PROVIDE NEW OPPORTUNITIES TO INTERVENE TO SUPPRESS CHRONIC INFLAMMATION DRIVEN BY SASP IN VIVO, THEREBY POTENTIATING HEALTHY AGING AND LONGEVITY.

SHOTGUN LIPIDOMICS AND ALTERATIONS IN SPHINGOLIPIDOMES IN ALZHEIMER'S DISEASES

CENTER FOR CANCER BIOLOGY RESEARCH

FUNCTION AND MECHANISM OF THE HIPPO-PI5P4K AXIS FOR GROWTH CONTROL - PROJECT SUMMARY OUR LONG-TERM GOAL IS TO DECIPHER THE MOLECULAR MECHANISMS UNDERLYING PI5P4K-DRIVEN GROWTH AND TO ELUCIDATE HOW THEIR DYSREGULATION CONTRIBUTES TO BREAST TUMORIGENESIS. IN THIS PROPOSAL, WE WILL SET OUT TO PROVIDE A COMPREHENSIVE UNDERSTANDING OF A NEW SIGNALING NETWORK THAT IS CRITICAL TO INTEGRATE METABOLIC INFORMATION AND DETERMINE HOW THE RELATED PROTEINS AND PATHWAYS INTERACT TO MAINTAIN CELLULAR HOMEOSTASIS. PHOSPHATIDYLINOSITOL-5-PHOSPHATE 4-KINASES (PI5P4KS) ARE A FAMILY OF NON-CANONICAL PHOSPHOINOSITIDE KINASES WHICH WE HAVE SHOWN TO BE CRITICAL FOR PRESERVING NUTRIENT AVAILABILITY AND FUEL GROWTH YET HOW THEY ARE REGULATED THEMSELVES IS LARGELY UNKNOWN. USING A COMPREHENSIVE IN VITRO KINASE SCREEN, WE HAVE DISCOVERED THE CORE KINASES OF THE HIPPO PATHWAY, MST1 AND MST2 AS DIRECT NEGATIVE REGULATORS OF THE PI5P4KS AS WELL AS THE LIPID SUBSTRATE OF THE PI5P4KS, PI-5-P TO INTERACT WITH THE HIPPO ADAPTOR PROTEIN MOB1 WHICH IN TURN EFFECTS THE ACTIVITY OF THE HIPPO DOWNSTREAM EFFECTOR YAP. THE HIPPO PATHWAY IS KNOWN TO BE IMPORTANT FOR DEVELOPMENT, GROWTH AND ORGANOGENESIS, AND DYSREGULATION OF THIS PATHWAY LEADS TO TUMOR PROGRESSION. WHILE PI5P4KS HAVE BEEN LINKED TO TUMOR GROWTH, EXACTLY HOW THE PI5P4KS ARE REGULATED IS POORLY UNDERSTOOD. FURTHERMORE, HOW THE PI5P4KS ACTUALLY COMMAND CELL GROWTH UNDER BOTH PHYSIOLOGICAL CONDITIONS AND PATHOLOGICAL CONTEXTS SUCH AS BREAST CANCER IS INCOMPLETE. TO ANSWER THESE IMPORTANT QUESTIONS, WE WILL PERFORM THE FOLLOWING AIMS: IN AIM 1 WE WILL USE VERY FEASIBLE BIOPHYSICAL AND STRUCTURAL BIOLOGY TO DETERMINE ATOMIC-RESOLUTION DETAILS OF THE INTERFACE BETWEEN PI-5-P AND MOB1 AND QUANTITATE HOW PI-5-P REGULATES MOB1 BINDING TO OTHER HIPPO PATHWAY COMPONENTS IN VITRO. IN AIM 2 WE WILL INVESTIGATE HOW THE ACTIVITY OF THE PI5P4KS CONTROL THE ACTIVATION OF THE MOB-LATS COMPLEX DOWNSTREAM OF MST1/2, EXPLORING ONE POTENTIAL MECHANISM FOR PI5P4K TUMORIGENIC PROPERTIES. IN AIM 3 WE WILL ESTABLISH THE BIOLOGICAL SIGNIFICANCE OF THE PI5P4K-HIPPO AXIS IN BREAST CANCER USING A PANEL OF TNBC CELL LINES, NOVEL MOUSE BREAST TUMOR MODELS, AND PATIENT BREAST TUMOR SAMPLES. THIS PROJECT SEEKS TO COMBINE QUANTITATIVE STRUCTURAL BIOLOGY WITH MOLECULAR CELL GENETICS TO DECIPHER HOW PI-5- P REGULATES THE HIPPO PATHWAY FOR BREAST CANCER CELL GROWTH. GIVEN THAT PI5P4KS ARE EXCITING ‘DRUGGABLE’ TARGETS FOR BREAST CANCER, CHARACTERIZING HOW PI5P4KS CONNECT TO THE HIPPO PATHWAY REVEALS A POSSIBLE STRATEGY FOR FUTURE ANTI-CANCER THERAPIES.

COLLAGEN FIBERS AND FRAGMENTS REGULATE PANCREATIC CANCER METABOLISM AND GROWTH. - PROJECT SUMMARY PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) IS CHARACTERIZED BY EXTENSIVE DESMOPLASIA, RESULTING IN OCCUPANCY OF MOST OF THE TUMOR MASS BY A DENSE STROMA CONSISTING OF FIBROBLASTS, MACROPHAGES, AND EXTRACELLULAR MATRIX (ECM). FIBRILLAR COLLAGENS, SUCH AS TYPE I COLLAGEN (COL I), ARE MAJOR ECM COMPONENTS AND WERE REPORTED TO EXERT POORLY UNDERSTOOD STIMULATORY AND INHIBITORY EFFECTS ON PDAC GROWTH AND MALIGNANCY. IT WAS ALSO OBSERVED THAT PATIENTS WHOSE STROMA IS FIBROLYTIC, WITH A HIGH CONTENT OF CLEAVED COLLAGEN FRAGMENTS GENERATED BY MATRIX METALLOPROTEASES (MMP), SHOW POOR OVERALL SURVIVAL AFTER RESECTION, WHEREAS PATIENTS WITH AN INERT STROMA, RICH IN INTACT COLLAGEN FIBERS, HAVE CONSIDERABLY IMPROVED SURVIVAL. THE MECHANISMS UNDERLYING THESE CLINICAL OBSERVATIONS AND THE TUMOR STIMULATORY OR INHIBITORY EFFECTS OF COL I WERE HERETOFORE UNKNOWN. USING A UNIQUE CULTURE SYSTEM, PRECLINICAL MODELS, AND CLINICAL SPECIMENS, WE DISCOVERED THAT INTACT (I) COL I FIBERS AND MMP- GENERATED CLEAVED (C) COL I FRAGMENTS HAVE DIAMETRICALLY OPPOSED EFFECTS ON PDAC METABOLISM AND GROWTH DUE TO THEIR DIFFERENTIAL INTERACTIONS WITH THE COLLAGEN RECEPTOR DDR1. WHEREAS CCOL I (AND OTHER MMP-CLEAVED COLLAGENS) ACTIVATES DDR1 AND ITS DOWNSTREAM EFFECTORS, WHICH INCLUDE TRANSCRIPTION FACTORS NF-KB AND NRF2, TO STIMULATE PDAC METABOLISM, MITOCHONDRIAL BIOGENESIS, AND RESPIRATION, ICOL I DOES NOT ACTIVATE DDR1 AND INSTEAD INDUCES ITS UBIQUITIN-DEPENDENT PROTEASOMAL DEGRADATION, THEREBY INHIBITING NRF2 ACTIVATION AND PDAC METABOLISM. OUR GOAL IS TO UNDERSTAND HOW MMP CLEAVAGE ENABLES THE BINDING OF CCOL I TO DDR1 AND DETERMINE IF BINDING TO DDR1 IS REQUIRED FOR INDUCTION OF ITS DEGRADATION BY ICOL I. TO FULLY UNDERSTAND HOW CCOL I AND DDR1 SIGNALING CONTROL PDAC METABOLISM AND GROWTH, AND WHY COL I ABLATION IS NOT AS INHIBITORY AS PREVENTION OF COL I CLEAVAGE, WE WILL TEST WHETHER THE DIFFERENTIAL EFFECTS OF CCOL I AND ICOL I ON DDR1 EXPRESSION AND ACTIVATION APPLY TO OTHER FIBRILLARY COLLAGENS (COL III) AND DEFINE THE UNDERLYING BIOCHEMICAL MECHANISMS THAT ENABLE DDR1 BINDING. NEXT, WE WILL INVESTIGATE HOW ICOL I TRIGGERS DDR1 UBIQUITINATION AND PROTEASOMAL DEGRADATION, AND THE ROLE OF THE NEWLY UNCOVERED DDR1 E3 LIGASE FBXW2 IN THIS PROCESS. THESE STUDIES WILL INCLUDE IDENTIFICATION OF THE MECHANISM BY WHICH ICOL I STIMULATES DDR1 UBIQUITINATION BY FBXW2 AND DEVELOPMENT OF LIGASE-RECRUITING TOOL COMPOUNDS THAT INDUCE DDR1 DEGRADATION, WHOSE BIOLOGICAL EFFECTS WILL BE EVALUATED IN CULTURED CELLS AND MOUSE MODELS. THIS WILL BE FOLLOWED BY STUDYING THE ROLE OF INDIVIDUAL MMP ISOZYMES, EXPRESSED IN PDAC CANCER CELLS, CANCER-ASSOCIATED FIBROBLASTS, AND MACROPHAGES, IN COLLAGEN REMODELING AND PDAC METABOLISM AND GROWTH. WE WILL DETERMINE THE SUITABILITY OF INDIVIDUAL COLLAGEN CLEAVING MMPS AS DRUG TARGETS FOR THE TREATMENT OF CCOL I ENRICHED PDAC. WE WILL ALSO DETERMINE THE MECHANISMS RESPONSIBLE FOR MMP INDUCTION IN THE PDAC MICROENVIRONMENT, AS THEY MAY PROVIDE ADDITIONAL THERAPEUTIC TARGETS. THE PROPOSED STUDIES WILL EMPLOY CUTTING- EDGE TECHNOLOGIES, INNOVATIVE ECM BASED PDAC CULTURES, NEW GENETICALLY MODIFIED MOUSE STRAINS THAT ALLOW THE EVALUATION OF BOTH PRIMARY AND LIVER METASTATIC PDAC AND CLINICAL SPECIMENS.

MECHANISMS OF CARDIOVASCULAR REMODELING

STRUCTURE AND FUNCTION OF BCAR1 AND BCAR3 ASSOCIATION IN BREAST CANCER MALIGNANCY

LEAD OPTIMIZATION OF NOVEL INHIBITORS OF TISSUE NON-SPECIFIC ALKALINE PHOSPHATASE

JIP MIMICS FOR THE TREATMENT OF DIABETES

APOPTOSIS-BASED CANCER DRUG DISCOVERY

NOVEL INSIGHTS IN THE REGULATION OF HIF1ALPHA STABILITY

MOLECULAR PATHOGENESIS OF AF10-REARRANGED LEUKEMIAS - PROJECT SUMMARY THERE HAS BEEN CONSIDERABLE PROGRESS IN THE LAST FEW YEARS IN THE CHARACTERIZATION OF MOLECULAR ALTERATIONS IN CHILDHOOD CANCER. MANY DEFINING GENOMIC ABNORMALITIES AS WELL AS TRANSCRIPTIONAL AND SIGNALING NETWORKS INVOLVED IN SPECIFIC SUB-TYPES OF PEDIATRIC CANCERS HAVE BEEN IDENTIFIED. HOWEVER, DESPITE THESE IMPRESSIVE ADVANCES, SAFE AND EFFECTIVE THERAPIES FOR MOST PEDIATRIC CANCERS ARE LACKING. LEUKEMIAS ARE THE LEADING CAUSE OF CANCER-LINKED MORTALITY IN CHILDREN AND FOR MANY PEDIATRIC LEUKEMIAS, THERAPIES THAT CAN SELECTIVELY ELIMINATE CANCER CELLS WITH FEW UNDESIRABLE SIDE EFFECTS HAVE BEEN ELUSIVE. THEREFORE, THERE IS AN URGENT CLINICALLY UNMET NEED TO DEVELOP NON-TOXIC AND TARGETED THERAPIES FOR PEDIATRIC LEUKEMIA. OUR PROPOSAL IS FOCUSED ON LEUKEMIAS WITH REARRANGEMENTS OF THE AF10 GENE. THESE REARRANGEMENTS ARE OBSERVED IN ACUTE MYELOID LEUKEMIA (AML), T-CELL ACUTE LYMPHOBLASTIC LEUKEMIA (T-ALL) AND OTHER SUB-TYPES OF LEUKEMIA IN CHILDREN AND ADULTS. AF10-REARRANGED (AF10-R) LEUKEMIAS CAN BE CONSIDERED PROTOTYPICAL MODELS OF PEDIATRIC CANCERS IN WHICH CHIMERIC FUSION ONCOPROTEINS OR ALTERED TRANSCRIPTION FACTORS DRIVE EXTENSIVE SELF- RENEWAL AND IMPAIRED DIFFERENTIATION OF CANCER CELLS. OUR RECENT ANALYSIS OF DATA FROM OVER 1,000 PEDIATRIC AML PATIENTS DEMONSTRATES THAT AF10 GENE FUSIONS ARE MUCH MORE COMMON THAN GENERALLY APPRECIATED (REPRESENTING 6.5% OF PEDIATRIC AML) AND CONSTITUTE ONE OF THE HIGHEST RISK-GROUPS IN TERMS OF RESISTANCE TO PRIMARY THERAPIES AND RELAPSE RATES. OUR DETAILED MECHANISTIC STUDIES DEMONSTRATE THAT THE AF10 FUSION PROTEIN CALM-AF10 ACTIVATES A NUMBER OF PATHWAYS LINKED TO STEM CELL SELF-RENEWAL AND INFLAMMATORY SIGNALING, MANY OF WHICH ARE INDEPENDENTLY ASSOCIATED WITH POOR OUTCOMES IN AML. IN THIS PROPOSAL, WE WILL USE A SERIES OF ORTHOGONAL METHODOLOGIES, INCLUDING PROTEOMIC, GENOMIC, EPIGENOMIC AND DETAILED MECHANISTIC STUDIES IN BOTH HUMAN AND MOUSE AML MODELS, TO DETERMINE HOW THE CALM-AF10 FUSION PROTEIN CAUSES LEUKEMOGENESIS. THE SUCCESSFUL EXECUTION OF THE PROPOSED AIMS WILL ADVANCE OUR EFFORTS TOWARDS FINDING NEWER TARGETED THERAPIES FOR AF10-R AML WITH FEWER UNWANTED TOXICITIES. RESULTS FROM OUR STUDIES ALSO HAVE THE POTENTIAL TO SIGNIFICANTLY INFLUENCE TREATMENT STRATEGIES IN SEVERAL OTHER SUB-TYPES OF PEDIATRIC LEUKEMIAS DRIVEN BY RELATED MECHANISMS.

THE P62/MEKK3 COMPLEX IN MTORC1 ACTIVATION

NOVEL PATHWAYS AND TECHNOLOGIES TOWARD BROAD-SPECTRUM THERAPEUTICS

PROTEIN KINASE CZ TARGETS IN INTESTINAL CANCER STEM CELLS

NON-CANONICAL PHOSPHATIDYLINOSITOL KINASES IN TRIPLE NEGATIVE BREAST CANCER

TARGETING ER+ BREAST CANCER THROUGH INDUCED VIRAL MIMICRY - PROJECT SUMMARY REPETITIVE ELEMENTS (RES) COMPOSE ~45% OF THE HUMAN GENOME AND ARE NORMALLY TRANSCRIPTIONALLY SILENCED IN SOMATIC CELLS, ALTHOUGH THE MECHANISM HAD REMAINED ELUSIVE. THROUGH A HIGH-CONTENT RNAI SCREEN, WE IDENTIFIED THE LARGELY UNCHARACTERIZED PROTEIN FBXO44 AS AN ESSENTIAL REPRESSOR OF RES IN BREAST CANCER CELLS. FBXO44 BOUND REPRESSIVE HISTONE H3 LYSINE 9 TRIMETHYLATED (H3K9ME3) NUCLEOSOMES AT THE REPLICATION FORK AND RECRUITED H3K9ME3 METHYLTRANSFERASE SUV39H1, UBIQUITIN LIGASE CRL4RBBP4/7, AND HISTONE DEACETYLASE AND CHROMATIN- REMODELING COMPLEX MI-2/NURD TO TRANSCRIPTIONALLY SILENCE RES POST-DNA REPLICATION. FBXO44/SUV39H1 INHIBITION TRANSCRIPTIONALLY REACTIVATED ENDOGENOUS RETROVIRUSES (ERVS) AND RETROTRANSPOSONS (E.G. ALU, LINE-1) IN BREAST CANCER CELLS, LEADING TO EXTENSIVE DNA REPLICATION STRESS AND STIMULATION OF RIG-I/MDA5-MAVS AND CGAS-STING INTRACELLULAR ANTIVIRAL PATHWAYS TO PROMOTE ENHANCED IMMUNOGENICITY AND DECREASED TUMORIGENICITY. IN SILICO ANALYSIS REVEALED THE FBXO44/SUV39H1 PATHWAY INVERSELY CORRELATED WITH DNA REPLICATION STRESS, ANTIVIRAL PATHWAYS, AND CYTOTOXIC T AND NATURAL KILLER (NK) CELL INFILTRATION IN HUMAN BREAST TUMORS. IMPORTANTLY, FBXO44/SUV39H1 WERE FOUND DISPENSABLE FOR RE SILENCING IN NORMAL CELLS AND THEIR INHIBITION HAD NO EFFECT ON H3K9ME3 LEVELS, DNA REPLICATION STRESS, OR VIABILITY, SUGGESTING A THERAPEUTIC WINDOW. OUR HYPOTHESIS IS THAT FBXO44/SUV39H1-MEDIATED RE SILENCING IS AN EPIGENETIC VULNERABILITY OF BREAST CANCER CELLS THAT COULD BE TARGETED TO INHIBIT TUMOR GROWTH/PROGRESSION AND ENHANCE THE EFFICACY OF CERTAIN ANTITUMOR THERAPIES THROUGH THE UNIQUE MECHANISM OF INDUCED VIRAL MIMICRY. IN THIS PROPOSAL, WE WILL EVALUATE 3 POTENTIAL THERAPEUTIC APPLICATIONS OF FBXO44/SUV39H1 PATHWAY TARGETING IN THE TREATMENT OF ESTROGEN RECEPTOR (ER)+ BREAST CANCERS BASED ON OUR PRELIMINARY DATA: 1) PREVENTION OF BONE METASTASIS RELAPSE THROUGH STIMULATION OF NK CELL RECOGNITION AND KILLING OF DORMANT BREAST CANCER CELLS; 2) ENHANCEMENT OF IMMUNE CHECKPOINT BLOCKADE (ICB) THERAPY THROUGH STIMULATION OF IFN SIGNALING AND INTRATUMORAL INFILTRATION OF CYTOTOXIC T CELLS; AND 3) SYNERGY WITH PARP INHIBITORS THROUGH INDUCTION OF DNA REPLICATION STRESS AND DOUBLE-STRAND BREAKS (DSBS) AT RES. THESE STUDIES COULD LEAD TO THE DEVELOPMENT OF A SAFE AND EFFECTIVE THERAPEUTIC APPROACH THAT SELECTIVELY INDUCES VIRAL MIMICRY IN ER+ BREAST CANCER CELLS TO PREVENT BONE METASTASIS RELAPSE AND ENHANCE THE EFFICACY OF ICB AND PARP INHIBITOR THERAPIES, UNDOUBTEDLY LEADING TO A SIGNIFICANT REDUCTION IN DISEASE MORTALITY.

TARGETING THE FBXO44/SUV39H1 PATHWAY IN CANCER - PROJECT SUMMARY REPETITIVE ELEMENTS (RES) COMPOSE ~45% OF THE HUMAN GENOME AND ARE NORMALLY TRANSCRIPTIONALLY SILENCED, ALTHOUGH THE MECHANISM HAS REMAINED ELUSIVE. THROUGH A HIGH-CONTENT RNAI SCREEN, WE IDENTIFIED FBXO44 AS AN ESSENTIAL REPRESSOR OF RES IN CANCER CELLS. FBXO44 BOUND REPRESSIVE H3K9ME3-MODIFIED NUCLEOSOMES AT THE REPLICATION FORK AND RECRUITED H3K9ME3 METHYLTRANSFERASE SUV39H1, UBIQUITIN LIGASE CRL4RBBP4/7, AND HISTONE DEACETYLASE AND CHROMATIN-REMODELING COMPLEX MI-2/NURD TO TRANSCRIPTIONALLY SILENCE RES POST-DNA REPLICATION. FBXO44/SUV39H1 INHIBITION TRANSCRIPTIONALLY ACTIVATED SATELLITE REPEATS AND ENDOGENOUS RETROVIRUSES AND RETROTRANSPOSONS IN CANCER CELLS, LEADING TO DNA REPLICATION STRESS AND STIMULATION OF MAVS AND STING INTRACELLULAR ANTIVIRAL PATHWAYS TO PROMOTE DECREASED TUMORIGENICITY AND ENHANCED IMMUNOTHERAPY RESPONSE. IN SILICO ANALYSIS REVEALED THAT FBXO44 EXPRESSION INVERSELY CORRELATED WITH DNA REPLICATION STRESS, ANTIVIRAL PATHWAYS, AND CYTOTOXIC T AND NATURAL KILLER (NK) CELL INFILTRATION IN HUMAN CANCERS. IMPORTANTLY, WE FOUND THAT FBXO44/SUV39H1 ARE DISPENSABLE FOR RE SILENCING IN NORMAL CELLS AND THEIR INHIBITION DID NOT AFFECT H3K9ME3 LEVELS AT RES OR CELL VIABILITY. OUR HYPOTHESIS IS THAT FBXO44/SUV39H1-MEDIATED RE ELEMENT SILENCING IS AN EPIGENETIC VULNERABILITY OF CANCER CELLS THAT COULD POTENTIALLY BE TARGETED TO INDUCE VIRAL MIMICRY RESPONSES THAT INHIBIT TUMOR GROWTH AND PROGRESSION AND ENHANCE THE EFFICACY OF CERTAIN CANCER THERAPIES. IN THIS PROPOSAL, WE WILL PERFORM PRECLINICAL STUDIES THAT EVALUATE TWO POTENTIAL THERAPEUTIC APPLICATIONS OF FBXO44/SUV39H1 PATHWAY TARGETING IN CANCER: 1) PREVENTION OF METASTATIC RELAPSE THROUGH STIMULATION OF INTRACELLULAR ANTIVIRAL PATHWAYS AND NK CELL RECOGNITION; AND 2) ENHANCEMENT OF PARP INHIBITOR EFFICACY THROUGH INDUCTION OF EXTENSIVE DNA REPLICATION STRESS AT RES. MOREOVER, WE WILL INVESTIGATE THE ROLE OF THE CRL4RBBP4/7 UBIQUITIN LIGASE IN FBXO44/SUV39H1-MEDIATED RE SILENCING AND EVALUATE ITS TARGETING FOR CANCER TREATMENT. THESE STUDIES COULD UNCOVER A TARGETABLE EPIGENETIC VULNERABILITY OF CANCER CELLS WHOSE INHIBITION INDUCES VIRAL MIMICRY TO PREVENT TUMOR GROWTH AND PROGRESSION AND ENHANCE THE EFFICACY OF CANCER THERAPEUTICS, UNDOUBTEDLY LEADING TO A SIGNIFICANT REDUCTION IN DISEASE MORTALITY.

GENETIC DISSECTION OF THE ROLE OF CHONDROITIN SULFATE IN CARTILAGE

SANFORD BISMARCK HEALTHY TRANSITIONS - SANFORD HEALTHY TRANSITION PROJECT IS LED BY THE SANFORD EAST DICKINSON CLINIC REPRESENTING AN ORGANIZED COUNCIL OF COMMUNITY PARTNERS. SANFORD AND PARTNERS SHARE IN THE MISSION OF INCREASING ACCESS AND REDUCING OBSTACLES REGARDING COMMUNITY SUPPORT AND BEHAVIORAL HEALTH SERVICES IN SOUTHWEST NORTH DAKOTA. THE SANFORD HEALTHY TRANSITION PROJECT PLANS TO INCREASE ACCESS TO TRANSITION SERVICES UTILIZING EVIDENCE-BASED PRACTICES FOR TRANSITION-AGED YOUTH (16-25) AND THEIR FAMILIES. SANFORD WILL PROVIDE A SUSTAINABLE MODEL OF SERVICE THAT WILL ALLOW FOR LONG-TERM ACCESS TO PROJECT SERVICES. WITH HELP FROM STATE AND LOCAL-COMMUNITY PARTNERS, SANFORD WILL IMPLEMENT A REFERRAL NETWORK TO PROVIDE ACCESS TO RELEVANT TRANSITION SERVICES FOR THEIR RELEVANT TARGET POPULATIONS. SERVICES WILL PRIMARILY IMPACT UNDERSERVED RURAL COMMUNITIES THAT HAVE LITTLE TO NO ACCESS TO THE CURRENT LEVEL OF CARE. GOALS AND OBJECTIVES GOAL 1. CREATE A SYSTEM OF CARE COORDINATION BETWEEN BEHAVIORAL HEALTH AND OTHER EXISTING SYSTEMS – MEDICAL, EDUCATION, AND LAW ENFORCEMENT. 1. ESTABLISHING MONTHLY ADVISORY COUNCIL MEETINGS TO DISCUSS PROJECT NEEDS. 2. IDENTIFYING AND SCREENING AT RISK YOUTH/YOUNG ADULTS WITH SERIOUS MENTAL ILLNESS AND/OR EMOTIONAL DISTURBANCE. 3. HIRING AND TRAINING CARE COORDINATORS AND CLINICIANS TO IMPLEMENT PROJECT FUNCTIONS. 4. HAND-OFF OF TRANSITIONED AGED YOUTH FROM YOUTH-BASED AGENCIES. 5. IMPLEMENT SUICIDE PREVENTION AND INTERVENTION PROGRAMS. GOAL 2. CREATE A SCHOOL-BASED YOUTH MENTORSHIP SUPPORT PROGRAM TO ENCOURAGE STAYING IN SCHOOL TO ATTAIN A HIGH SCHOOL DIPLOMA, BASED ON THE CHECK & CONNECT PROGRAM. 1. TRAIN BEHAVIORAL HEALTH PROVIDERS ON THE DELIVERY OF EVIDENCE-BASED PRACTICES. 2. CHECK & CONNECT EBT SITE VISIT. 3. RECRUITMENT AND TRAINING OF MENTORS TO ASSIST IN REFERRALS. 4. PROVIDING HIGH SCHOOL YOUTH MENTORSHIP SERVICES GOAL 3. CREATE AN INTEGRATED SYSTEM OF COMMUNITY SUPPORT FOR YOUNG ADULTS WHO ARE OUT OF HIGH SCHOOL TO SUCCESSFULLY MANAGE THE MULTIPLE AREAS OF HEALTHY ADULTHOOD – EDUCATION, VOCATIONAL, HEALTH AND BEHAVIORAL HEALTH, FINANCIAL, AND LIFESTYLE. 1. COMMUNICATE WITH ADVISORY COUNCIL MEMBERS AND LOCAL AGENCIES FOR A. HOUSING ASSISTANCE ADULT EDUCATION, VOCATIONAL REHABILITATION, TECHNICAL TRAINING, AND FINANCIAL MANAGEMENT SKILLS TRAINING 2. ADVANCED TRAINING FOR CARE COORDINATORS IN THE YOUTHBUILD EBT TO ASSIST IN PROVIDING THE ABOVE SERVICES.