Augusta University Research Institute Inc

COORDINATING AND BIOINFORMATICS UNIT FOR THE AMDCC/MMPC

AGE-INDUCED IMPAIRMENT OF NUTRIENT SIGNALING RESULTS IN BONE LOSS

IMMUNE CELL INTERACTIONS IN ATHEROSCLEROSIS

STRESS RELATED MECHANISMS OF HYPERTENSION RISK IN YOUTH

GEORGIA CARES

COORDINATING UNIT FOR DIACOMP

ENDOTHELIAL BARRIER PROTECTION AND REPAIR IN ACUTE LUNG INJURY

MANIPULATING NATURAL HOST IMMUNOREGULATION VIA IDO DURING VIRAL INFECTION

DAMAGE-ASSOCIATED MOLECULAR PATTERNS IN HYPERTENSION

THE ROLE OF ENDOTHELIN-1 IN SICKLE CELL DISEASE

THE TEDDY STUDY: GEORGIA/FLORIDA CLINICAL CENTER

REGULATION OF HEAT SHOCK FACTOR 1 IN ANIMAL MODELS

NCMHD SOUTHEASTERN EXPLORATORY SICKLE CELL CENTER OF EXCELLENCE

ENDOTHELIAL METABOLISM AND REDOX IMBALANCE IN VASCULAR DISEASE - PROJECT SUMMARY REDOX IMBALANCE BETWEEN REACTIVE OXYGEN SPECIES (ROS) AND REACTIVE NITROGEN SPECIES (RNS) (HIGH ROS AND LOW NITRIC OXIDE NO) IS A FUNDAMENTAL MECHANISM OF ENDOTHELIAL CELL (EC) DYSFUNCTION AND A DRIVER OF CARDIOVASCULAR DISEASES (CVD) SUCH AS ATHEROSCLEROSIS, HYPERTENSION AND PERIPHERAL ARTERIAL DISEASE (PAD). STRATEGIES TO INDIVIDUALLY MITIGATE ROS OR AMPLIFY NO HAVE EXPERIENCED LIMITED SUCCESS. THEREFORE, A GREATER UNDERSTANDING OF THE KEY FACTORS AND EVENTS THAT LEAD TO REDOX IMBALANCE IS THEREFORE ESSENTIAL TO DEVELOP MORE EFFECTIVE TREATMENTS FOR CARDIOVASCULAR DISEASE (CLASSICAL CONCEPT) WHICH REMAINS THE LEADING CAUSE OF MORBIDITY AND MORTALITY. IN ECS, AEROBIC GLYCOLYSIS (“WARBURG EFFECT”) HAS BEEN REGARDED AS THE DOMINANT METABOLIC PATHWAY SUPPORTING HOMEOSTATIC FUNCTIONS. AN EMERGING CONCEPT IS THAT THE PATHWAYS OF CELLULAR METABOLISM ARE NOT FIXED, AND THAT ECS CAN ALTER THEIR METABOLIC PATHWAYS OR “REPROGRAM” TO MEET DIFFERENT NEEDS. A MAJOR GAP IN OUR KNOWLEDGE IS HOW EC METABOLISM AND ROS/RNS BALANCE ARE ALTERED IN VASCULAR DISEASE AND HOW METABOLISM INFORMS FUNCTION. RECENT STUDIES HAVE SHOWN THAT AEROBIC GLYCOLYSIS IS INCREASED IN ECS OF ATHEROSCLEROTIC, DIABETIC AND ANGIOGENIC BLOOD VESSELS, HOWEVER, THE FUNCTIONAL CONSEQUENCES OF THIS DYNAMIC HAS YET TO BE ELUCIDATED. ENZYMES THAT PRODUCE ROS ARE NOT ONLY INFLUENCED BY METABOLISM, BUT MAY ALSO PROMOTE CHANGES IN METABOLISM. OUR PRELIMINARY DATA ADVANCE THE NOVEL HYPOTHESIS THAT IMBALANCE BETWEEN CHANGES IN EC METABOLISM AND ROS/RNS (REDOX BALANCE), INDUCED BY VARIOUS RISK FACTORS, IS A KEY DRIVER OF EC DYSFUNCTION AND AN UNDERLYING MECHANISM OF VASCULAR DISEASE. TO TEST THIS HYPOTHESIS, A PROGRAMMATIC APPROACH IS ESSENTIAL AS IT IS THE ONLY MECHANISM CAPABLE OF SUPPORTING THE RIGOR NEEDED TO INVESTIGATE THE COMPLEX INTERPLAY BETWEEN METABOLISM AND REDOX SIGNALING IN DIFFERENT VASCULAR BEDS AND DISEASE STATES. OUR PROGRAM HAS 3 WELL ALIGNED PROJECTS THAT BENEFIT FROM THE SYNERGY OF MODEL AND REAGENT SHARING AND DEDICATED CORES. PROJECT 1 IS FOCUSED ON THE HYPOTHESIS THAT DISTURBED COPPER (CU) METABOLISM IN ECS ARISES FROM DYSFUNCTION OF THE CU EXPORTER, ATP7A, LEADING TO EXCESSIVE PFKFB3/GLYCOLYSIS, ROS/NO IMBALANCE AND MITOCHONDRIAL ROS-EPIGENETIC REMODELING THAT DRIVES ENDMT AND ACCELERATED ATHEROSCLEROSIS. PROJECT 2 WILL TEST THE HYPOTHESIS THAT EC METABOLISM SHAPES EC FUNCTION VIA SEX-SPECIFIC, NOX1 AND LEPTIN-DEPENDENT MECHANISMS. PROJECT 3 TESTS THE HYPOTHESIS THAT ABILITY OF THE MITOCHONDRIAL DYNAMICS PROTEIN, DRP1, TO PROMOTE REPARATIVE ANGIOGENESIS VIA CROSSTALK BETWEEN EC MITOCHONDRIAL REDOX SIGNALING AND GLYCOLYSIS IS IMPAIRED IN DIABETES DUE TO EXCESS PFKFB3/GLYCOLYSIS-ROS SIGNALING WHICH INCREASES THE SEVERITY OF PAD. TO TEST THESE CONCEPTS DIRECTLY, WE HAVE DEVELOPED A STATE-OF-THE-ART APPROACH USING CRISPR/CAS9 TO GENERATE KNOCK-IN MICE ENABLING INDUCIBLE REDUCTION OF GLYCOLYSIS ONLY IN ECS, THAT WILL BE SHARED ACROSS PROJECTS TO ENHANCE SYNERGY. OUR PROGRAM WILL IDENTIFY KEY MECHANISMS THROUGH WHICH CHANGES IN METABOLISM IMPACT ROS/NO BALANCE, EC FUNCTION AND VASCULAR DISEASE.

COORDINATING UNIT FOR THE INNOVATIVE SCIENCE ACCELERATOR PROGRAM - THE GOAL OF THE NIDDK INNOVATIVE SCIENCE ACCELERATOR (ISAC) PROGRAM IS TO NOT ONLY PROVIDE FUNDING FOR HIGHLY INNOVATIVE HYPOTHESIS DRIVEN RESEARCH BUT TO DELIVER A PLATFORM FOR THE KUH RESEARCH COMMUNITY TO COLLABORATE, EXCHANGE IDEAS AND FIND RESOURCES THAT WILL ACCELERATE INNOVATION IN AREAS OF INTEREST TO THAT COMMUNITY. OUR LABORATORY HAS EXTENSIVE EXPERIENCE IN DEVELOPING THESE TYPES OF RESOURCES AND MANAGING THE ACTIVITIES OF NATIONAL CONSORTIA. WE ARE CURRENTLY THE COORDINATING UNITS (CU) FOR TWO NIDDK CONSORTIA, THE MOUSE METABOLIC PHENOTYPING CENTERS (MMPC) AND THE DIABETIC COMPLICATIONS CONSORTIUM (DIACOMP). BOTH CONSORTIA ARE NIDDK FUNDED INITIATIVES THAT PROVIDE AN ENVIRONMENT TO FOSTER COMMUNICATION AND COLLABORATION BETWEEN INVESTIGATOR COMMUNITIES INVOLVED IN METABOLIC AND DIABETIC COMPLICATIONS RESEARCH. EACH SPONSORS ANNUAL MEETINGS IN RELEVANT SCIENTIFIC AREAS, SOLICITS AND FUNDS PILOT PROJECTS IN HIGH IMPACT AREAS OF NIDDK RESEARCH, AND SUPPORTS WEBSITES TO SERVE THEIR RESPECTIVE SCIENTIFIC COMMUNITIES. OVER THE LAST 14 YEARS, OUR GROUP HAS DEVELOPED AN EXTENSIVE INFRASTRUCTURE TO SUCCESSFULLY MANAGED THE COORDINATING UNITS (CU) FOR THESE NIDDK FUNDED CONSORTIA (MMPC, 2006-PRESENT, DIACOMP, 2011-PRESENT). THE ISAC-CU WILL BE RESPONSIBLE FOR SUPPORTING ISAC INVESTIGATORS THROUGH THE USE OF OPPORTUNITY POOLS (FUNDING PROGRAMS), MANAGE/MAINTAIN THE WEBSITE/RESOURCES AND ADMINISTRATIVELY ORGANIZE THE MEETINGS AND WORKSHOPS SPONSORED BY ISAC TO ENGAGE THE GREATER KUH SCIENTIFIC COMMUNITY. THE ISAC-CU IS RESPONSIBLE FOR CREATING AND MAINTAINING THE ADMINISTRATIVE, SCIENTIFIC AND INFORMATICS INFRASTRUCTURE NECESSARY TO ORGANIZE AND FACILITATE ISAC’S OPERATIONS. THE GOAL OF THIS PROPOSAL IS TO PROVIDE THAT INFRASTRUCTURE. WE WILL BUILD UPON THE SUCCESS OF THE CURRENT CU INFRASTRUCTURE TO PROVIDE ISAC WITH A ROBUST AND COMPREHENSIVE SERVICE ORIENTED SOLUTION THAT SUPPORTS THE UNIQUE ASPECTS OF THIS CONSORTIUM.

IMPACT OF HIV-1 TAT PROTEIN ON COCAINE-DOPAMINE TRANSPORTER INTERACTION

APOPTOTIC GENE REGULATION IN RENAL PATHOLOGY

VASCULAR MACROPHAGES AND T CELLS IN ATHEROSCLEROSIS

ACUTE KIDNEY INJURY BY CISPLATIN AND RENOPROTECTIVE STRATEGIES

THE TEDDY STUDY: GEORGIA/FLORIDA CLINICAL CENTER

EXERCISE & OVERWEIGHT CHILDREN'S COGNITION

PRIDE-FUNCTIONAL AND APPLIED GENOMICS OF BLOOD DISORDERS

IMPLEMENTATION OF MEDICAL HOMES FOR EVIDENCE BASED CARE OF ADOLESCENTS AND ADULTS WITH SICKLE CELL DISEASE

GALECTIN-3: A MEDIATOR OF VASCULAR REMODELING IN PULMONARY ARTERIAL HYPERTENSION

CELLULAR MECHANISMS OF RETINAL ANGIOGENESIS

CALCULATOR FOR LENGTH OF USE OF BISPHOSPHONATES (CLUB) - OSTEOPOROTIC FRACTURES ARE A MAJOR PUBLIC HEALTH PROBLEM, ESPECIALLY FOR OLDER PERSONS. BISPHOSPHONATES (BPS) ARE AN INEXPENSIVE THERAPEUTIC CLASS OF DRUGS EFFECTIVE AT PREVENTING THESE FRACTURES. HOWEVER, THERE HAS BEEN A SUBSTANTIAL DECLINE IN USE OF BPS FOR OSTEOPOROSIS AT LEAST IN PART BECAUSE OF CONCERNS FOR RARE, ALBEIT POTENTIALLY SERIOUS SIDE EFFECTS, NAMELY ATYPICAL FEMORAL FRACTURES (AFF) AND BISPHOSPHONATE MEDICATION RELATED OSTEONECROSIS OF THE JAW (MRONJ). DURATION OF TREATMENT IS A RISK FACTOR FOR LONG-TERM SIDE EFFECTS FROM BPS, INCLUDING AFF, AND TO A LESSER EXTENT, MRONJ. TO ASSIST WITH DECISION-MAKING REGARDING DURATION OF USE OF BPS, THE FDA AND PROFESSIONAL SOCIETIES DEVELOPED GUIDELINES SUGGESTING THAT AFTER THREE TO FIVE YEARS OF ORAL OR THREE YEARS OF INTRAVENOUS BP USE, THAT REASSESSMENT OF FRACTURE RISK WITH POSSIBLE PROVISION OF A DRUG HOLIDAY BE DONE. HOWEVER, THESE RECOMMENDATIONS WERE BASED ON VERY LIMITED EVIDENCE LARGELY DERIVED FROM WHITE, HEALTHY, COMMUNITY-DWELLING POSTMENOPAUSAL WOMEN. MOREOVER, THE RISK ASSOCIATED WITH DRUG HOLIDAYS IS UNCERTAIN, AS EMERGING REPORTS SUGGEST THAT A NUMBER OF WOMEN HAVE SUSTAINED OSTEOPOROTIC FRACTURES WHILE OFF THERAPY. IT IS PARAMOUNT TO BETTER UNDERSTAND THE RISK-BENEFIT PROFILE OF LONG-TERM BP THERAPY AND DRUG HOLIDAYS, IN BOTH MEN AND WOMEN, ALL RACES AND ETHNICITIES, AND THOSE WITH SERIOUS COMORBIDITIES. TO ACCOMPLISH THIS, WE HAVE AMASSED AN EXPERT MULTIDISCIPLINARY TEAM TO STUDY LARGE AND DIVERSE PATIENT POPULATIONS RANGING FROM COMMUNITY DWELLING TO LONG-TERM CARE RESIDENTS FROM KAISER PERMANENTE (KP; NORTHERN CALIFORNIA, GEORGIA, AND MID-ATLANTIC REGIONS), HEALTH PARTNERS (HP; MINNESOTA), AND THE NATIONAL VETERANS HEALTH ADMINISTRATION (VHA), SPANNING 30 YEARS (1996 TO 2025) AND INCLUDING OVER HALF A MILLION OLDER U.S. ADULTS WHO INITIATED OSTEOPOROSIS TREATMENT WITH A BP. UTILIZING THIS POPULATION, WE WILL DETERMINE RISKS AND BENEFITS, HEALTH CARE COSTS AND COST EFFECTIVENESS OF UNINTERRUPTED CONTINUATION OF BPS VERSUS BP DRUG HOLIDAYS. THESE INFORMATION WILL BE UTILIZED TO DEVELOP A RISK FACTOR CALCULATOR TO DETERMINE IF A DRUG HOLIDAY SHOULD BE DONE (CALCULATOR FOR LENGTH OF USE OF BISPHOSPHONATES (CLUB)). AN EXPERT PANEL INCLUDING BOTH CLINICIAN AND CONSUMER VOICES WILL INFORM THE FINAL DESIGN OF CLUB. WE ANTICIPATE THAT CLUB WILL BE A CLINICALLY USEFUL POINT-OF-CARE TOOL TO GUIDE SHARED DECISION-MAKING ON BP USE AND OPTIMIZE CARE FOR OLDER PATIENTS AT RISK FOR FRACTURE.

IDO-EXPRESSING IMMUNOREGULATORY DENDRITIC CELLS

GENETIC ANALYSIS OF MYELOPROLIFERATIVE DISEASE

IMPROVING AWARENESS OF WOMEN WITH HYPERTENSION: ROAR (RURAL, OBESE, AT RISK) - MORE THAN 1 IN 3 WOMEN ARE LIVING WITH HEART DISEASE AND MANY ARE UNAWARE OF THE RISKS. HYPERTENSION IS A MAJOR MODIFIABLE RISK FACTOR FOR CARDIOVASCULAR DISEASE (CVD), AND ~47% OF ADULTS IN THE US HAVE HYPERTENSION. IN ~85% OF CASES, THE CAUSE OF HYPERTENSION IS UNKNOWN AND ONLY ~50% OF PATIENTS TAKING MEDICATION ACHIEVE BLOOD PRESSURE (BP) CONTROL TO RECOMMENDED LEVELS. A CRITICAL BARRIER TO LIMITING PREMATURE DEATH FROM CVD IS LACK OF AWARENESS SURROUNDING THE RISKS OF CVD. OUR LOCATION IN THE SOUTHEAST OF THE US WILL CONTRIBUTE TO OUR IMPACT. THE PREVALENCE OF HYPERTENSION AND OBESITY ARE HIGH IN GA, MS, AND SC. THE PREVALENCE OF HYPERTENSION AND ASSOCIATED RISK IS NOT EQUAL AMONG ALL POPULATIONS. INDEED, THE PREVALENCE OF HYPERTENSION IN BLACK PEOPLE IN THE US IS AMONG THE HIGHEST IN THE WORLD. OUR OVERALL GOAL IS TO TRANSFORM ACADEMIC AND COMMUNITY UNDERSTANDING OF SEX AS A BIOLOGICAL VARIABLE (SABV) IN THE CONSIDERATION OF HYPERTENSION. WE WILL ACCOMPLISH THIS THROUGH THE CREATION AND DISSEMINATION OF INNOVATIVE EDUCATIONAL CONTENT, THE DEVELOPMENT OF STRATEGIC PARTNERSHIPS, AND ENHANCED COMMUNITY ENGAGEMENT IN THE SOUTHEAST OF THE US. THIS SCORE IS DESIGNED TO PROVIDE NOVEL INSIGHT INTO THE MECHANISMS CONTROLLING BP IN FEMALES VS. MALES, DEVELOP A COMPREHENSIVE EDUCATIONAL PLATFORM ON SABV IN CVD EXTENDING FROM GRADE SCHOOL TO MEDICAL SCHOOL, AND INCREASE AWARENESS AND EDUCATION OF CVD AND THE RISKS OF UNCONTROLLED HYPERTENSION IN WOMEN ACROSS THE HEALTH SPAN THROUGH COMMUNITY OUTREACH. THIS SCORE HAS 4 OBJECTIVES. OBJECTIVE 1 IS TO CONDUCT CUTTING EDGE RESEARCH TO ADVANCE OUR UNDERSTANDING OF SEX-SPECIFIC MECHANISMS THAT CONTROL BP TO LIMIT CARDIOVASCULAR RISK. OBJECTIVE 2 IS TO DEVELOP A CULTURALLY DIVERSE RESEARCH PIPELINE OF BIOMEDICAL SCIENTISTS TRAINED IN THE FIELD OF SABV IN CVD. OBJECTIVE 3 IS TO ESTABLISH A COMMUNITY OUTREACH PIPELINE OF TRAINED SCIENTISTS TO INCREASE AWARENESS OF SABV AND PROMOTE OPTIMAL CARDIOVASCULAR HEALTHCARE FOR MEN AND WOMEN. OBJECTIVE 4 IS TO EMPOWER THE COMMUNITY TO SELF-MONITOR AND WORK TO IDENTIFY BARRIERS TO IMPROVING ACCESS TO HEALTHCARE. THE OUTCOMES OF OUR PROPOSED SCORE INCLUDE NOT ONLY ADVANCES IN SCIENCE AND TRAINING OF THE NEXT GENERATION OF LEADERS IN THE STUDY OF THE IMPACT OF BIOLOGICAL SEX ON THE DEVELOPMENT AND PROGRESSION OF CVD, BUT ALSO THE DEVELOPMENT OF A RANGE OF EDUCATIONAL CONTENT THAT WILL BE FULLY AVAILABLE TO THE CONSORTIUM. THIS WILL INCLUDE 1) THE DEVELOPMENT EDUCATIONAL MATERIALS BY MEDICAL ILLUSTRATORS FOR STUDENTS/TRAINEES AT ALL STAGES, THE COMMUNITY, AND PHYSICIANS, 2) ESTABLISHING A SABV MENTORING ACADEMY, 3) CREATING A SABV CURRICULUM AND WORKSHOP FOR STUDENTS, FELLOWS, AND JUNIOR INVESTIGATORS. THE IMPACT OF OUR WORK WILL BE TO INCREASE RECOGNITION AND AWARENESS OF THE RISKS OF CVD AND HYPERTENSION TO WOMEN ACROSS THEIR HEALTH SPAN AND BEST PRACTICES FOR SAVB EXPERIMENTAL DESIGN. OUR COMPREHENSIVE COMMUNITY OUTREACH PLAN WILL ALSO BE CRITICAL TO REACH A HIGHLY AT-RISK POPULATION IN THE SOUTHEAST OF THE US TO INCREASE ROUTINE CARDIOVASCULAR HEALTH MONITORING AND DECREASE THE OVERALL CVD BURDEN ON BOTH THE POPULATION AND THE HEALTHCARE SYSTEM.

SBIRT INITIATIVE FOR GEORGIA'S RURAL AND UNDERSERVED POPULATIONS

NEDDYLATION AND CARDIAC PROTEIN QUALITY CONTROL

NOVEL ROLE OF BSCL2 IN CARDIAC SUBSTRATE METABOLISM AND FUNCTION

NOVEL ROLE OF MAPK14 IN REGULATION OF VSMC CONTRACTILE PHENOTYPE

CENTER CORE GRANT FOR VISION RESEARCH

MCG MINORITY-BASED COMMUNITY CLINICAL ONCOLOGY PROGRAM

THE ADIPOCYTE PPARGAMA/ADIPONECTIN AXIS AND ALZHEIMER'S DISEASE

HYPOXIA AND CARDIAC STEM CELL HOMING

MECHANISMS FOR CARDIOVASCULAR CONTROL EARLY IN DIABETES

DEVELOPMENT OF A NOVEL SOLUBLE EPOXIDE HYDROLASE INHIBITOR AS A STRATEGY FOR TREATING NEUROPATHIC PAIN IN PATIENTS WITH SPINAL CORD INJURY

COOPERATIVE MULTICENTER REPRODUCTIVE MEDICINE NETWORK

THE ROLES OF SEMAPHORIN SIGNALING DURING MOUSE VALVULOSEPTAL DEVELOPMENT

GENE DISCOVERY IN FAMILIAL KERATOCONUS

ESTROGEN-MEDIATED IMPAIRMENTS OF VASCULAR HEALTH IN DIABETES

REDOX REGULATION OF CU IMPORTER CTR1 IN ANGIOGENESIS

EPIGENETIC BASIS OF OBESITY INDUCED CARDIOVASCULAR DISEASE AND TYPE 2 DIABETES

ROLE OF SKELETAL MUSCLE HEALTH ON POOR LIFESTYLE RELATED TYPE 2 DIABETES AND CARDIOVASCULAR DISEASE RISK

CYBERCORPS SCHOLARSHIPS FOR SERVICE: AUGUSTA UNIVERSITY DEVELOPING IMMERSIVE EXPERIENCES FOR NOVEL CYBERSECURITY EDUCATION

LEPTIN AND PERIPHERAL GLUCOSE METABOLISM

GAMMA GLOBIN INDUCTION: MOLECULAR AND CELL-BASED STRATEGIES

SIGMA-1 RECEPTOR PROVIDES NEUROPROTECTION AGAINST OPTIC NEUROPATHY

VITAMIN D METABOLISM AND FUNCTION IN THE CORNEA AND ANTERIOR SEGMENT

MECHANISMS AND CONSEQUENCES OF IMPAIRED GLUTATHIONE HOMEOSTASIS IN THE AGING LENS

FIT BODY AND SOUL: A LIFESTYLE INTERVENTION FOR DIABETES PREVENTION CONDUCTED THR

CHARACTERIZATION OF LEPTIN'S ANTIDEPRESSANT ACTIVITY

ADIPOSE TISSUE DISTRIBUTION DETERMINES MICROGLIAL REGULATION OF HIPPOCAMPAL PLASTICITY

PSYCHOSOCIAL STRESS, EPIGENETICS AND HEALTH DISPARITY IN HYPERTENSION

CLINICAL PHENOTYPING AND DISEASE SPECIFIC SAMPLING TO IDENTIFY NON-CODING RNAS FOR HUMAN THERAPEUTICS IN PAD

REASONS FOR POOR DURABILITY OF RESIN-DENTIN BONDS

A GENOME-WIDE METHYLATION STUDY ON ESSENTIAL HYPERTENSION

ROLE OF EPIGENETIC REPRESSION OF IRF8 IN TUMOR PROGRESSION/METASTASIS

EFFECT OF RANDOMIZED COCOA SUPPLEMENTATION ON INFLAMMAGING AND EPIGENETIC AGING - PROJECT SUMMARY/ABSTRACT COCOA PRODUCTS HAVE BECOME A WIDELY CONSUMED FOOD, WITH GROWING DEMAND ACROSS THE WORLD AND STEADILY INCREASING INTEREST IN ITS POTENTIAL HEALTH BENEFITS AND ANTI-AGING BIOLOGY. HOWEVER, THE ROLE OF COCOA CONSUMPTION ON EPIGENETIC AGING REMAINS ESSENTIALLY UNEXPLORED. THERE HAS BEEN COMPELLING EVIDENCE FOR THE ANTI- INFLAMMATORY ROLE OF COCOA AND COCOA-DERIVED FLAVANOLS BASED UPON IN VITRO AND ANIMAL MODELS. HOWEVER, A CRITICAL BARRIER OF RESEARCH PROGRESS REMAINS THE LACK OF DEFINITIVE LARGE-SCALE RANDOMIZED CONTROLLED TRIALS (RCTS). ACCELERATED EPIGENETIC AGING THAT COMPRISES DNA METHYLATION CHANGES HAS RECENTLY EMERGED AS A POWERFUL, NOVEL PREDICTOR FOR LIFESPAN AND HEALTH SPAN, DISEASE SUSCEPTIBILITY, MORBIDITY, AND MORTALITY RISK. FURTHER, INFLAMMAGING IS A HIGHLY SIGNIFICANT RISK FACTOR FOR BOTH MORBIDITY AND MORTALITY IN THE ELDERLY, AS MANY AGE-RELATED DISEASES INCLUDING CARDIOVASCULAR DISEASE (CVD) SHARE AN INFLAMMATORY PATHOGENESIS. AS SUCH, INTERVENTIONS TO SLOW OR MITIGATE EPIGENETIC AGING AND INFLAMMAGING ARE CRITICAL FOR PROMOTING HEALTHY AGING. THE COCOA SUPPLEMENT AND MULTIVITAMIN OUTCOMES STUDY (COSMOS) IS AN ONGOING RANDOMIZED, DOUBLE- BLIND, PLACEBO-CONTROLLED, 2X2 FACTORIAL RCT OF A HIGH-QUALITY COCOA EXTRACT SUPPLEMENT (CONTAINING 600 MG/D FLAVANOLS, INCLUDING 80 MG EPICATECHINS), AND A MULTIVITAMIN SUPPLEMENT TO REDUCE THE RISK OF CVD AND CANCER IN WOMEN AGED =65 YEARS AND MEN AGED =60 YEARS. COSMOS HAS RANDOMIZED 21,444 PARTICIPANTS INTO THE TRIAL, INCLUDING 2,006 PARTICIPANTS WITH BIO-SAMPLES ALREADY COLLECTED AT BASELINE, YEAR 1 AND YEAR 2 FOLLOW-UP. COSMOS IS THE ONLY LARGE RCT TESTING THE EFFECTS OF COCOA EXTRACT IN WOMEN AND MEN, AND FOR MULTIVITAMINS IN WOMEN. THUS, THIS PROPOSED ANCILLARY STUDY WILL BUILD UPON THE PARENT COSMOS TRIAL TO EXAMINE THE EFFECTS OF RANDOMIZED COCOA OR MULTIVITAMIN SUPPLEMENTATION ON ANTI-AGING (EPIGENETIC AGING AND INFLAMMAGING) IN 600 REPRESENTATIVE SUBJECTS WITH THREE BIO-SAMPLE COLLECTIONS AT BASELINE, YEAR 1, AND YEAR 2, AS WELL AS MEASURED CVD RISK FACTORS, AND EXAMINING THESE FINDINGS IN THE CONTEXT OF CVD OUTCOMES. WE WILL TEST OUR CENTRAL HYPOTHESES THAT COCOA SUPPLEMENTATION REDUCES EPIGENETIC AGING INDICES (E.G., AGEACCELHORVATH, AGEACCELHANNUM, AGEACCELGRIM, AGEACCELPHENO, AND DNAMTLADJAGE) AND INFLAMMAGING (SERUM HSCRP, IL- 1SS, IL-4, IL-6, AND TNF-A) OVER TIME. WE WILL ALSO DETERMINE TO WHAT EXTENT THE EFFECTS OF COCOA SUPPLEMENTATION ON INFLAMMAGING AND CVD RISK FACTORS ARE MEDIATED BY EPIGENETIC AGING. THE PURPORTED ANTI-AGING HEALTH BENEFITS OF COCOA-RICH PRODUCTS SUCH AS DARK CHOCOLATE HAVE POPULARIZED AND INCREASED CONSUMPTION OF COCOA-RICH PRODUCTS OVER THE PAST DECADE. MOREOVER, MULTIVITAMINS REMAIN THE MOST COMMON DIETARY SUPPLEMENT TAKEN BY AT LEAST ONE-THIRD OF OLDER US ADULTS. IT REMAINS OF CRITICAL PUBLIC HEALTH IMPORTANCE TO RIGOROUSLY TEST THE ANTI-AGING PROPERTIES OF THESE TWO SUPPLEMENTS VIA LARGE RCTS FOR MORE DEFINITIVE EVIDENCE OF THEIR POTENTIAL EFFICACY, AND TO APPROVE OR REFUTE THEIR POTENTIAL ANTI-AGING BENEFITS.

ANALYSIS OF NEURAL RETINA TRANSPORT FUNCTION

CHEMO-RADIO IMMUNOTHERAPY FOR PEDIATRIC BRAIN TUMORS

REPARATIVE EFFECT OF JUVENILE FACTORS IN AGING AND INJURY - AGING AND INJURY ARE AMONG THE MAJOR GLOBAL HEALTH PROBLEMS AND DEATH DUE TO INJURY INCREASES SHARPLY WITH AGE. AS HEMORRHAGE ACCOUNTS FOR ALMOST HALF OF ALL TRAUMA-RELATED DEATHS, THERE IS A NEED TO DEVELOP METHODS TO REDUCE THE ADVERSE EFFECTS OF AGING ON INJURY TO FACILITATE HEALTHY LIVING. IN THIS PROPOSAL, OUR OBJECTIVE IS TO ESTABLISH THAT CIRCULATORY FACTORS OF JUVENILE ORIGIN CAN IMPROVE OUTCOME FOLLOWING INJURY IN THE MATURE AND AGED ANIMALS. THE EXPERIMENTS PROPOSED IN THIS PROJECT ARE BASED UPON OUR FINDING THAT FOLLOWING HEMORRHAGIC SHOCK IN A MOUSE MODEL (HEMORRHAGIC SHOCK INJURY; HI) JUVENILE MICE HAVE A SURVIVAL ADVANTAGE COMPARED TO ADULT MICE. WE ALSO FOUND THAT EVS FROM THE PLASMA OF JUVENILE MICE IMPROVED ORGAN FUNCTION AND SURVIVAL FOLLOWING HI. BASED UPON THESE DATA OUR HYPOTHESIS IS THAT PLASMA FACTORS FROM JUVENILE MICE CAN RESTORE MITOCHONDRIAL FUNCTION, ALLEVIATE OXIDATIVE STRESS AND REDUCE ORGAN DYSFUNCTION AND DEATH IN MATURE AND OLD MICE SUBJECTED TO HI. WE WILL TEST OUR HYPOTHESIS BY DETERMINING THE PROTECTIVE EFFECT OF JUVENILE MICE-DERIVED EVS IN MATURE AND OLD MICE AND IDENTIFY POTENTIAL MECHANISMS BY WHICH JUVENILE PLASMA FACTORS EXERT SALUTARY EFFECT IN MATURE AND AGED MICE FOLLOWING HI. USING 5XFAD MICE WE WILL DETERMINE WHETHER JUVENILE EVS CAN REDUCE PATHOLOGY IN ALZHEIMER’S DISEASE, AN AGE ASSOCIATED NEURODEGENERATIVE DISEASE. OUR GOAL IS TO DEVELOP METHODS TO REVITALIZE THE AGING SYSTEM BY IDENTIFYING MOLECULAR FACTORS INVOLVED IN MATURATIONAL DEVELOPMENT. WE WILL USE A COMBINATION OF CELL BIOLOGICAL, BIOCHEMICAL AND GENOMIC TOOLS AND TECHNIQUES TO TEST THE HYPOTHESIS. WE EXPECT THAT OUR STUDIES WILL RESULT IN THE IDENTIFICATION OF JUVENILE PROTECTIVE FACTORS THAT CAN IMPROVE OUTCOME FOLLOWING HEMORRHAGIC SHOCK. THE PROPOSED RESEARCH IS RELEVANT TO THE PART OF NIH’S MISSION PERTAINING TO DEVELOPING FUNDAMENTAL KNOWLEDGE TO POTENTIALLY HELP REDUCE THE BURDENS OF HUMAN DISEASE. THE OUTCOME OF THIS RESEARCH WILL BE SIGNIFICANT BECAUSE THE FUNDAMENTAL KNOWLEDGE GAINED FROM THIS STUDY IS EXPECTED TO ADVANCE METHODS TO PROMOTE HEALTHY LIVING.

MODULATION OF VASCULAR EXTRACELLULAR SUPEROXIDE DISMUTASE

CALPAIN/TALIN/MLCP AXIS IN PULMONARY ENDOTHELIAL BARRIER REGULATION - PROJECT SUMMARY ACUTE LUNG INJURY (ALI) IS CHARACTERIZED BY LUNG VASCULAR ENDOTHELIAL CELL (EC) BARRIER COMPROMISE RESULTING IN PULMONARY EDEMA. WHILE BACTERIAL INFECTIONS INDUCED BY E.COLI OR STREPTOCOCCUS PNEUMONIAE (SPN), AND THEIR RESPECTIVE BACTERIA TOXIN LIPOPOLYSACCHARIDES (LPS) AND PNEUMOLYSIN (PLY) ARE THE MAJOR CAUSES FOR ALI, THE MOLECULAR MECHANISMS INVOLVED IN LPS- OR PLY-INDUCED ALI ARE ILL-DEFINED. THE SCIENTIFIC PREMISE COMES FROM OUR NOVEL PRELIMINARY DATA, WHICH SHOW THAT BOTH LPS AND PLY ACTIVATE ENDOPEPTIDASE CALPAIN IN HUMAN LUNG MICROVASCULAR ECS (HLMVECS) IN ERK-DEPENDENT MANNER AND THAT SPECIFIC CALPAIN INHIBITION PREVENTS LPS- AND PLY-INDUCED DISRUPTION OF EC BARRIER IN HLMVECS AND LPS-INDUCED PULMONARY EDEMA IN ALI. FURTHER, WE FOUND THAT ERK-MEDIATED CALPAIN PHOSPHORYLATION AT SER-50 AND CALPAIN ACTIVITY WERE MUCH HIGHER IN MURINE LUNG MICROVASCULAR ECS ISOLATED FROM LUNGS EXPOSED TO LPS, INDICATING THAT ERK-CALPAIN PATHWAY IS HIGHLY RELATED TO ALI. WE SHOW THAT LPS/PLY INDUCES TALIN CLEAVAGE INTO HEAD AND ROD DOMAIN AND TALIN PHOSPHORYLATION IN HLMVECS AND THAT OVEREXPRESSION OF CALPAIN CAUSES TALIN CLEAVAGE, AND RHO-MEDIATED INHIBITION OF MYOSIN LIGHT CHAIN PHOSPHATASE (MLCP). TALIN IS ACTIVATED THROUGH EITHER TALIN CLEAVAGE OR PHOSPHORYLATION. TALIN CLEAVAGE SEPARATES HEAD FROM ROD DOMAIN THUS REMOVING AUTO-INHIBITION AND STIMULATING TALIN HEAD BINDING TO INTEGRIN AND THUS INDUCES ACTIVATION OF FOCAL ADHESIONS (FAS), LEADING TO RHOA-MEDIATED MLCP INHIBITION AND INCREASED LUNG EC PERMEABILITY. WE FOUND THAT MLCP, WHICH OPPOSES EC BARRIER COMPROMISE IN ALI, IS DOWN- REGULATED IN LUNGS OF ARDS PATIENTS, HIGHLIGHTING THE IMPORTANCE OF MLCP INHIBITION IN HUMAN ALI. MLCP BINDS TALIN AND INDUCED TALIN DEPHOSPHORYLATION, WHICH MAY LEAD TO TALIN DEGRADATION FOLLOWED BY FAS DISASSEMBLY. INTERESTINGLY, MLCP DEPHOSPHORYLATES ANOTHER CALPAIN SUBSTRATE, EZRIN, LEADING TO ITS DEACTIVATION AND WITHDRAWAL FROM MEMBRANE CYTOSKELETON. ACTIVATION OF TALIN/EZRIN AXIS IS INVOLVED IN FA FORMATION AND OUR DATA SUGGEST THAT TALIN IS UPSTREAM OF EZRIN IN LPS-INDUCED EC CYTOSKELETAL REMODELING. IT WAS SHOWN THAT EZRIN DEPLETION DECREASES CALPAIN ACTIVITY SUGGESTING POSITIVE FEEDBACK EFFECT OF MLCP INHIBITION ON LPS/PLY-INDUCED CALPAIN ACTIVATION AND EC BARRIER COMPROMISE VIA EZRIN PHOSPHORYLATION/MEMBRANE ATTACHMENT. THESE DATA LED TO OUR NOVEL HYPOTHESIS THAT CALPAIN/MLCP CROSSTALK COORDINATES TALIN ACTIVATION LEADING TO LUNG EC BARRIER DISRUPTION IN ALI. TO TEST THIS HYPOTHESIS WE WILL EMPLOY UNIQUE MOLECULAR TOOLS SUCH AS MURINE LUNG-TARGETING GENE DELIVERY IN VIVO, EC-SPECIFIC INDUCIBLE CALPAIN AND MLCP KO MICE AND ERK INSENSITIVE KNOCKIN MOUSE (CAPN2S50A). WE WILL: (1) EVALUATE WHETHER ERK-MEDIATED CALPAIN ACTIVATION IS INVOLVED IN LUNG MICROVASCULAR EC BARRIER DISRUPTION AND CYTOSKELETAL REORGANIZATION IN ALI INDUCED BY BACTERIAL TOXINS (LPS, PLY), LIVE BACTERIA (E.COLI, SPN) AND IN SEPSIC CONDITIONS; (2) DETERMINE WHETHER GRAM- AND GRAM+ BACTERIA, THEIR TOXINS AND SEPSIS INDUCE TALIN ACTIVATION (CLEAVAGE/PHOSPHORYLATION) AND FA STRENGTHENING LEADING TO LUNG MICROVASCULAR EC BARRIER DISRUPTION; (3) EXAMINE WHETHER MLCP ACTIVITY AFFECTS CALPAIN ACTIVATION INDUCED BY LPS/E.COLI, PLY/SPN AND SEPSIS.

ORAL PATHOGENS AND DENDRITIC CELL SUBSETS

COPPER TRANSPORT PROTEIN AND INFLAMMATORY ANGIOGENESIS

PEDIATRIC AMBULATORY BLOOD PRESSURE TRAJECTORY AND BRAIN HEALTH IN MIDLIFE

NEURONAL HDAC9, SYNAPTIC PLASTICITY AND ALZHEIMER'S DISEASE - PROJECT SUMMARY ALZHEIMER’S DISEASE (AD) IS AN AGE-RELATED NEURODEGENERATIVE DISORDER THAT CAUSES MEMORY LOSS AND COGNITIVE DECLINE. SYNAPTIC DYSFUNCTION AND LOSS CORRELATES STRONGLY WITH COGNITIVE IMPAIRMENT IN AD. AGING IS THE LEADING RISK FACTOR FOR AD, AND EPIGENETIC MECHANISMS INVOLVING HISTONE DEACETYLASES (HDACS) PLAY AN IMPORTANT ROLE IN AGING AND AGE-RELATED NEURODEGENERATIVE DISORDERS. AMONG THE 11 ZINC-DEPENDENT HDACS, HDAC9 IS THE MOST ABUNDANT ISOFORM IN THE BRAIN, FOUND EXCLUSIVELY IN NEURONS. WE PROVIDE KEY PRELIMINARY DATA SHOWING THAT HDAC9 EXPRESSION IN THE HIPPOCAMPUS AND PREFRONTAL CORTEX (PFC) DIMINISHES WITH AGING IN WILD-TYPE MICE, AND THAT REDUCED HDAC9 EXPRESSION PRECEDES THE ONSET OF AMYLOID DEPOSITION IN THE APP/PS1 MOUSE MODEL OF AD. CONSISTENT WITH THESE PRECLINICAL FINDINGS, AD PATIENTS EXHIBITED DECREASED HDAC9 EXPRESSION IN THE DORSOLATERAL PFC. MOREOVER, GLOBAL OR HIPPOCAMPAL CA1-SPECIFIC DELETION OF HDAC9 INDUCES COGNITIVE IMPAIRMENT AND IMPAIRS SYNAPTIC PLASTICITY, WHILE HDAC9 OVEREXPRESSION PRODUCES COGNITIVE-ENHANCING EFFECTS. WE HYPOTHESIZE THAT REDUCED NEURONAL HDAC9 MEDIATES COGNITIVE DECLINE, SYNAPTIC DYSFUNCTION AND OTHER NEUROPATHOLOGIES ASSOCIATED WITH BRAIN AGING AND AD. TO TEST THIS HYPOTHESIS, WE PROPOSE THREE SPECIFIC AIMS. IN AIM 1, WE WILL TEST THE HYPOTHESIS THAT LOSS OF HDAC9 IN HIPPOCAMPAL AND PFC NEURONS MEDIATES AGE- AND AD-RELATED NEUROPATHOLOGY AND COGNITIVE IMPAIRMENT. IN AIM 2, WE WILL TEST THE HYPOTHESIS THAT THE HISTONE METHYLTRANSFERASE EZH2 [THE CATALYTIC COMPONENT OF THE POLYCOMB REPRESSIVE COMPLEX 2 (PRC2), WHICH CATALYZES REPRESSIVE H3K27ME3 MODIFICATIONS AT GENE PROMOTERS] EPIGENETICALLY SILENCES HDAC9 EXPRESSION IN THE HIPPOCAMPUS AND PFC DURING AGING AND IN AD. IN AIM 3, WE WILL TEST THE HYPOTHESIS THAT NEURONAL PENTRAXIN 2 (NP2), NERVE GROWTH FACTOR INDUCIBLE (VGF), AND BRAIN-DERIVED NEUROTROPHIC FACTOR (BDNF) MEDIATE THE DOWNSTREAM EFFECTS OF HDAC9 ON HIPPOCAMPAL SYNAPTIC PLASTICITY AND COGNITION. WE EXPECT THAT THE RESULTS WILL PROVIDE INSIGHT INTO MOLECULAR MECHANISMS UNDERLYING THE EPIGENETIC CONTROL OF GENES RELATED TO AGING AND AD AND OFFER POTENTIAL TARGETS FOR FUTURE THERAPEUTIC INTERVENTIONS.

RURAL COMMUNITIES OPIOID RESPONSE PROGRAM - MEDICATION ASSISTED TREATMENT ACCESS

THIS EDA INVESTMENT SUPPORTS THE GEORGIA AI MANUFACTURING (GA-AIM) COALITION, LED BY THE GEORGIA TECH RESEARCH CORPORATION, WITH INTEGRATING ARTIFICIAL INTELLIGENCE (AI) TECHNOLOGIES IN INDUSTRY BY ACCELERATING THE ADOPTION OF AI ACROSS THE STATE'S LEGACY INDUSTRIAL SECTORS. COALITION PROJECTS INCLUDE ESTABLISHING AN AI MANUFACTURING PILOT FACILITY AT GEORGIA TECH AND EXPAND JOB TRAINING OPPORTUNITIES IN AI FOR UNDERSERVED COMMUNITIES. THE GA-AIM COALITION AIMS TO SERVE AS A NATIONAL MODEL FOR HOW TO ACCELERATE THE TRANSITION TO AUTOMATION IN MANUFACTURING WHILE ENSURING THESE SYSTEMS COMPLEMENT RATHER THAN REPLACE EXISTING WORKERS. ONCE IMPLEMENTED, THE INVESTMENT WILL HELP DEVELOP AND STRENGTHEN REGIONAL INDUSTRY CLUSTERS - ALL WHILE EMBRACING ECONOMIC EQUITY, CREATING GOOD-PAYING JOBS, AND ENHANCING U.S. COMPETITIVENESS GLOBALLY.

THE ROLE OF CHD7 IN ACC NEURONS - A LIMITED FUNDAMENTAL UNDERSTANDING OF THE CELLULAR AND MOLECULAR MECHANISMS UNDERLYING COMPLEX BEHAVIORS POSES A MAJOR BARRIER FOR DEVELOPMENT OF EFFECTIVE CLINICAL APPLICATIONS TO TREAT MENTAL DISORDERS. THE LONG-TERM GOAL OF OUR RESEARCH IS TO SHED LIGHT ON THE GAPS IN KNOWLEDGE REGARDING THE MOLECULAR SIGNATURES AND SYNAPTIC PROPERTIES OF BRAIN NEURONAL CIRCUITS THAT CONTROL RESPONSES TO STRESS. EPIGENETIC GENE REGULATION HAS EMERGED AS A KEY MOLECULAR DRIVER UNDERLYING NEURONAL CIRCUIT DYNAMICS AND BEHAVIORAL CHANGES. OUR PRELIMINARY DATA SUGGEST THAT CHD7, A CHROMATIN-REMODELING FACTOR PRIMARILY EXPRESSED IN THE EMBRYONIC BRAIN, REMAINS ENRICHED IN EXCITATORY NEURONS WITHIN LAYER 2/3 OF THE ANTERIOR CINGULATE CORTEX (ACC) IN ADULT MICE. THE ACC, A REGION WITHIN THE MEDIAL PREFRONTAL CORTEX IN RODENTS, PLAYS CRITICAL ROLES IN PROCESSING MOOD-RELATED INFORMATION AND MODULATING ANXIETY-RELATED BEHAVIORS. WHILE CRITICAL ROLES FOR CHD7 DURING NEURAL DEVELOPMENT HAVE BEEN WELL- DOCUMENTED, ITS FUNCTION IN POSTMITOTIC NEURONS REMAINS UNCLEAR. OUR DATA SUGGEST THAT POSTMITOTIC DELETION OF CHD7 FROM ACC EXCITATORY NEURONS (REFERRED TO AS CHD7CKO) SIGNIFICANTLY REDUCED INNATE ANXIETY LEVELS. IN ADDITION, NEURONAL ACTIVITY IN THE ACC OF CHD7CKO MICE EXPOSED TO AN ENVIRONMENTAL STRESSOR WAS SIGNIFICANTLY LOWER THAN THAT IN WILD TYPE (WT) MICE. INTRIGUINGLY, THESE PHENOMENA WERE OBSERVED ONLY IN MALE (AND NOT FEMALE) MICE, INDICATING SEXUAL DIMORPHISM OF CHD7 FUNCTION. OUR DATA PROVIDE THE FIRST EVIDENCE SUGGESTING AN ESSENTIAL ROLE FOR CHD7 IN POSTMITOTIC ACC NEURONS IN REGULATING NEURONAL ACTIVITY AND INNATE ANXIETY. OUR PRIMARY OBJECTIVES ARE TO INTERROGATE THE ROLE OF CHD7 IN CONTROLLING A COMPLEX GENE NETWORK TO REGULATE THE SYNAPTIC ACTIVITY OF ACC NEURONS AND THEIR DOWNSTREAM TARGETS. IN THIS PROPOSAL, WE WILL FIRST DETERMINE HOW CHD7 REGULATES THE ACTIVITY OF ACC NEURONS AND THEIR DOWNSTREAM TARGETS IN THE BED NUCLEUS OF THE STRIA TERMINALIS (BNST). OUR PRELIMINARY DATA SUGGEST THAT CHD7 PLAYS A CRITICAL ROLE IN MAINTAINING PROPER NEURONAL ACTIVITY IN THE ACC-BNST PATHWAY. NEXT, WE WILL USE UNBIASED HIGH-THROUGHPUT APPROACHES TO DETERMINE HOW CHD7 CONTROLS A COMPLEX GENE NETWORK IN POSTMITOTIC NEURONS TO REGULATE ACTIVITY-DEPENDENT GENE TRANSCRIPTION. FINALLY, WE WILL INVESTIGATE HOW CHD7 ACTIVITY IS REGULATED BY AN INTERACTING PARTNER THAT IS INVOLVED IN G PROTEIN SIGNALING. IF SUCCESSFUL, OUR RESEARCH WILL SHED NEW LIGHT ON THE MOLECULAR UNDERPINNINGS AND NEUROPHYSIOLOGY OF NEURONAL CIRCUITS THAT RESPOND TO STRESS. SUCH INFORMATION WILL ULTIMATELY HELP DEFINE MECHANISMS UNDERLYING COMPLEX EMOTIONAL BEHAVIORS IN HUMANS.

MITOCHONDRIA-MEDIATED EFFECTS AND THERAPEUTIC POTENTIAL OF ATRIAL NATRIURETIC PEPTIDE IN SALT-SENSITIVE HYPERTENSION

A MOLECULAR EXAMINATION OF MRNA LOCALIZATION AND CELL POLARIZATION

CIRCADIAN ORIGINS OF VASCULAR DISEASE IN OBESITY

ROLE OF AGRP NEURONS IN CHRONIC STRESS-ACCELERATED BRAIN AGING AND PROGRESSION OF ALZHEIMER'S DISEASE - PROJECT SUMMARY CHRONIC STRESS IS INCREASINGLY BEING RECOGNIZED AS A RISK FACTOR FOR SPORADIC AD. DYSREGULATION OF THE HYPOTHALAMIC–PITUITARY–ADRENAL AXIS (HPA AXIS) IS COMMON IN AD PATIENTS. BY STIMULATING CORTICOTROPIN-RELEASING HORMONE (CRH) EXPRESSION AND GLUCOCORTICOID SECRETION, CHRONIC STRESS EXPOSURE EXACERBATES ASS AND TAU PATHOLOGIES AND COGNITIVE DECLINE. AGRP NEURONS, LOCATED IN THE ARCUATE NUCLEUS, CO-EXPRESS GABA AND SEND EXTENSIVE PROJECTIONS TO THE PARAVENTRICULAR NUCLEUS OF THE HYPOTHALAMUS. OUR RECENT STUDIES HAVE SHOWN THAT CHRONIC STRESS DECREASES FIRING RATES OF AGRP NEURONS, AND THAT STIMULATING AGRP NEURONS CAN REVERSE CHRONIC STRESS-INDUCED BEHAVIORAL DEFICITS, INCLUDING MEMORY IMPAIRMENT. HPA RESPONSES TO STRESS CAN ALSO BE DAMPENED BY STIMULATING AGRP NEURONS. IN ADDITION, WE PROVIDE NOVEL PRELIMINARY DATA SHOWING THAT AGING INDUCES SILENCING OF AGRP NEURONS. WE HYPOTHESIZE THAT CHRONIC STRESS AND AGING CONVERGE TO SILENCE AGRP NEURONS, WHICH LEADS TO REDUCED INHIBITORY INPUTS TO THE PVN. THIS WEAKENS NEGATIVE FEEDBACK AND DRIVES HPA HYPERACTIVITY, WHICH IN TURN ACCELERATES BRAIN AGING AND WORSENS AD PATHOLOGIES. TO TEST THIS HYPOTHESIS, WE PROPOSE THREE SPECIFIC AIMS. IN AIM 1, WE WILL IDENTIFY MOLECULAR MECHANISMS OF CHRONIC STRESS- AND AGING- INDUCED SILENCING OF AGRP NEURONS. IN AIM 2, WE WILL DETERMINE WHETHER CHRONIC STRESS-INDUCED INHIBITION OF AGRP NEURON ACTIVITY CONTRIBUTES TO OVERSTIMULATION OF THE HPA AXIS AND STRESS-EXACERBATED ASS AND TAU PATHOLOGIES. IN AIM 3, WE WILL DETERMINE WHETHER AGE-INDUCED SILENCING OF AGRP NEURONS ACCELERATES THE PROGRESSION OF ASS ACCUMULATION AND COGNITIVE DECLINE.

PBK: A NOVEL MEDIATOR OF VSMC PROLIFERATION AND VASCULAR REMODELING IN PAH - PROJECT SUMMARY PULMONARY ARTERIAL HYPERTENSION (PAH) IS A DEBILITATING AND EVENTUALLY LETHAL DISEASE THAT IS RESISTANT TO CURRENT THERAPEUTICS. A DEFINING CHARACTERISTIC OF PAH IS EXCESSIVE CELLULAR PROLIFERATION AND REMODELING OF PULMONARY ARTERIES (PA), THAT RESULTS IN PROGRESSIVE INCREASES IN PULMONARY VASCULAR RESISTANCE LEADING TO RIGHT VENTRICULAR FAILURE AND DEATH. PAH HAS A SURVIVAL TIME OF FIVE TO SEVEN YEARS POST DIAGNOSIS, AND MOST OF THE CURRENT THERAPIES FOR PAH ARE VASODILATORS, WHICH PROVIDE SYMPTOMATIC RELIEF, BUT DO NOT REVERSE PULMONARY VASCULAR REMODELING OR STOP DISEASE PROGRESSION. WE HAVE DISCOVERED A NOVEL GENE, PDZ-BINDING KINASE (PBK) THAT IS UPREGULATED IN HYPERTENSIVE PA. PBK IS A SERINE/THREONINE KINASE THAT IS OVEREXPRESSED IN A SUBSET OF AGGRESSIVE CANCERS. THE HYPERPROLIFERATIVE NATURE OF VASCULAR CELLS IN PAH SHARES MANY MECHANISMS WITH THAT OF CANCER CELLS, HOWEVER, THE THERAPEUTIC UTILITY OF TARGETING PBK IN PAH AND THE MECHANISMS BY WHICH PBK INFLUENCES PULMONARY VASCULAR REMODELING ARE NOT YET KNOWN AND ARE THE GOALS OF THIS PROPOSAL. IN PRELIMINARY EXPERIMENTS IN EXPERIMENTAL MODELS AND HUMAN PAH, WE SHOW THAT PBK IS ROBUSTLY UPREGULATED IN THE MEDIAL LAYER OF PA WHERE IT OVERLAPS WITH MARKERS OF SMOOTH MUSCLE CELLS. GAIN AND LOSS OF FUNCTION APPROACHES SHOW THAT PBK EXPRESSION REGULATES PULMONARY ARTERY SMOOTH MUSCLE CELL (PASMC) PROLIFERATION. IN EXPERIMENTAL RAT AND MOUSE MODELS OF PAH IN VIVO, WE FOUND THAT SELECTIVE INHIBITORS OF PBK IMPROVE PA REMODELING AND CARDIOPULMONARY FUNCTION. TO DETERMINE THE MECHANISMS UNDERLYING INCREASED EXPRESSION OF PBK, WE FOUND THAT THE TRANSCRIPTIONAL CO-ACTIVATOR, YES ASSOCIATED PROTEIN1 (YAP1) WAS UPREGULATED IN PAH AND INCREASED PBK PROMOTER ACTIVITY AND PBK PROTEIN EXPRESSION IN PASMC. WE EMPLOYED A PROXIMITY LIGATION APPROACH TO IDENTIFY NOVEL SUBSTRATES OF PBK WHICH REVEALED PROTEIN REGULATOR OF CYTOKINESIS 1 (PRC1) AS A BINDING PARTNER. PBK UPREGULATED PRC1 AND INDUCED PRC1 PHOSPHORYLATION AND CYTOKINESIS IN PASMC. THESE NOVEL PRELIMINARY DATA INFORM OUR CENTRAL HYPOTHESIS THAT YAP1 UPREGULATES PBK IN PASMC TO ENHANCE PROLIFERATION VIA PRC1 MEDIATED CYTOKINESIS. COLLECTIVELY THESE MECHANISMS CONTRIBUTE TO PATHOLOGIC PULMONARY VASCULAR REMODELING AND PAH. THIS HYPOTHESIS WILL BE TESTED USING INTEGRATED MOLECULAR, CELLULAR, GENETIC, IMAGING, AND TRANSLATIONAL PHARMACOLOGICAL APPROACHES IN MULTIPLE RODENT MODELS INCLUDING A PBK KO RAT. OUR LONG-TERM GOAL IS TO DEFINE THE KEY MECHANISMS BY WHICH PBK REGULATES PASMC PROLIFERATION TO ORCHESTRATE CHANGES IN ARTERIAL REMODELING, A HALLMARK OF PAH. AT THEIR CONCLUSION, THE PROPOSED STUDIES WILL MOVE THE FIELD FORWARD BY DEFINING NOVEL SIGNALING PATHWAYS IN PAH AND A NOVEL MECHANISM OF PASMC PROLIFERATION. THESE STUDIES WILL ALSO ADVANCE THE UTILITY OF NOVEL THERAPEUTIC APPROACHES TARGETING PBK AND CYTOKINESIS TO REDUCE PA REMODELING AND SUBSEQUENTLY IMPROVE THE MORBIDITY AND MORTALITY ASSOCIATED WITH PAH.

SYNAPTIC CIRCUITRY IN STROKE

TARGETING THE CD73-ADENOSINERGIC PATHWAY IN HEAD AND NECK CANCER - HEAD AND NECK SQUAMOUS CELL CARCINOMA (HNSC) RELATED TO TOBACCO AND ALCOHOL CONSUMPTION IS AGGRESSIVE AND RESPONDS POORLY TO AVAILABLE THERAPY. EVEN THE NEW PD-1/PD-L1 IMMUNE CHECKPOINT BLOCKADE IS INSUFFICIENT TO CONTROL PROGRESSION DUE TO OTHER HIGHLY IMMUNOSUPPRESSIVE CONDITIONS IN THE TUMOR MICROENVIRONMENT (TME). COMPELLING EVIDENCE IMPLICATES THE ECTO-5'-NUCLEOTIDASE (NT5E) CD73 AS ANOTHER IMPORTANT IMMUNE CHECKPOINT, AS HIGH CD73 EXPRESSION IN THE TME OF MANY TUMORS, INCLUDING HNSC, IS LINKED TO POOR PATIENT SURVIVAL. SEVERAL CONSISTENT TME CONDITIONS, SUCH AS HYPOXIA AND TREATMENT, CAUSE AN INCREASE IN EXTRACELLULAR (E)ATP, AND CD73 SERVES AT THE LAST, RATE-LIMITING STEP IN CONCERT WITH CD39 AND OTHER ECTOENZYMES, TO CONVERT EATP TO ADENOSINE (EADO). EADO ACTIVATES ADENOSINE RECEPTOR (AR) A2AR TO SUPPRESS T CELL ACTIVATION, WHILE A2BR PROMOTES IMMUNOSUPPRESSION VIA CANCER-ASSOCIATED FIBROBLASTS (CAF) AND MYELOID CELLS PRESENT IN THE TME. STUDIES, INCLUDING OUR OWN, SHOW THAT CD73 AND A2BR, AND TO A LESSER EXTENT A2AR, ARE EXPRESSED IN THE HIGHLY IMMUNOSUPPRESSIVE TME OF PROGRESSIVE HNSC OF BOTH CLINICAL AND PRECLINICAL SAMPLES, SUGGESTING THEIR ROLE IN THE IMMUNOSUPPRESSION. HOWEVER, THE CONTRIBUTIONS OF MOLECULES DOWNSTREAM OF CD73 ARE POORLY UNDERSTOOD. RECENT RAPID ADVANCES IN SPATIAL ANALYSIS OF THE TME HAVE REVEALED A CRITICAL ROLE OF IMMUNE EFFECTOR TO TARGET CELL DISTANCE IN GENERATING PRODUCTIVE ANTITUMOR IMMUNITY WITH IMPACT ON CLINICAL OUTCOMES. BECAUSE EADO IS SHORT-LIVED, THE SPATIAL RELATIONSHIP BETWEEN CD73 ON ONE HAND AND A2AR OR A2BR ON THE OTHER HAND, IS LIKELY TO AFFECT THE SPECIFIC AR ACTIVATED BY EADO. WE DISCOVERED IN HNSC CLINICAL SPECIMENS THAT A2AR IS RATHER LIMITED AND LOCATED REMOTELY FROM CD73, WHICH SUPPORTS THE HYPOTHESIS THAT THE MAJOR IMMUNOSUPPRESSIVE IMPACT OF CD73-AR IMMUNE CHECKPOINT IS MEDIATED PRIMARILY THROUGH THE CD73-A2BR AXIS. OUR PLAN IS TO: (1) DISSECT HOW THE CD73-A2BR PATHWAY REGULATES THE IMMUNE LANDSCAPE IN THE TME OF ECTOPIC PRECLINICAL HNSC MODELS DURING PROGRESSION AND TREATMENT; (2) DEVELOP EFFECTIVE CD73-ADO-A2BR ICB REGIMENS IN PRECLINICAL MODELS OF ORTHOTOPIC AND IN PRIMARY CARCINOGEN-INDUCED TONGUE HNSC; (3) ASSESS THE RELATIONSHIPS BETWEEN HNSC PATIENT PERIPHERAL BLOOD MYELOID PRECURSORS, LEVELS AND DISTRIBUTION OF CD73 AND A2BR ON MYELOID AND CAF SUBSETS IN THE TME, PATIENTS’ THREE-YEAR POST-OPERATIVE OUTCOMES. IMPACT: THE PROPOSED STUDY WILL IDENTIFY CD73-A2BR-DEPENDENT MECHANISMS OF IMMUNOSUPPRESSION IN HNSC PRECLINICAL MODELS AND DEVELOP TRANSLATABLE THERAPEUTIC INTERVENTION. THE RESULTS WILL PROVIDE HIGHLY VALUABLE INSIGHTS INTO THE EFFECTS OF CD73-A2BR AXIS ON DISEASE PROGRESSION AND THE THERAPEUTIC POTENTIAL OF BLOCKING IT.

THE ANTI-ANGIOGENIC VEGF165B AND VEGFR1 SIGNALING IN PERIPHERAL ARTERY DISEASE

WRESTLING STRESS: ROLE OF UFM1 MODIFICATION IN PATHOLOGICAL CARDIAC REMODELING

VASCULAR SMOOTH MUSCLE PROTEIN QUALITY CONTROL AND AORTIC ANEURYSM FORMATION - PROJECT SUMMARY ABDOMINAL AORTIC ANEURYSM (AAA) IS A DEVASTATING DISEASE CARRYING HIGH MORBIDITY AND MORTALITY DUE TO THE HIGH LIKELIHOOD OF FATAL DISSECTION AND RUPTURE. THERE ARE CURRENTLY NO PROVEN PHARMACEUTICAL TREATMENTS TO PREVENT AAA PROGRESSION. VSMC DEGENERATION CONTRIBUTES LARGELY TO AAA PATHOGENESIS, BUT THE MECHANISM REMAINS ELUSIVE. THE UBIQUITIN PROTEASOME SYSTEM (UPS) SERVES AS AN ESSENTIAL PROTEIN QUALITY CONTROL MECHANISM BY DEGRADING MISFOLDED PROTEINS AND SURPLUS NORMAL PROTEINS. WHILE PROTEOTOXICITY RESULTING FROM INSUFFICIENT UPS FUNCTION HAS BEEN WIDELY ACCEPTED AS AN IMPORTANT MECHANISM FOR MULTIPLE DEGENERATIVE HUMAN CONDITIONS, THE IMPLICATION OF UPS DYSFUNCTION IN AAA IS COMPLETELY UNKNOWN. OUR PATHWAY ANALYSIS IN HUMAN AAA TISSUES REVEALED AN ASSOCIATION OF UPS FUNCTION WITH AAA. IN A MOUSE MODEL OF AAA, ACCUMULATION OF UBIQUITINATED PROTEINS, A HALLMARK OF IMPAIRED UPS PERFORMANCE, PRECEDES VSMC DEGENERATION AND AAA FORMATION AND IS EXACERBATED WITH DISEASE PROGRESSION. THIS SUGGESTS THAT INADEQUATE UPS PERFORMANCE MAY ACT AS A NOVEL MECHANISM UNDERLYING AAA ETIOLOGY. MYOCD IS A MASTER SWITCH OF VSMC CONTRACTILE GENE PROGRAM. HOW MYOCD IS REGULATED AND FUNCTIONS IN AAA IS UNKNOWN. BULK RNA-SEQ IN VSMCS SHOWED AN ENRICHMENT OF UPS- RELATED PATHWAYS IN THE TOP MYOCD-UPREGULATED GENE PROGRAMS BESIDES THOSE RELEVANT TO VSMC CONTRACTION. FORCED EXPRESSION OF MYOCD IMPROVED UPS PERFORMANCE WHILE SUPPRESSING VSMC DEGENERATION IN CULTURED VSMCS AND VSMC-SPECIFIC MYOCD TRANSGENIC (TG) MICE AT THE EARLY STAGE OF AAA FORMATION. MYOCD INCREASED THE EXPRESSION OF NFE2L1 AND KLHL3, TWO KEY PLAYERS OF UPS FUNCTION. THESE LINES OF EVIDENCE SUGGEST A NOVEL ROLE OF MYOCD IN UPS FUNCTION. PHOSPHODIESTERASES (PDES), BY CATALYZING THE HYDROLYSIS OF CAMP AND CGMP TO SPECIFICALLY MODULATE CYCLIC NUCLEOTIDE SIGNALING, PLAY CRITICAL ROLES IN VSMC PATHOPHYSIOLOGY AND ARE PROVEN DRUG TARGETS FOR MULTIPLE HUMAN DISEASES. WE FOUND THAT PDE10A WAS THE MOST INDUCED PDE FAMILY MEMBER DURING AAA FORMATION, WHILE MYOCD PROTEIN EXPRESSION WAS SUPPRESSED. INHIBITION OF PDE10A INCREASED MYOCD PROTEIN, IMPROVED UPS PERFORMANCE, AND SUPPRESSED AAA FORMATION. THESE EXCITING PRELIMINARY FINDINGS SUPPORT A NOVEL HYPOTHESIS THAT DOWNREGULATION OF MYOCD PROTEIN BY PDE10A IMPAIRS UPS PERFORMANCE, LEADING TO VSMC DEGENERATION AND AAA. WE PROPOSE THREE AIMS TO TEST THIS HYPOTHESIS. AIM 1 WILL USE NOVEL VSMC-SPECIFIC MYOCD KNOCKOUT AND TG MICE TO DETERMINE THE FUNCTION OF MYOCD IN ANGII-INDUCED AAA MODEL. AIM 2 WILL USE A NOVEL UPS REPORTER MOUSE LINE AND BIOCHEMICAL ASSAYS TO DETERMINE THE IMPORTANCE OF UPS FUNCTION IN MYOCD-REGULATED AAA AND HOW MYOCD MODULATES UPS FUNCTION. AIM 3 WILL DETERMINE HOW PDE10 PROMOTES MYOCD PROTEIN DEGRADATION AND PDE10A FUNCTIONS IN UPS PERFORMANCE AND AAA FORMATION. THIS PROPOSAL WILL ADDRESS FOR THE FIRST TIME THE IMPORTANCE OF UPS PERFORMANCE IN VSMC DEGENERATION AND AAA FORMATION, AND ELUCIDATE A NOVEL REGULATORY CASCADE COMPRISING A DRUGGABLE UPSTREAM MODULATOR (PDE10A) AND A DOWNSTREAM EFFECTOR (MYOCD) IN SAFEGUARDING UPS PERFORMANCE AND PROTEOSTASIS.

REGULATION AND FUNCTION OF MAMMALIAN HSF4 IN VIVO

FUNCTION AND REGULATION OF TSPAN2 IN VASCULAR DISEASE

TACE AND CLOCK MECHANISMS IN AGING AND VASCULAR STIFFENING

INNOVATIVE APPROACHES TO TREAT DUCHENNE MUSCULAR DYSTROPHY USING IPSC-DERIVED MUSCLE PROGENITORS

GENETICS OF MULLERIAN DEVELOPMENT

MYELOID GLYCOLYSIS IN PATHOLOGICAL OCULAR ANGIOGENESIS

PERSISTENT STAT5 SIGNALING IN POLYFUNCTIONAL CD4 T CELLS AND ITS APPLICATION IN ADOPTIVE T CELL THERAPY - PROJECT SUMMARY/ABSTRACT RECENT ADVANCES IN ADOPTIVE T-CELL THERAPY (ACT), ESPECIALLY CD19-TARGETING CHIMERIC ANTIGEN RECEPTOR (CAR) T-CELL THERAPY (CD19CART), HAVE HIGHLIGHTED THE POTENTIAL OF IMMUNOTHERAPY TO ACHIEVE DURABLE AND CURATIVE PATIENT OUTCOMES. HOWEVER, EVEN FOR THE WELL-DEVELOPED CD19CART, MANY PATIENTS HAVE FAILED TO RESPOND TO THE TREATMENT OR SUCCUMBED TO LATE RELAPSE. MOREOVER, SO FAR ACT IN GENERAL HAS NOT BEEN EFFECTIVE IN TREATING MOST SOLID TUMORS. THE MAJOR BARRIERS TO EFFECTIVE ACT INCLUDE DEFICIENCIES IN DONOR T CELL EXPANSION, PERSISTENCE AND TUMOR- INFILTRATION, AS WELL AS LOSS OF T-CELL EFFECTOR FUNCTION - A PROCESS TERMED EXHAUSTION. THERE IS INCREASING DEMAND FOR NOVEL STRATEGIES THAT CAN OVERCOME THESE BARRIERS TO IMPROVE THE EFFICACY OF ACT. THE PRESENCE OF CD4 T CELLS WITH A POLYFUNCTIONAL PHENOTYPE, CHARACTERIZED BY CONCOMITANT PRODUCTION OF MULTIPLE INFLAMMATORY CYTOKINES, HAS BEEN ASSOCIATED WITH FAVORABLE THERAPEUTIC OUTCOMES IN PRECLINICAL MODELS AND CANCER PATIENTS. HOWEVER, HOW TO GENERATE POLYFUNCTIONAL CD4 (PFCD4) T CELLS SUITABLE FOR ACT REMAINS ELUSIVE. WE PREVIOUSLY REPORTED THAT CD4 T CELLS EXPOSED TO INTERLEUKIN 7 (IL7) DURING ANTIGENIC STIMULATION CAN ACQUIRE POLYFUNCTIONALITY IN A STAT5-DEPENDENT MANNER. OUR RECENT WORK DEMONSTRATED THAT ECTOPIC EXPRESSION OF A CONSTITUTIVELY ACTIVE FORM OF STAT5 (CASTAT5) IN CD4 T CELLS INDUCES GENOME- WIDE TRANSCRIPTIONAL AND EPIGENETIC REMODELING IN TUMOR-SPECIFIC CD4 T CELLS, ENDOWING THESE CELLS POLYFUNCTIONALITY, EXHAUSTION-RESISTANCE AND TUMOR-INFILTRATING CAPABILITY. IMPORTANTLY, CASTAT5 CAN MARKEDLY IMPROVE THE EXPANSION AND PERSISTENCE OF CD19CAR T CELLS, RESULTING IN HIGH CURE RATE IN MICE WITH ADVANCED LYMPHOMA. THESE NEW FINDINGS FORM THE BASIS OF OUR CENTRAL HYPOTHESIS THAT CASTAT5-INDUCED EPIGENETIC REMODELING ENDOWS CD4 T CELLS POLYFUNCTIONALITY AND EXHAUSTION-RESISTANCE, AND THESE FEATURES EQUIP CD4 T CELLS WITH SUPERIOR HELPER ACTIVITIES TO ORCHESTRATE POTENT ANTITUMOR IMMUNE RESPONSES. THIS CROSS-DISCIPLINARY MULTI- PI PROJECT WILL EMPLOY OUR WELL-CHARACTERIZED ACT MODEL SYSTEMS IN CONJUNCTION WITH CUTTING-EDGE EPIGENOMICS AND SINGLE CELL RNA SEQUENCING TECHNOLOGIES TO ADVANCE OUR UNDERSTANDING OF THE ONTOGENY, HETEROGENEITY, REGULATORY CIRCUITRY AND MECHANISM OF ACTION OF PFCD4 T CELLS. SPECIFICALLY, WE WILL INVESTIGATE HOW PERSISTENT STAT5 SIGNALING INDUCES AND MAINTAINS PFCD4 T CELLS (AIM 1), HOW CASTAT5-INDUCED PFCD4 T CELLS POTENTIATE ACT (AIM 2), AND THE POTENTIAL UTILITY OF CASTAT5 IN HUMAN CD19CAR T CELLS (AIM 3). SUCCESSFUL COMPLETION OF THIS PROJECT WILL ESTABLISH CASTAT5 AS A CLINICALLY APPLICABLE STRATEGY TO OVERCOME THE BARRIERS IMPEDING EFFECTIVE ACT INCLUDING CAR T CELL THERAPY.

TRANSLUMBOSACRAL NEUROMODULATION THERAPY FOR FECAL INCONTINENCE: RANDOMIZED TRIAL

ASTROCYTIC OSMR/JAK/STAT SIGNALING IN AD - THE ROLE OF REACTIVE ASTROCYTES IN ALZHEIMER’S DISEASE (AD) PATHOGENESIS REMAINS LARGELY UNDERSTUDIED. LITTLE IS KNOWN ABOUT HOW ASTROCYTES CHANGE THEIR FUNCTIONS/PROPERTIES UNDER DIFFERENT REACTIVE STATES AND WHAT CONSEQUENCES SUCH CHANGES CAUSE UNDER PATHOLOGICAL CONDITIONS. THE JAK/STAT PATHWAY IS A KEY PLAYER IN INDUCING ASTROCYTE REACTIVITY IN RESPONSE TO PROINFLAMMATORY CYTOKINES. ACTIVE JAK/STAT SIGNALING HAS BEEN OBSERVED IN HUMAN AD BRAINS AND AD ANIMAL MODELS, AND PLAYS A VITAL ROLE IN PROMOTING AD-RELATED PATHOLOGY AND COGNITIVE DEFICITS IN AD MODEL MICE. HOWEVER, HOW THE JAK/STAT PATHWAY IS ACTIVATED IN ASTROCYTES DURING AD PROGRESSION AND HOW AD-RELATED ASTROCYTE REACTIVITY AFFECTS OTHER BRAIN CELLS TO PROMOTE AD PATHOLOGY REMAIN UNCLEAR. OSM RECEPTOR SS (OSMRSS), ENCODED BY THE OSMR GENE, IS A KEY UPSTREAM ACTIVATOR OF THE JAK/STAT PATHWAY IN RESPONSE TO STIMULATION BY ONCOSTATIN M (OSM), A MEMBER OF THE IL-6 FAMILY OF CYTOKINES. SIGNIFICANTLY, OSMR HAS BEEN REVEALED AS A PROMINENT DISEASE-ASSOCIATED ASTROCYTE (DAA) MARKER, WHILE GENES ENCODING OTHER JAK/STAT UPSTREAM ACTIVATORS ARE NOT SPECIFICALLY ASSOCIATED WITH DAAS, SUGGESTING THAT OSMRSS PLAYS A UNIQUE ROLE IN AD-RELATED ACTIVATION OF THE JAK/STAT PATHWAY IN ASTROCYTES. OUR FUNCTIONAL TESTS SHOWED THAT ACTIVATION OF OSMRSS INCREASED EXPRESSION OF MULTIPLE DAA MARKERS IN ASTROCYTES, WHEREAS ASTROCYTIC DELETION OF OSMR (REFERRED TO AS OSMRCKO), OR BLOCKADE OF OSMRSS SIGNALING USING AN OSM NEUTRALIZING ANTIBODY, SIGNIFICANTLY REDUCED ASS-INDUCED EXPRESSION OF DAA MARKERS. THESE DATA SUGGEST THAT OSMRSS PLAYS A CRUCIAL ROLE IN INITIATING AD-RELATED ASTROCYTE REACTIVITY. MOREOVER, TREATMENT WITH THE OSM NEUTRALIZING ANTIBODY ATTENUATED ASS DEPOSITION IN APPNL-G-F KNOCK-IN MICE AND IMPROVED THEIR COGNITIVE PERFORMANCE, SUPPORTING A ROLE OF OSM/OSMRSS IN MODULATING THESE AD-RELEVANT PROCESSES. TO FURTHER SUPPORT THE DISEASE RELEVANCE OF OSMRSS IN AD, WE FOUND THAT ITS LEVEL WAS ELEVATED IN POSTMORTEM AD BRAINS. COLLECTIVELY, OUR PRELIMINARY DATA REVEAL A NOVEL OSMRSS/JAK/STAT AXIS THAT PLAYS A CRUCIAL ROLE IN INITIATING ASTROCYTE REACTIVITY AND PROMOTING AD-RELATED PATHOLOGY AND COGNITIVE DEFICITS. OUR CENTRAL HYPOTHESES ARE THAT ACTIVATION OF THE OSMRSS/JAK/STAT AXIS INDUCES AD-RELATED ASTROCYTE REACTIVITY TO DRIVE AD PROGRESSION AND THAT OSM/OSMRSS SIGNALING REPRESENTS AN ATTRACTIVE TARGET FOR AD THERAPY. WE WILL TEST THESE HYPOTHESES IN THREE AIMS. IN AIM 1, WE WILL EXAMINE THE MOLECULAR FEATURES OF OSMRSS-INITIATED JAK/STAT SIGNALING AND DETERMINE ITS ROLE IN INDUCING AD-RELATED ASTROCYTE DIFFERENTIATION AND HETEROGENEITY. IN AIM 2, WE WILL EXAMINE HOW OSMRSS/JAK/STAT-INDUCED ASTROCYTE REACTIVITY LEADS TO MULTI-FACETED FUNCTIONAL IMPAIRMENT OF ASTROCYTES AS WELL AS NEURONS AND MICROGLIA. IN AIM 3, WE WILL TEST WHETHER BLOCKING OSM/OSMRSS SIGNALING EFFECTIVELY AMELIORATES AD-RELATED PATHOLOGICAL AND COGNITIVE DEFICITS. SUCCESSFULLY ACCOMPLISHING THE PROPOSED STUDIES WILL REVEAL FUNDAMENTAL INFORMATION ABOUT THE HETEROGENEITY AND FUNCTIONAL IMPACT OF AD-RELATED ASTROCYTE REACTIVITY AND OFFER PRECLINICAL INSIGHT INTO TARGETING OSM/OSMRSS SIGNALING FOR IMPROVEMENT OF AD-RELATED NEUROPATHOLOGY AND COGNITIVE DEFICITS.

NOVEL MECHANISMS OF MUSCLE AND BONE LOSS WITH HIV INFECTION, ANTIRETROVIRAL THERAPY, AND AGING. - OUR PROPOSAL IS IN DIRECT RESPONSE TO THE SPECIAL FUNDING OPPORTUNITY ANNOUNCEMENT (FOA) PAR-21-068 “MULTIDISCIPLINARY STUDIES OF HIV/AIDS AND AGING (R01)”. A FRAILTY PHENOTYPE IS FREQUENTLY OBSERVED IN HIV PATIENTS ON LONG TERM ANTIRETROVIRAL THERAPY (ART), AND BOTH OSTEOPOROSIS AND SARCOPENIA ARE NOW RECOGNIZED AS CO-MORBIDITIES AMONG OLDER PEOPLE WITH HIV. LOSS OF MUSCLE MASS AND STRENGTH ARE IN TURN ASSOCIATED WITH POOR HEALTH OUTCOMES RANGING FROM FALLS AND FRACTURES TO ACCELERATED DISEASE PROGRESSION AND INCREASED MORTALITY. OUR GOAL IS TO ADDRESS THIS PROBLEM BY PROVIDING CRITICAL, NEW INFORMATION ON THE CELLULAR AND MOLECULAR MECHANISMS UNDERLYING MUSCULOSKELETAL DYSFUNCTION WITH HIV INFECTION AND ART, AND THEREBY IMPROVE SCIENTIFIC KNOWLEDGE, TECHNICAL CAPABILITY, AND EVENTUALLY CLINICAL PRACTICE. RECENT STUDIES IDENTIFY THE ARYL HYDROCARBON RECEPTOR (AHR) AS PLAYING A KEY ROLE IN REGULATING ORGANISMAL AGING AND LIFESPAN. OUR GROUP HAS FOUND THAT AHR ACTIVATION CAN INDUCE SENESCENCE IN BONE MARROW STEM CELLS, AND OTHERS HAVE OBSERVED THAT AHR OVEREXPRESSION INDUCES MUSCLE ATROPHY. UNITING THESE OBSERVATIONS, OUR CENTRAL HYPOTHESIS IS THAT MUSCLE- AND BONE-SPECIFIC AHR ACTIVATION ARE KEY DRIVERS OF MUSCLE AND BONE LOSS IN PATIENTS WITH HIV ON ART. OUR PRELIMINARY DATA PROVIDE A STRONG RATIONALE FOR THIS HYPOTHESIS AND INDICATE THAT 1) MARKERS OF MUSCLE ATROPHY, BONE LOSS, AND AHR ACTIVATION ARE INCREASED IN OUR MOUSE MODEL OF HIV INFECTION, 2) AHR IS HIGHLY EXPRESSED IN MUSCLE AND BONE, AND TARGETED KNOCKOUT OF AHR IN THESE TISSUES INCREASES LEAN MASS AND TRABECULAR BONE MASS, 3) PHARMACOLOGICAL INHIBITION OF AHR INCREASES MUSCLE STRENGTH AND MARKERS OF BONE FORMATION IN MICE, AND 4) THE ANTIRETROVIRAL EMTRICITABINE (FTC) INCREASES AHR ACTIVATION AND SENESCENCE IN MUSCLE CELLS AND THESE EFFECTS ARE ATTENUATED BY AHR SILENCING. SPECIFIC AIM 1 TESTS THE HYPOTHESIS THAT AHR ACTIVATION IS A KEY FACTOR DRIVING MUSCLE AND BONE LOSS WITH AGING AND HIV INFECTION. SPECIFIC AIM 2 TESTS THE HYPOTHESIS THAT AHR ACTIVATION IS A KEY FACTOR DRIVING MUSCLE AND BONE LOSS WITH AGING AND ANTIRETROVIRAL THERAPY. OUR EXPECTED OUTCOMES INCLUDE 1) DEFINING THE ROLE OF AHR IN SKELETAL MUSCLE AND BONE WITH HIV INFECTION AND ART SO THAT IT CAN BE TARGETED THERAPEUTICALLY, AND 2) CHARACTERIZING THE IMPACT OF AGING AND ART ON AHR ACTIVATION IN SKELETAL MUSCLE AND BONE. IN THE FUTURE THIS KNOWLEDGE MAY BE CRITICAL IN THE DIAGNOSIS, TREATMENT AND MANAGEMENT OF VULNERABLE PATIENT POPULATIONS DEBILITATED BY THE VAST ARRAY OF HIV- AND AGE-INDUCED PATHOLOGIES. ULTIMATELY, THESE DATA WILL ENABLE CLINICIANS TO IMPROVE DISEASE OUTCOMES AND, CONSEQUENTLY, PUBLIC HEALTH.

GUT METABOLITES, T CELLS, AND SALT-SENSITIVE HYPERTENSION - SUMMARY HYPERTENSION IS A PRIMARY MODIFIABLE RISK FACTOR FOR CARDIOVASCULAR, CEREBROVASCULAR, AND RENAL DISEASE, AND IS THE LARGEST INDIVIDUAL CONTRIBUTING FACTOR TO DISEASE AND MORTALITY IN THE WORLD. SALT-SENSITIVE HYPERTENSIVE INDIVIDUALS, WHO COMPRISE 30-50% OF THE HYPERTENSIVE POPULATION, HAVE GREATER MORTALITY THAN SUBJECTS WITH SALT- RESISTANT HYPERTENSION AND EXHIBIT RENAL END-ORGAN DAMAGE. IMMUNITY AND INFLAMMATION ARE IMPLICATED IN HYPERTENSION AND RENAL DAMAGE IN HUMANS AND EXPERIMENTAL ANIMAL, BUT THE MECHANISMS TRIGGERING IMMUNITY IN HYPERTENSION ARE NOT UNDERSTOOD. DIETARY COMPONENTS OTHER THAN SALT CAN ALSO PLAY AN IMPORTANT ROLE IN THE DEVELOPMENT OF CARDIOVASCULAR DISEASE AND HYPERTENSION. INTERESTINGLY, AN INVERSE RELATIONSHIP HAS BEEN DEMONSTRATED BETWEEN PLANT PROTEIN INTAKE AND BLOOD PRESSURE, ULTIMATELY ASSOCIATING HEALTH BENEFITS WITH GREATER PLANT PROTEIN CONSUMPTION. WE RECENTLY DISCOVERED A NOVEL LINK BETWEEN DIETARY PROTEIN INTAKE, IMMUNE- ACTIVATION, AND HYPERTENSION IN THE DAHL SALT-SENSITIVE (SS) RAT, A RODENT MODEL OF HUMAN DISEASE. THE EXPERIMENTS IN THIS PROPOSAL WILL TEST THE OVERARCHING HYPOTHESIS THAT FREE RADICAL PRODUCTION FROM PHAGOCYTIC NOX2 IN INFILTRATING CD4+ T CELLS IN THE KIDNEY OF DAHL SS LEADS TO AN INAPPROPRIATE ELEVATION OF RENAL VASCULAR RESISTANCE (RVR), A REDUCTION IN GLOMERULAR FILTRATION RATE (GFR), THE RETENTION OF SODIUM, AND THE FURTHER DEVELOPMENT OF HYPERTENSION FOLLOWING HIGH SALT FEEDING. AS A COROLLARY TO THIS HYPOTHESIS, WE PROPOSE THAT THE MICROBIAL METABOLITE CARNITINE, RELEASED BY CONSUMPTION OF ANIMAL-BASED DIETS, UPREGULATES NOX2 IN T CELLS AND AMPLIFIES SALT-SENSITIVE HYPERTENSION. IN CONTRAST, THE METABOLITE PROPIONATE, RELEASED FROM CONSUMPTION OF GRAIN- BASED DIETS, DOWNREGULATES NOX2 AND DIMINISHES THE FULL AMPLITUDE OF SALT-SENSITIVE HYPERTENSION. THE HYPOTHESIS WILL BE ADDRESSED IN TWO SPECIFIC AIMS. AIM 1 WILL ADDRESS THE MECHANISMS WHEREBY ALTERATIONS IN DIETARY PROTEIN SOURCE AFFECT THE RELEASE OF METABOLITES FROM THE GUT MICROBIOTA AND DETERMINE THE INFLUENCE OF THESE METABOLITES ON NOX2 IN T CELLS AND IN THE DEVELOPMENT OF SALT-SENSITIVE HYPERTENSION. AIM 2 WILL ADDRESS THE ROLE OF PHAGOCYTIC NOX2 IN CD4+ T CELLS AS A MEDIATOR OF INAPPROPRIATELY INCREASED RENAL VASCULAR RESISTANCE AND THE DEVELOPMENT OF SALT-SENSITIVE HYPERTENSION IN DAHL SS BY ADOPTIVE TRANSFER OF EITHER WILD TYPE OR NOX2- DEFICIENT CD4+ T CELLS INTO SS RATS LACKING T CELLS. THIS WORK WILL TRANSFORM THE UNDERSTANDING OF SALT-SENSITIVE HYPERTENSION BY UTILIZING NOVEL ANIMAL MODELS AND APPROACHES TO DEMONSTRATE THE MECHANISMS WHEREBY DIETARY PROTEIN INTAKE MODIFIES IMMUNE MECHANISMS WHICH SERVE TO AMPLIFY DISEASE SEVERITY. THESE STUDIES SHOULD REVEAL NEW PARADIGMS AND PROVIDE INSIGHT WITH THE POTENTIAL TO TRANSFORM CLINICAL/THERAPEUTIC APPROACHES FOR THE TREATMENT OF SALT-SENSITIVE HYPERTENSION.

NOTCH1/MIR-322 AXIS IN STEM CELL MEDIATED VASCULAR REPAIR

EFFECTS OF VITAMIN D AND OMEGA-3 SUPPLEMENTATION ON TELOMERES IN VITAL

MITOCHONDRIAL QUALITY CONTROL IN THE HEART - PROJECT SUMMARY ADULT CARDIOMYOCYTES (CMS) ARE CRITICALLY DEPENDENT ON THE PROPER FUNCTION OF MITOCHONDRIA TO SUPPLY THE FUEL FOR CONTRACTION (ATP) AND TO REGULATE OTHER ESSENTIAL CELLULAR PROCESSES. MITOCHONDRIAL HEALTH IS MAINTAINED BY A SELECTIVE FORM OF AUTOPHAGY, TERMED MITOPHAGY, WHICH REMOVES UBIQUITIN (UB)-LABELED DAMAGED MITOCHONDRIA. OUR CURRENT UNDERSTANDING OF MITOPHAGY IS BASED ON THE ACTIONS OF TWO KEY PROTEINS, THE UB LIGASE PARKIN AND KINASE PINK1, WHICH MARK DYSFUNCTIONAL MITOCHONDRIA FOR DEGRADATION VIA A PHOSPHORYLATED UB CHAIN. SURPRISINGLY, ABLATION OF PARKIN FAILED TO IMPACT HOMEOSTATIC MITOPHAGY OR CARDIAC FUNCTION IN BOTH YOUNG AND AGEING MICE, SUGGESTING THE EXISTENCE OF YET TO BE IDENTIFIED, PARKIN-INDEPENDENT MECHANISMS CONTROL MITOCHONDRIAL TURNOVER IN CMS. GIVEN ROBUST MITOPHAGIC ACTIVITY IN ADULT HEARTS AND THE DELETERIOUS IMPACT OF IMPAIRED MITOPHAGY IN VARIOUS CARDIAC DISEASES, THERE IS A GROWING NEED TO IDENTIFY NOVEL, PARKIN-INDEPENDENT REGULATORS OF MITOPHAGY IN THE HEART. IN PRELIMINARY DATA FOR THIS PROPOSAL, WE DOCUMENT A NEW MECHANISM LINKING CULLIN-RING UB LIGASE 5 (CRL5) WITH MITOCHONDRIAL QUALITY CONTROL IN THE HEART. CRL5 IS A MULTI-PROTEIN COMPLEX COMPRISED OF THE RING BOX PROTEIN RBX2, SCAFFOLD PROTEIN CULLLIN 5 (CUL5), ADAPTOR PROTEINS ELONGIN B/C, AND VARIOUS SUBSTRATE RECEPTOR PROTEINS. ACTIVATION OF CRL5 REQUIRES THE NEDDYLATION OF CUL5, A PROCESS THAT CONJUGATES THE SMALL UB-LIKE PROTEIN, NEDD8 TO TARGET PROTEINS. BY CONTROLLING THE TURNOVER OF ITS PROTEIN SUBSTRATES, CRL5 PARTICIPATES IN SEVERAL BIOLOGICAL PROCESSES AND HUMAN DISEASES, BUT A ROLE IN THE HEART IS NOT YET KNOWN. INHIBITION OF NEDDYLATION (UPSTREAM SIGNALING THAT GOVERNS CRL5 ACTIVITY), ROBUSTLY SUPPRESSED MITOCHONDRIAL UBIQUITINATION AND MITOPHAGY, AND MICE DEFICIENT IN NEDDYLATION DEVELOP HEART FAILURE DUE TO IMPAIRED MITOPHAGY AND MITOCHONDRIAL DYSFUNCTION. PROTEOMICS ANALYSIS IDENTIFIES RBX2 AND CUL5 ARE ASSOCIATED WITH MITOCHONDRIA IN HOMEOSTASIS; AND THEIR RECRUITMENT TO MITOCHONDRIA IS UPREGULATED FOLLOWING MITOCHONDRIA DAMAGE. RBX2 IS REQUIRED FOR MITOCHONDRIAL UBIQUITINATION AND TURNOVER, AND ITS ACTIONS IN MITOCHONDRIA ARE INDEPENDENT OF PARKIN BUT ARE RECAPITULATED BY CUL5. MOREOVER, DELETION OF RBX2 IN ADULT MICE PROVOKES ACCUMULATION OF DAMAGED MITOCHONDRIA AND LEADS TO HEART FAILURE. THESE FINDINGS SUPPORT OUR CENTRAL HYPOTHESIS THAT RBX2-CRL5 UB LIGASE MEDIATES MITOCHONDRIAL UBIQUITINATION AND MITOPHAGY TO REGULATE CARDIAC MYOCYTE FUNCTION AND SURVIVAL IN HOMEOSTASIS AND UNDER STRESS. THREE AIMS ARE PROPOSED. AIM 1 WILL DEFINE THE ROLE OF RBX2-CRL5 IN MITOCHONDRIAL TURNOVER IN THE HEALTHY NORMAL HEART USING RBX2- AND CUL5- DEFICIENT MICE. AIM 2 WILL IDENTIFY THE MOLECULAR UNDERPINNINGS OF CRL5-MEDIATED MITOPHAGY BY IDENTIFYING ITS SUBSTRATE RECEPTORS, SUBSTRATES AND SCAFFOLD PROTEINS IN MITOCHONDRIA AND BY INVESTIGATING THE INTERPLAY BETWEEN CRL5 AND PINK1. AIM 3 WILL TEST THE FEASIBILITY OF MODULATION OF RBX2 TO IMPROVE MITOPHAGY AND CARDIAC INJURY IN PRESSURE- OVERLOADED HEARTS. THIS STUDY IS SIGNIFICANT AS IT WILL IDENTIFY CRL5 AS A MITOCHONDRIAL UB LIGASE TO MAINTAIN MITOCHONDRIAL INTEGRITY AND THUS PROVIDE POTENTIAL THERAPEUTIC TARGETS FOR TREATMENT OF HEART FAILURE.

PULSED LASER THERAPY IN AN ASYMPTOMATIC HEAD INJURY-ACCELERATED ALZHEIMER'S MODEL - ABSTRACT TRAUMATIC BRAIN INJURY (TBI) IS A SIGNIFICANT CAUSE OF DEATH AND DISABILITY IN THE US. THE TRUE EXTENT OF TBI MAY BE UNDERESTIMATED, WITH MANY CASES OF ASYMPTOMATIC TBI GOING UNDIAGNOSED, OFTEN REFERRED TO AS A “SILENT EPIDEMIC.” RECENT META-ANALYSES SUGGEST THAT HEAD INJURIES INCREASE THE RISK OF DEMENTIA AND ALZHEIMER'S DISEASE (AD). AS A RESULT, THERE IS AN URGENT NEED FOR STRATEGIES TO PREVENT TBI-INDUCED AD DEMENTIA AND RESEARCH TO UNCOVER ITS UNDERLYING MECHANISMS. NOTABLY, TBI OFTEN RESULTS IN SIGNIFICANT DAMAGE TO THE BLOOD-BRAIN BARRIER (BBB). EVEN A SINGLE MILD TBI CAN LEAD TO PERSISTENT BBB BREAKDOWN. ADDITIONALLY, TBI COMMONLY DAMAGES BRAIN MICROVASCULAR ENDOTHELIAL CELLS, CRUCIAL COMPONENTS OF THE BBB. BOTH CEREBROVASCULAR DYSFUNCTION AND BBB DAMAGE ARE EARLY AND CRITICAL EVENTS IN AD PATHOLOGY. THEREFORE, IT IS REASONABLE TO HYPOTHESIZE THAT TBI-RELATED BBB BREAKDOWN MAY CONTRIBUTE TO THE AGGRAVATED PROGRESSION OF AD. HOWEVER, THE CAUSAL RELATIONSHIP BETWEEN DISRUPTED ENDOTHELIAL CELL AND BBB FUNCTION CAUSED BY TBI AND THE ACCELERATED AD PROGRESSION REMAINS UNCLEAR. IN THIS STUDY, WE PROPOSE NONINVASIVE LASER THERAPY AS A POTENTIAL THERAPEUTIC APPROACH FOR BRAIN PROTECTION IN A MILD CLOSED-HEAD INJURY (CHI)-ACCELERATED AD DEMENTIA MODEL. TO TEST OUR OVERALL HYPOTHESIS, AIM 1 WILL DETERMINE THE IMPACT OF PULSED LASER THERAPY ON CHI-ACCELERATED AD PATHOGENESIS IN A NOVEL AD RAT MODEL AND ELUCIDATE THE ROLE OF DDR-SENESCENT SIGNALING IN THIS PROCESS. AIM 2 WILL TEST THE HYPOTHESIS THAT ENDOTHELIAL CELL SENESCENCE AND BBB DAMAGE REPRESENT SIGNIFICANT PATHOLOGICAL LINKS BETWEEN TBI AND AD. ADDITIONALLY, WE WILL ESTABLISH A MECHANISTIC ANGLE TO DETERMINE WHETHER H2S SIGNALING MOLECULE IS ASSOCIATED WITH LASER PROTECTION IN CHI-AD. AIM 3 WILL EXAMINE THE HYPOTHESIS THAT LASER THERAPY CAN TARGET ION-CONTAINING CYSTATHIONINE-Β-SYNTHASE/H2S SYSTEM TO ENHANCE DNA/VASCULAR REPAIR AND FUNCTIONAL NEUROVASCULAR COUPLING DURING CHI-AD PATHOGENESIS. THE PROPOSED RESEARCH IS INNOVATIVE BOTH TECHNICALLY AND MECHANISTICALLY. THE STUDIES HAVE THE POTENTIAL FOR PROFOUND IMPACT AS THEY AIM TO ELUCIDATE THE MECHANISMS UNDERLYING TBI-ACCELERATED AD PROGRESSION, IDENTIFY NOVEL TARGET FOR LASER THERAPY IN CHI-AD, AND ASSESS THE EFFICACY AND UTILITY OF THIS EMERGING NONINVASIVE THERAPY IN THE EARLY STAGES OF TBI-AD PROGRESSION.

DECIPHERING THE MECHANISMS AND CELLULAR ROLES OF MONOMER-DRIVEN ACTIN DYNAMICS

GPCR ANTEROGRADE TRAFFICKING

ROLE OF SMOOTH MUSCLE CALPONIN IN VASCULAR PATHOBIOLOGY

NOVEL MECHANISMS OF HIV-ASSOCIATED PULMONARY VASCULAR DISEASE - PROJECT SUMMARY THANKS TO THE ADVENT OF COMBINATION ANTIRETROVIRAL THERAPY (CART), LIFE EXPECTANCY HAS DRAMATICALLY INCREASED IN PEOPLE WITH HIV (PWH) AND THE SPECTRUM OF DISEASES RELATED TO HIV HAS SHIFTED TOWARDS NON-INFECTIOUS CHRONIC ILLNESSES INCLUDING CARDIOPULMONARY DISORDERS. PULMONARY VASCULAR DISEASE (PVD) IS AMONGST THE MOST DEVASTATING CARDIOVASCULAR COMPLICATIONS OF HIV. HOWEVER, THE ETIOPATHOLOGY OF PULMONARY VASCULAR REMODELING, THE CRITICAL STRUCTURAL ALTERATION AND PATHOLOGICAL FEATURE FOR PVD, HAS YET TO BE DETERMINED. HEREIN, WE WILL TEST WHETHER HIV-DERIVED PROTEINS, WHICH REMAIN IN CIRCULATION DESPITE CART AND WELL-CONTROLLED VIREMIA, CONTRIBUTE TO THE DEVELOPMENT OF PVD. WE PROVIDE NOVEL EXCITING PRELIMINARY DATA IN TWO MOUSE MODELS OF HIV SUPPORTING A ROLE FOR CD4+ T CELLS, HIV-TAT, AND PODOCAN-LIKE PROTEIN 1 (PODNL1) IN PVD. WE DEMONSTRATE THAT EXPRESSION OF HIV-DERIVED PROTEINS IN TG26 MICE PROMOTES PVD AS SHOWN BY REMODELED PULMONARY ARTERIES (PAS), INCREASED RIGHT VENTRICULAR SYSTOLIC PRESSURE AND RIGHT VENTRICLE HYPERTROPHY, DECREASED PULMONARY ARTERY ACCELERATION TIME (PAAT) AND RATIO OF PAAT/PULMONARY EJECTION TIME, AS WELL AS INCREASED CIRCULATING INTERLEUKIN-1Α (IL-1Α) LEVELS IN MALE AND FEMALE MICE. WE ALSO REPORT THAT THESE CARDIOPULMONARY ALTERATIONS ARE EXACERBATED WITH AGING. REMARKABLY, WE SHOW THAT BONE MARROW TRANSPLANT (BMT) FROM TG26 TO WILD TYPE (WT) MICE FULLY RECAPITULATE THE CARDIOPULMONARY PHENOTYPE OF TG26 MICE, WHICH SUPPORTS A ROLE FOR VIRAL PROTEINS DERIVED FROM HEMATOPOIETIC CELLS IN PVD. USING RNA SEQ ANALYSIS, WE IDENTIFIED A NEW MARKER OF PVD, PODNL1. ITS LEVELS ARE INCREASED IN THE LUNGS OF BOTH INTACT AND BMT TG26 MICE, BUT MORE IMPORTANTLY IN THE LUNGS OF PWH ON CART. WE DEMONSTRATED THAT OVEREXPRESSION OF PODNL1 IN VASCULAR SMOOTH MUSCLE CELLS (VSMCS) INCREASES COLLAGEN LEVELS AND THAT OF PCNA, A MARKER OF CELL PROLIFERATION. USING A NEWLY DEVELOPED “T CELL- BLOOD VESSEL” CO-CULTURE SYSTEM, WE SHOW THAT EXPOSURE OF DISCARDED HUMAN AORTA AND PULMONARY ARTERY SPECIMENS TO TG26 T CELLS ELEVATE PODNL1 EXPRESSION. REMARKABLY, WE REPORT THAT HIV-TAT IS THE ONLY VIRAL PROTEIN DETECTABLE IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) OBTAINED FROM PWH ON CART AND SHOW THAT CHRONIC TREATMENT OF MICE WITH TAT REPRODUCES THE CARDIOPULMONARY COMPLICATIONS OF INTACT AND BMT TG26 MICE. STRIKINGLY, WE PROVIDE PRELIMINARY EVIDENCE OF HIGH IL-1Α LEVELS IN PBMCS FROM PWH AND FOUND HIGHER IL-1Α LEVELS IN CD4+ THAN IN CD8+ T CELLS ISOLATED FROM TG26 MICE. TOGETHER, THESE EXCITING AND NOVEL FINDINGS FORM THE CORE HYPOTHESIS OF THIS PROPOSAL THAT CD4+ T CELLS EXPRESSING TAT PROMOTE PVD VIA IL1Α-INDUCED, PODNL1-MEDIATED PULMONARY ARTERY REMODELING. WE WILL TEST THIS HYPOTHESIS IN THREE AIMS. IN AIM 1, WE WILL TEST WHETHER HIV-ENCODED PROTEINS ACCELERATE PVD VIA PODNL1-DEPENDENT MECHANISMS. IN AIM 2, WE WILL INVESTIGATE WHETHER HIV-ASSOCIATED PVD INVOLVES CD4+ T CELL-MEDIATED, IL-1Α-INDUCED PVD, WHILE AIM 3 WILL TEST THE HYPOTHESIS THAT HIV-DERIVED TAT PROTEIN PROMOTES PODNL1 PRODUCTION AND MEDIATES THE DEVELOPMENT OF HIV-PVD. WE ANTICIPATE THAT THIS PROPOSAL WILL LEAD TO THE IDENTIFICATION OF NEW THERAPEUTIC TARGETS FOR THE PREVENTION OF HIV-ASSOCIATED PVD.

PFKFB3 IN VASCULAR REMODELING

PHASE I STUDY OF PANOBINOSTAT IN ADULTS WITH SICKLE CELL DISEASE: NOVEL APPROACH TO RECRUITMENT AND RETENTION - ABSTRACT ALTHOUGH AN ORPHAN DISEASE IN THE US WITH ~110,000 AFFECTED INDIVIDUALS, SICKLE CELL DISEASE (SCD) POSES A MAJOR MEDICAL-ECONOMIC PROBLEM, WITH HOSPITALIZATION COSTS OF >$1 BILLION ANNUALLY. PATIENTS WITH SCD SUFFER FREQUENT VASO-OCCLUSIVE PAIN EPISODES DUE TO MICROVASCULAR OCCLUSION, HEMOLYTIC ANEMIA, AND A CHRONIC INFLAMMATORY STATE PRECIPITATED BY ISCHEMIA-REPERFUSION INJURY, CULMINATING IN ORGAN FAILURE AND A SHORTENED LIFE EXPECTANCY. THERE ARE ONLY FOUR FDA-APPROVED DISEASE-MODIFYING AGENTS FOR SCD, INCLUDING HYDROXYUREA (HU) IN 1998, ENDARI® (L-GLUTAMINE) IN 2017 AND MORE RECENTLY ADAKEVO® (CRIZANLIZUMAB) AND OXBRYTA® (VOXELOTOR) IN 2019. HU PRIMARILY ACTS BY REVERSING THE PERINATAL SWITCH FROM FETAL TO ADULT HEMOGLOBIN (HB) EXPRESSION; INCREASING THE FETAL HEMOGLOBIN (HB F) CONTENT IN RED BLOOD CELLS (RBCS) EXERTS A POTENT ANTI-SICKLING EFFECT. LONG-TERM BENEFICIAL EFFECTS OF HU ON SURVIVAL AND PRESERVATION OF ORGAN FUNCTION WAS DOCUMENTED IN MULTI- CENTER STUDY OF HYDROXYUREA, BASED ON A 17-YEAR FOLLOW-UP, AND MANY OTHER STUDIES IN ADULTS AND CHILDREN WITH SCD; DESPITE THESE BENEFITS, HU IS UNDERUTILIZED IN SCD. WHILE THE REASONS FOR UNDERUTILIZATION OF HU ARE BEING INVESTIGATED IN EIGHT CENTERS IN AN NHLBI-FUNDED IMPLEMENTATION STUDY, CLINICAL EXPERIENCE SUGGESTS PATIENTS HAVE CONCERNS ABOUT SIDE EFFECTS, FERTILITY, AND CARCINOGENICITY RELATED TO HU, THUS CREATING AN UNMET NEED FOR ADDITIONAL DISEASE-MODIFYING ANTI-SWITCHING THERAPIES. HISTONE DEACETYLASE INHIBITORS (HDACI) HAVE BEEN SHOWN TO INDUCE HB F EXPRESSION IN ERYTHROID CULTURES, AND IN PRECLINICAL STUDIES IN MOUSE MODELS OF SCD, PRIMARILY BY INHIBITING EPIGENETIC SILENCING OF THE FETAL () GLOBIN GENES VIA ACETYLATION OF HISTONES AND OPENING -GLOBIN GENES CHROMATIN TO ACTIVATE TRANSCRIPTION. WE PREVIOUSLY CONDUCTED AN INVESTIGATOR-INITIATED PHASE I TRIAL OF THE PAN- HDACI PANOBINOSTAT IN ADULTS WITH SCD. NINE PATIENTS WERE TREATED WITH PANOBINOSTAT 10 MG MWF, IN THREE COHORTS ON DIFFERENT DOSING SCHEDULES. THE DRUG WAS WELL TOLERATED WITHOUT TOXICITIES OR SIDE EFFECTS. A SLIGHT INCREASE IN HB F AND F-CELLS WAS NOTED IN THE 10 MG MWF CONTINUOUS TREATMENT DOSE. TO TEST THE EFFICACY OF PANOBINOSTAT, WE PROPOSE TO COMPLETE THREE SPECIFIC AIMS: 1) COMPLETE A PHASE I TRIAL WITH ESCALATING PANOBINOSTAT DOSES (18 TOTAL PATIENTS AT 15 MG OR 20 MG MWF ON INTERMITTENT OR CONTINUOUS TREATMENT). THE PRIMARY END-POINT IS TO ESTABLISH THE SAFETY AND TOLERABILITY OF PANOBINOSTAT IN SCD PATIENTS. SECONDARY ENDPOINTS WILL INCLUDE MONITORING CHANGES IN HB F AND F-CELLS. 2) INITIATE A NOVEL INTERVENTION TO SUPPORT STUDY ADHERENCE, MOTIVATIONAL INTERVIEWING, WHICH HAS BEEN SHOWN TO BE EFFECTIVE IN HELPING INDIVIDUALS OVERCOME BARRIERS TO MEDICATION AND TREATMENT ADHERENCE IN A VARIETY OF SETTINGS. 3) DEFINE BIOMARKERS PREDICTIVE OF PANOBINOSTAT TREATMENT RESPONSE. THE ACETYLATION OF NON-HISTONE PROTEINS INCLUDING NFKB, STAT, AND P53 AND INFLAMMATORY MARKERS WILL BE INVESTIGATED. FURTHERMORE, GENOME-WIDE BIOMARKERS OF HISTONE ACETYLATION WILL BE DISCOVERED BY CHIPSEQ AND DIFFERENTIALLY EXPRESSED GENES BY RNASEQ. THIS STUDY WILL ESTABLISH THE SAFETY OF PANOBINOSTAT IN SCD AND WILL PROVIDE PROOF-OF-CONCEPT FOR TARGETING MULTIPLE PATHWAYS TO EXPAND TREATMENT OPTIONS.

CU UPTAKE TRANSPORTER AS A DISTURBED FLOW SENSOR IN VASCULAR INFLAMMATORY DISEASE - PROJECT SUMMARY THE AIM OF THIS GRANT IS TO ELUCIDATE THE NOVEL ROLE OF ENDOTHELIAL CU UPTAKE TRANSPORTER CTR1 AS A “MECHANOSENSOR” THAT PROMOTES CU-DEPENDENT INFLAMMATION AND CUPROPTOSIS INVOLVED IN ATHEROSCLEROSIS. OXIDATIVE STRESS, INFLAMMATION, AND MITOCHONDRIAL (MITO) DYSFUNCTION IN ENDOTHELIAL CELLS (ECS) CONTRIBUTES TO ATHEROSCLEROSIS, PREDOMINANTLY OCCURRING IN ARTERIAL REGIONS EXPOSED TO DISTURBED BLOOD FLOW (D- FLOW), WHILE THOSE IN THE STABLE LAMINAR FLOW (S-FLOW) REGIONS ARE PROTECTED. THE MECHANISMS BY WHICH D-FLOW AND S-FLOW REGULATE ATHEROGENESIS ARE STILL POORLY UNDERSTOOD. COPPER (CU), AN ESSENTIAL MICRONUTRIENT, IS GREATLY INCREASED IN HUMAN ATHEROSCLEROTIC PLAQUES, WHILE CU PROMOTES, AND CU CHELATION INHIBITS ATHEROSCLEROSIS IN MICE VIA UNKNOWN MECHANISMS. THE BIOAVAILABILITY OF INTRACELLULAR CU IS TIGHTLY CONTROLLED BY CU TRANSPORT AND CU CHAPERONE PROTEINS INCLUDING CU UPTAKE TRANSPORTER CTR1 AND CYTOSOLIC CU CHAPERONE ATOX1 THAT ALSO FUNCTIONS AS A CU-DEPENDENT TRANSCRIPTION FACTOR TO UPREGULATE INFLAMMATORY GENE EXPRESSION. RECENT EVIDENCE REVEALS THAT EXCESS CU INDUCES A NEW TYPE OF PROGRAMMED CELL DEATH, TERMED “CUPROPTOSIS”, CHARACTERIZED BY DECREASE IN LIPOYLATED TCA CYCLE PROTEINS AND FE-S CLUSTER PROTEINS, RESULTING IN MITO DYSFUNCTION. HOWEVER, THE ROLES OF CU AND ENDOTHELIAL CTR1 IN D-FLOW-INDUCED MECHANO-SIGNALING, INFLAMMATION, MITO DYSFUNCTION AND ANY INVOLVEMENT OF CUPROPTOSIS ARE ENTIRELY UNKNOWN. OUR PRELIMINARY DATA ARE CONSISTENT WITH THE HYPOTHESIS THAT ENDOTHELIAL CTR1 FUNCIONS AS A NOVEL DISTURBED FLOW “MECHANOSENSOR” THAT ORCHESTRATES CYTOSOLIC CU- MEDIATED ATOX1 NUCLEAR TRANSLOCATION VIA ROS-DEPENDENT ACETYLATION LEADING TO INFLAMMATION (EARLY PHASE) AS WELL AS MITO CU ACCUMULATION FOLLOWING CTR1 BINDING TO MITOCU TRANSPORTER, LEADING TO CUPROPTOSIS (LATE PHASE), WHICH CONTRIBUTES TO ATHEROSCLEROSIS. AIM 1 WILL ESTABLISH THE ROLE OF CTR1 AS A D-FLOW SENSOR TO DRIVE CU- AND ROS-DEPENDENT ATOX1 NUCLEAR TRANSLOCATION AND INFLAMMATION AND ADDRESS MOLECULAR MECHANISMS IN CULTURED ECS. AIM 2 WILL DETERMINE WHETHER D-FLOW INDUCES MITO DYSFUNCTION AND CUPROPTOSIS VIA INCREASING MITOCU FOLLOWING CTR1 BINDING TO MITOCU TRANSPORTER SLC25A3 IN ECS. AIM 3 WILL DETERMINE THE ROLE OF ENDOTHELIAL CTR1 IN VASCULAR INFLAMMATION, CUPROPTOSIS AND FLOW-DEPENDENT ATHEROSCLEROSIS AND ADDRESS UNDERLYING MECHANISMS IN VIVO. WE WILL USE INDUCIBLE EC-SPECIFIC CTR1-/- OR -ATOX1-/- OR -SLC25A3-/- MICE OR NEWLY DEVELOPED CRISPR/CAS9-GENERATED “ACETYLATION DEAD” ATOX1 KNOCK-IN MUTANT MICE WITH HIGH FAT DIET OR PARTIAL CAROTID LIGATION TO INDUCE ATHEROSCLEROSIS. WE WILL ALSO USE COMPARTMENT-SPECIFIC REDOX-SENSITIVE BIOSENSORS; BIOTIN-LABELED CYSOH TRAPPING PROBE; SCRNASEQ AND SCATACSEQ; NEWLY DEVELOPED, HIGHLY SPECIFIC MITO-TARGETED CU-DEPLETING NANOPARTICLE (MITOCDN); HIGHLY INNOVATIVE ICP-MASS SPEC, X-RAY FLUORESCENCE MICROSCOPY AND MITO-TARGETED CU FLUORESCENCE PROBES TO MEASURE CU LEVELS IN CELLS OR TISSUES. OUR PROPOSAL WILL PROVIDE NEW INSIGHTS INTO ENDOTHELIAL CTR1 AS A POTENTIAL THERAPEUTIC TARGET FOR TREATMENT OF FLOW- AND CU- DEPENDENT ATHEROSCLEROSIS.

TRANSCRIPTIONAL CONTROL OF MYOCARDIN AND THE MYOCARDOME

EARLY LIFE STRESS, DAMAGE ASSOCIATED MOLECULAR PATTERNS, AND CARDIOVASCULAR RISK

A MULTIFACETED APPROACH TO TARGET OBESITY

DARC, INFLAMMAGING AND ALZHEIMER'S DISEASE - PROJECT SUMMARY CHRONIC INFLAMMATION OF THE BRAIN OCCURS WITH AGING AND PLAYS AN IMPORTANT ROLE IN AGE-RELATED COGNITIVE DECLINE AND ALZHEIMER’S DISEASE (AD) PROGRESSION. PROINFLAMMATORY CHEMOKINES NOT ONLY BIND TO THEIR COGNATE RECEPTORS, BUT ALSO TO “ATYPICAL” CHEMOKINE RECEPTORS (ACKR) THAT LACK INTRACELLULAR SIGNALING DOMAINS AND PRINCIPALLY SERVE TO BUFFER OR CONTROL CHEMOKINE BIOAVAILABILITY. THE DUFFY ANTIGEN RECEPTOR FOR CHEMOKINES (DARC) IS A MEMBER OF THE ACKR SUBFAMILY. DARC IS HIGHLY EXPRESSED ON RED BLOOD CELLS (RBCS), AND TO A LESSER EXTENT IN OTHER CELL TYPES, WHERE IT FUNCTIONS AS A CHEMOKINE ‘BUFFER-SINK’, SEQUESTERING PROINFLAMMATORY CHEMOKINES. THERE ARE TWO DOMINANT DARC ALLELES IN HUMANS, FYB (PREDOMINANT IN EUROPEAN CAUCASIANS) AND FYA (HYPOFUNCTIONAL ALLELE PREVALENT IN ASIANS). A MUTATION IN THE PROMOTER REGION OF THE FYB ALLELE ABOLISHES THE EXPRESSION OF DARC IN RBC. NOTABLY, APPROXIMATELY 70% OF AFRICAN AMERICANS HARBOR THIS MUTATION, WHICH IS ASSOCIATED WITH A PRO- INFLAMMATORY PHENOTYPE. WE PROVIDE NOVEL PRELIMINARY DATA SHOWING THAT BOTH GLOBAL DARC KNOCKOUT MICE, AND MICE LACKING DARC SELECTIVELY IN BLOOD CELLS, EXHIBIT COGNITIVE IMPAIRMENT AND INCREASED LEVELS OF PROINFLAMMATORY MEDIATORS ASSOCIATED WITH AD. MOREOVER, DARC IS DOWNREGULATED IN THE BRAIN WITH AGING AND IN AD MICE, AND OVEREXPRESSION OF MOUSE DARC IN THE BRAIN ALLEVIATED COGNITIVE IMPAIRMENT IN 5XFAD MICE, SUPPORTING THAT DARC PROTECTS AGAINST AD-RELATED COGNITIVE DECLINE. THESE NOVEL FINDINGS LEAD US TO HYPOTHESIZE THAT DARC PLAYS AN IMPORTANT ROLE IN REGULATING NEUROINFLAMMATION, COGNITIVE DYSFUNCTION AND AD PATHOLOGIES. TO TEST THIS HYPOTHESIS, WE PROPOSE THREE SPECIFIC AIMS. IN AIM 1, WE WILL DETERMINE IF SELECTIVE ABLATION OF DARC IN BLOOD CELLS ACCELERATES AGE-RELATED NEUROINFLAMMATION AND COGNITIVE DECLINE AND EXACERBATES NEUROPATHOLOGICAL AND COGNITIVE PHENOTYPES OF AD MICE. IN AIM 2, WE WILL DETERMINE IF ABLATION OR OVEREXPRESSION OF MOUSE DARC IN FOREBRAIN GLUTAMATERGIC NEURONS MODULATES NEURONAL VULNERABILITY TO DEGENERATION, AΒ/TAU PATHOLOGIES, AND AGING- AND AD-ASSOCIATED COGNITIVE IMPAIRMENT. IN AIM 3, WE WILL DETERMINE IF DARC-FYA AND FYB ISOFORMS DIFFER IN THEIR ABILITY TO SEQUESTER CHEMOKINES AND AΒ AND THEIR PROTECTIVE EFFECTS AGAINST NEUROINFLAMMATION AND COGNITIVE IMPAIRMENT.

REGULATION OF ANTIBODY SECRETING CELL (ASC) HOMEOSTASIS BY UFBP1.

NOVEL BIOMARKERS FOR THE CLINICAL MANAGEMENT OF BLADDER CANCER

SMC MACROPINOCYTOSIS: A NOVEL TARGET IN ATHEROSCLEROTIC VASCULAR DISEASE - PROJECT SUMMARY ATHEROSCLEROSIS IS A LEADING CAUSE OF MORBIDITY AND MORTALITY GLOBALLY. RECENTLY, CELL LINEAGE TRACING, SINGLE-CELL RNA SEQUENCING AND HUMAN GENOMIC STUDIES HAVE BEEN INTEGRATED TO DEMONSTRATE THAT A) MAJORITY OF PLAQUE FOAM CELLS ARE OF VASCULAR SMOOTH MUSCLE CELL (SMC) ORIGIN AND B) SMC CAN UNDERGO A FATE SWITCH TO TRANSITIONAL, MULTIPOTENTIAL CELLS THAT CAN ADOPT PLAQUE ALTERING PHENOTYPES. ALTHOUGH THESE RESULTS IDENTIFY SMC AS POTENTIAL THERAPEUTIC TARGETS, MOST CURRENT TREATMENTS FOR ATHEROSCLEROSIS HAVE LITTLE DIRECT IMPACT ON SMC. THE ENDOCYTIC PROCESSES BY WHICH SMC TAKE UP LIPIDS AND BECOME FOAM CELLS IN THE ARTERIAL WALL ARE NOT CLEARLY DEFINED. IN ADDITION, THE MECHANISMS UNDERLYING SMC PHENOTYPIC SWITCHING IN THE ARTERIAL WALL REMAIN LARGELY UNKNOWN. USING A COMBINATION OF HIGH-RESOLUTION IMAGING, 3D CELL RECONSTRUCTION, AND LDL IMMUNOLABELING, WE HAVE RECENTLY DEMONSTRATED THAT FOAM CELLS (LINEAGE UNKNOWN) IN HUMAN AND MURINE ATHEROSCLEROTIC PLAQUES INTERNALIZE LDL VIA MACROPINOCYTOSIS. NOVEL PRELIMINARY DATA USING SMC LINEAGE TRACING IDENTIFIES SMC SUBSETS PERFORMING MACROPINOCYTOSIS IN THE ARTERIAL WALL IN VIVO. STIMULATION OF MACROPINOCYTOSIS OF LDL IN SMC PROMOTES DEDIFFERENTIATION AND PHENOTYPE SWITCHING INTO PLAQUE-PROMOTING PHENOTYPES IN VITRO. FURTHER, PHARMACOLOGICAL BLOCKADE OF MACROPINOCYTOSIS USING THE NA+/H+ EXCHANGER 1 (NHE1) BLOCKER, EIPA, INHIBITS LDL UPTAKE IN ATHEROSCLEROTIC ARTERIES AND ABROGATES ATHEROSCLEROSIS DEVELOPMENT IN MULTIPLE MURINE MODELS OF ATHEROSCLEROSIS. PRELIMINARY DATA ALSO SHOW THAT THE MATRICELLULAR PROTEIN THROMBOSPONDIN-1 (TSP1) STIMULATES MACROPINOCYTOSIS VIA CD47 IN SMC, LEADING TO FOAM CELL FORMATION AND PHENOTYPIC SWITCHING, AND GLOBAL CD47-/- MICE ARE PROTECTED FROM ATHEROSCLEROSIS. BASED ON THESE OBSERVATIONS, WE HYPOTHESIZE THAT SMC MACROPINOCYTOSIS DRIVES ATHEROSCLEROSIS THROUGH FOAM CELL FORMATION AND REGULATION OF SMC PHENOTYPIC SWITCHING. THE HYPOTHESIS WILL BE TESTED VIA THE FOLLOWING SPECIFIC AIMS: AIM 1: SMC INTERNALIZE LDL VIA MACROPINOCYTOSIS IN ATHEROSCLEROTIC ARTERIES. AIM 2: SMC-SPECIFIC INHIBITION OF MACROPINOCYTOSIS ATTENUATES TRANSDIFFERENTIATION OF SMC INTO A PLAQUE- PROMOTING PHENOTYPE AND INHIBITS ATHEROSCLEROSIS. AIM 3: TSP1 VIA CD47 STIMULATES SMC MACROPINOCYTOSIS, PROMOTES SMC PHENOTYPIC SWITCHING AND CONTRIBUTES TO ATHEROSCLEROSIS DEVELOPMENT. THE PROPOSAL WILL EMPLOY SMC-SPECIFIC KNOCKOUT MICE (NHE1-/-, CD47-/- AND CD36-/-), PRIMARY HUMAN AND MURINE AORTIC SMC, A VASCULAR SMC-RESTRICTED CRE DRIVER MOUSE MODEL (ITGA8-CREERT2+/-; MTMG+/-), AND OTHER GENETIC TOOLS TO TEST THE HYPOTHESIS. MULTIPLE COMPLEMENTARY TECHNIQUES WILL BE USED TO STUDY SMC MACROPINOCYTOSIS IN VITRO (PHARMACOLOGICAL, GENETIC, FLUORESCENCE/SEM IMAGING) AND IN VIVO (TEM/IEM MICROSCOPY, SMC-SPECIFIC NHE1 KNOCKOUT MICE). AT THEIR CONCLUSION, THE PROPOSED STUDIES WILL DEFINE KEY MECHANISM(S) PROMOTING SMC FOAM CELL FORMATION AND PHENOTYPIC SWITCHING AND POTENTIALLY IDENTIFY NEW TARGETS FOR THERAPEUTIC INTERVENTIONS OF ATHEROSCLEROSIS.

NOVEL MECHANISTIC PATHWAYS OF CARDIOVASCULAR DISEASE IN OBESITY

NEUROPROTECTIVE AND NEUROGENIC ACTIONS OF E2 AND SERMS

EFFICACY-TO-EFFECTIVENESS TRANSITION OF AN EDUCATIONAL PROGRAM TO INCREASE COLORE

MOLECULAR BASIS OF INFLAMMATION IN RETINA, AND NOVEL STRATEGIES FOR LIMITING IT

INFLAMMATION AND RETINOPATHY OF PREMATURITY

ROLE OF IDO MALIGNANCY

COORDINATING UNIT FOR THE NATIONAL CENTERS FOR METABOLIC PHENOTYPING IN LIVE MODELS OF OBESITY AND DIABETES (MPMOD) - THE NATIONAL CENTERS FOR METABOLIC PHENOTYPING IN LIVE MODELS OF OBESITY AND DIABETES (MPMOD) WILL BE A MULTI-CENTER INITIATIVE FUNDED BY THE NIH TO PROVIDE STATE-OF-THE-ART TECHNOLOGIES TO INVESTIGATORS FOR A FEE, WITH THEIR SERVICES INCLUDING CHARACTERIZATION OF MOUSE METABOLISM, BLOOD COMPOSITION (INCLUDING HORMONES), ENERGY BALANCE, EATING AND EXERCISE, ORGAN FUNCTION AND MORPHOLOGY, AND PHYSIOLOGY. THE MPMOD COORDINATING UNIT (CU) WILL BE RESPONSIBLE FOR SUPPORTING MPMOD BY CREATING/MAINTAINING THE WEBSITE, ADMINISTRATIVELY ORGANIZE THE MEETINGS AND WORKSHOPS SPONSORED BY THE MPMOD TO ENGAGE THE GREATER MOUSE METABOLISM SCIENTIFIC COMMUNITY AS WELL AS DEVELOP THE VIBRANT PROGRAM TO ESTABLISH A P&F FUNDING PROGRAM AND OUTREACH TO UNDERSERVED RESEARCHERS AND INSTITUTIONS. IN ADDITION, THE CU WILL BE RESPONSIBLE FOR MANAGING AND ORGANIZING THE ACTIVITIES OF THE MPMOD CENTER SITES TO PROVIDE QUALITY PHENOTYPING SERVICES TO THE SCIENTIFIC COMMUNITY. THE CU IS RESPONSIBLE FOR CREATING AND MAINTAINING THE ADMINISTRATIVE, SCIENTIFIC AND INFORMATICS INFRASTRUCTURE NECESSARY TO ORGANIZE AND FACILITATE THEIR OPERATIONS. THE GOAL OF THIS PROPOSAL IS TO PROVIDE THAT INFRASTRUCTURE. OVER THE LAST 15 YEARS, OUR GROUP HAS SUCCESSFULLY MANAGED AND CREATED THE INFRASTRUCTURE AS THE CU FOR THE NIDDK MOUSE METABOLIC PHENOTYPING CENTERS (MMPC). THE MISSION OF THE MMPC WAS SIMILAR TO MPMOD, ALLOWING US TO BUILD UPON THE SUCCESS OF THE ESTABLISHED MMPC CU INFRASTRUCTURE TO PROVIDE MPMOD WITH A ROBUST AND COMPREHENSIVE SERVICE ORIENTED SOLUTION THAT HAS BEEN TESTED OVER TIME AND SUPPORTS THE UNIQUE ASPECTS OF THIS CONSORTIUM.

EXPLOITING THE IMMUNOMODULATORY EFFECTS OF SULINDAC AND NOVEL NON-COX INHIBITORY DERIVATIVES FOR CANCER TREATMENT

BETA AMYLOID-ADRENERGIC RECEPTOR INTERACTION IN ALZHEIMER'S DISEASE

MECHANISM OF CARDIOVASCULAR DISEASE IN PREMENOPAUSAL WOMEN - PROJECT SUMMARY SUBOPTIMAL NUTRITION IS THE LEADING RISK FACTOR FOR DEATH AND DISABILITY WORLDWIDE AND ACCOUNTS FOR ACCOUNTS FOR MORE THAN 45% OF CARDIOVASCULAR DEATH IN THE US. DIETARY RISKS AFFECT PEOPLE REGARDLESS OF AGE, SEX, AND SOCIODEMOGRAPHIC DEVELOPMENT. HOWEVER, STUDIES INVESTIGATING THE CARDIOVASCULAR CONSEQUENCES OF SUBOPTIMAL DIET IN PREMENOPAUSAL WOMEN REMAIN SCARCE. NOTABLY, ALTHOUGH COMPELLING RECENT EVIDENCE INDICATES THAT WOMEN OF REPRODUCTIVE AGE ARE MORE SALT SENSITIVE AND PRONE TO OBESITY-ASSOCIATED CARDIOVASCULAR DISEASE (CVD) THAN MEN, THE MECHANISMS WHEREBY EXCESS SALT CONSUMPTION AND OBESITY NEGATE THE PREMENOPAUSAL ADVANTAGE FOR HYPERTENSION REMAIN UNKNOWN. IN PRELIMINARY DATA FOR THIS APPLICATION, WE PROVIDE NEW EVIDENCE INVOLVING THE STEROID HORMONE PROGESTERONE, THE ADIPOKINE LEPTIN, AS WELL AS THE “ADRENAL-ALDOSTERONE – ENDOTHELIUM-MINERALOCORTICOID RECEPTOR (MR) AXIS” IN BOTH SALT SENSITIVITY AND OBESITY RELATED-CVD IN PREMENOPAUSAL WOMEN. WE IDENTIFIED FOR THE FIRST TIME A MOUSE MODEL OF ENDOGENOUS SALT SENSITIVITY, THE BALB/C MOUSE, WHICH REPRODUCES THE HUMAN PHENOTYPE AND EXHIBITS A HIGHER SALT-SENSITIVITY IN FEMALES THAN MALES. WE PROVIDE DATA PRESENTING LACK OF ALDOSTERONE SUPPRESSION, MR OVERACTIVATION AND INCREASED ADRENAL LEPTIN RECEPTOR EXPRESSION AS POTENTIAL CONTRIBUTORS TO THE SEX-SPECIFIC ELEVATION IN BLOOD PRESSURE IN FEMALES FED A HIGH SALT DIET. CONCOMITANTLY, WE IDENTIFIED LEPTIN AS A NEW DIRECT REGULATOR OF ADRENAL-ALDOSTERONE PRODUCTION AND PRESENTED LEPTIN-MEDIATED ALDOSTERONE PRODUCTION AND MR ACTIVATION AS MAJOR CONTRIBUTORS TO OBESITY-ASSOCIATED VASCULAR DYSFUNCTION AND HYPERTENSION IN FEMALES. SUBSEQUENTLY, WE SHOW THAT ARTERIES FROM FEMALES ARE MORE PRONE TO ALDOSTERONE-MEDIATED ENDOTHELIAL DYSFUNCTION THAN THAT OF MALES AND THAT BOTH WOMEN AND FEMALE MICE EXHIBIT HIGHER EXPRESSION OF THE ENDOTHELIAL MR (ECMR) THAN MEN AND MALE ANIMALS. REMARKABLY, WE FOUND THAT ENDOTHELIAL PROGESTERONE RECEPTOR ACTIVATION UPREGULATES ECMR IN FEMALES. LASTLY, WE SHOW THAT SALT SENSITIVE FEMALE BALB/C MICE HAVE A 3-FOLD HIGHER EXPRESSION OF ECMR THAN FEMALE MICE ON THE C57BL/6 BACKGROUND, WHICH ARE KNOWN TO BE SALT-RESISTANT. TAKEN TOGETHER, THESE EXCITING AND NOVEL FINDINGS INFORM THE CORE HYPOTHESIS OF THIS PROPOSAL: PROGESTERONE-INDUCED ECMR EXPRESSION AND LEPTIN-MEDIATED ADRENAL ALDOSTERONE PRODUCTION COOPERATE TO ABOLISH THE PROTECTIVE EFFECTS OF FEMALE SEX HORMONES AND PREDISPOSE FEMALES OF REPRODUCTIVE AGE TO DIET-ASSOCIATED CVD. WE WILL TEST THIS HYPOTHESIS IN THREE AIMS. IN AIM 1 WE WILL INVESTIGATE THE SPECIFIC CONTRIBUTION OF ADRENAL LEPTIN RECEPTOR TO SALT AND OBESITY ASSOCIATED CVD, WHILE IN AIM 2 WE WILL DETERMINE WHETHER PROGESTERONE CONTRIBUTES TO SALT AND OBESITY-ASSOCIATED CVD VIA INCREASING ECMR EXPRESSION. FINALLY, IN AIM 3, WE WILL INVESTIGATE USING DISCARDED HUMAN TISSUES WHETHER DIFFERENCES IN ECMR LEVELS ARE RESPONSIBLE FOR SEX, STRAIN AND RACIAL DIFFERENCES IN SALT-SENSITIVITY VIA INCREASING ENDOTHELIAL ENACA ACTIVITY. WE ANTICIPATE THAT THIS PROPOSAL WILL LEAD TO THE IDENTIFICATION OF MECHANISMS PREDISPOSING PREMENOPAUSAL WOMEN TO DIET- ASSOCIATED CVD AND OPEN NEW AVENUES FOR TREATMENT.

METABOLIC DETERMINANTS OF CARDIOVASCULAR DYSFUNCTION IN OBESITY

REGULATION AND FUNCTION OF SRF IN VASCULAR PATHIOBIOLOGY

THE IMPACT OF BLOOD PRESSURE VARIABILITY ON NEUROVASCULAR FUNCTION - INTRAVASCULAR PRESSURE DRIVES PERFUSION, WHICH IS CRITICAL FOR OPTIMAL NEURONAL FUNCTION. HIGH BLOOD PRESSURE (HYPERTENSION), HOWEVER, IS A RISK FACTOR FOR COGNITIVE DECLINE. EMERGING EVIDENCE IDENTIFIES INCREASED BLOOD PRESSURE VARIABILITY (IBPV), BEFORE THE DEVELOPMENT OF HYPERTENSION, AS A STRONG PREDICTOR FOR VASCULAR COGNITIVE IMPAIRMENT AND DEMENTIA. THE MECHANISM WHEREBY IBPV MEDIATES COGNITIVE DECLINE IS UNKNOWN AND IS THE SUBJECT OF THIS NOVEL PROPOSAL. THE MYOGENIC RESPONSE OF CEREBRAL ARTERIOLES PROTECTS THE BRAIN FROM BLOOD PRESSURE FLUCTUATIONS THAT COULD CAUSE HYPER- OR HYPOPERFUSION. MECHANOSENSORY MECHANISMS ARE ESSENTIAL IN THIS PROCESS, BUT THE IMPACT OF CHRONIC BLOOD PRESSURE ELEVATIONS AT THE LEVEL OF THE NEUROVASCULAR UNIT HAS NOT BEEN PREVIOUSLY DESCRIBED. FOR EXAMPLE, MECHANOSENSITIVE CA2+-PERMEABLE CATION CHANNELS ARE EXPRESSED ON ENDOTHELIAL CELLS AND ASTROCYTES. OUR EXCITING PRELIMINARY DATA DEMONSTRATE THAT INCREASED INTRAVASCULAR PRESSURE SIGNIFICANTLY INCREASED ASTROCYTE CA2+ IN A A PROCESS THAT IS ENHANCED IN HYPERTENSION. ASTROCYTE CA2+ DYSREGULATION IS OFTEN OBSERVED IN NEURODEGENERATIVE DISEASES SUGGESTING IT MAY UNDERLIE CELLULAR PROCESSES THAT CONTRIBUTE TO THE LOSS OF HOMEOSTATIC FUNCTION AND TRANSITION INTO REACTIVE ASTROCYTES. BECAUSE ABERRANT BLOOD PRESSURE FLUCTUATIONS ARE AN EARLY PREDICTOR OF HYPERTENSION, WE WILL EXPLORE THE CELLULAR MECHANISMS BY WHICH INTERMITTENT INCREASES IN ARTERIAL PRESSURE CONTRIBUTE TO COGNITIVE DECLINE. SPECIFICALLY, WE WILL TEST THE CENTRAL HYPOTHESIS THAT CHRONIC IBPV AMPLIFIES MECHANO-DRIVEN CA2+ INCREASES AT THE NVU, WHICH IMPAIRS ASTROCYTE HOMEOSTASIS, DECREASES PERFUSION, AND CAUSES COGNITIVE DECLINE. STUDIES WILL BE CONDUCTED IN A NOVEL MURINE MODEL OF CHRONIC INCREASED BLOOD PRESSURE VARIABILITY INDUCED BY PULSATILE ANGIOTENSIN II INFUSION COUPLED WITH CONTINUOUS BLOOD PRESSURE MEASUREMENT IN CONSCIOUS MICE. AIMS 1-3 WILL TEST THE FOLLOWING HYPOTHESES: 1) THAT IBPV IMPAIRS VASCULAR FUNCTION AND CAUSES CEREBRAL HYPOPERFUSION; 2) THAT INCREASED IBPV ENHANCES MYOGENIC- INDUCED INCREASES IN ASTROCYTE CA2+AND SHIFTS ASTROCYTES TOWARD A PRO-INFLAMMATORY/REACTIVE PHENOTYPE; AND 3) THAT IBPV COMPROMISES SENSORY-EVOKED INCREASES IN CEREBRAL BLOOD FLOW, CONTRIBUTING TO NEURONAL DYSFUNCTION. USING IN VIVO AND EX VIVO APPROACHES, WE WILL LINK MACROSCOPIC CARDIOVASCULAR VARIABLES TO MICROSCOPIC CELLULAR EVENTS AT THE NEUROVASCULAR UNIT AND ASSESS HOW IBPV PROGRESSIVELY IMPAIRS VASCULAR, GLIAL AND NEURONAL FUNCTION. A LONGITUDINAL APPROACH WILL DETERMINE THE RELATIONSHIP BETWEEN BLOOD PRESSURE FLUCTUATIONS AND ABERRANT CA2+ DYNAMICS IN ASTROCYTES, ENDOTHELIAL CELLS AND NEURONS. PHARMACOLOGICAL, MOLECULAR, AND GENETIC TOOLS WILL BE USED TO IDENTIFY THE CELLULAR PATHWAYS UNDERLYING THE LOSS OF FUNCTION AT THE NEUROVASCULAR UNIT. FINDINGS FROM THIS INNOVATIVE APPLICATION WILL ESTABLISH IBPV AS A KEY DRIVER AND PREDICTOR OF COGNITIVE DECLINE, INTRODUCE A NOVEL MURINE MODEL TO INVESTIGATE THE IMPACT OF IBPV ON BRAIN (AND MULTI-ORGAN) FUNCTION, AND IDENTIFY CELLULAR AND MOLECULAR TARGETS OF PRESSURE-INDUCED VASCULAR AND ASTROCYTE DYSFUNCTION LEADING TO COMPROMISED CEREBRAL PERFUSION AND ULTIMATELY, NEURONAL DYSFUNCTION.

MOLECULAR MECHANISMS UNDERLYING THE ESTABLISHMENT OF CELL POLARITY. - A FUNDAMENTAL FEATURE OF EUKARYOTIC LIFE IS THE ESTABLISHMENT AND MAINTENANCE OF CELLULAR POLARITY. MOLECULAR MOTORS HELP TO ESTABLISH POLARITY BY TRANSPORTING MRNAS, PROTEINS, VESICLES, AND ORGANELLES TO SPECIFIC SITES WITHIN THE CELL. A VARIETY OF ORGANISMS FROM THE SINGLE-CELLED YEAST TO HUMANS USE MRNA LOCALIZATION COUPLED WITH TRANSLATIONAL REGULATION AS A WAY TO ASYMMETRICALLY SORT PROTEINS. THE PREVALENCE OF THIS PHENOMENON IS BEST ILLUSTRATED IN DEVELOPING EMBRYOS, NEURONS AND EPITHELIAL, IN WHICH THOUSANDS OF MRNAS ARE SPATIALLY LOCALIZED. WHEN THIS PROCESS IS COMPROMISED, IT CAN RESULT IN DEVELOPMENTAL AND NEUROLOGICAL DISORDERS. DESPITE THE IMPORTANCE OF THIS TOPIC AND THE PREVALENCE OF THIS PHENOMENON, WE LACK A MECHANISTIC UNDERSTANDING OF MRNA LOCALIZATION. A CRITICAL GAP IN OUR UNDERSTANDING PERTAINS TO HOW MRNAS DESTINED FOR LOCALIZATION ARE RECOGNIZED BY THE CELL AND DISTINGUISHED FROM NON-LOCALIZING MRNAS. IN ADDITION TO SEQUENCES PRESENT WITHIN LOCALIZING MRNAS, THE PROTEINS THAT BIND THESE MRNAS ARE KEY TO THEIR CELLULAR FATE. THIS COMPLEX OF PROTEINS IS RESPONSIBLE FOR LINKING LOCALIZING MRNAS WITH MOLECULAR MOTORS AND FOR REGULATING THEIR TRANSLATION. HOWEVER, IDENTIFYING THESE CRITICAL PROTEINS HAS PROVEN TO BE EXTREMELY CHALLENGING. BY ITS VERY NATURE, THE PROCESS OF MRNA LOCALIZATION IS HIGHLY DYNAMIC. CONSEQUENTLY, THE PROTEIN-PROTEIN AND PROTEIN-RNA INTERACTIONS REQUIRED FOR ASSEMBLING LOCALIZING MESSENGER RIBONUCLEOPROTEIN (MRNP) PARTICLES ARE WEAK AND TRANSIENT. THIS HAS MADE THEIR IDENTIFICATION USING CLASSICAL BIOCHEMICAL APPROACHES ALMOST IMPOSSIBLE; THE COMPLEX FALLS APART DURING THE PURIFICATION STEP. IN THIS APPLICATION, WE PROPOSE NOVEL STRATEGIES TO ADDRESS THIS CRITICAL KNOWLEDGE GAP. THE MODEL WE PROPOSE TO USE FOR THESE STUDIES IS THE WELL-CHARACTERIZED DROSOPHILA MELANOGASTER EGG CHAMBER. IN OBJECTIVE 1 OF THIS APPLICATION, WE PROPOSE TO USE PROXIMITY BIOTIN LIGATION TO DEFINE THE CORE COMPONENTS OF TRANSPORT PARTICLES AND TO EXAMINE THE CONSERVATION OF THESE FACTORS BETWEEN FLIES AND MAMMALS. IN OBJECTIVE 2, WE PROPOSE TO TEST THE HYPOTHESIS THAT GRANULAR STRUCTURES REFERRED TO AS P BODIES COORDINATE THE LOCALIZATION OF MRNAS WITH THEIR TRANSLATIONAL REGULATION.

NEXT GEN VIROTHERAPY FOR GBM - ABSTRACT: THE OVERALL GOAL OF THIS APPLICATION IS TO DEVELOP A NOTCH BLOCKING STRATEGY IN COMBINATION WITH OHSV THAT CAN BE SAFELY DELIVERED TO INTRACRANIAL GBM TO ENHANCE THERAPEUTIC EFFICACY WITHOUT NEUROLOGIC TOXICITY. IN THE NORMAL BRAIN, NOTCH SIGNALING PLAYS A SIGNIFICANT ROLE IN MEMORY PROCESSING AND ADULT NEUROGENESIS. IN GLIOBLASTOMA (GBM) NOTCH SIGNALING HAS ALSO BEEN IMPLICATED IN THE DEVELOPMENT OF RESISTANCE TO CHEMOTHERAPY AND RADIATION, AND CONTRIBUTES TO THE DISMAL SURVIVAL ASSOCIATED WITH GBM, A DISEASE ASSOCIATED WITH A LESS THAN 2 YEAR MEDIAN SURVIVAL DESPITE TREATMENT WITH SURGERY, RADIATION, TEMOZOLOMIDE, AND WITH TUMOR TREATING FIELD DEVICE. THE OVERALL GOAL OF THIS APPLICATION IS TO UNDERSTAND THE IMPACT OF SPECIFIC NOTCH LIGAND MEDIATED NOTCH ACTIVATION ON OHSV THERAPY FOR BRAIN TUMORS. SINCE NOTCH SIGNALING PLAYS A SIGNIFICANT ROLE IN THE BRAIN IN MEMORY PROCESSING AND ADULT NEUROGENESIS, WE WILL ALSO EVALUATE THE IMPACT OF BLOCKING SPECIFIC LIGANDS ON MEMORY DEVELOPMENT AND SAFETY FOR INTRACRANIAL USAGE. IN OUR PRELIMINARY RESULTS WE HAVE IDENTIFIED THAT OHSV INFECTION INDUCES NOTCH ACTIVATION IN TUMOR AND TUMOR ASSOCIATED MACROPHAGES (TAM). NOTCH ACTIVATION OF TAMS RESULTS IN INDUCTION OF CCL2 THAT RECRUITS MONOCYTIC MDSCS TO INFECTED TUMORS. WHILE OHSV TREATMENT AWAKENS ANTI-TUMOR EFFICACY, THESE MONOCYTIC MDSCS LIMIT THE IMMUNOTHERAPEUTIC BENEFIT BY EDUCATING AN IMMUNE-SUPPRESSIVE ENVIRONMENT IN TUMORS. IT HAS BEEN SHOWN THAT DIFFERENT NOTCH LIGANDS HAVE DIFFERENT EFFECTS ON ANTI-TUMOR IMMUNITY. FOR EXAMPLE, DLL1-MEDIATED NOTCH ACTIVATION IS IMPORTANT FOR T CELL MATURATION INTO MEMORY CELLS AND ITS OVEREXPRESSION AUGMENTS T CELL ACTIVITY AND ANTI-TUMOR IMMUNITY. WHILE JAG1, AND TO A LESSER EXTENT JAG2, INDUCE PD-1 AND SUPPRESS T CELL IMMUNITY. THUS, WE HYPOTHESIZE THAT BLOCKADE OF JAG1 MEDIATED SIGNALING SHOULD ENHANCE VIROTHERAPY INDUCED ANTI-TUMOR IMMUNITY, AND ITS TRANSIENT EXPRESSION BY AN OHSV WOULD NOT CREATE A NEUROLOGIC MEMORY DEFICIT IN MICE. SINCE, MEMBRANE BOUND LIGANDS CAN ACTIVATE NOTCH SIGNALING AND SOLUBLE MONOMERIC LIGANDS CAN INHIBIT NOTCH SIGNALING, HERE WE WILL TEST THE EFFECT OF BLOCKING INDIVIDUAL NOTCH LIGAND MEDIATED NOTCH ACTIVATION ON OHSV EFFICACY AND ANTI-TUMOR IMMUNITY (AIM 1). IN AIM 2 WE WILL CREATE AN OHSV THAT CAN EFFECTIVELY BLOCK VIRUS INDUCED LIGAND SPECIFIC NOTCH SIGNALING TO AUGMENT VIRUS INDUCED ANTI-TUMOR IMMUNITY. THIS VIRUS WILL ALSO BE EVALUATED FOR SAFETY, SENSITIVITY TO ACV, AND EFFECT ON MOUSE BEHAVIOR AND MEMORY. FURTHER WE HAVE FOUND THAT COMBINATION OF AN OHSV WITH IRRADIATION SYNERGISTICALLY ACTIVATES NOTCH SIGNALING. IN AIM 3 WE WILL EVALUATE THE EFFECT OF THIS VIRUS IN COMBINATION WITH IRRADIATION.

REGULATION AND ROLE OF LEPTIN IN PREECLAMPSIA - PREECLAMPSIA (PE) IS A HYPERTENSIVE DISEASE IN PREGNANCY THAT IS A LEADING CAUSE OF ADVERSE MATERNAL AND FETAL OUTCOMES IN THE US. DECADES OF CLINICAL STUDIES DEMONSTRATE THAT LEVELS OF THE ADIPOKINE LEPTIN INAPPROPRIATELY INCREASE, INDEPENDENT OF BODY MASS, IN PE PATIENTS. HOWEVER, WHETHER LEPTIN PLAYS A ROLE IN THE CARDIOVASCULAR AND FETAL OUTCOMES OF PE IS UNKNOWN. WE RECENTLY ESTABLISHED A MOUSE MODEL OF LEPTIN-INDUCED PE, IN WHICH EXOGENOUS LEPTIN ADMINISTRATION INDUCES THE CLINICAL CHARACTERISTICS OF PE IN ENDOTHELIAL DYSFUNCTION, HYPERTENSION, PLACENTAL MITOCHONDRIAL DYSFUNCTION AND FETAL GROWTH RESTRICTION WHEN GIVEN IN MID-LATE GESTATION PREGNANT MICE. THE GOAL OF THIS PROPOSAL IS TO ADDRESS A CRITICAL GAP IN KNOWLEDGE REGARDING THE STIMULI UPREGULATING LEPTIN IN PE AND TO INVESTIGATE MECHANISMS VIA WHICH LEPTIN INDUCES ADVERSE MATERNAL AND FETAL ADAPTATIONS. OUR CENTRAL HYPOTHESIS IS THAT SFLT-1 INDUCES HYPERLEPTINEMIA IN PE, WHICH PROMOTES HYPERTENSION AND FETAL GROWTH RESTRICTION VIA ENDOTHELIAL DYSFUNCTION. PLACENTAL ISCHEMIA IS A KEY INITIATING EVENT IN PE AND INDUCES AN ANTI-ANGIOGENIC MILIEU, MOST NOTABLY INCREASING SOLUBLE FMS LIKE TYROSINE KINASE-1 (SFLT-1), THE SOLUBLE FORM OF VASCULAR ENDOTHELIAL GROWTH FACTOR RECEPTOR 1 (VEGFR1). WE SHOW NOVEL PRELIMINARY DATA THAT SFLT-1 INCREASES LEPTIN PRODUCTION IN BOTH HUMANS AND MICE. WE ALSO DEMONSTRATE THAT PLACENTAL ISCHEMIA, INDUCED IN THE REDUCED UTERINE PERFUSION PRESSURE (RUPP) MOUSE MODEL OF PE, INCREASES CIRCULATING LEPTIN AS WELL AS SFLT-1. WE WILL TEST IN AIM 1 WHETHER PLACENTAL ISCHEMIA INCREASES LEPTIN LEVELS VIA SFLT-1, WHICH MEDIATES PLACENTAL ISCHEMIA-INDUCED ENDOTHELIAL AND PLACENTAL DYSFUNCTION. IN THIS AIM, WE PROPOSE THAT THE RUPP MOUSE DEVELOPS ENDOTHELIAL DYSFUNCTION, HYPERTENSION, PLACENTAL MITOCHONDRIAL DYSFUNCTION AND FETAL GROWTH RESTRICTION, CHARACTERISTICS OF PE, VIA LEPTIN-MEDIATED MECHANISMS. WE WILL ADDITIONALLY INVESTIGATE WHETHER SFLT-1 SEQUESTRATION OF VEGF REDUCES VEGF RECEPTOR SIGNALING AND PROMOTES LEPTIN PRODUCTION IN PE. ADDITIONAL INNOVATIVE PRELIMINARY DATA SHOWS THAT ENDOTHELIAL LEPTIN RECEPTOR ACTIVATION INCREASES THE PRODUCTION OF ENDOTHELIN CONVERTING ENZYME-1 (ECE-1) AND ENDOTHELIN-1 (ET-1) IN PREGNANT MICE. WE FURTHER DEMONSTRATE THAT LEPTIN UPREGULATES ENDOTHELIAL MINERALOCORTICOID RECEPTOR EXPRESSION, WHICH WE HAVE PUBLISHED DECREASES ECE-1 EXPRESSION. THEREFORE, IN AIM 2 WE WILL TEST WHETHER LEPTIN INDUCES PE CHARACTERISTICS VIA ENDOTHELIAL ET-1-MEDIATED ENDOTHELIAL DYSFUNCTION. WE WILL UTILIZE MICE WITH ENDOTHELIAL MINERALOCORTICOID RECEPTOR DELETION AS WELL AS ENDOTHELIAL LEPTIN RECEPTOR KNOCKOUT MICE TO DETERMINE WHETHER LEPTIN-INDUCED ET-1 EXPRESSION BY THESE ENDOTHELIAL PATHWAYS LEADS TO PE CHARACTERISTICS IN PREGNANT MICE. WE WILL FURTHER DETERMINE WHETHER ECE-1 INDUCES VASCULAR ENDOTHELIAL DYSFUNCTION AND WHETHER ECE-1 OR ET-1 RECEPTOR ANTAGONISM ABLATES LEPTIN-INDUCED PE. COLLECTIVELY, THE RESULTS OF AIM 1 AND 2 WILL SIGNIFICANTLY MOVE FORWARD THE FIELD OF LEPTIN IN PE AND WILL GIVE PRECLINICAL EVIDENCE IF REGULATORY OR DOWNSTREAM MECHANISMS OF LEPTIN IN PE ARE POTENTIAL THERAPEUTICS TO IMPROVE CLINICAL CARE OF PE PATIENTS.

MECHANISMS OF MYELOPEROXIDASE AND NOX4 INTERACTIONS IN ABDOMINAL AORTIC ANEURYSM

BIOPHYSICAL MECHANISMS OF HYPEROXIA-INDUCED LUNG INJURY

PERIVASCULAR ADIPOSE TISSUE AND VASCULAR REMODELING

THE BIOLOGICAL CLOCK, THE ENDOTHELIUM, AND VASCULAR REMODELING

PROMOTING ASTHMA WELLNESS IN RURAL COMMUNITIES

MOLECULAR AND TEMPORAL DISSECTION OF HABIT LEARNING

MOLECULAR CONTROL OF VASCULAR SMOOTH MUSCLE REPROGRAMMING IN ARTERIOVENOUS FISTULA MATURATION - PROJECT SUMMARY A SURGICALLY CREATED ARTERIOVENOUS FISTULA (AVF) BETWEEN AN ARTERY AND VEIN IS NOW THE PREFERRED APPROACH TO PROVIDE A VASCULAR ACCESS FOR LIFE-SAVING HEMODIALYSIS IN CHRONIC KIDNEY DISEASE CKD) PATIENTS. HOWEVER, NEARLY 60% OF AVFS FAIL TO MATURE INTO A CLINICALLY USEFUL CONDUIT DUE TO INSUFFICIENT OUTWARD REMODELING AND FLOW CAPACITY, OCCLUSIVE NEOINTIMAL HYPERPLASIA, AND/OR FIBROTIC STENOSIS. CURRENTLY, THERE ARE NO THERAPIES THAT CAN IMPROVE AVF EARLY MATURATION FAILURE BY ENHANCING AVF OUTWARD REMODELING, LARGELY DUE TO OUR NASCENT UNDERSTANDING OF THE MECHANISMS UNDERLYING VEIN ADAPTATIONS TO AVF HEMODYNAMIC STRESSES. ACUTE INCREASES IN SHEAR STRESS AND PULSATILE PRESSURE IN THE VEIN ARE NORMALIZED BY RAPID DILATION FOLLOWED BY WALL THICKENING. VENOUS SMOOTH MUSCLE CELLS (VSMCS) ARE THE PREDOMINANT CELLS SENSING VESSEL WALL STRETCH IN RESPONSE TO INCREASED FLOW VOLUME AND BLOOD PRESSURE. A SIGNIFICANT BARRIER TO PROGRESS IS A DEFICIT IN OUR KNOWLEDGE OF THE MECHANISMS BY WHICH VSMCS RESPOND TO ARTERIAL HEMODYNAMICS IN EARLY AVF ADAPTATION. STRONG EVIDENCE FROM BOTH AVF MOUSE MODELS AND HUMAN SAMPLE STUDIES DEMONSTRATE, FOR THE FIRST TIME, A ROLE FOR DIFFERENTIATED VSMCS IN AVF OUTWARD REMODELING AND MATURATION. THIS IS FURTHER SUPPORTED BY NEW PRELIMINARY DATA FROM OUR CLINICALLY RELEVANT 5/6-NEPHRECTOMY CKD AVF MOUSE MODEL. WE FURTHER SHOW EARLY AVF MATURATION INVOLVES VSMCS REPROGRAMMING FROM A QUIESCENT TO A PREVIOUSLY UNCHARACTERIZED PROLIFERATIVE, SYNTHETIC STATE THAT SURPRISINGLY RETAINS DIFFERENTIATED CONTRACTILE PROPERTIES. MYOCARDIN RELATED TRANSCRIPTION FACTOR (TF) A AND B (MRTFA AND B, MRTFS) RESPOND TO CYCLIC STRETCH BY TRANSACTIVATING MULTIPLE GENE PROGRAMS. VSMC-DEFICIENCY OF MRTFS IMPAIRS AVF MATURATION WITH REDUCED AVF WALL THICKNESS. BEYOND THE CONTRACTILE GENE PROGRAM, MRTFA UPREGULATES NOVEL TARGET GENES (MMP2 AND ATF3) TO FACILITATE MATRIX REMODELING AND CELL PROLIFERATION. THIS SUGGESTS THAT MRTFS ACT AS NODAL TFS OF VSMC REPROGRAMMING. CAMK2 IS A MAJOR SIGNAL TRANSDUCER POISED TO INTEGRATE STRETCH-INDUCED VASCULAR REMODELING. PRELIMINARY DATA SHOW GROWTH FACTORS INDUCE NUCLEAR INTERACTION OF CAMK2 AND MRTFA IN CULTURED VSMCS. VSMC-DEFICIENCY OF A MAJOR VSMC CAMK2 ISOFORM, CAMK2D, PHENOCOPIES LOSS OF VSMC MRTFS, SUGGESTING THAT CAMK2D TRANSDUCES A SIGNAL(S) FROM AVF WALL STRESS TO TRIGGER MRTF TRANSACTIVITY. THESE PRELIMINARY DATA SUPPORT OUR CENTRAL HYPOTHESIS THAT SUCCESSFUL AVF ADAPTATION AND MATURATION INVOLVES CAMK2/MRTFS-DEPENDENT VSMC REPROGRAMMING TO A PROLIFERATIVE, MATRIX ORGANIZING, AND CONTRACTILE PHENOTYPE. AIM1 WILL ELUCIDATE MECHANISMS OF VSMC-DEPENDENT AVF ADAPTIVE REMODELING AND MATURATION USING ITGA8CREERT2CONFETTI REPORTER, SINGLE NUCLEUS (SN) ATAC/RNA-SEQ, AND SPATIAL OMICS TO DETERMINE VSMC CLONAL EXPANSION AND TRANSCRIPTOMICS UNDERLYING AVF MATURATION. AIM 2 WILL USE NOVEL VSMC-SPECIFIC MRTFS AND CAMK2D KNOCKOUT MICE TO ELUCIDATE THE MECHANISTIC ROLE OF CAMK2D/MRTFS IN AVF MATURATION. SUCCESSFUL COMPLETION OF THESE STUDIES WILL PROVIDE NOVEL INSIGHTS INTO, AND POTENTIAL THERAPEUTIC TARGETS FOR, AVF MATURATION FAILURE ATTRIBUTABLE TO INADEQUATE VEIN ADAPTION IN HUMANS.

ALPHA2 ADRENERGIC SIGNALING IN AD PATHOGENESIS

REGULATION OF SORLA BY BETA-ARRESTIN2

ENAC-? MEDIATES LUNG FLUID CLEARANCE AND CAPILLARY BARRIER FUNCTION IN PNEUMONIA

THE NOVEL SMOOTH MUSCLE-SPECIFIC LNCRNA CARMN IS A CRITICAL REGULATOR OF SMOOTH MUSCLE PHENOTYPE

OSTEOCYTE ENERGY METABOLISM IN AGING - PROJECT SUMMARY AGING IS ASSOCIATED WITH NEGATIVE EFFECTS ON THE SKELETAL SYSTEM, LEADING TO IMPAIRED BONE HEALTH, INCREASED FRAILTY, AND RISK OF MUSCULOSKELETAL DISORDERS. HOWEVER, OUR CURRENT UNDERSTANDING OF THE MECHANISMS BY WHICH AGING AFFECTS SKELETAL HEALTH IS LIMITED. OSTEOCYTES ARE THE MOST NUMEROUS AND LONG-LIVED CELLS IN BONE AND PLAY KEY ROLES IN MAINTAINING BONE MASS BY RESPONDING TO ANABOLIC SIGNALS SUCH AS MECHANICAL LOADING. THIS RESPONSE TO LOADING IS IMPAIRED IN AGED BONE, BY UNKNOWN MECHANISMS, LEADING TO DERANGEMENTS IN BONE HOMEOSTASIS. ENERGY METABOLISM IS DISRUPTED IN MANY CELLS AND TISSUES WITH AGING, HOWEVER REGULATION OF ENERGY METABOLISM IN OSTEOCYTES AND HOW THIS IS AFFECTED DURING AGING AND BY MECHANICAL LOADING REMAINS UNDEFINED. OUR PRELIMINARY FINDINGS HAVE IDENTIFIED THAT ANABOLIC MECHANICAL LOADING REGULATES ENERGY METABOLISM IN OSTEOCYTES IN VITRO AND IN VIVO. SPECIFICALLY, FATTY ACID SS-OXIDATION IS UPREGULATED IN RESPONSE TO STRAIN, AS ARE THE KEY PROMOTERS OF SS-OXIDATION, PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR DELTA (PPARD) AND CARNITINE PALMITOYLTRANSFERASE 1 (CPT1). DELETION OF PPARD IN OSTEOCYTES IN VIVO INHIBITED SS-OXIDATION AND DECREASED BONE MASS IN FEMALE MICE BUT NOT MALES. FURTHERMORE, SS-OXIDATION IS DECREASED IN AGED MOUSE BONE COMPARED TO YOUNG ANIMALS, AND PHARMACOLOGICAL ACTIVATION OF PPARD IN AGING MICE IMPROVES BONE HEALTH. THESE FINDINGS SUGGEST IMPORTANT FUNCTIONS OF OSTEOCYTE ENERGY METABOLISM, AND SS-OXIDATION IN PARTICULAR, IN THE EFFECTS OF AGING AND MECHANICAL LOADING ON BONE. IN THIS PROPOSAL, WE WILL DETERMINE THE ROLE OF OSTEOCYTE SS-OXIDATION IN THE RESPONSE TO MECHANICAL STIMULATION AND THE MAINTENANCE OF BONE HEALTH WITH AGING. FOR THESE STUDIES, WE HAVE GENERATED MICE WITH TARGETED DELETION OF PPARD IN OSTEOCYTES AND WILL USE THESE MICE AND EX VIVO ISOLATED OSTEOCYTES FROM YOUNG AND AGED MICE TO DETERMINE THE FUNCTION OF SS-OXIDATION IN OSTEOCYTES. IN AIM 1, WE WILL DETERMINE THE ROLE OF OSTEOCYTE SS-OXIDATION IN REGULATING BONE HEALTH IN YOUNG AND AGED MICE. IN AIM 2, WE WILL DETERMINE HOW AGING AFFECTS THE METABOLIC RESPONSE OF OSTEOCYTES TO MECHANICAL LOADING AND THE ROLE OF PPARD-DRIVEN SS-OXIDATION IN THIS PROCESS. IN AIM 3, WE WILL EXAMINE WHETHER ACTIVATION OF PPARD TO INCREASE SS-OXIDATION IN OSTEOCYTES CAN IMPROVE BONE HEALTH IN AGED MICE. THE FINDINGS FROM THE PROPOSED STUDIES WILL MARKEDLY INCREASE THE KNOWLEDGE OF OSTEOCYTE ENERGY METABOLISM AND DEFINE THE ROLE OF PPARD-DRIVEN SS-OXIDATION IN OSTEOCYTES AND ITS FUNCTION DURING AGING AND UNDER MECHANICAL LOADING.

LEPTIN IN HIV ASSOCIATED VASCULAR DISEASES

ELUCIDATING THE ROLE OF CULLIN 3 IN THE HEART - PROJECT SUMMARY CARDIOMYOCYTE (CM) MITOCHONDRIA PLAY A CRUCIAL ROLE IN ENERGY PRODUCTION AND OTHER ESSENTIAL CELLULAR PROCESSES. DISRUPTIONS IN MITOCHONDRIAL HOMEOSTASIS CONTRIBUTE TO CARDIOMYOPATHIES AND HEART FAILURE, WHICH REMAIN SIGNIFICANT GLOBAL HEALTH CHALLENGES. THEREFORE, IT IS VITAL TO IDENTIFY NOVEL REGULATORS OF MITOCHONDRIAL HOMEOSTASIS IN THE HEART TO REDUCE CARDIAC DYSFUNCTION-RELATED MORTALITY. PROPER CONTROL OF MITOCHONDRIAL BIOGENESIS AND TURNOVER IS ESSENTIAL FOR MAINTAINING A HEALTHY MITOCHONDRIAL POPULATION IN CMS. RECENT RESEARCH HAS HIGHLIGHTED THE IMPORTANCE OF MITOCHONDRIAL DNA TRANSCRIPTION IN MITOCHONDRIAL BIOGENESIS, AS WELL AS THE ROLE OF UBIQUITIN (UB) LIGASE PARKIN IN MITOPHAGY. HOWEVER, THERE ARE SIGNIFICANT KNOWLEDGE GAPS REGARDING THE UPSTREAM SIGNALING INVOLVED IN MITOCHONDRIAL DNA TRANSCRIPTION AND OTHER MITOCHONDRIAL UB LIGASES CRUCIAL FOR MITOPHAGY IN CMS. CULLIN 3-RING UB LIGASE (CRL3) BELONGS TO THE MULTI-SUBUNIT CULLIN-RING UB LIGASES (CRLS) FAMILY. IN THE CRL3 COMPLEX, CULLIN 3 SERVES AS A SCAFFOLD FOR INTERACTING WITH THE RING-BOX PROTEIN RBX1 AND ONE OF THE SUBSTRATE RECEPTORS. THE ASSEMBLY OF CRL3 REQUIRES THE CONJUGATION OF CULLIN 3 BY THE UB-LIKE PROTEIN NEDD8 (NEDDYLATION). KNOWN AS A CYTOSOLIC UB LIGASE, CRL3 CONTROLS THE DEGRADATION OF NUMEROUS PROTEINS AND PARTICIPATES IN MULTIPLE PATHOPHYSIOLOGICAL PROCESSES SUCH AS ORGANISMAL DEVELOPMENT, TUMOR GROWTH, VASCULAR INTEGRITY, AND RENAL SODIUM TRANSPORT. MOREOVER, MUTATIONS IN CULLIN 3 WERE SHOWN TO CAUSE FAMILIAL HYPERKALEMIC HYPERTENSION, AUTISM SPECTRUM DISORDERS, AND EPILEPSY. DESPITE THE INCREASINGLY RECOGNIZED IMPORTANCE OF CRL3 IN HEALTH AND DISEASE, ITS ROLE IN MITOCHONDRIA AND THE HEART REMAINS LARGELY UNKNOWN. IN SILICO ANALYSES HAVE REVEALED AN INTRICATE INTERACTION BETWEEN CRLS AND MITOCHONDRIAL HOMEOSTASIS. DISTURBANCES IN CRLS ADVERSELY AFFECT MITOCHONDRIAL INTEGRITY AND FUNCTION IN CMS, AND DEPLETION OF CULLIN 3 INHIBITS MITOCHONDRIAL TURNOVER AND RESPIRATION CAPACITY, SUGGESTING A POTENTIAL ROLE FOR CRL3 IN MAINTAINING MITOCHONDRIAL HOMEOSTASIS. NOTABLY, NEDDYLATION OF CULLIN 3 IS DOWNREGULATED DURING CARDIAC DEVELOPMENT BUT UPREGULATED IN HUMAN FAILING HEARTS AND MOUSE HEARTS UNDERGOING PATHOLOGICAL REMODELING. HENCE, THE GOAL OF THIS PROJECT IS TO INVESTIGATE THE PATHOPHYSIOLOGICAL SIGNIFICANCE OF CULLIN 3 IN THE HEART AND TO ELUCIDATE THE MOLECULAR MECHANISMS BY WHICH CULLIN 3 REGULATES MITOCHONDRIAL HOMEOSTASIS USING A SERIES OF NEWLY GENERATED, GENETICALLY ENGINEERED MOUSE MODELS AND ADENOVIRUS-ASSOCIATED VIRUSES. THREE INTERCONNECTED YET INDEPENDENT AIMS ARE PROPOSED: AIM 1 WILL ASSESS THE IMPORTANCE OF NEDDYLATED CULLIN 3 IN CARDIAC MATURATION AND MITOCHONDRIAL BIOGENESIS, AIM 2 WILL ESTABLISH THE FUNCTIONAL SIGNIFICANCE OF CULLIN 3 IN MITOCHONDRIAL TURNOVER IN THE ADULT HEART, AND AIM 3 WILL INVESTIGATE THE IMPACT OF CULLIN 3 MUTATIONS AND DEFICIENCY ON CARDIAC REMODELING WHILE EXPLORING THE POTENTIAL OF TARGETING CULLIN 3 TO PREVENT STRESS-INDUCED CARDIAC DAMAGE. SUCCESSFUL COMPLETION OF THESE EXPERIMENTS WILL EXPAND AND DEEPEN OUR UNDERSTANDING OF HOW MITOCHONDRIAL HOMEOSTASIS IS MAINTAINED IN THE DEVELOPING, HOMEOSTATIC, AND REMODELING HEART.

CD14 AND SALT-SENSITIVE HYPERTENSION - SUMMARY HYPERTENSION IS A PRIMARY MODIFIABLE RISK FACTOR FOR CARDIOVASCULAR, CEREBROVASCULAR, AND RENAL DISEASE, AND IS THE LARGEST INDIVIDUAL CONTRIBUTING FACTOR TO DISEASE AND MORTALITY IN THE WORLD. SALT-SENSITIVE HYPERTENSIVE INDIVIDUALS, WHO COMPRISE 30-50% OF THE HYPERTENSIVE POPULATION, HAVE GREATER MORTALITY THAN SUBJECTS WITH SALT- RESISTANT HYPERTENSION AND EXHIBIT RENAL END-ORGAN DAMAGE. WE RECENTLY DISCOVERED A NOVEL ANTI-HYPERTENSIVE MECHANISM THAT IS MEDIATED BY CLUSTER OF DIFFERENTIATION 14 (CD14) AND ENHANCED IN FEMALES RELATIVE TO MALES. CD14 IS A CO-RECEPTOR WITH TOLL LIKE RECEPTOR 4 (TLR4). INTERESTINGLY, THE EXPRESSION OF CD14 IN LEUKOCYTES AND LEVEL OF PLASMA CD14 PROTEIN ARE INCREASED IN CARDIOVASCULAR DISEASE AND HYPERTENSION IN HUMANS. WE PROPOSE THAT CD14 SIGNALING IN MACROPHAGES IN THE KIDNEY OPPOSES THE EFFECTS OF TLR4 AND LEADS TO THE ATTENUATED RELEASE OF FREE RADICALS AND THE PROINFLAMMATORY CYTOKINE IL-1SS, BLUNTED T CELL ACTIVATION, AND ATTENUATED SALT-SENSITIVE HYPERTENSION. THIS PROJECT IS BASED UPON OUR UNIQUE (AND SOMEWHAT SURPRISING) OBSERVATION THAT GENETIC DELETION OF CD14, WHICH IS UPREGULATED IN MACROPHAGES IN THE KIDNEY OF DAHL SALT-SENSITIVE (SS) RATS FED HIGH SALT, LEADS TO INCREASED RELEASE OF THE PROINFLAMMATORY CYTOKINE IL-1SS AND REACTIVE OXYGEN SPECIES. REMARKABLY, WE OBSERVED THAT FEMALE DAHL SS RATS LACKING CD14 (SSCD14-/-) EXHIBIT AMPLIFIED SALT-SENSITIVE HYPERTENSION AND RENAL DAMAGE COMPARED TO WILD TYPE LITTERMATES. FURTHER STUDY DEMONSTRATED THAT THE EFFECTS OF CD14 DELETION ARE DEPENDENT ON HEMATOPOIETIC CELLS, AMPLIFIED IN FEMALES, AND ELIMINATED IN RATS LACKING PHAGOCYTIC NADPH OXIDASE 2 (NOX2). THESE EXCITING DATA INDICATE AN UNEXPECTED, SEX-DEPENDENT ROLE OF CD14 IN THE DEVELOPMENT OF SALT-SENSITIVE HYPERTENSION AND RENAL DAMAGE. THIS PROPOSAL WILL TEST THE HYPOTHESIS THAT THE ANTI-HYPERTENSIVE EFFECTS OF CD14 IN MACROPHAGES IN THE KIDNEY LEAD TO DECREASED ACTIVATION OF T CELLS IN THE KIDNEY, ATTENUATED SALT-SENSITIVE HYPERTENSION, AND DECREASED RENAL END-ORGAN DAMAGE. THE HYPOTHESIS WILL BE ADDRESSED IN THREE SPECIFIC AIMS. AIM 1 WILL ADDRESS THE HYPOTHESIS THAT THE PROTECTIVE EFFECTS OF CD14 AGAINST SALT-SENSITIVE HYPERTENSION AND ASSOCIATED RENAL DAMAGE INVOLVE INHIBITION OF TLR4-MEDIATED SIGNALING IN MACROPHAGES. AIM 2 WILL ADDRESS THE HYPOTHESIS THAT ELEVATED NOX2 IN MACROPHAGES MEDIATE ELEVATED BLOOD PRESSURE IN DAHL SS RATS FED HIGH SALT, AN EFFECT ATTENUATED IN FEMALES BY CD14 AND AMPLIFIED IN MALES BY TLR4. AIM 3 WILL ADDRESS THE HYPOTHESIS THAT THE PRO-HYPERTENSIVE EFFECTS OF TLR4 AND THE ANTI-HYPERTENSIVE EFFECTS OF CD14 ARE MEDIATED BY CORRESPONDING CHANGES IN PRESSURE NATRIURESIS-DIURESIS AND INTRARENAL HEMODYNAMICS IN A PROCESS DEPENDENT UPON FREE RADICALS RELEASED FROM NOX2 IN MACROPHAGES AND T CELLS. THE PROPOSAL EMPLOYS IN VITRO AND IN VIVO APPROACHES TO ADDRESS THE HYPOTHESIS USING UNIQUE, GENETICALLY-ENGINEERED RAT STRAINS DEVELOPED FOR THIS GRANT, NOVEL IMMUNE CELL TRANSFER APPROACHES, AND STATE-OF-THE ART METHODOLOGY TO ASSESS HEMODYNAMIC VARIABLES IN CONSCIOUS RATS.

IMMUNOMETABOLIC REGULATION AFTER CNS INJURY - TITLE: IMMUNOMETABOLIC REGULATION AFTER CNS INJURY PROGRESSIVE NEURODEGENERATION IS A SEQUELA OF TRAUMATIC BRAIN INJURY (TBI), WITH ~2% OF THE POPULATION LIVING WITH CHRONIC NEUROLOGICAL DEFICITS, INCLUDING COGNITIVE IMPAIRMENT AND DEMENTIA, CAUSED BY A PRIOR HEAD INJURY. EPIDEMIOLOGICAL DATA INDICATE A HISTORY OF TBI IS THE LEADING EXTRINSIC CAUSE OF DEMENTIA, INCLUDING A HEIGHTENED RISK OF DEVELOPING ALZHEIMER’S DISEASE AND RELATED DEMENTIAS (AD/ADRD); HOWEVER, THE MECHANISMS LINKING TBI WITH SUBSEQUENT NEURODEGENERATION INCLUDING DUE TO VASCULAR CONTRIBUTION TO COGNITIVE IMPAIRMENT AND DEMENTIA (VCID) ARE UNDEFINED. DESPITE IMMUNE PRIVILEGE WITHIN THE CNS, DEVELOPMENT OF A COORDINATED SERIES OF SPATIALLY- AND TEMPORALLY-REGULATED CEREBRAL IMMUNE RESPONSES CORRELATES WITH PREMATURE COGNITIVE AGING, NEURODEGENERATION AND VCID AFTER TBI. OUR OBJECTIVE IS TO TEST THE OVERARCHING HYPOTHESIS THAT CEREBRAL METABOLIC DYSREGULATION ACTIVATES MENINGEAL INNATE LYMPHOID CELLS (ILCS) TO PERPETUATE A CHRONIC, PRO-INFLAMMATORY CASCADE THAT CULMINATES IN NEURODEGENERATION AND POOR COGNITIVE OUTCOMES. SPECIFICALLY, WE PROPOSE THAT REDUCED ACTIVITY OF THE ENERGETIC SENSOR, 5’-AMP-ACTIVATED PROTEIN KINASE (AMPK), INDUCES A SENESCENCE-ASSOCIATED SECRETORY PHENOTYPE (SASP) WITHIN ASTROCYTES TO EXPAND PRO-INFLAMMATORY ILCS, RECRUIT PERIPHERAL IMMUNE CELLS, AND DRIVE POST-TRAUMATIC DEMENTIA. AIM 1 WILL TEST THE HYPOTHESIS THAT ASTROCYTE-SPECIFIC AMPK ACTIVATION LIMITS NEURODEGENERATION. STUDIES WILL INCORPORATE ADVANCED GENETIC MODELS, NEUROIMAGING, AND SPATIAL PROTEOGENOMICS TO LONGITUDINALLY DEFINE REGION- AND CELL TYPE-SPECIFIC NEURODEGENERATIVE CHANGES AFTER TBI. AIM 2 WILL TEST THE HYPOTHESIS THAT ASTROCYTE-SPECIFIC AMPK ACTIVATION RESTRAINS PRO-INFLAMMATORY MENINGEAL ILC EXPANSION. STUDIES WILL FUNCTIONALLY IMPLICATE MENINGEAL ILCS AS CRITICAL MEDIATORS OF CEREBRAL IMMUNITY AND PROGRESSIVE NEURODEGENERATION, INCLUDING CEREBRAL ATROPHY AND GREY/WHITE MATTER LOSS, AFTER TBI. AIM 3 WILL TEST THE HYPOTHESIS THAT REGULATORY ILC2 REDUCE THE PROGRESSIVE NEURODEGENERATIVE PROFILE TO ATTENUATE CHRONIC NEUROLOGICAL INJURY AFTER TBI. STUDIES WILL DEMONSTRATE THE THERAPEUTIC POTENTIAL OF TARGETING MENINGEAL ILCS TO RESTRAIN NEURODEGENERATION. EXPECTED OUTCOMES: OUR MECHANISTIC STUDIES WILL SHOW ACUTE CEREBRAL METABOLIC DERANGEMENTS INITIATE A DELETERIOUS, PRO-INFLAMMATORY CASCADE THAT CULMINATES IN NEURODEGENERATION. SPECIFICALLY, OUR CONCEPTUALLY INNOVATIVE STUDIES WILL IDENTIFY ASTROCYTE SENESCENCE AS A CELLULAR CONVERGENCE POINT THAT INTEGRATES CEREBRAL METABOLISM AND CHRONIC INFLAMMATION. WE ALSO WILL ELUCIDATE A NOVEL ROUTE OF CELL-CELL COMMUNICATION WHEREBY ASTROCYTES COORDINATE PERIPHERAL IMMUNITY VIA REGULATION OF MENINGEAL ILCS. TOGETHER, OUR STUDIES WILL SHOW HOW LOCAL PATHOLOGICAL CHANGES WITHIN THE CNS ARE TRANSLATED INTO CONTEXT-SPECIFIC IMMUNE RESPONSES THAT CULMINATE IN POST-TRAUMATIC DEMENTIA. CLINICAL SIGNIFICANCE: PROGRESSIVE NEURODEGENERATION CAUSES CHRONIC COGNITIVE DYSFUNCTION TO WORSEN PATIENT QUALITY OF LIFE. BY DEMONSTRATING THE FEASIBILITY AND EFFICACY OF REGULATORY ILC2 AS A NOVEL, CELL- BASED THERAPY TO RESTRAIN CHRONIC INFLAMMATION, LIMIT NEURODEGENERATION, AND IMPROVE LONG-TERM NEUROCOGNITIVE OUTCOMES, OUR STUDIES WILL ESTABLISH A CONCEPTUAL FRAMEWORK FOR THERAPEUTIC DEVELOPMENT TO TREAT POST-TRAUMATIC DEMENTIA, AD, AND RELATED PROGRESSIVE DEMENTIAS.

MITOCHONDRIA DYNAMICS PROTEIN DRP1 IN ROS SIGNALING, ENDOTHELIAL METABOLISM AND ANGIOGENESIS - THE AIM OF THIS GRANT IS TO ELUCIDATE THE ROLE OF MITOCHONDRIAL DYNAMICS PROTEIN DRP1 AS A NOVEL REDOX SENSOR THAT TRANSMITS VEGF-DERIVED H2O2 SIGNALING TO ENHANCE ANGIOGENESIS VIA REGULATION OF ENDOTHELIAL CELL (EC) GLYCOLYSIS. THE INDUCTION OF NEW BLOOD VESSELS IS CRITICAL FOR TISSUE REPAIR IN RESPONSE TO INJURY SUCH AS PERIPHERAL ARTERIAL DISEASE (PAD), WHICH IS IMPAIRED IN DIABETES. REACTIVE OXYGEN SPECIES (ROS) SUCH AS H2O2 DERIVED FROM NADPH OXIDASE (NOX) AND MITOCHONDRIA AT NORMAL LEVEL ACT AS SIGNALING MOLECULES TO PROMOTE VEGF-INDUCED ANGIOGENESIS IN ENDOTHELIAL CELLS (ECS) AND REPARATIVE NEOVASCULARIZATION. HOWEVER, IT REMAINS UNCLEAR “HOW DIFFUSIBLE H2O2 SIGNAL CAN BE SPECIFICALLY TRANSMITTED TO PROMOTE THERAPEUTIC ANGIOGENESIS”. SIGNALING FUNCTION OF ROS IS MAINLY THROUGH OXIDATION OF REACTIVE CYS RESIDUES TO GENERATE “CYSTEINE SULFENIC ACID (CYS-OH)” (SULFENYLATION) WHICH IS INVOLVED IN DISULFIDE BOND FORMATION WITH TARGET PROTEIN AND REDOX SIGNALING. IN ADDITION, ECS UTILIZE GLYCOLYSIS AS A MAJOR SOURCE OF ATP TO PROMOTE ANGIOGENESIS. HOWEVER, THE MECHANISTIC LINK BETWEEN NOX-MITOCHONDRIAL ROS (MITOROS)/REDOX SIGNALING AND EC METABOLISM (GLYCOLYSIS) IN VEGF-INDUCED ANGIOGENESIS IS ENTIRELY UNKNOWN. DRP1 GTPASE IS KEY REGULATOR OF MITOCHONDRIAL (MITO) FISSION VIA ITS POST TRANSLATIONAL MODIFICATION, BUT ITS ROLE IN ROS DEPENDENT VEGFR2 SIGNALING AND ANGIOGENESIS IN ECS AND IN VIVO HAS NEVER BEEN REPORTED. OUR PRELIMINARY DATA ARE CONSISTENT WITH THE HYPOTHESIS THAT VEGF INDUCES SULFENYLATION OF DRP1 VIA NOX-DERIVED H2O2, WHICH DRIVES MITO FISSION-MITOROS AXIS THAT PROMOTES OXIDATIVE ACTIVATION OF KEY METABOLIC ENZYME AMPK VIA DISULFIDE BOND FORMATION (EARLY PHASE) AS WELL AS PFKFB3 EXPRESSION (LATE PHASE) IN ECS. THIS IN TURN ENHANCES ENDOTHELIAL GLYCOLYSIS AND ANGIOGENESIS REQUIRED FOR RESTORING NEOVASCULARIZATION IN ISCHEMIC VASCULAR DISEASE. AIM1 WILL CHARACTERIZE THE VEGF-INDUCED DRP1 SULFENYLATION AND ESTABLISH ITS ROLE IN ROS-DEPENDENT ANGIOGENIC RESPONSES IN ECS. AIM2 WILL DETERMINE THE MOLECULAR MECHANISM BY WHICH VEGF-INDUCED DRP1 SULFENYLATION PROMOTES GLYCOLYSIS VIA MITOCHONDRIAL ROS-DEPENDENT MANNER IN ECS. AIM 3 WILL DETERMINE THE FUNCTIONAL ROLE OF ENDOTHELIAL DRP1 IN ROS-DEPENDENT REPARATIVE NEOVASCULARIZATION AND ADDRESS UNDERLYING MECHANISMS IN VIVO USING ANIMAL MODEL OF PAD (HINDLIMB ISCHEMIA MODEL). WE WILL ALSO ADDRESS HOW DIABETES -INDUCED EXCESS ROS IMPAIR ANGIOGENESIS IN ECS AND IN VIVO BY FOCUSING ON DRP1 PHOSPHORYLATION AT S616, BUT NOT DRP1-CYSOH. WE WILL USE VARIOUS INNOVATIVE REAGENTS, METHODS AND MICE INCLUDING BIOTIN-LABELLED CYS-OH TRAPPING PROBE; BIFC-BASED PROTEIN-PROTEIN INTERACTION IN SITU; REAL-TIME IMAGING OF CYTOSOL- AND MITOROS USING REDOX-SENSITIVE BIOSENSORS; NEWLY DEVELOPED EC-SPECIFIC DRP1-/- MICE AND CRISPR/CAS9-GENERATED “REDOX DEAD” CYS OXIDATION-DEFECTIVE DRP1 OR AMPK KNOCK-IN MUTANT MICE. OUR PROPOSAL WILL PROVIDE NOVEL MECHANISTIC INSIGHTS INTO CYS OXIDIZED MITOCHONDRIAL FISSION PROTEIN DRP1 THAT ORCHESTRATES NOX/MITO ROS SIGNALING AND GLYCOLYSIS AS A POTENTIAL THERAPEUTIC TARGET FOR TREATMENT OF ISCHEMIC CARDIOVASCULAR DISEASES.

THE IMPACT OF A TOBACCO CONTROL INTERVENTION IN AFRICAN-AMERICAN FAMILIES

AUGUSTA AREA COMPREHENSIVE OFFENDER REENTRY PROGRAM (AACORP)

SERVING UNDERSERVED POPULATIONS IN RURAL SETTING: TARGETING TREATMENT AND ADOLESCENT RESILIENCE TOGETHER (SUPRSTART) - THE "SERVING UNDERSERVED POPULATIONS IN RURAL SETTINGS: TARGETING TREATMENT AND ADOLESCENT RESILIENCE TOGETHER" (SUPRSTART) PROJECT WILL BRING EVIDENCE-BASED MENTAL HEALTH SERVICES TO SEVEN RURAL, UNDERSERVED GEORGIA COUNTIES BY TRAINING THERAPISTS IN TRAUMA-FOCUSED COGNITIVE BEHAVIORAL THERAPY (TF-CBT) AND TELEHEALTH PROVISION. IT WILL EXPAND CHILD MALTREATMENT (CM) PREVENTION AWARENESS BY TRAINING 760 COMMUNITY LEADERS IN TWO EVIDENCE-INFORMED PROGRAMS. CM IS ASSOCIATED WITH AN INCREASED RISK OF DELETERIOUS OUTCOMES SUCH AS POST-TRAUMATIC STRESS, MAJOR DEPRESSION, SUBSTANCE USE, AND REVICTIMIZATION. IT IS CRITICALLY IMPORTANT THAT ABUSED CHILDREN HAVE ACCESS TO EVIDENCE-BASED TREATMENT. YET, FAMILIES IN THE SEVEN TARGET COUNTIES MUST TRAVEL IN EXCESS OF 27 MILES TO OBTAIN SERVICES. SUPRSTART WILL BRING TELEHEALTH TF-CBT SERVICES TO YOUTH VICTIMS AGES 3-18 YEARS IN BURKE COUNTY, AND THE TOOMBS CIRCUIT, CIRCUMVENTING MAJOR GEOGRAPHIC BARRIERS TO CARE. AMONG YOUTH SEEKING SERVICES IN THE REGION, MORE THAN 60% ARE UNDER 12 YEARS OF AGE, AND ALMOST 70% IDENTIFY AS FEMALE. FIFTY PERCENT OF YOUTH ARE AFRICAN AMERICAN. IMPORTANTLY, THE INCREASED RECOGNITION OF CM IMPACT ON HEALTH HAS LED TO STATE-LEVEL EFFORTS ENFORCING EVIDENCE-INFORMED PREVENTION TRAINING AMONG ADULTS IN CLOSE CONTACT WITH YOUTH TO IMPROVE RECOGNITION OF CM OCCURRENCES IN COMMUNITY SETTINGS. HOWEVER, LESS THAN 5% OF ADULTS IN THE RURAL GEORGIA COUNTIES ARE TRAINED TO DATE. SUPRSTART WILL ADDRESS CM SERVICE GAPS IN BURKE COUNTY AND THE TOOMBS CIRCUIT BY 1) EXPANDING THE NETWORK OF TRAINED THERAPISTS IN TELEHEALTH TF-CBT, TF-CBT FOR COMMERCIALLY SEXUALLY EXPLOITED YOUTH, AND TREATMENT FOR YOUTH VICTIMS STRUGGLING WITH SUICIDAL IDEATION; 2) CREATING A SUSTAINABLE TELEHEALTH DELIVERY MODEL USING LOCAL GEORGIA FAMILY CONNECTION PARTNERSHIP COUNTY OFFICES, AND 3) EXPANDING THE DISSEMINATION OF THE EVIDENCE-INFORMED STEWARDS OF CHILDREN AND CONNECTIONS MATTER CM PREVENTION CURRICULA TO PEDIATRIC RESIDENTS AT THE MEDICAL COLLEGE OF GEORGIA, AND COMMUNITY LEADERS ACROSS THE SEVEN COUNTIES. THESE GOALS WILL BE MET UNDER THE GUIDANCE OF AN ADVISORY COMMITTEE COMPRISED OF LOCAL AND NATIONAL EXPERTS. IN YEARS 1 AND 2, WE WILL TRAIN TWO COHORTS OF 15 COMMUNITY THERAPISTS EACH IN TF_CBT, AND TELEHEALTH DELIVERY. IN YEAR 3, THESE COHORTS WILL RECEIVE ADVANCED TRAINING, TF-CBT+, TO ADDRESS COMPLEX TRAUMA AMONG SEXUALLY EXPLOITED YOUTH. ADDITIONALLY, THERAPISTS WILL RECEIVE TRAINING IN A TRAUMA-INFORMED SELF-HARM RISK REDUCTION MODEL (SAFETY-A) IN YEARS' 2 AND 3. WE ANTICIPATE AN ANNUAL INCREASE IN YOUTH TO BE SERVED VIA SUPRSTART: YEAR 1 N=20, YEAR 2 N=30, YEAR 3 N=40, YEAR 4 N=50, YEAR 5 N=60, FOR A TOTAL OF 200 YOUTH. EACH YEAR, AT LEAST 12 PEDIATRIC RESIDENTS (TOTAL 60) AND 20 COMMUNITY LEADERS PER COUNTY (TOTAL 700) WILL RECEIVE PREVENTION TRAINING. THE ADVISORY COMMITTEE WILL PERFORM CONTINUOUS PROGRAM EVALUATION ACROSS IMPLEMENTATION AND SERVICE DELIVERY GOALS TO ENSURE SEAMLESS TELEHEALTH TREATMENT PROVISION IN THESE RURAL REGIONS, AND POSITIVE HEALTH OUTCOMES AMONG YOUTH.

TCR ENGINEERING OF HUMAN T CELLS FOR IMMUNOTHERAPY OF HEPATOCELLULAR CARCINOMA

SEX DISPARITIES IN ALDOSTERONE-DEPENDENT RENAL NA+ TRANSPORT AND BLOOD PRESSURE CONTROL - HYPERTENSION REMAINS A MAJOR PUBLIC HEALTH CHALLENGE WORLDWIDE DESPITE THE ABUNDANCE OF AVAILABLE THERAPEUTIC OPTIONS. INAPPROPRIATE THERAPY AND PATIENT NON-COMPLIANCE ARE THE MAIN FACTORS ACCOUNTING FOR POOR BLOOD PRESSURE (BP) CONTROL. IDENTIFICATION OF THE MECHANISMS RESPONSIBLE FOR ELEVATION OF BP IN SPECIFIC PATIENT COHORTS IS CRITICAL TO EFFECTIVELY COMBAT HYPERTENSION AND RELATED DISEASES. EXCESSIVE SODIUM RETENTION VIA THE EPITHELIAL NA- CHANNEL (ENAC) HAMPERS THE ABILITY OF THE KIDNEY TO MAKE PRECISE ADJUSTMENTS TO SODIUM BALANCE AND IS A COMMON PATHOGENIC DETERMINANT OF HYPERTENSION. CUMULATIVE EVIDENCE SUGGESTS THAT BIOLOGICAL SEX IS A PIVOTAL COVARIATE AFFECTING THE DEVELOPMENT AND PATHOPHYSIOLOGY OF HYPERTENSION IN THE HUMAN POPULATION AND ANIMAL MODELS. OUR PROPOSAL INTRODUCES A NOVEL CONCEPT THAT REGULATION OF NA- REABSORPTION IN THE KIDNEY AND RENAL BP CONTROL RELY ON SEX-SPECIFIC MECHANISMS GOVERNING ENAC ACTIVITY IN THE ALDOSTERONE-SENSITIVE DISTAL NEPHRON. WE AND OTHERS HAVE PREVIOUSLY SHOWN THAT ALDOSTERONE ANTAGONISM WITH MINERALOCORTICOID RECEPTOR (MR) INHIBITORS IS INSUFFICIENT TO REDUCE EXCESSIVE RENAL ENAC ACTIVITY AND FAILS TO EFFECTIVELY ATTENUATE BP IN MALE RODENTS WITH ANGIOTENSIN II (ANGIL) DEPENDENT HYPERTENSION. OUR PILOT EXPERIMENTS REVEAL THAT HYPERTENSIVE FEMALE RATS WITH SALT-SENSITIVE AND ANGIL-DEPENDENT HYPERTENSION BENEFIT FROM MR ANTAGONISM AND EXHIBIT A STRONGER BP REDUCTION, WHEN COMPARED TO MALES. MR BLOCKADE IS ALSO MUCH MORE POTENT AT DECREASING RENAL ENAC ACTIVITY IN ANG II-INFUSED FEMALE RATS THAN IN MALES. OUR FINDINGS STRONGLY SUGGEST THAT SENSITIVITY OF RENAL ENAC TO ALDOSTERONE IS AN UNRECOGNIZED SEX-SPECIFIC MECHANISM OF CHRONIC BP REGULATION. THE PROPOSAL IS BUILT AROUND THE CENTRAL HYPOTHESIS THAT HYPERSENSITIVITY OF ENAC TO ALDOSTERONE ACCOUNTS FOR A PREVALENT CONTRIBUTION OF ALDOSTERONE TO RENAL BP CONTROL IN HYPERTENSIVE FEMALE RATS, WHEN COMPARED TO MALES. WE PROPOSE TO TEST THE HYPOTHESIS WITH TWO COMPLEMENTARY SPECIFIC AIMS ADDRESSING A CLINICALLY RELEVANT PROBLEM OF ADEQUATE BLOOD PRESSURE CONTROL AT THE MOLECULAR, CELLULAR {AIM 1) AND INTEGRATIVE {AIM 2) LEVELS. AIM 1 WILL TEST THE HYPOTHESIS THAT SENSITIVITY OF RENAL ENAC TO ALDOSTERONE IS GREATER IN HYPERTENSIVE FEMALE RATS THAN IN MALES. OUR HYPOTHESIS PREDICTS THAT FEMALE SEX STEROIDS POSITIVELY MODULATE MR-DEPENDENT SIGNALING IN THE KIDNEY TO AUGMENT THE RESPONSE OF ENAC TO ALDOSTERONE IN HYPERTENSIVE FEMALES. AIM 2 WILL TEST THE HYPOTHESIS THAT ALDOSTERONE DOMINATES RENAL BP CONTROL IN HYPERTENSIVE FEMALE, BUT NOT IN MALE, RATS. OUR HYPOTHESIS PREDICTS THAT ALDOSTERONE-MR AXIS IS THE PRIMARY DRIVER OF HYPERTENSION AND RENAL DAMAGE IN FEMALES AND MR ANTAGONISM WILL BE DRAMATICALLY MORE EFFECTIVE AT REDUCING BP AND RENAL INJURY IN HYPERTENSIVE FEMALES THAN IN MALES. SUCCESSFUL COMPLETION OF THIS PROPOSAL WILL DIFFERENTIATE THE PATHOPHYSIOLOGICAL MECHANISMS GOVERNING NA- RETENTION IN HYPERTENSIVE MALES AND FEMALES AND PROVIDE THE MISSING PRE-CLINICAL EVIDENCE TO OPTIMIZE THE USE OF MR ANTAGONISTS IN HYPERTENSIVE PATIENTS OF BOTH SEXES.

MECHANISMS OF LENS EPITHELIUM FIBROSIS AND ITS RELEVANCE TO POSTERIOR CAPSULE OPACIFICATION - ABSTRACT CATARACT SURGERY IS ASSOCIATED WITH A HIGH RISK OF POSTERIOR CAPSULE OPACIFICATION (PCO), A FORM OF SECONDARY CATARACT, THE FORMATION OF WHICH IS STILL POORLY UNDERSTOOD. HOWEVER, PCO IS GENERALLY CONSIDERED A CHRONIC DISEASE AND A CONSEQUENCE OF THE LEC FIBROTIC PROCESS, ATTRIBUTED IN PART TO THE EPITHELIAL-MESENCHYMAL TRANSITION (EMT) IN WHICH TGFSS MEDIATED SIGNALING IS ONE OF THE KEY PLAYERS. RECENTLY, WE AND OTHERS HAVE SUGGESTED THAT CANONICAL WNT/SS-CATENIN SIGNALING IS ALSO A PIVOTAL PLAYER IN LEC FIBROSIS AND THE PATHOGENESIS OF PCO. TO TEST THE WNT SIGNALING IN CHRONIC CONDITIONS, WE CONDUCTED A 6-MONTH-KINETIC RABBIT CATARACT SURGERY STUDY AND FOUND A PROFOUND ACTIVATION OF THE CANONICAL WNT SIGNALING DURING THE COURSE OF PCO FORMATION, ASSOCIATED WITH ELEVATED OXIDATION AND CHRONIC INFLAMMATION. WE POSTULATE THAT LENS EPITHELIAL CELLS (LECS) UNDERGO SELF- REGULATORY SIGNALING TRANSDUCTION TO ADAPT TO THE NEW MICROENVIRONMENT AFTER CATARACT SURGERY, AND THE CANONICAL WNT/SS-CATENIN PLAYS A PIVOTAL ROLE IN THIS PROCESS. OUR INITIAL STUDY IDENTIFIES WNT3 AS A HIGHLY EXPRESSED AND UPREGULATED WNT-FAMILY GENE IN HUMAN AND MOUSE LECS AFTER CATARACT SURGERY. HOWEVER, THE ROLE OF WNT3 SIGNALING MEDIATED LEC FIBROSIS AND PCO FORMATION IS STILL UNCLEAR; THE UNDERLYING MECHANISMS OF HOW LENS EPITHELIAL CELLS REGULATE THE WNT SIGNALING REMAIN TO BE ADDRESSED. WE PLAN TO UNRAVEL THESE PUZZLES WITH A BASIS OF SUPPORT FROM SEVERAL KEY FINDINGS OF OUR PILOT STUDIES, E.G., OUR DISCOVERY THAT LECS PACKAGE WNT3 INTO EXOSOMES, A TYPE OF EXTRACELLULAR VESICLE (EVS), AND CONDUCT SIGNALING TRANSDUCTION IN AN ENDOCRINE FASHION. IMPORTANTLY, INFLAMMATORY CYTOKINES, SUCH AS TNFA, CAN DRASTICALLY PROMOTE EXOSOME BIOGENESIS AND WNT3 SECRETION. AIM 1 WILL TEST HOW LENS EPITHELIAL CELLS REGULATE WNT3 PACKAGING AND EXOSOME BIOGENESIS. WE NEED TO UNDERSTAND HOW LECS MANAGE TO PROMOTE EXOSOME BIOGENESIS AND WNT3 SECRETION UNDER THE CERTAIN STIMULI, E.G., INFLAMMATORY CYTOKINES AND OXIDATIVE STRESS. AIM 2 WILL DELINEATE THE ROLE OF WNT3 IN LEC PROLIFERATION, MIGRATION, AND DIFFERENTIATION USING IN VITRO CELL CULTURE, PRIMARY CELL CULTURE, EX VIVO EXPLANT CULTURE, AND IN VIVO MOUSE MODELS. AIM3 STUDIES THE LECS’ GROWTH MICROENVIRONMENT AND THE WNT3 MEDIATED AUTOCRINE SIGNALING BY USING HUMAN TISSUE WITH AND WITHOUT A HISTORY OF CATARACT SURGERY. HUMAN TISSUE STUDIES WILL OFFER A FINAL CHECKPOINT OF RESEARCH AFTER STUDIES FROM CELL CULTURE AND ANIMAL MODELS. IN SUMMARY, SUBSTANTIAL PILOT DATA FORM THE BASIS OF OUR PROPOSAL, ONE WHICH WILL UNRAVEL A NOVEL LECS SELF-REGULATORY PATHWAY THAT IS CLOSELY REMINISCENT OF THE PATHOLOGICAL AND PHYSIOLOGICAL CONDITIONS OF CATARACT SURGERY.

THE ROLES OF LIM-HOMEODOMAIN TRANSCRIPTION FACTORS IN RETINAL DEVELOPMENT

GPCR SIGNALING COMPLEXES IN LIVING CELLS

ROLE OF VAMP1 IN SYNAPTIC TRANSMISSION AND ALZHEIMER'S DEMENTIA

LIPID MACROPINOCYTOSIS: A NOVEL TARGET IN ATHEROSCLEROTIC CARDIOVASCULAR DISEASE

A TWO AMINO ACID SWITCH IN THE ANTI-ANGIOGENIC VEGF165B ISOFORM CREATES A NOVEL AGENT FOR THERAPEUTIC ANGIOGENESIS IN PERIPHERAL ARTERY DISEASE

ALDOSTERONE CONTRIBUTION TO LEPTIN-MEDIATED HYPERTENSION IN OBESE FEMALES

GENETICS OF DELAYED PUBERTY.

CHOLINESTERASE INHIBITORS, AXONAL TRANSPORT, AND MEMORY

ADIPONECTIN SIGNALING IN MOOD REGULATION

GLYCOSAMINOGLYCAN TARGETING TREATMENT FOR PROSTATIC HYPERPLASIA AND FIBROSIS - BENIGN PROSTATIC HYPERPLASIA (BPH) IS AN AGE-RELATED PROGRESSIVE PATHOLOGIC CONDITION IN AGING MEN. MORE THAN 50% OF PATIENTS WITH BPH HAVE LOWER URINARY TRACT SYMPTOMS (LUTS), A CONDITION AFFECTING STORAGE AND VOIDING. LUTS IS EVALUATED IN THE CLINIC USING A VALIDATED SCORING SYSTEM, IPSS (INTERNATIONAL PROSTATE SYMPTOM SCORE). HIGH IPSS DRIVES MEDICAL AND SURGICAL TREATMENTS. HOWEVER, THESE TREATMENTS HAVE VARIABLE EFFICACY AND UNWANTED SIDE EFFECTS. THE PATHOBIOLOGY OF BPH IS PROGRESSIVE HYPERPLASIA AND FIBROSIS IN THE TRANSITIONAL ZONE (TZ) OF THE PROSTATE WITH CHRONIC INFLAMMATION AS THE MAJOR CONTRIBUTOR. HYALURONIC ACID (HA), A NON-SULFATED GLYCOSAMINOGLYCAN, (GAG) IS CAUSALLY LINKED TO INFLAMMATORY AND FIBROTIC DISEASES. WE DISCOVERED THAT HA AND ASSOCIATED FAMILY MEMBERS ARE HIGHLY ELEVATED IN BPH TISSUES FROM PATIENTS WITH HIGH IPSS, REGARDLESS OF THE PROSTATE VOLUME. THE OVERALL OBJECTIVE OF THIS STUDY IS TO EXAMINE THE CLINICAL SIGNIFICANCE OF ELEVATED HA IN BPH PATHOLOGY AND ASSOCIATED SYMPTOMS. FURTHER, TO TEST THE EFFICACY OF AN ORALLY BIOAVAILABLE DRUG THAT INHIBITS HA SYNTHESIS. THIS DRUG HAS AN EXCELLENT SAFETY PROFILE FOR PROLONGED USE. CONSISTENT WITH HA’S ROLE IN INFLAMMATION, TISSUES FROM PATIENTS WITH HIGH IPSS SHOWED HEAVY IMMUNE CELL INFILTRATION IN A DENSE MATRIX, AND RELATED MITOGENIC AND FIBROSIS HALLMARKS. MOUSE MODELS OF INFLAMMATION-INDUCED PROSTATE HYPERPLASIA AND FIBROSIS MIRRORED THE PATTERN OF HA EXPRESSION, AND IMMUNE CELL INFILTRATION. INITIAL STUDIES SHOWED EFFICACY OF THE HA-INHIBITING DRUG IN A MOUSE MODEL. THE PROJECT WILL TEST THE HYPOTHESIS THAT HA MEDIATES INFLAMMATION-INDUCED PROSTATIC HYPERPLASIA AND FIBROSIS. FURTHERMORE, ITS INHIBITOR EFFECTIVELY ABROGATES THIS PATHOLOGY AND IMPROVES SYMPTOMS, WITH LITTLE TOXICITY. FURTHER, IPSS CAN STRATIFY PATIENTS WITH BPH TO THE HA-INHIBITING TREATMENT. WE WILL CHARACTERIZE HA AND THE RELATED MOLECULES FROM BPH TISSUES, INVESTIGATE WHY HA IS ELEVATED IN BPH TISSUES AND DETERMINE HA FUNCTIONS IN EX-VIVO CULTURES (AIM 1). EFFICACY OF THE HA INHIBITOR TO PREVENT/TREAT INFLAMMATION- INDUCED HYPERPLASIA AND FIBROSIS WILL BE EXAMINED IN MOUSE MODELS, ALONG WITH THE EVALUATION OF SYMPTOMS, TOXICITY, AND TISSUE/CELL ANALYSES (AIM 2). CLINICAL SIGNIFICANCE OF HA AND ASSOCIATED FAMILY WILL BE EVALUATED IN BPH SPECIMENS TO DETERMINE IF THEIR LEVELS CORRELATE WITH IPSS, WHILE ADJUSTING FOR CLINICAL PARAMETERS (AIM 3). IMPACT: THIS STUDY SHOULD ESTABLISH HA AS A CRITICAL MEDIATOR OF INFLAMMATION-INDUCED PROSTATIC HYPERPLASIA, FIBROSIS AND ASSOCIATED VOIDING SYMPTOMS. TESTING THE EFFICACY OF THE HA-SYNTHESIS INHIBITOR IN MULTIPLE MODELS AND EVALUATING IF IPSS CAN STRATIFY PATIENTS WITH BPH TO THIS TREATMENT SHOULD AID IN THE REPURPOSING OF THE DRUG TO TREAT BPH-RELATED SYMPTOMS.

REGULATION OF ENDOGENOUS ANTIOXIDANT SYSTEMS IN DIABETIC RETINOPATHY

FLOW-INDUCED CORONARY VASOSPASM IN DIABETIC PATIENTS

UNCOVERING THE PHYSIOLOGICAL ROLE OF FUNCTIONAL HYPEREMIA - NEURONAL ACTIVATION LEADS TO INCREASES IN BLOOD FLOW TO THE REGION. SINCE ITS DISCOVERY IN THE 19TH CENTURY, THIS PHENOMENON – TERMED FUNCTIONAL HYPEREMIA – HAS BEEN THOUGHT TO PROVIDE INCREASED ENERGY NUTRIENTS TO SUSTAIN THE INCREASED NEURAL ACTIVITY. IMPAIRED FUNCTIONAL HYPEREMIA IS SEEN IN MANY NEURODEGENERATIVE DISEASES INCLUDING ALZHEIMER'S DISEASE (AD). HOWEVER, THESE DISEASES ALSO MANIFEST REDUCED BASELINE FLOW LEVELS, MAKING IT DIFFICULT TO DETERMINE THE IMPORTANCE OF FUNCTIONAL HYPEREMIA PER SE IN SUSTAINING HEALTHY NEURONAL FUNCTION. FUNCTIONAL HYPEREMIA ALSO FORMS THE BASIS OF MANY IMAGING TECHNIQUES (SUCH AS FMRI), THAT TAKE ADVANTAGE OF THE SPATIALLY LOCALIZED BLOOD FLOW INCREASE TO INFER THE LOCATION OF NEURAL ACTIVITY FROM VASCULAR/METABOLIC MEASURES. DESPITE THE WIDESPREAD IMPORTANCE OF UNDERSTANDING FUNCTIONAL HYPEREMIA FOR NEUROSCIENCE, THE IMPACTS OF ELIMINATING ONLY THE ACTIVITY-INDUCED INCREASE IN BLOOD FLOW – WITHOUT ALTERING BASELINE FLOW LEVELS OR THE ACTIVITY OF NEURONS AND OTHER CORTICAL CELLS – ARE STILL UNKNOWN. THIS PROPOSAL WILL DETERMINE HOW NEURONAL ACTIVITY AND NEURO-METABOLISM ARE AFFECTED IN HEALTH AND IN ALZHEIMER'S DISEASE WHEN FUNCTIONAL HYPEREMIA IS BLOCKED. WE RECENTLY DEVELOPED A MODEL SYSTEM TO BLOCK FUNCTIONAL HYPEREMIA USING OPTOGENETICS. TO OUR SURPRISE, WE FOUND THAT SENSORY-EVOKED NEURONAL RESPONSES WERE NOT DIMINISHED WHEN FUNCTIONAL HYPEREMIA WAS BLOCKED. IN AIM 1 WE WILL BUILD ON THIS PRELIMINARY DATA BY STUDYING WHAT ASPECTS OF NEURAL RESPONSES TO SENSORY STIMULI ARE ALTERED BY THE LOSS OF FUNCTIONAL HYPEREMIA. TWO- PHOTON CALCIUM IMAGING WILL BE USED IN MOUSE PRIMARY VISUAL CORTEX TO QUANTIFY HOW THE RESPONSE AMPLITUDE AND SELECTIVITY TO STIMULUS ATTRIBUTES (ORIENTATION SELECTIVITY) OF EXCITATORY AND INHIBITORY NEURONS ARE AFFECTED. USING ELECTROPHYSIOLOGY, WE WILL DETERMINE IF TEMPORALLY PRECISE ASPECTS OF NEURONAL ACTIVITY, SUCH AS SPIKE TIMING AND NETWORK SYNCHRONY (I.E. GAMMA OSCILLATIONS) ARE ALTERED. OUR WORKING HYPOTHESIS IS THAT BLOCKING FUNCTIONAL HYPEREMIA IMPAIRS THE CELLULAR MACHINERY INVOLVED IN GENERATING ACTION POTENTIALS (SUCH AS RESTORING ION GRADIENTS). HOWEVER, THESE CONSEQUENCES MAY NOT INITIALLY APPEAR AS REDUCED RESPONSE LEVELS, BUT RATHER AS ALTERATIONS IN SPIKE TIMING, EXCITATORY/INHIBITORY BALANCE, NETWORK SYNCHRONY, AND INFORMATION ENCODING. WE WILL ALSO DETERMINE IF HEALTHY YOUNG BRAINS HAVE THE CAPACITY TO BUFFER THE LOSS OF FUNCTIONAL HYPEREMIA IN WAYS THAT A DISEASED BRAIN CANNOT BY BLOCKING FUNCTIONAL HYPEREMIA IN A MOUSE MODEL OF AD. THIS WILL ALSO SHED LIGHT ON THE RELATIVE IMPORTANCE OF REDUCED FUNCTIONAL HYPEREMIA VERSUS BASELINE FLOW LEVELS IN AD PATHOLOGY. IN AIM 2 WE WILL STUDY HOW NEURONAL METABOLISM IS AFFECTED BY BLOCKING FUNCTIONAL HYPEREMIA. WE WILL RECORD THE CONCENTRATIONS OF OXYGEN, GLUCOSE, LACTATE AND ATP IN THE TISSUE TO DETERMINE HOW BLOCKING FUNCTIONAL HYPEREMIA AFFECTS THE LEVELS OF THESE METABOLITES AND IF IT LEADS TO ALTERED METABOLIC PROCESSING IN NEURONS. WE WILL ALSO QUANTIFY HOW THE VASCULATURE REACTS TO TEMPORARY REDUCTIONS IN BLOOD FLOW. THIS PROPOSAL WILL DEFINE THE ROLE FUNCTIONAL HYPEREMIA PLAYS IN MAINTAINING THE MOMENT-TO-MOMENT METABOLIC NEEDS OF NEURONS.

CLINICALLY UNSCREENED VASCULO-GLIAL-NEURONAL COUPLING IS CRITICAL FOR PHYSIOLOGICAL BRAIN FUNCTION

ROLE OF HIPPO-YAP PATHWAY IN SMOOTH MUSCLE PHENOTYPIC MODULATION

CONVENTIONAL AND UNCONVENTIONAL GPCR-G PROTEIN COUPLING - PROJECT SUMMARY/ABSTRACT G PROTEIN-COUPLED RECEPTORS (GPCRS) ARE IMPORTANT TARGETS OF HORMONES, NEUROTRANSMITTERS AND APPROXIMATELY ONE-THIRD OF FDA-APPROVED DRUGS. THESE RECEPTORS SIGNAL BY COUPLING TO TRANSDUCER PROTEINS FROM FOUR FAMILIES OF HETEROTRIMERIC G PROTEINS AND A FAMILY OF FOUR ARRESTINS. RECENT STRUCTURAL AND FUNCTIONAL STUDIES HAVE DEFINED A CONVENTIONAL ALLOSTERIC MECHANISM WHEREBY LIGAND BINDING TO RECEPTORS PROMOTES ACTIVATION OF G PROTEINS AND ARRESTINS. HOWEVER, THERE IS EXTENSIVE FUNCTIONAL DIVERSITY ACROSS HUNDREDS OF GPCRS AND SIXTEEN G PROTEINS, AND MANY EXAMPLES OF PHYSIOLOGY AND PHARMACOLOGY DO NOT CONFORM TO THE CONVENTIONAL MODEL. FOR EXAMPLE, WE HAVE FOUND SIGNIFICANT VARIABILITY IN THE MECHANISM WHEREBY GPCRS SELECT APPROPRIATE G PROTEINS, AND SOME RECEPTORS THAT RECOGNIZE UNCONVENTIONAL G PROTEIN DETERMINANTS FOR SELECTIVITY. WE HAVE ALSO FOUND SOME RECEPTORS THAT PARADOXICALLY INHIBIT G PROTEIN SIGNALING INSTEAD OF THE USUAL ACTIVATION, A TYPE OF GPCR-G PROTEIN COUPLING THAT IS UNPRODUCTIVE. FINALLY, WE HAVE FOUND RECEPTORS THAT ASSOCIATE WITH G PROTEINS PRIOR TO LIGAND BINDING AND ON AVERAGE RELEASE G PROTEINS AFTER ACTIVATION, A TYPE OF INVERSE COUPLING THAT CAN EXPLAIN THE UNUSAL PHARMACOLOGY OF SUCH RECEPTORS. A COMPLETE UNDERSTANDING OF GPCR-MEDIATED SIGNALING WILL REQUIRE A BETTER UNDERSTANDING OF THESE UNCONVENTIONAL GPCR-G PROTEIN COUPLING MECHANISMS. IN ADDITION, IT WILL BE NECESSARY TO TRANSLATE MOLECULAR STUDIES CARRIED OUT WITH OVEREXPRESSED MOLECULES IN MODEL CELLS TO ENDOGENOUSLY-EXPRESSED GPCRS AND G PROTEINS IN THEIR NATIVE CONTEXT, WHERE EXPRESSION LEVELS, STOICHIOMETRY AND SUBCELLULAR COMPARTMENTALIZATION MORE CLOSELY MATCH THE IN VIVO SITUATION. ACCORDINGLY, WE WILL CARRY OUT STUDIES TO MAP UNCONVENTIONAL SELECTIVITY DETERMINANTS, DETERMINE THE PREVALENCE AND PHYSIOLOGICAL SIGNIFICANCE OF UNPRODUCTIVE AND INVERSE GPCR-G PROTEIN COUPLING, AND TO BETTER UNDERSTAND THE CONFORMATIONAL DYNAMICS OF GPCRS IN CELL MEMBRANES, AND HOW THIS CONFORMATIONAL LANDSCAPE CHANGES AS RECEPTORS BIND LIGANDS AND INTERACT WITH TRANSDUCER MOLECULES.

LONG-RANGE FUNCTION OF THE ERV-9 LTR IN REGULATING GLOBIN GENE SWITCHING

ROLE OF THE RENIN-ANGIOTENSIN SYSTEM IN SEXUAL DIMORPHISMS IN THE DEVELOPMENT OF

ROLE OF HEAT SHOCK TRANSCRIPTION FACTORS (HSFS) IN HEMATOLOGICAL MALIGNANCIES - ABSTRACT AMONG THE HUMAN HEMATOLOGICAL MALIGNANCIES, ACUTE LEUKEMIA TYPES INCLUDING ACUTE MYELOID LEUKEMIA (AML) ACCOUNTS FOR 4% OF ALL CANCER DEATHS WORLDWIDE AND 6% OF CANCER IN THE US POPULATION. AML IS THE SECOND MOST COMMON TYPE OF DIAGNOSED LEUKEMIA IN PEDIATRIC AND ADULT POPULATIONS BUT MOST FREQUENTLY OCCURS IN ADULTS ACCOUNTING FOR ABOUT 30% OF ALL LEUKEMIA CASES. INDUCTIVE THERAPY GENERALLY INVOLVES INTENSIVE AND GENOTOXIC CHEMOTHERAPY THAT CAN LEAD TO HIGH REMISSION RATES (>80%), BUT THE LONG-TERM SURVIVAL RATES FOR AML SUBTYPES ARE ONLY 30-50%. INDEED, THE PROGNOSIS OF PRIMARY RESISTANT AND RELAPSED LEUKEMIA IN PEDIATRIC AND ELDERLY POPULATIONS REMAINS VERY POOR. CURRENTLY, THERE IS A STRONG RATIONALE FOR IMMUNE INTERVENTION IN CANCERS, INCLUDING HEMATOLOGICAL MALIGNANCIES, BUT RECENT CLINICAL TRIALS HAVE DEMONSTRATED THAT THE BENEFITS OF IMMUNOTHERAPY ARE RELEGATED TO A SMALL FRACTION (~20%) OF CANCER PATIENTS, INCLUDING THOSE WITH AML. THIS PROMPTS THE NEED FOR GREATER UNDERSTANDING OF THE MECHANISMS UNDERLYING LEUKEMIC CELL PLASTICITY AND ADAPTATION, AS WELL AS DEVELOPING NOVEL THERAPEUTIC APPROACHES TO ACHIEVE MORE EFFECTIVE AND SELECTIVE CURE RATES FOR PRIMARY DRUG-RESISTANT AND RELAPSED LEUKEMIA. HSF1, AS THE MASTER ACTIVATOR OF THE CLASSICAL HEAT SHOCK RESPONSE AND GUARDIAN OF THE PROTEOME, HAS BEEN IMPLICATED IN THE PATHOGENESIS OF CANCER. OUR PRECLINICAL STUDIES HAVE REVEALED THE EXISTENCE OF A COORDINATED, HSF1-DEPENDENT PROTEIN HOMEOSTATIC AND METABOLIC PROGRAM THAT, WHEN INACTIVATED, CAN LEAD TO CANCER REGRESSION AND, IN PARTICULAR, EFFECTIVE LEUKEMIA INHIBITION. ENCOURAGINGLY, OUR RECENT STUDIES UNVEILED A HIGHLY NOVEL AND CLINICALLY SIGNIFICANT ROLE FOR HSF1 INHIBITION IN METABOLIC REPROGRAMMING AND ENHANCEMENT OF ANTI-TUMOR T CELL IMMUNITY. THIS MAY PROVIDE A NEW APPROACH TO IMPROVE LEUKEMIA TREATMENT BY IMPROVING THE ANTI-TUMOR IMMUNE CAPACITY TARGETING HSF1 ACTIVITY. WE PROPOSE THAT BY INHIBITING SUPPORTIVE NON- ONCOGENE ADDICTION PATHWAYS, INTERFERING WITH TUMOR-PROMOTING METABOLIC REPROGRAMMING, AND IMPROVING THE PREDICTED POWER OF ANTI-TUMOR IMMUNITY THROUGH TARGETING OF HSF1 ACTIVITY, WE CAN DEVELOP A VALID STRATEGY FOR THERAPEUTIC INTERVENTIONS IN LEUKEMIA. OUR EXPERIMENTAL STRATEGY ENTAILS THE FOLLOWING TWO MAJOR APPROACHES: 1. DETERMINE THE IMPACT OF HSF1 DELETION ON AML INDUCTION AND EXPLORE ITS THERAPEUTIC POTENTIAL FOR ADVANCED CHEMOTHERAPY-RESISTANT AML, AND 2. INVESTIGATE THE THERAPEUTIC IMPACT OF HSF1 DELETION ON IMPROVED MHC- RESTRICTED TCR OR CHIMERIC ANTIGEN RECEPTOR (CAR)-T CELL-BASED IMMUNOTHERAPY FOR PRIMARY AND DRUG RESISTANT/RELAPSED HUMAN AMLS. IN SUMMARY, THE LONG-TERM TRANSLATIONAL GOAL OF THE PROJECT IS TO TEST THE POTENTIAL OF HSF1 TARGETING IN HUMAN AML. IT WILL ALSO PROVIDE PROOF-OF-CONCEPT FOR TARGETING HSF1-MEDIATED METABOLIC PROGRAMS FOR IMMUNOTHERAPEUTIC APPLICATION OF CHEMOTHERAPY-RESISTANT HEMATOLOGICAL MALIGNANCIES.

UNDERSTANDING MECHANISMS OF RESISTANCE TO ANTI-ANGIOGENIC TREATMENTS

VISUALIZATION OF INDIVIDUAL FOXP3+ T CELLS DURING AN ONSET AND PROGRESSION OF AUT

DC EXOSOME THERAPY TO RESOLVE INFLAMMATORY BONE LOSS AND ORAL INFECTION - PROJECT ABSTRACT PERIODONTITIS (PD) IS A BONE DEGENERATIVE INFLAMMATORY DISEASE THAT CONTRIBUTES TO INCREASED RISK OF OTHER DISEASES WITH MORE SERIOUS MORTALITY AND MORBIDITY PROFILES. WE PROPOSE HERE A NOVEL IMMUNE THERAPEUTIC STRATEGY FOR PD, USING CELLULAR NANO-PARTICLES, EXOSOMES (EXO) FROM DENDRITIC CELLS. DENDRITIC CELLS ARE THE “DIRECTORS” OF THE ADAPTIVE IMMUNE RESPONSE, AS SUCH, THEIR IMMUNE FUNCTIONS CAN BE EXPLOITED TO TREAT CHRONIC IMMUNE-MEDIATED DISEASES SUCH AS CANCER AND AUTOIMMUNE DISEASES. THIS IS FIRST STUDY TO OUR KNOWLEDGE TO EMPLOY PHENOTYPICALLY STABLE EXOSOMES FROM DENDRITIC CELLS TO TREAT EXPERIMENTAL PD IN MICE. OUR STRATEGY CONSISTS OF “REPROGRAMING” IMMUNE CELLS IN THE PERIODONTIUM RESPONSIBLE FOR DESTRUCTIVE INFLAMMATORY BONE LOSS USING CUSTOM DC EXO. OUR TACTIC IS TO INJECT DC EXO DIRECTLY INTO THE GINGIVA, TO TARGET ANTI-INFLAMMATORY PATHWAYS IN SPECIFIC IMMUNE CELLS, TO PREVENT/RESOLVE ALVEOLAR BONE LOSS. THE OVERALL OBJECTIVE OF THESE STUDIES TO ACQUIRE A BETTER UNDERSTANDING OF THE IMMUNOBIOLOGY OF DC-DERIVED EXO IN VIVO AND IN VITRO IN THE MURINE MODEL OF EXPERIMENTAL PD, AND RELATE THESE FINDINGS TO EXO ISOLATED FROM HUMANS WITH PD. OUR AIMS ARE TO TEST THE HYPOTHESES THAT: AIM 1. EXOGENOUS DCEXO DELIVERED TO THE PD LESION PERSIST IN SITU AND REPROGRAM RECIPIENT DCS AND T CELLS INVOLVED IN EXPERIMENTAL PD IN MICE IN VIVO; AIM 2. DC EXO PRESERVE MOLECULAR CARGO AND REPROGRAM ACCEPTOR DCS AND T CELLS THROUGH RECEPTOR BINDING AND ENDOCYTOSIS; AIM 3. ENDOGENOUS EXO FROM HUMANS WITH PD CONTAIN MOLECULAR CARGO OF DIRECT RELEVANCE TO EXO THERAPY AND PD PATHOGENESIS. FUTURE GOALS INCLUDE: 1. THE DEVELOPMENT OF DC EXO THERAPY FOR PD IN HUMANS, 2. ASSESSMENT OF SALIVARY EXO MOLECULAR CARGO FOR DIAGNOSTIC POTENTIAL IN PD AND OTHER ORAL DISEASES

BALANCE BETWEEN EFFECTOR AND REGULATORY CD4 T CELLS IN SJOGREN'S SYNDROME

THE INFLAMMATORY CYTOKINES, MCP-1 AND TGF-BETA, MEDIATE RENAL AUTOREGULATORY IMPA

REMOTE ISCHEMIC CONDITIONING PROMOTES CEREBROVASCULAR RECOVERY AFTER INTRACEREBRAL HEMORRHAGE

MICRORNA FUNCTION DURING NEURONAL REPROGRAMMING IN TREATING SPINAL CORD INJURY

THERAPEUTIC TARGETING OF MONOACYLGLYCEROL LIPASE AFTER TRAUMATIC BRAIN INJURY

ENOS-ACTIN INTERACTION AND OXYGEN IN LUNG ENDOTHELIUM

ROLE OF MYD88-5 IN THE PATHOGENESIS OF PARKINSON'S DISEASE

BRAIN AROMATASE IN NEUROLOGICAL FUNCTION AND DISEASE

AUGUSTA SPAN 2 - THROMBOLYSIS AND ENDOVASCULAR THROMBECTOMY (ET), REMAIN THE ONLY TWO FDA-APPROVED THERAPIES FOR ACUTE ISCHEMIC STROKE. DESPITE THE EFFICACY OF ET, 50% OF THE PATIENTS REMAIN DISABLED AT 3 MONTHS. ADJUNCTIVE THERAPIES TO THROMBOLYSIS AND ET ARE NEEDED THAT PROVIDE “BRIDGING NEUROPROTECTION” AND IMPROVE COLLATERAL BLOOD FLOW. SINCE PRESENCE OF COLLATERALS IS A MAJOR PREDICTOR OF OUTCOME WITH ET, A PROMISING NEW THERAPEUTIC AVENUE IS DEVELOPMENT OF “COLLATERAL THERAPEUTICS. TRANSLATION FROM THE BENCH (RODENT STROKE MODELS) TO THE BEDSIDE HAS BEEN PLAGUED BY FAILURE. THE NINDS STROKE PRECLINICAL ASSESSMENT NETWORK (SPAN) HAS BEEN AN ATTEMPT TO BRIDGE THIS TRANSLATIONAL CHASM USING A MULTI-SITE PRECLINICAL RANDOMIZED CLINICAL TRIAL (PRCT) NETWORK MODELED AFTER HUMAN CLINICAL TRIALS. THE MEDICAL COLLEGE OF GEORGIA/AUGUSTA UNIVERSITY WAS ONE OF THE 6 TESTING CENTERS IN SPAN 1. WE WERE A HIGH PERFORMING CENTER AS DEMONSTRATED BY OUR CONSISTENT INFARCT SIZE IN THE MIDDLE CEREBRAL ARTERY (MCA) OCCLUSION MODEL. SPAN 1 WAS SUCCESSFUL AND DEMONSTRATED FEASIBILITY WITH EXCELLENT PROTOCOL ADHERENCE, EXCELLENT DATA COMPLETION AND LOW RATES OF ANIMAL LOSS IN A LARGE, MULTI-SITE PRECLINICAL NETWORK THE SPECIFIC AIMS FOR SPAN 2 INCLUDE: AIM 1. TEST UP TO 8 SELECTED DRUGS/INTERVENTIONS IN PARALLEL, IN COLLABORATION WITH THE COORDINATING CENTER, NINDS AND THE OTHER TESTING LABORATORIES IN THE SPAN NETWORK, IN A MULTISITE PRCT DESIGN TO SELECT THE BEST CEREBROPROTECTIVE AGENT(S) FOR CLINICAL TRIAL IN ACUTE STROKE. AIM 2. ASSESS SHORT AND LONG-TERM FUNCTIONAL OUTCOMES AFTER EXPERIMENTAL STROKE, WITH A SPECIFIC EMPHASIS ON NEUROIMAGING AND BEHAVIORAL TESTING. AIM 3. AS PART OF THE SPAN NETWORK, PROPOSE NEW IDEAS FOR SUB-STUDIES AND MANUSCRIPTS AND DEVELOP A TRAINING PIPELINE FOR THE NEXT GENERATION OF PRECLINICAL STROKE INVESTIGATORS. EXPECTED IMPACT: WE WILL ADVANCE THE MOST EFFECTIVE DRUG/INTERVENTION(S) INTO HUMAN CLINICAL TRIAL.

ROLE OF LIPOXYGENASE PATHWAY IN EARLY MICROVASCULAR DYSFUNCTION DURING DIABETIC R

EPIGENETIC REGULATION OF HDAC9 IN OBESITY AND ATHEROSCLEROSIS

MECHANISMS OF SUBCLINICAL RENAL INJURY IN FEMALES FOLLOWING AKI: IMPLICATIONS FOR ADVERSE PREGNANCY OUTCOMES - RECENT LARGE-SCALE CLINICAL STUDIES REPORT THAT WOMEN WITH A HISTORY OF ACUTE KIDNEY INJURY (AKI) HAVE ABNORMALLY HIGH RATES OF ADVERSE MATERNAL AND FETAL OUTCOMES DURING PREGNANCY, DESPITE CLINICAL EVIDENCE OF RENAL RECOVERY PRIOR TO CONCEPTION AS DEFINED BY MEASUREMENT OF SERUM CREATININE. WE ESTABLISHED A PREGNANCY POST-AKI MODEL IN SPRAGUE DAWLEY RATS USING ISCHEMIA REPERFUSION (IR) AS AN EXPERIMENTAL MODEL OF AKI WHICH RECAPITULATES MANY OF THE CLINICAL FINDINGS, INCLUDING FETAL GROWTH RESTRICTION. THE GOAL OF THIS PROPOSAL IS TO ADDRESS A CRITICAL GAP IN KNOWLEDGE REGARDING THE MECHANISMS BY WHICH AKI PREDISPOSES FEMALES TO ADVERSE OUTCOMES IN PREGNANCY. OUR CENTRAL HYPOTHESIS IS THAT AKI PRIOR TO CONCEPTION IMPAIRS THE RENAL, HEMODYNAMIC AND IMMUNE ADAPTATIONS REQUIRED FOR A HEALTHY PREGNANCY BY DECREASING NITRIC OXIDE (NO) BIOAVAILABILITY. NORMAL PREGNANCY IS CHARACTERIZED BY PROFOUND ADAPTATIONS IN ALMOST EVERY ORGAN SYSTEM TO MEET THE DEMANDS OF THE FETUS WHILE MAINTAINING THE PHYSIOLOGICAL NEEDS OF THE MOTHER. NO IS A CENTRAL MEDIATOR OF THE RENAL AND CARDIOVASCULAR ADAPTATIONS IN HEALTHY PREGNANCY, AND DECREASES IN NO BIOAVAILABILITY LEAD TO ADVERSE MATERNAL AND FETAL OUTCOMES, INCLUDING LOW BIRTH WEIGHT. WE HAVE ALSO SHOWN THAT NO SYNTHASE (NOS) IS REQUIRED FOR FEMALES TO INCREASE T REGULATORY CELLS (TREGS), AND FAILURE TO EXPAND TREGS IN PREGNANCY INDUCES RENAL AND VASCULAR DYSFUNCTION IN THE MOTHER AND PROMOTES FETAL GROWTH RESTRICTION. THERE IS GROWING EVIDENCE THAT THE RENAL NOS SYSTEM IS IMPAIRED FOLLOWING AKI IN MALES. THE IMPACT OF AKI ON NO/NOS IN FEMALES IS UNKNOWN. THE IMPACT OF AKI ON PHYSIOLOGICAL ADAPTATIONS TO PREGNANCY, INCLUDING INCREASES IN NO BIOAVAILABILITY ARE UNKNOWN. OUR HYPOTHESIS IS SUPPORTED BY STRONG PRELIMINARY DATA SHOWING AKI PRIOR TO PREGNANCY 1) DECREASES RENAL NOS EXPRESSION PRIOR TO CONCEPTION AND IN PREGNANCY, 2) RESULTS IN SUBCLINICAL INJURY PRIOR TO PREGNANCY AND RENAL INJURY IN PREGNANCY, 3) IMPAIRS PLASMA VOLUME EXPANSION, 4) IMPAIRS VASCULAR FUNCTION IN PREGNANCY, AND 5) DECREASES TREG EXPANSION IN PREGNANCY. AIM 1 WILL TEST THE HYPOTHESIS THAT AKI INDUCES REDUCTIONS IN NO BIOAVAILABILITY THAT ARE EXACERBATED IN PREGNANCY RESULTING IN RENAL AND VASCULAR DYSFUNCTION. WE PROPOSE THAT AKI RESULTS IN THE FAILURE TO APPROPRIATELY INCREASE NO-MEDIATED HEMODYNAMIC ADAPTATIONS AND LOSS OF NO-MEDIATED PLASMA VOLUME EXPANSION LEADING TO POOR MATERNAL AND FETAL OUTCOMES IN PREGNANCY. AIM 2 WILL TEST THE HYPOTHESIS THAT FAILURE TO UPREGULATE TREGS IN PREGNANCY CONTRIBUTES TO ADVERSE PREGNANCY OUTCOMES POST-AKI. WE WILL DETERMINE HOW AKI PRIOR TO PREGNANCY IMPACTS TREGS AND IF INCREASING TREGS DURING PREGNANCY IMPROVES FETAL GROWTH AND MATERNAL OUTCOMES. WE PROPOSE THAT AKI RESULTS IN THE FAILURE TO INCREASE NO WHICH IS REQUIRED FOR TREG EXPANSION, CONTRIBUTING TO FURTHER DECREASES IN NO. RESULTS WILL PROVIDE A CRITICALLY NEEDED PRE-CLINICAL FOUNDATION TO ELUCIDATE THE MECHANISMS UNDERLYING POOR PREGNANCY OUTCOMES AFTER AKI, GIVE EVIDENCE FOR IMPROVED PRE- AND PERINATAL CARE GUIDELINES, AND POTENTIALLY IDENTIFY NOVEL THERAPEUTIC TARGETS FOR CLINICAL TRIALS.

EPIGENETIC REGULATION OF THE METABOLIC SHIFT IN MAMMALIAN PERINATAL HEARTS - ABSTRACT: DURING THE TRANSITION TO EXTRA-UTERINE LIFE, MAMMALIAN HEARTS UNDERGO A CRUCIAL METABOLIC SHIFT THAT IS ESSENTIAL FOR NEONATAL HEARTS TO ADAPT TO NORMOXIC CONDITIONS AND THE INCREASED CARDIAC WORKLOAD EXPERIENCED AFTER BIRTH. PROPER METABOLIC TRANSITION IS ESSENTIAL FOR ESTABLISHING NORMAL HEART PHYSIOLOGY. THE LONG-TERM GOAL OF THIS PROJECT IS TO REVEAL THE MOLECULAR AND CELLULAR MECHANISMS THAT REGULATE THIS CARDIAC METABOLIC SHIFT USING MOUSE MODELS AND ULTIMATELY TRANSLATE OUR DISCOVERIES INTO CLINICAL APPLICATIONS ADDRESSING INBORN CARDIOMYOPATHIES. MONO-UBIQUITINATION OF HISTONE H2A K119 (H2AK119UB) IS A MAJOR POST-TRANSLATIONAL MODIFICATION OF HISTONE H2A, OCCURRING IN 5-15% OF TOTAL H2A IN MAMMALIAN CELLS. H2AK119UB IN THE PROMOTER REGION INHIBITS RNA POLYMERASE II ELONGATION AND ACTS AS A REPRESSIVE EPIGENETIC MECHANISM TO REGULATE KEY DEVELOPMENTAL PROGRAMS FROM DROSOPHILA TO MAMMALS. THE PRECISE LEVEL OF H2AK119UB IS DYNAMICALLY CONTROLLED BY THE BALANCE BETWEEN UBIQUITIN LIGASES AND DEUBIQUITINASES. USP16 IS A HISTONE H2AK119UB-SPECIFIC DEUBIQUITINASE THAT DE- REPRESSES GENE TRANSCRIPTION. TO TEST THE ROLE OF USP16 IN HEARTS, WE SPECIFICALLY DELETED USP16 IN THE MYOCARDIUM. ALL MUTANT MICE DIED WITHIN 3 DAYS AFTER BIRTH AND DISPLAYED SEVERE MYOCARDIAL WALL ANOMALIES. OUR SUBSEQUENT TWO-HYBRID AND BIOCHEMICAL ANALYSES REVEALED THAT NUCLEAR RESPIRATORY FACTOR 1 (NRF1) IS A NOVEL USP16 INTERACTION PARTNER. NRF1 IS A NUCLEAR TRANSCRIPTION FACTOR THAT ACTIVATES EXPRESSION OF THE VAST MAJORITY OF NUCLEAR GENES ENCODING SUBUNITS OF MITOCHONDRIAL OXIDATIVE PHOSPHORYLATION (OXPHOS) COMPLEXES. IN SUPPORT OF THE FUNCTIONAL SIGNIFICANCE OF THE USP16-NRF1 INTERACTION, OUR MRNA-SEQUENCING ANALYSIS REVEALED THAT THE PATHWAY INVOLVED IN OXPHOS WAS MOST SIGNIFICANTLY DOWNREGULATED BY MYOCARDIAL DELETION OF USP16. WE PROPOSE OUR CENTRAL HYPOTHESIS THAT USP16 INTERACTS WITH NRF1 TO UPREGULATE EXPRESSION OF NUCLEAR OXPHOS GENES, SUPPORTING THE METABOLIC SHIFT IN PERINATAL HEARTS. IN AIM 1, WE WILL TEST HOW DELETION OF USP16 AFFECTS THE METABOLIC SHIFT IN PERINATAL HEARTS. IN AIM 2, WE WILL TEST IF OXPHOS GENES ARE MAJOR DIRECT REGULATORY TARGETS OF USP16 IN PERINATAL HEARTS USING HIGH-THROUGHPUT APPROACHES. IN AIM 3, WE WILL TEST THE ROLE OF NRF1 IN LOADING USP16 ONTO ITS TARGET SITES. THIS PROJECT WILL FILL A MAJOR KNOWLEDGE GAP REGARDING THE BASIC MOLECULAR MECHANISM UNDERLYING THE CARDIAC METABOLIC SHIFT, WHICH IS CRITICAL FOR THE PROPER TRANSITION TO EXTRA-UTERINE LIFE. OUR RESEARCH WILL CONTRIBUTE TO THE DEVELOPMENT OF NOVEL CLINICAL APPLICATIONS AIMED AT OXPHOS-ASSOCIATED HEART DEFECTS.

ER-TO-GOLGI TRAFFIC AND SIGNAL REGULATION OF GPCRS

DIABETIC RETINOPATHY: AR1 AS A NOVEL THERAPEUTIC TARGET

SUPPRESSING THE CO-CHAPERONE UNC45A REPROGRAMS THE IMMUNOSUPPRESSIVE TUMOR MICROENVIRONMENT TO FIGHT CANCER - ABSTRACT BREAST CANCER REMAINS ONE OF THE LEADING CAUSES OF DEATH AMONG WOMEN IN THE US. WHEN AVAILABLE, TARGETED THERAPIES HAVE SIGNIFICANTLY IMPROVED THE OUTCOMES FOR BREAST CANCER PATIENTS. UNFORTUNATELY, RESISTANCE TO THERAPIES AND RELAPSES ARE COMMON, ESPECIALLY AMONG PATIENTS WITH TRIPLE-NEGATIVE BREAST CANCER (TNBC), A PARTICULARLY AGGRESSIVE DISEASE FOR WHICH LIMITED TARGET THERAPY OPTIONS ARE AVAILABLE. IT IS BECOMING CLEAR THAT THE IMMUNOSUPPRESSIVE TUMOR MICROENVIRONMENT (TME) IS A MAJOR OBSTACLE TO SUCCESSFULLY TREATING AGGRESSIVE BREAST CANCERS. TUMOR GROWTH IS ASSOCIATED WITH THE ACCUMULATION OF IMMUNOSUPPRESSIVE CELLS IN THE TME, CAUSING CYTOTOXIC T-CELL DYSFUNCTION AND TUMOR ESCAPE. FURTHERMORE, TUMOR CELLS ACTIVATE ESCAPE MECHANISMS SUCH AS TRIGGERING THE EXPRESSION OF THE IMMUNE CHECKPOINT PD-L1, WHICH REDUCES CD8+ T CELL KILLING. INDEED, IMMUNOTHERAPEUTIC AGENTS TARGETING PD-L1, AND OTHER IMMUNE CHECKPOINT MOLECULES, HAVE SHOWN REMARKABLE EFFICACY IN METASTATIC TUMORS REFRACTORY TO STANDARD PROTOCOLS. UNFORTUNATELY, ONLY A SMALL SUBSET OF PATIENTS RESPONDS TO THESE IMMUNOTHERAPIES. THEREFORE, THERE IS AN URGENT NEED FOR IMPROVED THERAPEUTIC STRATEGIES. DISCOVERING NEW IMMUNOTHERAPEUTIC TARGETS AND INNOVATIVE COMBINATIONS WITH EXISTING THERAPIES ARE ATTRACTIVE OPTIONS FOR EXPANDING THE BENEFITS OF ENGAGING THE IMMUNE SYSTEM AGAINST CANCER. WE DISCOVERED THAT DURING BREAST TUMORIGENESIS, THE HSP90 CO-CHAPERONE UNC45A BECOMES MORE NUCLEAR AND ESSENTIAL FOR CANCER CELL DIVISION BUT REMAINS DISPENSABLE FOR NORMAL CELL PROLIFERATION. FURTHER, WE FOUND THAT SUPPRESSING UNC45A IN MOUSE SYNGENEIC TNBC TUMORS TRIGGERS A DRASTIC REPROGRAMMING OF THE IMMUNE-SUPPRESSIVE TME INTO A MORE IMMUNE-RESPONSIVE ONE. IN ADDITION, LOSS OF UNC45A HAMPERS THE ABILITY OF CANCER CELLS TO REPAIR IONIZING RADIATION (IR)-INDUCED DNA DAMAGE AND SENSITIZES IN VIVO TNBC MODELS TO RADIATION THERAPY. IN THIS APPLICATION, WE PROPOSE THAT UNC45A IS A PROMISING NOVEL IMMUNOTHERAPEUTIC TARGET AND THAT ITS INHIBITION, IN COMBINATION WITH IR THERAPY, WILL REDUCE THE TNBC BURDEN. OUR SPECIFIC AIMS ARE 1. TO CHARACTERIZE TME REPROGRAMMING CAUSED BY LOSS OF UNC45A IN MALIGNANT CELLS; 2. TO DEFINE THE MOLECULAR MECHANISM BY WHICH UNC45A PROMOTES AN IMMUNOSUPPRESSIVE TME; 3. TO DETERMINE THE THERAPEUTIC POTENTIAL OF UNC45A INHIBITION IN COMBINATION WITH IR THERAPY.

ATF3 REGULATION OF HISTONE ACETYLATION IN GENOME MAINTENANCE

A NOVEL THERAPEUTIC STRATEGY TO ERADICATE BREAST CANCER THROUGH HSP90 INHIBITION AND REDUCED IMMUNE TOLERANCE - ABSTRACT: IMMUNOTHERAPY IS BECOMING A PILLAR OF CANCER TREATMENT, AND MANY RESPONSIVE PATIENTS HAVE EXPERIENCED DURABLE, OR EVEN CURATIVE, OUTCOMES. FOR REASONS THAT REMAIN UNCLEAR, HOWEVER, ONLY A MINORITY OF CANCER PATIENTS BENEFIT FROM IMMUNE CHECKPOINT BLOCKADE (ICB) THERAPIES. EXPANDING THIS REMARKABLE ACHIEVEMENT TO MOST CANCER PATIENTS IS BEING ACTIVELY SOUGHT THROUGH MULTIPLE AVENUES, INCLUDING COMBINED IMMUNOTHERAPY, SUCH AS BLOCKING BOTH PD-1 AND CTLA-4. IN ADDITION, PROMISING RESULTS HAVE BEEN OBSERVED WHEN ICB THERAPIES WERE COMBINED WITH AVAILABLE CHEMOTHERAPIES THAT POTENTIATE THE IMMUNE-MEDIATED ANTI-TUMOR RESPONSE. THIS OPENS THE POSSIBILITY THAT THE USE OF SMALL MOLECULES COULD RESTORE THE IMMUNE SYSTEM’S RECOGNITION OF CANCER CELLS AS A FOREIGN ENTITY AND THUS WOULD POTENTIATE IMMUNOTHERAPEUTIC PROGRESS. IN THIS CONTEXT, WE HAVE IDENTIFIED A CYCLIC PEPTIDE, ENNA, AS A NOVEL INHIBITOR OF HEAT SHOCK PROTEIN 90 (HSP90), WITH A POTENT ABILITY TO UNLEASH THE IMMUNE SYSTEM AGAINST TUMOR CELLS. THIS DISCOVERY STEMMED FROM OUR EFFORT TO FIND NEW HSP90 INHIBITORS THAT CIRCUMVENT KNOWN SIDE EFFECTS THAT HAVE HAMPERED THE CLINICAL PROGRESS OF FIRST-GENERATION INHIBITORS. IN PARTICULAR, THIS COMPOUND DOES NOT INDUCE A HEAT SHOCK RESPONSE, WHICH HAD REDUCED THE EFFICACY OF EARLY INHIBITORS THROUGH ACTIVATION OF PRO-SURVIVAL MECHANISMS. ENNA INDUCES IMMUNOGENIC CANCER CELL DEATH, PROMOTES TUMOR IMMUNE CELL INFILTRATION, AND UNLEASHES A POWERFUL T CELL-MEDIATED IMMUNE RESPONSE, RESULTING IN HIGHLY EFFICACIOUS TUMOR KILLING IN A SYNGENEIC MOUSE MODEL. MOLECULARLY, ENNA INTERFERES WITH SEVERAL ONCOGENIC PATHWAYS AND REDUCES THE PROTEIN LEVEL OF THE PROGRAMMED CELL DEATH LIGAND-1 (PD-L1), A KEY MEDIATOR OF TUMOR- INDUCED IMMUNE TOLERANCE. WE THEREFORE PROPOSE THAT ENNA IS A PROMISING ANTI-TUMOR AGENT TARGETING HSP90 THROUGH A NOVEL MECHANISM OF ACTION INVOLVING CANCER CELL TOXICITY THAT INCREASES ITS IMMUNOGENICITY AND MODULATION OF THE TUMOR MICROENVIRONMENT TO REDUCE IMMUNOTOLERANCE. IN AIM 1, WE WILL PERFORM PRECLINICAL DEVELOPMENT OF ENNA AS A DRUG TO DETERMINE ITS TOXICITY AND THE POTENTIAL IMPACT OF ENNA ON THE IMMUNE SYSTEM OF MICE. WE WILL ALSO CHARACTERIZE THE ENNA-HSP90 INTERACTION THROUGH MUTATIONAL AND BIOCHEMICAL ANALYSES. IN AIM 2, WE WILL DETERMINE HOW INHIBITION OF HSP90 BY ENNA INTERFERES WITH PD-L1 CHAPERONING AND FUNCTION. IN AIM 3, WE WILL DEFINE IMMUNE CELL MECHANISMS UNDERLYING ENNA’S ANTI-TUMOR EFFECT. COMBINING ENNA WITH ANTI-CTLA-4 WILL BE TESTED, AND COMPREHENSIVE PROFILING OF IMMUNE CELLS INVOLVED IN THE ANTI-TUMOR ACTIVITY WILL BE IMPLEMENTED. IN AIM 4, WE WILL TEST THE IMPORTANCE OF ENNA- INDUCED CANCER CELL AUTOPHAGY AND HSP90 CELL SURFACE EXPOSURE IN IMMUNE-DEPENDENT TUMOR ERADICATION. IF SUCCESSFUL, THESE STUDIES WILL SHED LIGHT ON THE ROLE OF THE HSP90 IN PROMOTING IMMUNE TOLERANCE AND WILL PROVIDE AN INNOVATIVE APPROACH TO POTENTIATE IMMUNOTHERAPY USING A NOVEL HSP90 INHIBITOR.

PTEN, TREGS AND MDSCS IN THE TUMOR MICROENVIRONMENT

CHEMOTHERAPY-INDUCED MDSCS AND ANTITUMOR IMMUNITY

REGULATION OF EXOSOME RELEASE AND ITS ROLE IN ACUTE KIDNEY INJURY.

IDO-EXPRESSING PLASMACYTOID DENDRITIC CELLS AND TUMORS

MECHANISMS UNDERLYING THE SUSCEPTIBILITY AND SEVERITY OF ACUTE KIDNEY INJURY

DRUG DISCOVERY FOR COGNITIVE IMPAIRMENT ASSOCIATED WITH DRUGS OF ABUSE

BACTERIA-DERIVED XENOBIOTICS IN COLON CANCER PREVENTION: LINK TO GPR109A AND COLONIC KETOGENESIS - PROJECT SUMMARY: CHEMICAL CARCINOGENS AND XENOBIOTICS IN THE DIET PLAY A SIGNIFICANT ROLE IN COLON CANCER (CC) FORMATION. THE INTESTINAL TRACT, INCLUDING THE COLON, IS OFTEN THE FIRST SITE THAT IS EXPOSED TO THESE CARCINOGENS AND XENOBIOTICS. CC IS A COMMON MALIGNANCY IN MEN AND WOMEN, AND P53 MUTATION (P53D) OR INACTIVATION IS OBSERVED IN ~60% OF CC AND ~90% OF ALL HUMAN CANCERS. AS SUCH, THE DEVELOPMENT OF THERAPEUTIC STRATEGIES THAT WILL REJUVENATE WILD-TYPE (WT) P53 FUNCTION OR TARGET P53D SHOULD HAVE PROFOUND IMPLICATIONS IN THE PREVENTION AND TREATMENT OF CC. SIMILARLY, A HIGH FREQUENCY (40-60%) OF KRAS MUTATION (KRASD) IS OBSERVED IN ORGANISMS EXPOSED TO ENVIRONMENTAL CHEMICAL CONTAMINANTS, AND ~40-45% OF CC IS ASSOCIATED WITH KRASD. WE FOUND A RECIPROCAL ASSOCIATION BETWEEN GPR109A EXPRESSION AND KRASD CC. GPR109A IS A GAI-COUPLED RECEPTOR FOR THE BACTERIAL FERMENTATION PRODUCT BUTYRATE (BTR), THE KETONE BODY B-HYDROXYBUTYRATE (BHB), AND THE VITAMIN NIACIN (NA). ABOUT 85% OF CC SHOWS A REDUCED EXPRESSION OF GPR109A. GPR109A AGONISTS NA IS THE PRECURSOR OF NAD+ AND P53 IS AN NAD+-DEPENDENT MOLECULE AND NA DEFICIENCY IMPAIRS P53 FUNCTION. THEREFORE, GPR109A COULD BE THE MOLECULAR TARGET OF KRASD AND IT PLAYS AN IMPORTANT ROLE IN THE REGULATION OF P53 SIGNALING. PHARMACOLOGIC DOSES OF NA, A VITAMIN AND ALSO AN FDA-APPROVED DRUG FOR DYSLIPIDEMIA, IS TAKEN BY MILLIONS OF AMERICANS. THOUGH THE ROLE OF NA IN DECREASING LDL AND INCREASING HDL IS WELL RECOGNIZED, LITTLE IS KNOWN ABOUT ITS ANTI-TUMOR POTENTIAL. SIMILARLY, THE LINK BETWEEN COMMENSAL BACTERIA AND CC PREVENTION IS KNOWN FOR A LONG TIME, BUT THE MECHANISMS UNDERLYING THIS LINK ARE NOT WELL UNDERSTOOD. BTR, THE PROMINENT BACTERIAL METABOLITE IN THE COLON, IS THE ENDOGENOUS AGONIST FOR GPR109A IN COLONIC EPITHELIA. BHB IS THE NATURAL AGONIST FOR GPR109A IN IMMUNE CELLS PRESENT IN THE LAMINA PROPRIA AND THIS AGONIST IS SYNTHESIZED IN COLONIC EPITHELIA USING THE BACTERIAL METABOLITE BTR VIA KETOGENESIS. THIS HIGHLIGHTS THE BIOLOGICAL SIGNIFICANCE OF THE COLON AS A KETOGENIC ORGAN. IN ADDITION, COMMENSAL BACTERIA CONTROL GPR109A EXPRESSION AND KETOGENESIS IN THE COLON VIA THEIR XENOBIOTIC METABOLITES (BUTYRATE, INDOLE DERIVATIVES, 1,3-DIAMINOPROPANE, AND REUTERIN). THIS OFFERS A NOVEL MOLECULAR LINK BETWEEN COMMENSAL BACTERIA AND CC PREVENTION. OUR CENTRAL HYPOTHESIS IS THAT ACTIVATION OF GPR109A IN COLONIC EPITHELIA AND IMMUNE CELLS LEAD TO SUSTAINED ACTIVATION OF P53 AND ANTI- TUMOR IMMUNITY. WE WILL TEST THIS HYPOTHESIS WITH THREE SPECIFIC AIMS: (AIM 1) DETERMINE THE ROLE OF THE GPR109A- P53 AXIS IN KRASD, CHEMICAL, AND DIETARY CARCINOGEN-INDUCED CC; (AIM 2) ELUCIDATE THE MOLECULAR MECHANISM(S) OF GPR109A AGONISTS-INDUCED P53 ACTIVATION AND TEST IF GPR109A AGONISTS CAN BE USED AS NOVEL THERAPEUTIC DRUGS TO TARGET P53D CC; AND (AIM 3) DEMONSTRATE THAT COMMENSAL BACTERIAL METABOLITES MAINTAIN WTP53 FUNCTION AND PREVENT THE IMMUNE SYSTEM FROM UNDUE ACTIVATION IN COLON VIA GPR109A WITH AN OBLIGATORY INVOLVEMENT OF COLONIC KETOGENESIS. THESE STUDIES WILL INVOLVE THE USE OF GENETICALLY MODIFIED MOUSE LINES WITH COLON-SPECIFIC DELETION OF NOT ONLY GPR109A AND P53, BUT ALSO THE RATE-LIMITING ENZYME IN KETOGENESIS, HMGCS2.

LOW-LEVEL LASER THERAPY IN GLOBAL CEREBRAL ISCHEMIA

DIETARY COMBINATION FOR PREVENTION OF METASTATIC RENAL CELL CARCINOMA