The Cleveland Clinic Foundation

ASTHMA INFLAMMATION RESEARCH (AIR)

IFNS AND CYTOKINES: SIGNALING AND ACTION

COBRE GRANT

PULMONARY VASCULAR DISEASE PHENOMICS PROGRAM (PVDOMICS) DATA COORDINATING CENTER

ACUTE HUMORAL REJECTION OF RENAL ALLOGRAFTS

ALCOHOL AND TISSUE INJURY FROM MECHANISMS TO TREATMENTS

CLEVELAND ALZHEIMERS DISEASE RESEARCH CENTER - PROJECT SUMMARY THE CLEVELAND ALZHEIMER'S DISEASE RESEARCH CENTER (CADRC) IS A COLLABORATIVE EFFORT OF PHYSICIANS AND INVESTIGATORS FROM CASE WESTERN RESERVE UNIVERSITY (CWRU), CLEVELAND CLINIC FOUNDATION (CCF), METROHEALTH SYSTEM (MHS), UNIVERSITY HOSPITALS (UH), AND THE LOUIS STOKES CLEVELAND VA MEDICAL CENTER (LSCVAMC), TO FOSTER EXCELLENCE IN RESEARCH AND FACILITATE DISCOVERY AS AN ESTABLISHED NATIONAL INSTITUTE ON AGING (NIA) FUNDED ALZHEIMER'S DISEASE RESEARCH CENTER. THE CADRC REPRESENTS A RICH CLINICAL AND RESEARCH COMMUNITY AND AN ESTIMATED 220,000 OHIOANS WHO SUFFER FROM ALZHEIMER'S DISEASE (AD) OR AD-RELATED DEMENTIAS (ADRD). THE CADRC HAS 8 CORES AND A RESEARCH EDUCATION COMPONENT DESIGNED TO CREATE THE FOUNDATION THAT WILL ENHANCE THE RESEARCH EFFORTS OF THE NORTHEAST OHIO AD/ADRD RESEARCH COMMUNITY, AS WELL AS ADD UNIQUE VALUE TO THE NATIONAL ALZHEIMER'S DISEASE RESEARCH CENTERS (ADRC) PROGRAM AND OTHER NATIONAL AND INTERNATIONAL RESEARCH PROGRAMS. THE CADRC IS FOCUSED ON PARTICIPANTS THAT WILL HELP US UNDERSTAND THE PATHOBIOLOGY OF CLINICAL AND PATHOLOGICAL HETEROGENEITY OBSERVED IN DEMENTIA INCLUDING ATYPICAL AND AMNESTIC AD, DEMENTIA WITH LEWY BODIES, COGNITIVELY NORMAL INDIVIDUALS WITH DIFFERENT LEVELS OF GENETIC RISK FOR AD, AND DIVERSITY OF PARTICIPANT POPULATIONS (CLINICAL AND OUTREACH, RECRUITMENT, AND ENGAGEMENT CORES). TO SUPPORT THE CADRC GOALS, THE FOCUS WILL BE ON DEEP PHENOTYPING OF PARTICIPANTS WITH LONGITUDINAL AND SYSTEMATIC COGNITIVE, BEHAVIORAL AND MOTOR CHARACTERIZATION (CLINICAL CORE), GENETIC AND BIOFLUID BIOMARKER COLLECTION AND ANALYSIS (BIOMARKER CORE), IMAGING (NEUROIMAGING CORE), AND AUTOPSY AFTER DEATH (NEUROPATHOLOGY CORE). RESULTS WILL BE SHARED WITH THE RESEARCH COMMUNITY IN A TIMELY AND REGULAR MANNER TO ALLOW OTHER INVESTIGATORS TO BENEFIT FROM THE CADRC EFFORTS (DATA MANAGEMENT AND STATISTICS CORE). IN ADDITION, THE CADRC WILL TRAIN THE NEXT GENERATION OF INVESTIGATORS UTILIZING A RIGOROUS AND WELL-DESIGNED RESEARCH EDUCATION COMPONENT, SUPPORT TRANSLATION OF NEW LABORATORY FINDINGS THROUGH THE TRANSLATIONAL THERAPEUTICS CORE, AND SUPPORT HIGH RISK/HIGH GAIN PROJECTS THROUGH THE DEVELOPMENTAL PROGRAM AS A PART OF THE ADMINISTRATIVE CORE. THE ULTIMATE GOALS ARE TO ADVANCE THE PRECISION MEDICINE APPROACH TO DEMENTIA DIAGNOSIS AND TREATMENT, SUPPORT THE DEVELOPMENT OF EARLY STAGE INVESTIGATORS, ASSIST ALL STAGES OF INVESTIGATORS WITH TOOLS FOR PERFORMING HUMAN-BASED RESEARCH, AND BETTER ENGAGE UNDERREPRESENTED POPULATIONS IN AD/ADRD RESEARCH.

GUT MICROBIOTA AND CARDIOMETABOLIC DISEASES

THE CLEVELAND CLINIC INNOVATION ACCELERATOR

PATHOBIOLOGY OF ASTHMA

CELL ADHESION AND SIGNALING IN BLOOD AND VASCULAR CELLS - PROJECT SUMMARY THIS APPLICATION HAS AS ITS THEME THE INTEGRINS, THEIR REGULATION AND THEIR CONTRIBUTION TO THE FUNCTIONAL RESPONSES OF BLOOD AND VASCULAR CELLS. THE INTEGRINS OF FOCUS ARE AMSS2 (MAC-1), AIIBSS3, AVSS3, AND A5SS1 BUT FINDINGS SHOULD APPLY TO BROADLY INTEGRINS. THE CELLS OF EMPHASIS ARE VASCULAR CELLS- ENDOTHELIAL CELLS, SMOOTH MUSCLE CELLS AND PERICYTES- AND BLOOD CELLS- LEUKOCYTES AND PLATELETS. ON THE BLOOD CELLS, THE CONJUGATION OF MAC-1 ON LEUKOCYTES AND GPIB ON PLATELETS WILL BE CONSIDERED. MAJOR EMPHASIS WILL BE PLACED ON THE MOLECULES THAT REGULATE INTEGRIN FUNCTION- KINDLINS, TALIN AND PAXILLIN- TO DETERMINE HOW THEY COLLABORATE TO REGULATE INTEGRIN ACTIVATION. THE FUNCTION OF THESE CYTOSKELETAL PROTEINS INDEPENDENT OF INTEGRIN ACTIVATING ACTIVITY WILL ALSO BE DISSECTED. THE PROGRAM CONSISTS OF THREE PROJECTS, EACH DIRECTED BY AN ACCOMPLISHED FACULTY MEMBER AT THEIR HOME INSTITUTIONS, CLEVELAND CLINIC, UNIVERSITY HOSPITALS OF CLEVELAND AND CASE WESTERN RESERVE UNIVERSITY WHICH ARE ALL CLOSELY LOCATED AND GOVERNED BY INTERINSTITUTIONAL AGREEMENTS. DR. EDWARD F. PLOW, PH.D. WILL SERVE AS PROGRAM DIRECTOR AND LEAD PROJECT 1. THIS PROJECT DEALS WITH THE MECHANISMS BY WHICH KINDLIN-2 REGULATES BOTH INTEGRIN-DEPENDENT AND INDEPENDENT RESPONSES OF BLOOD VESSEL CELLS. MOLECULAR, CELLULAR AND UNIQUE MOUSE MODELS ARE ALL BROUGHT TO BEAR TO DETERMINE HOW KINDLIN-2 SERVES AS A MASTER REGULATOR OF VASCULAR CELL RESPONSES. IN PROJECT 2, DR. JUN QIN WILL USE HIGH RESOLUTION STRUCTURAL APPROACHES IN COMBINATION WITH MUTAGENESIS AND CELLULAR STUDIES TO DETERMINE HOW TALIN REGULATE INTEGRIN ACTIVATION AND COOPERATES WITH KINDLINS AND PAXILLIN TO GAIN SUCH NOVEL INSIGHTS. HE WILL DETERMINE HOW TALIN INTERACTS WITH ACTIN TO CONTROL ORGANIZATION OF THE CYTOSKELETON. DR. DANIEL SIMON, M.D. WILL LEAD PROJECT 3 AND WILL CONSIDER HOW ENGAGEMENT OF INTEGRIN MAC-1 ON LEUKOCYTES AND GPIB ON PLATELETS REGULATES THE PARTICIPATION OF THESE CELLS IN INFLAMMATION AND THROMBOSIS. HIS STUDIES RANGE FROM BASIC STRUCTURAL APPROACHES TO TRANSLATIONAL STUDIES IN MICE AND TO HUMANS TO PROVIDE INSIGHTS INTO THEIR THROMBOTIC AND INFLAMMATORY CONTRIBUTIONS TO SYSTEMIC LUPUS ERYTHEMATOSUS. THE PROGRAM IS SUPPORTED BY TWO SCIENTIFIC CORES, PROTEIN EXPRESSION AND PURIFICATION (CORE B), AND ANIMAL MODELS AND TISSUE ANALYSIS (CORE C) AS WELL AS BY AN ADMINISTRATIVE CORE (AC1). A COMMON OBJECTIVE OF THE PROGRAM IS TO CONTINUE AND CREATE NEW COLLABORATIONS AMONG THE PROJECTS AND THEIR LEADERS TO RESOLVE THE STRUCTURAL AND BIOLOGICAL MECHANISMS THAT REGULATE THE FUNCTIONS OF INTEGRINS IN BLOOD AND VASCULAR CELLS. THE INFORMATION DERIVED FROM THESE STUDIES WILL PROVIDE INSIGHTS INTO BIOLOGICALLY IMPORTANT RESPONSES REGULATED BY INTEGRINS AND THEIR ACTIVATION THAT ARE RELEVANT TO THROMBOSIS AND CARDIOVASCULAR DISEASES.

FRMI OF THE PERSON IDENTITY NETWORK: AGING AND APOE

ATRIAL FIBRILLATION POST-GWAS: MECHANISMS TO TREATMENT - OVERALL COMPONENT PROJECT/SUMMARY ABSTRACT ATRIAL FIBRILLATION (AF), THE MOST COMMON CARDIAC ARRHYTHMIA, AFFLICTS THE U.S. AND WORLD WITH INCREASING PREVALENCE. AF INCIDENCE, PROGRESSION TO PERSISTENT AF, AND AF COMPLICATIONS, INCLUDING STROKE, ARE FED BY INCREASING OBESITY AND AGE. CURRENT THERAPIES ARE LIMITED BY RISKS AND LIMITED EFFICACY, WORSE AS AF PROGRESSES, BUT NO NEW PHARMACOLOGIC AGENTS HAVE BEEN APPROVED FOR AF IN >10 YEARS. WITH IDENTIFICATION OF >100 GENETIC LOCI THAT PREDISPOSE TO AF RISK IN GENOME-WIDE ASSOCIATION STUDIES (GWAS), THE HOPE HAS BEEN THAT GENETICS WOULD YIELD NOVEL THERAPEUTIC TARGETS. HOWEVER, EVEN FOR THE TOP LOCUS ON CHR. 4Q25 NEAR PITX2, A GENE INVOLVED IN FORMATION OF PULMONARY VEINS, THE TARGET OF AF ABLATION, MECHANISMS LINKING VARIANTS TO AF REMAIN ELUSIVE. GENETIC FINDINGS HAVE SO FAR FAILED TO YIELD CLINICALLY ACTIONABLE RESULTS. TO FILL THESE GAPS, WE SEEK TO GO BEYOND GWAS FINDINGS TO IDENTIFY DIRECT GENOMIC MECHANISMS UNDERLYING AF AND BETTER UNDERSTAND THEIR INTERACTIONS WITH ENVIRONMENT, COMORBIDITIES OR CELL STRESSORS. OUR LONG-TERM GOAL IS TO USE GENOMIC FINDINGS TO PERSONALIZE PREVENTIVE AND THERAPEUTIC STRATEGIES FOR AF. OUR OVERALL P01 THEME IS TO TRANSLATE AF GENETIC DISCOVERIES TOWARDS THE BEDSIDE, FOCUSING ON GENES TO MECHANISMS, GENES TO DRUGS, AND INTERACTIONS OF GENES WITH METABOLISM AND ENVIRONMENT. WE BUILD ON STRONG PRELIMINARY DATA AND COALESCE UNIQUE HUMAN ATRIAL TISSUE BIOREPOSITORY AND GENOMIC DATA RESOURCES, NOVEL CELL AND ANIMAL MODELS, AND COMPLEMENTARY EXPERTISE FROM OUR MULTIDISCIPLINARY TEAM WITH A STRONG COLLABORATION HISTORY. OUR CENTRAL HYPOTHESIS IS THAT GENOMIC MECHANISTIC DISCOVERIES IN AF CELLULAR AND ANIMAL MODELS WILL TRANSLATE TO HUMAN THERAPIES. OUR THEMATIC AIMS INCLUDE: 1) IDENTIFY CAUSAL GENES AND FUNCTIONAL MECHANISMS WITH A GOAL TOWARDS IDENTIFICATION OF NEW THERAPEUTIC APPROACHES FOR AF; 2) INVESTIGATE METABOLIC AND INFLAMMATORY MECHANISMS, IMPLICATED BY GENOMICS STUDIES TO BE IMPORTANT IN AF PATHOPHYSIOLOGY, TO IDENTIFY NEW THERAPEUTIC TARGETS FOR AF PREVENTION AND TREATMENT; AND 3) IDENTIFY CANDIDATE NOVEL DRUGS FOR AF AND DEVELOP A PIPELINE FOR IN VITRO AND IN VIVO FUNCTIONAL TESTING OF CANDIDATE THERAPIES. PROJECT 1 GENES TO FUNCTION WILL DETERMINE CAUSAL GENES, VARIANTS AND MECHANISMS UNDERLYING TWO AF GWAS LOCI. PROJECT 2 GENES AND METABOLISM WILL STUDY THE CONTRIBUTION OF MITOCHONDRIAL DYSFUNCTION TO AF ONSET AND PROGRESSION. EARLY STAGE INVESTIGATOR PROJECT GENES AND NUTRITION BUILDS ON NOVEL ASSOCIATIONS OF AF WITH TRIMETHYLAMINE N-OXIDE (TMAO), PRODUCED BY GUT MICROBIOTA FROM PRECURSORS SUCH AS CHOLINE FOUND IN EGGS, MEATS AND CHEESES. PROJECT 4 GENES TO OMICS-INFORMED DRUGS WILL IDENTIFY MECHANISMS AND REPURPOSABLE DRUGS TO PREVENT AF PROGRESSION. PROJECTS ARE SUPPORTED BY 4 CORES PROVIDING ADMINISTRATION, ENGINEERED HEART TISSUE AND ATRIAL PHENOTYPING, ELECTROPHYSIOLOGY, AND NETWORK AND SYSTEMS BIOLOGY ANALYTICS SUPPORT THAT SYNERGIZE DISCOVERY AND TRANSLATION IN AF AND INCREASE THE SCOPE AND IMPACT OF EACH PROJECT. ALL P01 COMPONENTS AIM TO BRIDGE BASIC RESEARCH IN AF TOWARDS CLINICAL UTILITY, THEREBY ADVANCING GENOMIC DATA AND RESEARCH TOWARDS THE BEDSIDE TO HELP OUR PATIENTS SUFFERING FROM ATRIAL FIBRILLATION.

SELF REGULATING CONTINUOUS FLOW TOTAL ARTIFICIAL HEART

GUT FLORA METABOLISM OF DIETARY PHOSPHATIDYLCHOLINE AND CARDIOVASCULAR DISEASE

SEX-BASED DIFFERENCES IN GLIOMA

NASH PROJECT

ALLIANCE OF RANDOMIZED TRIALS OF MEDICINE VS METABOLIC SURGERY IN TYPE 2 DIABETES - (ARMMS-T2D)

MOLECULAR DISSECTION OF CYTOKINE CROSSTALK IN THE TUMOR MICROENVIRONMENT - PROJECT SUMMARY DESPITE RECOGNITION OF THE BROAD CONSEQUENCES OF INFLAMMATION IN CANCER BIOLOGY, THE MECHANISTIC IMPACT ON THE TUMOR LANDSCAPE REMAINS INCOMPLETELY UNDERSTOOD. INDEED, INNATE AND ADAPTIVE IMMUNE FUNCTIONS IN CANCER CAN BE BENEFICIAL OR DETRIMENTAL AND THE OPPOSING ROLES HIGHLIGHT THE GAP OF KNOWLEDGE IN OUR UNDERSTANDING OF HOW INFLAMMATION SCULPTS THE TUMOR MICROENVIRONMENT (TME). THIS PROGRAM PROJECT WILL ADDRESS THIS GAP OF KNOWLEDGE BY DEFINING AND DELINEATING HOW CYTOKINES MODULATE THE FUNCTIONS OF THE MULTIPLE CELL TYPES COMPOSING THE TUMOR MICROENVIRONMENT. OUR PREVIOUS WORK HAS REVEALED THE POTENTIAL FOR THESE INFLAMMATORY CYTOKINES TO REGULATE A SPECTRUM OF CANCER CELL PHENOTYPES, INCLUDING THEIR SELF-RENEWAL AND CELLULAR HIERARCHY OR STEMNESS, THAT ARE ASSOCIATED WITH THE EPITHELIAL-MESENCHYMAL TRANSITION (EMT). MOREOVER, THESE PHENOTYPES ARE COMMONLY ASSOCIATED WITH CANCER PROGRESSION THROUGH MODULATION OF DIFFERENTIATION POTENTIAL, CELL-CELL INTERACTIONS AND MOBILITY, FIBROSIS, AND SENSITIVITY TO MULTIPLE THERAPEUTIC MODALITIES. THE PROGRAM IS NOW CENTERED ON TWO MAJOR THEMES. THE FIRST IS TO DEFINE THE SIGNALING MECHANISMS THAT GOVERN HOW CYTOKINES (TYPE I IFNS, IL- 17,TGFSS) MODULATE (BOTH POSITIVELY AND NEGATIVELY) THE EMT PROCESS. THE CELLULAR TARGETS INCLUDE STEM-LIKE TUMOR CELLS AS WELL AS THE NON-TUMOR DERIVED POPULATIONS, INCLUDING FIBROBLASTS AND IMMUNE CELLS. THE SECOND THEME RELATES THESE CELL-SPECIFIC EMT RESPONSES TO SPECIFIC EFFECTS ON METASTASIS, FIBROSIS, AND RESISTANCE TO MULTIPLE THERAPEUTIC STRATEGIES. OUR MAJOR GOAL IS TO PARLAY OUR IMPROVED UNDERSTANDING OF CYTOKINE EFFECTS IN THE TME INTO SPECIFIC IMPROVEMENTS IN CANCER THERAPY. COLLECTIVELY THE THREE PROJECTS IN THE APPLICATION WILL TEST THE FOLLOWING OVERARCHING HYPOTHESIS: CYTOKINE SIGNALS HAVE DISTINCT AND SOMETIMES CONFLICTING MECHANISTIC ROLES IN CANCER PROGRESSION THOUGH ALTERATIONS IN EMT AND CANCER STEM CELL DEVELOPMENT. SUCH MECHANISMS LEAD TO CRITICAL PHENOTYPIC PROPERTIES RESPONSIBLE FOR CONTINUOUS METASTATIC SPREAD AND RESISTANCE TO MULTIPLE THERAPEUTIC MODALITIES (CHEMOTHERAPY, IMMUNE THERAPY). THIS HYPOTHESIS WILL BE TESTED BY (1) DEFINING THE SIGNALING EVENTS INITIATED BY TGFSS, IL-17, AND/OR TYPE I IFNS AND THE ENDPOINT CHANGES IN SPECIFIC GENE EXPRESSION THAT ARE CAUSALLY LINKED WITH CONTROL OF TME AND CANCER CELL PHENOTYPES, (2) DETERMINATION OF HOW THESE SPECIFIC SIGNALING PATHWAYS AND GENE EXPRESSION EVENTS ARE MECHANISTICALLY RESPONSIBLE FOR ACQUISITION OF THERAPEUTIC RESISTANCE AND (3) EVALUATION OF THE DISTINCT CELL TYPE CONTRIBUTIONS TO TUMOR PHENOTYPES AND THERAPEUTIC RESISTANCE, WITH EMPHASIS ON TUMOR CELL INTRINSIC MECHANISMS, IMMUNE CELL INFILTRATES AND ACTIVITIES, AND STROMAL CELL CONTROL OF TUMOR ACCESS.

PROGNOSIS AND PREDICTORS OF ACL RECONSTRUCTION: A MULTICENTER COHORT STUDY

CYCLE-AD: RANDOMIZED CONTROLLED TRIAL TO ASSESS THE EFFICACY OF INDOOR CYCLING IN SLOWING DISEASE PROGRESSION IN HEALTHY OLDER PERSONS AT GENETIC RISK FOR ALZHEIMER?S DISEASE - PROJECT SUMMARY/ABSTRACT THE APOLIPOPROTEIN E EPSILON 4 (APOE 4) ALLELE IS THE MOST IMPORTANT GENETIC RISK FACTOR FOR LATE ONSET ALZHEIMER'S DISEASE (AD). A RECENT REVIEW BY THE WORLD HEALTH ORGANIZATION HIGHLIGHTED THE POTENTIAL PROTECTIVE ROLE OF PHYSICAL ACTIVITY AND EXERCISE AGAINST COGNITIVE DECLINE, ALL-CAUSE DEMENTIA, AD, AND VASCULAR DEMENTIA IN HEALTHY INDIVIDUALS. IN AN 18-MONTH LONGITUDINAL OBSERVATIONAL STUDY, WE SHOWED THAT SEDENTARY 4 CARRIERS EXPERIENCE SIGNIFICANT DECLINES IN EPISODIC MEMORY AND HIPPOCAMPAL VOLUME COMPARED TO 4 CARRIERS WHO ENGAGED IN MODERATE PA. IMPORTANTLY, AMONG 4 NON-CARRIERS, NO SIGNIFICANT LONGITUDINAL CHANGES IN COGNITION AND BRAIN IMAGING WERE OBSERVED WHETHER THE NON-CARRIERS WERE SEDENTARY OR ENGAGED IN MODERATE PA, SUGGESTING THAT PA HAS A SPECIFIC NEUROPROTECTIVE ROLE IN DELAYING THE PROGRESSION OF AD IN 4 CARRIERS. BASED ON OUR RESULTS, A PRAGMATIC, RANDOMIZED CONTROLLED TRIAL WITH BLINDED CLINICAL AND IMAGING OUTCOMES IS PROPOSED TO DETERMINE THE IMPACT OF A HOME BASED, HIGH INTENSITY EXERCISE INTERVENTION IN HEALTHY, COGNITIVELY INTACT 4 CARRIERS BETWEEN THE AGES OF 65 AND 80 YEARS. THE CYCLE-AD (CYCLING TO CEASE OR LIMIT THE EFFECTS OF ALZHEIMER'S DISEASE) TRIAL WILL RECRUIT OTHERWISE HEALTHY SEDENTARY CARRIERS RANDOMIZED TO ONE OF TWO GROUPS (N=75 EACH): 1) AN INDOOR CYCLING (IC) GROUP THAT PARTICIPATES IN HIGH-INTENSITY INTERVAL TRAINING (HIIT; 60-90% OF HEART RATE RESERVE) IN THEIR HOME VIA THE COMMERCIALLY AVAILABLE PELOTON® CYCLING SYSTEM OR 2) A USUAL AND CUSTOMARY CARE (UCC) GROUP, IN WHICH PARTICIPANTS ENGAGE IN THEIR HABITUAL LEVEL OF PA. WE HYPOTHESIZE THAT AN 18-MONTH HIGH-INTENSITY AEROBIC EXERCISE REGIMEN WILL SLOW AD-RELATED DISEASE PROGRESSION IN SEDENTARY ELDERS AT GENETIC RISK FOR AD. PARTICIPANTS IN THE INTERVENTION GROUP WILL ENGAGE IN EXERCISE 3X/WEEK (MINIMUM 90 MINUTES/WEEK) FOR 18 MONTHS. PRIMARY OUTCOME MEASURES, OBTAINED AT STUDY ENTRY AND AT 18 MONTHS, WILL INCLUDE COMPREHENSIVE COGNITIVE TESTING AND BRAIN MR IMAGING TO ASSESS DISEASE PROGRESSION AND A COMPREHENSIVE PA/FITNESS ASSESSMENT TO MEASURE THE DEGREE OF CHANGE IN PHYSICAL FITNESS DUE TO HIGH INTENSITY AEROBIC EXERCISE. THE OVERALL GOAL OF THE CYCLE-AD TRIAL IS TO DETERMINE THE ROLE OF LONG-TERM, HIGH INTENSITY EXERCISE IN SLOWING OR DELAYING THE ONSET OF COGNITIVE AND AD-RELATED BRAIN CHANGES IN 4 CARRIERS. SUCCESSFUL TRANSLATION AND DEMONSTRATION OF THE EFFECTIVENESS OF A SCALABLE HOME-BASED EXERCISE INTERVENTION CAPABLE OF SLOWING OR DELAYING DISEASE ONSET WILL TRANSFORM AD TREATMENT, IMPROVE PATIENT OUTCOMES AND QUALITY OF LIFE, AND REDUCE HEALTH CARE COSTS.

POST-TRANSPLANT INFLAMMATORY RESPONSE

CENTRAL VEIN SIGN: A DIAGNOSTIC BIOMARKER IN MULTIPLE SCLEROSIS

MOLECULAR BASIS OF ILK SIGNALING IN CELL ADHESION

TOWARDS PRECISION IMMUNO-ONCOLOGY: UNRAVELING THE GENOMIC DETERMINANTS AND MECHANISMS UNDERLYING IMMUNOTHERAPY EFFICACY AND RESISTANCE

NEI CENTER CORE GRANT FOR VISION RESEARCH

PATHOGENESIS OF NEUROLOGICAL DISABILITY IN PRIMARY DISEASES OF MYELIN

CLEVELAND CLINIC CARDIOTHORACIC COLLABORATIVE CLINICAL CENTER (C6)

GENOMIC AND MICROENVIRONMENTAL DETERMINANTS, TEMPORAL DYNAMICS, AND TREATMENT EFFICACY OF RADIATION-BASED COMBINATION THERAPIES - PROJECT SUMMARY OVERALL SECTION OUR ROBIN CENTER FOCUSES ON ELUCIDATING THE GENOMIC AND MICROENVIRONMENTAL DETERMINANTS, AND TEMPORAL DYNAMICS UNDERLYING EFFICACY OF RADIATION-BASED COMBINATION THERAPIES. RADIOTHERAPY (RT), ALONE OR IN COMBINATION WITH OTHER TREATMENTS, IS USED TO TREAT ABOUT TWO-THIRDS OF ALL CANCER PATIENTS. DESPITE THE WIDESPREAD USE OF RADIATION THERAPY IN ONCOLOGY, OUR UNDERSTANDING OF THE MECHANISMS DRIVING RESPONSE AND RESISTANCE REMAINS POOR. OUR LONG-TERM GOAL IS TO UNDERSTAND THE MECHANISMS THAT UNDERLIE EFFICACY AND RESISTANCE OF RADIATION-BASED THERAPIES. NEW EFFORTS TO IMPROVE TREATMENT FOR MANY CANCER TYPES NOW FOCUS ON USING COMBINATION THERAPIES IN WHICH RADIATION IS USED WITH SYSTEMIC AGENTS, HIGHLIGHTING THE URGENT NEED TO UNDERSTAND THE DRIVERS OF EFFICACY. AMONG THE MOST PROMISING NEW BIOLOGICS BEING STUDIED FOR USE WITH RADIATION ARE ANTIBODY-DRUG CONJUGATES (ADC) AND IMMUNE CHECKPOINT INHIBITORS (ICI). WE WILL USE AN INNOVATIVE MOLECULAR CHARACTERIZATION TRIAL TESTING RADIATION PLUS ADC IN BLADDER CANCER AND RADIATION PLUS ICI IN HEAD AND NECK CANCER TO CHARACTERIZE THE MECHANISTIC DRIVERS UNDERLYING THESE NEXT GENERATION RT-BASED COMBINATIONS. THE CENTRAL HYPOTHESIS OF THIS U54 APPLICATION IS THAT SPECIFIC GENETIC AND IMMUNOLOGIC MECHANISMS UNDERLIE SENSITIVITY AND RESISTANCE TO RADIATION-BASED COMBINATION THERAPIES. WE WILL ADDRESS THESE QUESTIONS THROUGH 3 SPECIFIC AIMS. IN AIM 1, WE WILL WORK TO UNDERSTAND THE MOLECULAR MECHANISMS THAT UNDERLIE EFFICACY OF TREATMENT WITH RADIATION PLUS ADC. HERE, OUR WORKING HYPOTHESIS IS THAT SPECIFIC GENETIC AND IMMUNOLOGIC EVENTS UNDERLIE RESPONSE TO RT PLUS SACITUZUMAB GOVITECAN (SG) TREATMENT. WE WILL LEVERAGE OUR MOLECULAR CHARACTERIZATION TRIAL (PART A) INVESTIGATING THE USE OF RT AND SACITUZUMAB FOR BLADDER PRESERVATION THERAPY. WE WILL DETERMINE THE DIFFERENTIAL MOLECULAR EFFECTS WITH STANDARD-OF-CARE RT + CISPLATIN VERSUS RT + SG. IN AIM 2, WE WILL IMPROVE IDENTIFICATION OF PATIENTS WHO ARE SENSITIVE OR RESISTANT TO RT-BASED THERAPIES BASED ON NEW INSIGHTS INTO TRANSCRIPTIONAL DYNAMICS AND TEMPORAL REPROGRAMMING DURING TREATMENT WITH RADIATION-BASED THERAPIES. HERE, WE WILL LEVERAGE OUR MOLECULAR CHARACTERIZATION TRIAL TREATING HEAD AND NECK SQUAMOUS CELL CARCINOMA (HNSCC) OR BLADDER CANCER PATIENTS WITH RT + CHEMOTHERAPY VERSUS RT + SG OR ICI. WE WILL BUILD ON RECENT EXPERIMENTAL AND CLINICAL BREAKTHROUGHS LED BY OUR RESEARCH GROUPS, WHICH HAVE IDENTIFIED HIGHLY REFINED GENE EXPRESSION PROGRAMS ASSOCIATED WITH RT SENSITIVITY AND DELTA RADIOMICS. IN AIM 3, WE WILL IDENTIFY THE DIFFERENTIAL MECHANISMS UNDERLYING THE ANTI-TUMOR ACTIVITIES OF RT + CISPLATIN VERSUS RT + IMMUNE CHECKPOINT BLOCKADE. HERE, USING OUR HEAD AND NECK TRIAL (PART B), WE WILL UNCOVER THE UNIQUE GENETIC AND IMMUNOLOGIC FACTORS THAT GOVERN RESPONSE TO RT WHEN COMBINED WITH THESE TWO CLASSES OF AGENTS. WE WILL ELUCIDATE THE DIFFERENTIAL MOLECULAR EFFECTS OF THE TWO APPROACHES, IMMUNE REPROGRAMMING, AND MECHANISMS OF ACQUIRED RESISTANCE. OUR STUDIES WILL HELP BUILD A FOUNDATION TO OPTIMIZE MULTIMODAL, RADIATION- BASED DEFINITIVE TREATMENT STRATEGIES.

MOLECULAR BASIS OF KSHV ONCOGENESIS

FUNCTIONAL GENOMICS OF ATRIAL FIBRILLATION IN HUMAN ATRIA

BEAR-MOON: A TWO ARM NONINFERIORITY BLINDED RANDOMIZED CLINICAL TRIAL COMPARING ACL REPAIR WITH BEAR DEVICE VS. STANDARD OF CARE AUTOGRAFT PATELLAR TENDON ACL RECONSTRUCTION

RANDOMIZED TRIAL OF ASSISTED AMBULATION TO IMPROVE HEALTH OUTCOMES FOR OLDER MEDICAL INPATIENTS - PROJECT SUMMARY/ABSTRACT FOR OLDER ADULTS, PROLONGED HOSPITALIZATION CAN LEAD TO A DEVASTATING LOSS OF MOBILITY AND INDEPENDENCE. EACH YEAR, 12 MILLION ADULTS OVER THE AGE OF 65 ARE HOSPITALIZED, AND 30% ARE DISCHARGED TO A POST-ACUTE CARE FACILITY. ONE OF THE RISKS OF HOSPITALIZATION IS BED REST, WHICH IS ASSOCIATED WITH A NUMBER OF HOSPITAL-ACQUIRED COMPLICATIONS, INCLUDING FALLS, DELIRIUM, VENOUS THROMBOSIS AND SKIN BREAKDOWN. HOSPITAL MOBILITY PROGRAMS ATTEMPT TO AMBULATE PATIENTS UP TO THREE TIMES DAILY, BUT THIS WORK IS GENERALLY ASSIGNED TO NURSES, WHO HAVE MANY COMPETING AND OFTEN MORE PRESSING TASKS. CONSEQUENTLY, AMBULATING PATIENTS IS THE MOST FREQUENTLY OVERLOOKED NURSING DUTY. THIS PROBLEM HAS BEEN EXACERBATED BY THE COVID-19 PANDEMIC AND THE RESULTING NURSING SHORTAGE. SMALL STUDIES HAVE EXAMINED THE BENEFITS OF MOBILITY TECHNICIANS (MTS), WHOSE SOLE JOB IS TO SAFELY AMBULATE PATIENTS. THESE STUDIES HAVE DEMONSTRATED THAT MTS CAN INCREASE STEPS TAKEN, BUT THEY ARE TOO SMALL TO PROVE THE IMPACT OF MTS ON OTHER OUTCOMES, SUCH AS WHETHER PATIENTS HAVE IN-HOSPITAL COMPLICATIONS OR WHETHER THEY CAN GO HOME INSTEAD OF TO A POST-ACUTE CARE FACILITY. HOSPITALS ARE HESITANT TO ADOPT MT PROGRAMS BECAUSE THEY PERCEIVE THEM TO BE EXPENSIVE AND UNPROVEN. WE PROPOSE TO CONDUCT A LARGE RANDOMIZED TRIAL TO TEST THE IMPACT OF MTS ON SHORT AND INTERMEDIATE TERM OUTCOMES FOR 3000 PATIENTS AGED 65 YEARS AND OLDER AT 5 HOSPITALS IN 2 HEALTH SYSTEMS. PATIENTS WILL BE RANDOMIZED TO RECEIVE SUPERVISED AMBULATION UP TO 3 TIMES DAILY WITH A MT OR TO RECEIVE USUAL CARE. ALL PARTICIPANTS WILL WEAR AN ACCELEROMETER ON THEIR WRIST TO TRACK THEIR MOVEMENT THROUGHOUT THE HOSPITAL STAY. THE STUDY HAS 3 AIMS. FIRST, WE WILL COMPARE THE MOBILITY OF PATIENTS AT DISCHARGE (OR 10 DAYS) TO ASSESS THE IMPACT OF THE MTS ON THIS OUTCOME. WE ARE PARTICULARLY INTERESTED IN WHETHER THE USE OF MTS WILL INCREASE THE PROPORTION OF PATIENTS WHO CAN GO HOME VS. POST-ACUTE CARE, AND WHETHER THE IMPROVEMENTS IN MOBILITY ARE SUSTAINED AT 30 DAYS. SECOND, WE WILL USE PREDICTIVE MODELING TO IDENTIFY WHICH PATIENTS ARE MOST LIKELY TO BENEFIT FROM THIS INTERVENTION. THIRD, WE WILL ASSESS THE IMPACT OF THE INTERVENTION ON OVERALL COSTS ASSOCIATED WITH THE EPISODE OF CARE, INCLUDING INPATIENT COSTS AND THE 30 DAYS AFTER DISCHARGE. THIS INFORMATION WILL BE IMPORTANT TO CONVINCE HEALTH SYSTEMS TO ADOPT THIS APPROACH.

ENSURING PATIENTS' INFORMED ACCESS TO NONINVASIVE PRENATAL TESTING

ACTIVATION OF BLOOD AND ENDOTHELIAL CELL A5B3 INTEGRIN

DEFINING THE VIRAL PTMOME: TOWARDS THE DEVELOPMENT OF NOVEL ANTIVIRAL APPROACHES - PROJECT SUMMARY OVER THE PAST SEVERAL DECADES, THE TRADITIONAL APPROACH TO COMBATING VIRAL INFECTIOUS DISEASES HAS BEEN TO TARGET THE VIRUS ITSELF, IN MOST CASES BY EITHER BLOCKING VIRUS-ENCODED ENZYMES THAT ARE REQUIRED FOR VIRAL REPLICATION, OR BY PREVENTING THE VIRUS FROM ENTERING HOST CELLS. ONE OF THE MAJOR CAVEATS OF THESE APPROACHES HAS BEEN THE ABILITY OF THE VIRUS TO READILY MUTATE AND THEREBY BECOME RESISTANT TO THESE CLASSICAL TYPES OF ANTIVIRAL THERAPIES. IN FACT, THIS IS A SERIOUS PROBLEM FOR THE THERAPY OF RNA VIRUS INFECTIONS, SUCH AS HIV OR INFLUENZA VIRUS, WHICH ARE KNOWN TO RAPIDLY MUTATE AND THEREBY ESCAPE ANTIVIRAL DRUGS. ADDITIONALLY, TRADITIONAL ANTIVIRAL APPROACHES ARE DESIGNED TO TARGET A SPECIFIC VIRUS, AND THEREFORE ARE INEFFECTIVE AGAINST ANY NEW VIRUS THAT MAY EMERGE IN THE FUTURE, AND IT IS IMPOSSIBLE TO PREDICT WHAT VIRUS WILL CAUSE THE NEXT OUTBREAK OR PANDEMIC. THEREFORE, THERE IS THE URGENT NEED TO DEVELOP NEW WAYS FOR TARGETING VIRAL PATHOGENS, WHICH WILL REQUIRE CREATIVE AND INNOVATIVE RESEARCH. LIKE HUMAN PROTEINS, VIRAL PROTEINS ROBUSTLY UNDERGO POSTTRANSLATIONAL MODIFICATIONS (PTMS) FOR THEIR REGULATION AND PROPER FUNCTIONING IN THE VIRUS LIFE CYCLE. IN MOST CASES, VIRAL PTMS ARE DYNAMICALLY REGULATED BY HUMAN ENZYMES, SUCH AS KINASES/PHOSPHATASES, UBIQUITIN E3 LIGASES/DEUBIQUITINATING ENZYMES, OR ACETYL TRANSFERASES/DEACETYLASES. THUS, CELLULAR ENZYMES PLAY AN IMPORTANT ROLE IN CONTROLLING THE ABILITY OF THE VIRUS TO REPLICATE AND TO CAUSE DISEASE. THIS PROPOSAL’S OVERARCHING GOAL IS TO COMPREHENSIVELY MAP THE ‘VIRAL PTMOME’ TO IDENTIFY THE PTMS THAT ARE ESSENTIAL FOR VIRUS REPLICATION AND PATHOGENESIS. WE WILL COMBINE PROTEOMICS SCREENS AND MOLECULAR VIROLOGY APPROACHES INCLUDING REVERSE GENETICS TECHNIQUES WITH CUTTING- EDGE MOLECULAR, BIOCHEMICAL AND BIOPHYSICAL STUDIES. THIS WILL ALLOW US TO IDENTIFY AND CHARACTERIZE VIRAL PTMS AND THE RESPONSIBLE HOST MODIFYING ENZYMES, AS WELL AS TO DETERMINE THEIR ROLES FOR EFFECTIVE VIRAL REPLICATION AND PATHOGENESIS. THIS POWERFUL APPROACH, COMBINED WITH COLLABORATIVE STUDIES TO DESIGN AND TEST CHEMICAL INHIBITORS TO BLOCK THE ENZYMES THAT REGULATE CRITICAL VIRAL PTMS, WILL NOT ONLY PROVIDE UNIQUE MECHANISTIC INSIGHT INTO HOST CONTROL OF VIRUS REPLICATION BUT WILL ALSO LAY THE GROUNDWORK FOR DEVELOPING NEW ANTIVIRALS FOR A RANGE OF EMERGING VIRAL INFECTIOUS DISEASES.

ACTIVATION OF THE BETA-3 INTEGRINS: ROLE OF THE KINDLINS

THERAPEUTIC IMPLICATIONS OF MOLECULAR DEFECTS IN BONE MARROW FAILURE

ER-TO-GOLGI TRANSPORT OF COAGULATION FACTORS V AND VIII

DEEP CEREBELLAR ELECTRICAL STIMULATION FOR POST-STROKE MOTOR RECOVERY

CILIA ASSEMBLY AND TRANSPORT IN THE VERTEBRATE RETINA

ASSESSING NOVEL BIOMARKERS FOR COGNITIVE FLUCTUATIONS IN DLB - AGE-RELATED DISEASES, SUCH AS ALZHEIMER DISEASE (AD) AND DEMENTIA WITH LEWY BODIES (DLB), ARE DEFINING PUBLIC HEALTH CONCERNS OF THE 21ST CENTURY AND ARE THE LEADING CAUSE OF DISABILITY WORLDWIDE. NOVEL DISEASE MODIFYING THERAPIES HAVE OPENED A NEW HORIZON FOR MANAGING THESE PROGRESSIVE DISEASES. HOWEVER CLINICAL TRIALS ASSESSING THESE THERAPEUTIC MEDICATIONS ARE BEDEVILED BY SMALL EFFECT SIZES AND SUBSTANTIAL INTERSUBJECT VARIABILITY THAT CHALLENGES EVALUATION OF THEIR EFFECTIVENESS. TO MEET THIS CHALLENGE, ADDITIONAL FACTORS MEDIATING VARIABILITY IN COGNITIVE PERFORMANCE ARE A KEY KNOWLEDGE GAP TO OVERCOME. COGNITIVE FLUCTUATIONS (CFS) ARE INTERMITTENT EPISODES OF DECREASED ALERTNESS, ATTENTION, AND RESPONSIVENESS THAT OCCUR IN SOME PATIENTS WITH DEMENTIA. THE UNPREDICTABLE NATURE OF THESE EVENTS POSES A MAJOR BURDEN FOR PATIENTS AND CAREGIVERS, AS WELL AS COMPLICATING THE MEASUREMENT OF TREATMENT EFFICACY IN CLINICAL TRIALS, PARTICULARLY WHEN THE TREATMENT IS INTENDED TO IMPROVE COGNITION. UNFORTUNATELY, METHODS TO TRACK CFS EFFECTIVELY AND THE PATHOPHYSIOLOGY OF THIS CONDITION ARE POORLY UNDERSTOOD. IN THIS PROPOSAL, WE AIM TO VALIDATE NOVEL BIOMARKERS THAT CAPTURES THE MOST RELEVANT FEATURES OF COGNITIVE FLUCTUATIONS IN AD AND DLB. TOWARDS THIS GOAL, WE WILL PROVIDE A MULTIMODAL CHARACTERIZATION OF THE FREQUENCY AND SEVERITY OF CFS AND THEIR LONGITUDINAL VARIABILITY AT ANNUAL VISITS. THE DEMENTIA WITH LEWY BODY CONSORTIUM (DLBC) COHORT PRESENTS A GOLD STANDARD COHORT FOR DLB BIOMARKER DIAGNOSIS (CSF Α-SYNUCLEIN, AGGREGATION ASSAY), WHILE THE ALZHEIMER’S DISEASE RESEARCH CENTERS (ADRC) PRESENTS COHORTS FOR AD DIAGNOSIS (CSF A-BETA42,40, T-TAU, AND P-TAU181) ALONG WITH AUTOPSY CONFIRMATION. IN THESE COHORTS, WE WILL OBTAIN BOTH CLINICAL (FLUCTUATION SCALES, COGNITIVE VARIABILITY) AND MULTIPLE OBJECTIVE DIGITAL BIOMARKERS (ACTIGRAPHY, EEG, SLEEP, RESTING STATE FUNCTIONAL MRI) TO EVALUATE THE FREQUENCY AND SEVERITY OF CFS AT BASELINE AND LONGITUDINAL VARIABILITY ANNUALLY OVER FOUR YEARS. THIS PROJECT WILL ENABLE US TO HELP DEVELOP APPROPRIATE BIOMARKERS THAT CAPTURE THE MOST RELEVANT FEATURES THAT CAN BE SCALED TO TRACK CFS EFFECTIVELY IN LARGE CLINICAL TRIALS. FURTHER, THIS PROJECT WILL HELP US DEVELOP A MODEL OF THE UNDERLYING NEURAL FEATURES OF CFS TO ENABLE FUTURE TARGETED THERAPIES.

THE CYCLICAL LOWER-EXTREMITY EXERCISE FOR PARKINSON'S TRIAL

NOVEL APPROACHES TO IMPROVE PREDICTION OF CANCER-ASSOCIATED THROMBOSIS

CORNEAL EPITHELIAL GROWTH FACTORS AND RECEPTORS

CLEVELAND ALZHEIMER'S DISEASE RESEARCH CENTER

IMAGING POST-TRAUMATIC OSTEOARTHRITIS 10-YEARS AFTER ACL RECONSTRUCTION: A MULTICENTER COHORT STUDY WITH QUANTITATIVE MRI

ANTIVIRAL ACTIONS OF INTERFERONS

DESIGNING INDUCTION THERAPIES TO TARGET MEMORY T CELLS IN HIGH RISK RECIPIENTS

NITRIC OXIDE PRODUCTION AND REACTIONS IN THE LUNG

REPRODUCIBILITY IN SIMULATION-BASED PREDICTION OF NATURAL KNEE MECHANICS

MULTI-OMIC BIOMARKERS FOR NEUROPATHIC PAIN SECONDARY TO CHEMOTHERAPY

CLINICAL STUDY OF VASCULAR PLAQUE DETERMINATION FOR STROKE RISK ASSESSMENT

1/2 POMALIDOMIDE FOR BLEEDING IN PATIENTS WITH HEREDITARY HEMORRHAGIC TELANGIECTASIA (HHT)

EFFECT OF LIPIDS ON VASCULAR GRAFT HEALING

MR FINGERPRINTING FOR EPILEPSY

KSHV EPIGENETIC REGULATION

ENDOPHENOTYPE NETWORK-BASED APPROACHES TO PREDICTION AND POPULATION-BASED VALIDATION OF IN SILICO DRUG REPURPOSING FOR ALZHEIMER?S DISEASE

TREM2 GENOTYPE-INFORMED DRUG REPURPOSING AND COMBINATION THERAPY DESIGN FOR ALZHEIMER?S DISEASE - PROJECT SUMMARY THE CUMULATIVE EVIDENCE INDICATES NEURO-INFLAMMATION PLAY CRUCIAL ROLES IN ALZHEIMER’S DISEASE (AD) AND ANTI- INFLAMMATORY AGENTS (I.E., MICROGLIA-TARGETED THERAPIES) SHOW POTENTIAL TREATMENTS FOR AD. THE TREATMENT WINDOW FOR MICROGLIA-TARGETED THERAPIES MAY AT LEAST BE OPEN LATER IN AD DUE TO THE NATURE OF MICROGLIAL BIOLOGY. IF AMYLOID-SS IS THE TRIGGER FOR AD, AND TAU IS THE EXECUTIONER, THEN MICROGLIA ARE ACCELERATORS OF DISEASE PROGRESSION. MICROGLIA-TARGETED APPROACHES HAVE A BETTER CHANCE OF SUCCESS IN MILD TO MODERATE DISEASE COMPARED TO ANTI-AMYLOID THERAPIES THAT HAVE TO PREVENT THE TRIGGER EARLY IN AD. AMONG THE MICROGLIAL GENES, TRIGGERING RECEPTOR EXPRESSED ON MYELOID CELLS 2 (TREM2) HAS RECEIVED MUCH ATTENTION BECAUSE A HEMIZYGOUS R47H VARIANT OF TREM2 (TREM2R47H) INCREASES THE AD RISK BY 2-7 FOLDS IN VARIOUS POPULATIONS, INCLUDING AFRICAN AMERICAN. OUR PRELIMINARY SINGLE-NUCLEUS RNA-SEQUENCING (SNRNASEQ) STUDY IDENTIFIED DISEASE-ASSOCIATED MICROGLIA (DAM) WITH ENHANCED AD PRO-INFLAMMATORY SIGNATURES (I.E., ACTIVATION OF AKT-SIGNALING) ASSOCIATED WITH THE TREM2R47H VARIANT AND PHARMACOLOGICAL AKT INHIBITION REVERSED THE TREM2R47H INDUCED INFLAMMATION, SHOWING PROOF-OF-CONCEPT OF TREM2R47H TARGETED DRUG DISCOVERY IN AD. USING MULTIMODAL SNRNASEQ ANALYSIS, WE IDENTIFIED TWO APPROVED ANTI-INFLAMMATORY ASTHMA DRUGS (FLUTICASONE AND MOMETASONE) THAT WERE PREDICTED TO MODULATE DAM MOLECULAR NETWORKS ARE ASSOCIATED WITH A REDUCED INCIDENCE OF AD IN ELECTRONIC HEALTH RECORD (EHR) DATA OF 7 MILLION PATIENTS. WE THEREFORE POSIT THAT UNDERSTANDING HOW THE AD-LINKED MUTATIONS (I.E., TREM2R47H) ENHANCE MICROGLIAL TOXICITY COULD LEAD TO UNDERSTANDING HOW MICROGLIA BECOME MALADAPTIVE/TOXIC AND DEVELOPMENT OF MICROGLIA-TARGETED THERAPEUTIC STRATEGY FOR LATE-ONSET SPORADIC AD IN GENERAL. THIS APPLICATION CALLS FOR NOVEL MICROGLIA-TARGETED DRUG REPURPOSING AND COMBINATION THERAPIES FOR AD USING OUR WELL-ESTABLISHED MULTI-OMICS AND DEEP LEARNING-BASED EHR APPROACHES, AND FUNCTIONAL OBSERVATIONS IN AD PATIENT-INDUCED PLURIPOTENT STEM CELLS (IPSCS), CEREBRAL ORGANOIDS, AND MOUSE MODELS, WITH THREE SPECIFIC AIMS. AIM 1 WILL TEST THE TREM2R47H INFORMED MICROGLIA-TARGETED THERAPEUTIC HYPOTHESIS FOR IDENTIFYING CELL TYPE-SPECIFIC MOLECULAR DRIVERS/NETWORKS, REPURPOSABLE DRUGS, AND COMBINATION THERAPIES FOR AD USING MULTI-OMICS EVIDENCE AGGREGATION. THESE ANALYSES WILL LEVERAGE OUR EXISTING SNRNASEQ DATA (N = 24 TREM2R47H AND N = 23 COMMON VARIANT (CV)-TREM2 WITH MATCHED AGE, AD PATHOLOGY AND APOE GENOTYPES) AND PUBLIC GENOMIC DATA FROM THE AD KNOWLEDGE PORTAL. AIM 2 WILL IDENTIFY REPURPOSABLE DRUGS AND COMBINATION THERAPIES USING HIGH-THROUGHPUT EHR-BASED HYPOTHESIS GENERATION AND SEQUENTIAL DEEP LEARNING-BASED PROPENSITY SCORE MATCHING APPROACHES. WE WILL LEVERAGE DE-IDENTIFIED EHRS FROM THE INSIGHT NETWORKS (~11 MILLION PATIENTS ACROSS NEW YORK CITY’S 5 HEALTH SYSTEMS AND THE GREATER METROPOLITAN AREA). AIM 3 WILL SCREEN, TEST AND VALIDATE DRUGS USING AD PATIENT- DERIVED IPSC LINES CARRYING TREM2R47H IN CONJUNCTION WITH CEREBRAL ORGANOID AND MOUSE MODELS. THE SUCCESSFUL COMPLETION OF THIS PROJECT WILL OFFER A VIABLE STRATEGY TO MOVE AD DRUG REPURPOSING FROM BENCH TO BEDSIDE RAPIDLY. 1

REGULATION OF CHOROIDAL NEOVASCULARIZATION IN SORSBY'S FUNDUS DYSTROPHY

THE MOLECULAR BASIS OF LIVER INDUCED RETINAL BLOOD VESSEL PLASTICITY.

SFTSV NONSTRUCTURAL PROTEIN NSS-MEDIATED IMMUNOPATHOGENESIS

ALZHEIMER'S MULTIOME DATA REPURPOSING: ARTIFICIAL INTELLIGENCE, NETWORK MEDICINE, AND THERAPEUTICS DISCOVERY - PROJECT SUMMARY PREDISPOSITION TO AD INVOLVES A COMPLEX, POLYGENIC, AND PLEIOTROPIC GENETIC ARCHITECTURE; FURTHERMORE, THERE ARE NO DISEASE MODIFYING TREATMENTS THAT SLOW THE NEURODEGENERATIVE PROCESS FOR AD. TRADITIONAL REDUCTIONIST PARADIGMS OVERLOOK THE INHERENT COMPLEXITY OF AD AND HAVE OFTEN LED TO TREATMENTS THAT ARE LACK OF CLINICAL BENEFITS OR FRAUGHT WITH ADVERSE EFFECTS. EXISTING MULTI-OMICS DATA RESOURCES, INCLUDING GENETICS, GENOMICS, TRANSCRIPTOMICS, INTERACTOMICS (PROTEIN-PROTEIN INTERACTIONS AND CHROMATIN INTERACTIONS), HAVE NOT YET BEEN FULLY UTILIZED AND INTEGRATED TO EXPLORE THE PATHOBIOLOGY AND DRUG DISCOVERY FOR AD. UNDERSTANDING AD GENETICS AND GENOMICS FROM THE POINT-OF-VIEW OF HOW CELLULAR SYSTEMS AND MOLECULAR INTERACTOME PERTURBATIONS UNDERLIE THE DISEASE (TERMED DISEASE MODULE) IS THE ESSENCE OF NETWORK MEDICINE. SYSTEMATIC IDENTIFICATION AND CHARACTERIZATION OF NOVEL UNDERLYING PATHOGENESIS AND DISEASE MODULE, WILL SERVE AS A FOUNDATION FOR IDENTIFYING AND VALIDATING NOVEL RISK GENES AND DRUG TARGETS IN AD. GIVEN OUR PRELIMINARY RESULTS, WE POSIT THAT A GENOME- WIDE, MULTIMODAL ARTIFICIAL INTELLIGENCE (AI) FRAMEWORK TO IDENTIFY NEW RISK GENES AND NETWORKS FROM HUMAN GENOME/EXOME SEQUENCING AND MULTI-OMICS FINDINGS ENABLE A MORE COMPLETE MECHANISTIC UNDERSTANDING OF AD PATHOGENESIS AND THE RAPID DEVELOPMENT OF TARGETED THERAPEUTIC INTERVENTION FOR AD WITH GREAT SUCCESS. AIM 1 WILL DETERMINE WHETHER RARE CODING AND NON-CODING VARIANTS BY WHOLE-GENOME/EXOME SEQUENCING (WGS/WES) ARE ENRICHED IN PROTEIN-FUNCTIONAL AND GENE-REGULATORY REGIONS USING SEQUENCE AND STRUCTURE-BASED DEEP LEARNING MODELS. THESE ANALYSES WILL ASSEMBLE WGS/WES AND CLINICAL DATA FROM ALZHEIMER'S DISEASE SEQUENCING PROJECT (ADSP), PUBLICLY AVAILABLE PROTEIN STRUCTURE (I.E., PROTEIN-PROTEIN INTERFACES, PROTEIN-LIGAND BINDING SITES, POST-TRANSLATIONAL MODIFICATIONS) AND SEQUENCE (EXPRESSION QUANTITATIVE TRAIT LOCUS [EQTLS], HISTONE-QTLS, AND TRANSCRIPTION FACTOR BINDING-QTLS) INFORMATION FROM THE PDB DATABASE, GTEX, NIH ROADMAP, FANTOM5, PSYCHENCODE, AND NIH 4D NUCLEOME. AIM 2 WILL DETERMINE WHETHER GWAS COMMON VARIANTS LINKED TO AD PATHOBIOLOGY AND ENDOPHENOTYPES ARE ENRICHED IN GENE REGULATORY NETWORKS IN A CELL-TYPE SPECIFIC MANNER USING A BAYESIAN FRAMEWORK. WE WILL VALIDATE RISK GENE AND NETWORK FINDINGS USING WGS/WES AND PROTEIN PANEL EXPRESSION DATA FROM OUR EXISTING COHORTS: THE CLEVELAND CLINIC LOU RUVO CENTER FOR BRAIN HEALTH AGING AND NEURODEGENERATIVE DISEASE BIOBANK (CBH-BIOBANK) AND THE CLEVELAND ALZHEIMER'S DISEASE RESEARCH CENTER (CADRC). AIM 3 WILL TEST THE HYPOTHESIS THAT RISK GENES AND NETWORKS CAN BE MODULATED VIA IN SILICO DRUG REPURPOSING, POPULATION-BASED VALIDATION, AND FUNCTIONAL TEST, TO IDENTIFY CANDIDATE AGENTS AND DRUG COMBINATIONS THAT WILL MODIFY AD. THE SUCCESSFUL COMPLETION OF THIS PROJECT WILL OFFER CAPABLE AND INTELLIGENT COMPUTER-BASED TOOLBOXES THAT ENABLE SEARCHING, SHARING, VISUALIZING, QUERYING, AND ANALYZING GENETICS, GENOMICS, AND MULTI-OMICS PROFILING DATA FOR GENOME-INFORMED THERAPEUTIC DISCOVERIES FOR AD AND OTHER NEURODEGENERATIVE DISEASE IF BROADLY APPLIED.

EXPLOITING ECOLOGY AND EVOLUTION TO PREVENT THERAPY RESISTANCE IN EGFR-DRIVEN LUNG CANCER

REGULATION OF HOST INNATE IMMUNITY AGAINST VIRAL INFECTION

FORECASTING LUNG TRANSPLANT BENEFIT: A DYNAMIC RISK MODELING APPROACH

DEVELOPMENT OF AN LMIC-ADAPTED THERMOCOAGULATION PROTOTYPE FOR THE TREATMENT OF CERVICAL PRE-CANCER.

COMBINED TRAINING PROGRAM IN DIGESTIVE DISEASE SCIENCES

MULTI-VENDOR MULTI-SITE NOVEL ACCELERATED MRI RELAXOMETRY

REGULATION OF B CELLS IN THE CNS

REFERENCE MODELS FOR MULTI-LAYER TISSUE STRUCTURES

FAILURE WITH CONTINUITY AND ITS RELATION TO ROTATOR CUFF REPAIR CLINICAL OUTCOMES

A NOMOGRAM TO PREDICT SEIZURE OUTCOMES AFTER RESECTIVE EPILEPSY SURGERY

MECHANISM OF APOA1 LIPIDATION BY ABCA1 IN HDL BIOGENESIS

VARIATION IN TUMOR-ASSOCIATED IMMUNE PROFILES AND COLORECTAL CANCER OUTCOMES - CONSIDERABLE VARIABILITY IN TUMOR-ASSOCIATED IMMUNE RESPONSES EXISTS ACROSS RACIAL/ETHNIC POPULATIONS. THESE VARIATIONS MAY EXPLAIN PART OF THE OBSERVED DISPARITIES IN RESPONSE TO CANCER THERAPIES, PARTICULARLY IMMUNOTHERAPY, AND TREATMENT OUTCOMES. IN COLORECTAL CANCER (CRC), THE INTENSITY AND COMPOSITION OF TUMOR INFILTRATING LYMPHOCYTES (TIL) ARE ESTABLISHED PROGNOSTIC AND PREDICTIVE INDICATORS. HOWEVER, FACTORS CONTRIBUTING TO THE DIVERSITY OF TIL RESPONSES OBSERVED AMONG CRCS REMAIN LARGELY UNKNOWN, AND THE INFLUENCE OF RACE/ETHNICITY AND GENETIC ANCESTRY HAVE BEEN UNDEREXPLORED. IN A RECENT STUDY COMPARING CRCS FROM AFRICAN AMERICANS AND NON-HISPANIC WHITES, DIFFERENCES IN LYMPHOCYTIC REACTIONS WERE OBSERVED TO PARTIALLY EXPLAIN THE SURVIVAL DISPARITY BETWEEN THE TWO GROUPS. NO DATA IS AVAILABLE FOR OTHER RACIAL/ETHNIC GROUPS. PRIOR RESEARCH HAS ALSO BEEN LIMITED BY RELYING SOLELY ON SELF-REPORTED RACE/ETHNICITY, A SIGNIFICANT LIMITATION. STUDIES SHOW THAT SELF-REPORT DOES NOT FULLY OR ACCURATELY REFLECT THE GENETIC DIVERSITY PRESENT IN ADMIXED MINORITY POPULATIONS. WE HYPOTHESIZE THAT ANCESTRAL GENETIC ARCHITECTURE IS IMPORTANT FOR SHAPING IMMUNE-RELATED DETERMINANTS OF CRC OUTCOMES GIVEN THE DIFFERENTIAL EFFICIENCY OF IMMUNE FUNCTION OBSERVED ACROSS RACIAL/ETHNIC GROUPS. STUDIES IN THE GENERTICALLY ADMIXED LATINX POPULATION OFFER NOTABLE ADVANTAGES INCLUDING A UNIQUE OPPORTUNITY TO SIMULTANEOUSLY TEASE OUT THE CONTRIBUTIONS OF MULTIPLE ANCESTRAL BACKGROUNDS (E.G. AFRICAN, EUROPEAN, INDIGENOUS AMERICAN) TO VARIABILITY IN IMMUNE FUNCTION. HERE, WE WILL TEST THE HYPOTHESIS THAT GENETIC ANCESTRY IS INDEPENDENTLY ASSOCIATED WITH DIFFERENCES IN TUMOR-ASSOCIATED T CELL PROFILES THAT CONTRIBUTE TO CRC OUTCOME DISPARITIES (I.E. OBSERVED ACROSS POPULATIONS DEFINED BY ETHNICITY AND BY GENETIC ANCESTRY) USING EXISTING RESOURCES FROM THE HISPANIC COLORECTAL CANCER STUDY, THE PUERTO RICO BIOBANK, THE TOTAL CANCER CARE PROTOCOL, AND THE MOLECULAR EPIDEMIOLOGY OF COLORECTAL CANCER STUDY . WE WILL ADDRESS THREE AIMS: (1) QUANTIFY CRC-ASSOCIATED T CELL PROFILES IN LATINXS FROM DIVERSE GENETIC ANCESTRAL BACKGROUNDS USING DNA- AND PROTEIN-BASED APPROACHES; (2) INVESTIGATE THE INDEPENDENT ASSOCIATIONS OF GENETIC ANCESTRY, EPIDEMIOLOGIC FACTORS, AND CLINICAL VARIABLES WITH T CELL PROFILES IN THE TUMOR MICROENVIRONMENT OF LATINX CRCS; AND (3) COMPARE CRC-ASSOCIATED T CELL PROFILES BETWEEN LATINX AND NHW POPULATIONS. THIS STUDY IS UNIQUE IN LEVERAGING THE ANCESTRAL DIVERSITY OF LATINOS TO UNDERSTAND THE RELATIONSHIPS BETWEEN RACE/ETHNICITY, GERMLINE GENETICS, TUMOR IMMUNOBIOLOGY, AND CANCER DISPARITIES. RESULTS WILL PROVIDE NEW AVENUES FOR UNDERSTANDING IMMUNOLOGICAL FACTORS CONTRIBUTING TO DISPROPORTIONATE TREATMENT RESPONSE AND MORTALITY IN DIVERSE POPULATIONS OF PATIENTS WITH CRC.

TROOP READINESS EVALUATION WITH AN AUGMENTED REALITY RETURN-TO-DUTY (READY) SYSTEM: AN OBJECTIVE METHOD OF EVALUATING SERVICE MEMBER PERFORMANCE FOLLOWING MTBI

SOFTWARE FOR PRACTICAL ANNOTATION AND EXCHANGE OF VIRTUAL ANATOMY

PRIMARY IMMUNOPREVENTION OF TRIPLE-NEGATIVE BREAST CANCER

A COGNITIVE RISK CALCULATOR AND SCREENING TOOL FOR PRIMARY CARE SETTINGS - PROJECT SUMMARY/ABSTRACT THE NUMBER OF PEOPLE WITH ALZHEIMER’S DISEASE IS EXPECTED TO EXCEED 88 MILLION BY 2050. EARLY DETECTION OF ALZHEIMER’S DISEASE AND RELATED DISORDERS SYMPTOMS ARE IMPERATIVE FOR EFFORTS TO DEVELOP DISEASE MODIFYING THERAPIES OR INTERVENTIONS, AND ARE IMPORTANT FOR OPTIMAL PATIENT CARE. THE OBJECTIVE OF THIS PROPOSAL IS TO PROVIDE AN ACCURATE, FEASIBLE, AND LOW-COST PROCESS TO FACILITATE COGNITIVE SCREENING IN POPULATION HEALTH. THE AIMS OF THE R61 PROJECT ARE TO DEVELOP AND VALIDATE TWO COMPLEMENTARY, AUTOMATED TOOLS TO IDENTIFY AND SCREEN PATIENTS AT HIGH RISK FOR COGNITIVE DECLINE. THE FIRST IS A RISK CALCULATOR THAT USES ROUTINE CLINICAL DATA TO ESTIMATE PATIENTS’ RISK FOR COGNITIVE DECLINE. PATIENTS 60 YEARS AND OLDER WILL UNDERGO FORMAL COGNITIVE TESTING, AND FIVE PRIOR YEARS OF ROUTINELY-COLLECTED ELECTRONIC HEALTH RECORD DATA WILL BE USED TO MODEL COGNITIVE STATUS. THE COGNITIVE RISK CALCULATOR WILL BE VALIDATED IN TWO HISTORICAL SAMPLES AND INTEGRATED INTO THE ELECTRONIC MEDICAL RECORD TO HELP PROVIDERS IDENTIFY VULNERABLE PATIENTS. THE SECOND TOOL IS A VALIDATED IPAD-BASED SCREENING TOOL, THE BRIEF ASSESSMENT OF COGNITIVE HEALTH (BACH) THAT PATIENTS CAN COMPLETE INDEPENDENTLY BEFORE, DURING, OR AFTER THEIR DOCTOR APPOINTMENT. PATIENTS WILL COMPLETE THE BACH, THE MONTREAL COGNITIVE ASSESSMENT TEST (A PAPER-AND-PENCIL SCREENING TOOL), AND FORMAL NEUROPSYCHOLOGICAL TESTING AT TWO TIME POINTS. WE HYPOTHESIZE THE BACH WILL BE MORE SENSITIVE TO COGNITIVE CHANGE OVER TIME THAN THE ESTABLISHED SCREENING TOOL. THE BACH IS ALREADY INTEGRATED INTO THE EPIC ELECTRONIC MEDICAL RECORD PLATFORM; COGNITIVE SCORE AND SCREENING RESULTS FOR DEPRESSION, STRESS, AND SLEEP DISRUPTION ARE AUTOMATICALLY RECORDED IN THE PATIENT’S MEDICAL RECORD FOR PROVIDERS TO DOCUMENT AND ACT UPON. THE R33 PROJECT IS A PHASE IV PRAGMATIC CLINICAL TRIAL TO GAUGE THE UPTAKE AND UTILITY OF THESE TOOLS IN PRIMARY CARE PRACTICES. SIX INTERNAL MEDICINE CLINICS WILL BE RANDOMIZED OVER TIME TO THE ACTIVE STUDY ARM (RISK CALCULATOR AND BACH). THIS WILL PERMIT COMPARISONS OF STANDARD OF CARE SCREENING PRACTICES WITH SCREENING ACTIVITIES FOLLOWING INTRODUCTION OF THE COGNITIVE RISK CALCULATOR AND BACH. WE WILL USE A MIXED-METHODS APPROACH TO DETERMINE OPTIMAL IMPLEMENTATION STRATEGIES ACROSS A VARIETY OF PRACTICE MODELS. IMPACT: THE EHR RISK-CALCULATOR AND BACH HAVE THE CAPACITY TO IMPROVE DETECTION OF COGNITIVE DECLINE ON A POPULATION HEALTH SCALE. THE PROJECT ALSO EXAMINES HOW REGIONAL DIFFERENCES IN SOCIOECONOMIC ADVANTAGE RELATE TO HEALTH DISPARITIES MEASURED IN TERMS OF COGNITIVE RISK AS WELL AS SCREENING ATTITUDES AND BEHAVIORS ON THE PART OF BOTH PROVIDERS AND PATIENTS.

TRANSCRIPTIONAL AND NON-TRANSCRIPTIONAL FUNCTIONS OF IRF3 IN ALD

SERIAL EVALUATION OF CARDIOPROTECTIVE EFFECTS OF EXERCISE TRAINING IN HEART FAILURE USING CARDIAC DIFFUSION TENSOR MRI

TESTING THE EFFECTIVENESS OF INDIVIDUALIZED DISEASE PREVENTION FOR MIDDLE-AGED ADULTS

TOWARDS SELECTIVE ANDROGEN DEPRIVATION BY TARGETING ANDROGEN ACTIVATION OF SRF

VISTA, A NOVEL CHECKPOINT THAT SUPPRESSES ANTI-TUMOR T CELL RESPONSES

IDENTIFYING NOVEL PARKINSON'S DISEASE GENES EXPLORING UNDERSTUDIED LATINO POPULATIONS

CLEVELAND COPE-AKI CLINICAL CENTER - PROJECT SUMMARY ACUTE KIDNEY INJURY (AKI) IS A FREQUENT AND DEBILITATING COMPLICATION IN HOSPITALIZED PATIENTS; HOWEVER, AKI SURVIVORS AFTER HOSPITAL DISCHARGE OFTEN RECEIVE FRAGMENTED HEALTH CARE. THIS INCREASES THE LIKELIHOOD FOR THE DEVELOPMENT OF LONG-TERM MAJOR ADVERSE KIDNEY EVENTS (MAKE), DEFINED AS INCIDENT OR PROGRESSIVE CHRONIC KIDNEY DISEASE, END-STAGE KIDNEY DISEASE AND ALL-CAUSE MORTALITY. PATIENTS WHO EXPERIENCE SEVERE AKI (STAGES 2 AND 3 PER THE KDIGO CLASSIFICATION) ARE AT EVEN HIGHER RISK FOR POOR OUTCOMES INCLUDING MAKE, DUE TO HIGHER PROPENSITY FOR PERSISTENT DAMAGE MANIFESTED AS LOWER RENAL FUNCTION AND PROTEINURIA. IN ADDITION, AKI SURVIVORS REPORT POOR QUALITY OF LIFE AND EXHIBIT FRAILTY FOR AT LEAST A YEAR AFTER THE AKI INCIDENT. TO OVERCOME THE GAPS IN SEVERE AKI MANAGEMENT, WE HYPOTHESIZE THAT SCREENING AND INTENSIVE MONITORING AND MANAGEMENT OF BLOOD PRESSURE AND PROTEINURIA VIA RENIN-ANGIOTENSIN SYSTEM BLOCKADE AND VOLUME OPTIMIZATION (FOLLOWING KDIGO GUIDELINES) REDUCES THE RATE OF MAKE WITHIN 2 YEARS, IMPROVES PATIENT-REPORTED OUTCOMES, AND REDUCES HEALTHCARE COSTS. THUS, CLEVELAND CLINIC AND METROHEALTH SYSTEM HAVE DEVELOPED THE CLEVELAND COPE-AKI CLINICAL CENTER TRIAL TO COMPARE THE USUAL STANDARD OF CARE WITH AN INTENSIVE, MANAGED CARE PATHWAY, CALLED CHAMPION CARE PATHWAY. THE CHAMPION CARE PATHWAY TEAM WILL COMPRISE A VIRTUAL/REMOTE NURSE-NAVIGATOR UNDER THE SUPERVISION OF A NEPHROLOGIST AND SUPPORTED BY A PATIENT ADVISORY COUNCIL. THE STRUCTURED LONGITUDINAL OUTPATIENT CARE PATHWAY WILL BE FACILITATED BY MULTI-LEVEL REMOTE PATIENT MONITORING, TARGETING THE RISK FACTORS FOR MAKE IN AKI STAGES 2 AND 3 SURVIVORS. PATIENTS ENROLLED TO THE CHAMPION CARE PATHWAY ARM WILL BE EMPOWERED TO BE ACTIVE PARTICIPANTS IN THEIR HEALTH MANAGEMENT BY PROVIDING THEIR OWN BLOOD PRESSURE, WEIGHT, KIDNEY FUNCTION, AND ALBUMINURIA DATA FOR THE FIRST THREE MONTHS AFTER THEIR HOSPITAL DISCHARGE AND AT PREDEFINED TIME INTERVALS THEREAFTER FOR 2 YEARS POST DISCHARGE. THIS DATA WILL ALLOW THE CHAMPION CARE TEAM TO MITIGATE CHANGING HEALTH STATUS EPISODES IN AN OUTPATIENT SETTING. THE PLANNED ADAPTIVE TRIAL DESIGN WILL ALLOW THE FOCUS OF THE INTERVENTION TO BE DIRECTED TOWARD THE PATIENTS MOST AT RISK FOR POST-DISCHARGE KIDNEY-SPECIFIC INCIDENTS. THUS, THE SPECIFIC AIMS OF THE CLEVELAND COPE-AKI CLINICAL CENTER ARE TO DETERMINE THE IMPACT OF THE CHAMPION CARE PATHWAY ON 1) TIME TO MAKE BY 2 YEARS 2) PATIENT-REPORTED OUTCOMES AND 3) COST EFFECTIVENESS. ANALYSES WILL DETERMINE WHETHER LONG-TERM CONTINUITY OF CARE REDUCES MORBIDITY AND MORTALITY FOR AKI STAGES 2 AND 3 PATIENTS, REDUCES HEALTHCARE COSTS AND IMPROVES THE WELL-BEING OF LONG-TERM AKI STAGES 2 AND 3 SURVIVORS. IN ADDITION, THE CLEVELAND COPE-AKI CLINICAL CENTER WILL WORK COLLABORATIVELY AND CLOSELY WITH THE SCIENTIFIC DATA AND RESEARCH CENTER ALONG WITH OTHER PARTICIPATING CLINICAL CENTERS TO DEVELOP COMMON POLICIES AND PROTOCOLS DESIGNED FOR STUDY INTERVENTIONS AND PERIODIC BIOSPECIMEN SAMPLING FOR CONTRIBUTION TO THE ENTIRE COPE-AKI CONSORTIUM.

NEVADA EXPLORATORY ALZHEIMER'S DISEASE RESEARCH CENTER (NVEADRC)

PROSPECTIVE EVALUATION OF OUTCOMES IN CIRRHOSIS OF DIFFERENT ETIOLOGIES: IMPACT OF HIV INFECTION AND SIMVASTATIN THERAPY - CHRONIC LIVER DISEASE, PRIMARILY CIRRHOSIS, REMAINS THE 6TH LEADING CAUSE OF DEATH IN ADULTS YOUNGER THAN 65Y IN THE UNITED STATES. DESPITE ADVANCES IN DIAGNOSTICS AND THERAPIES, MORTALITY IN CIRRHOSIS HAS NOT CHANGED SIGNIFICANTLY OVER THE LAST 40Y AND REMAINS A MAJOR SIGNIFICANT PUBLIC HEALTH BURDEN. WE AND OTHERS HAVE USED MODELING AND DATABASE EVALUATIONS TO SHOW THAT ALCOHOL RELATED LIVER DISEASE (ALD) AND NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD) ARE THE 2 MAJOR CAUSES OF CIRRHOSIS IN THE UNITED STATES. TREATING THE UNDERLYING ETIOLOGY OF CIRRHOSIS MAY HELP FIBROSIS REGRESS BUT WHETHER CIRRHOSIS IS REVERSIBLE IS NOT YET ESTABLISHED. WHETHER FIBROSIS PROGRESSES ONCE A DIAGNOSIS OF CIRRHOSIS IS ESTABLISHED AND IF SUCH A PROGRESSION IS RELATED TO DECOMPENSATION OR HEPATOCELLULAR CARCINOMA (HCC) ARE ALSO NOT KNOWN. OF THE VARIOUS COMPLICATIONS OF CIRRHOSIS, SARCOPENIA AND PHYSICAL FRAILTY DUE TO IMPAIRED CONTRACTILE FUNCTION ARE FREQUENT, PROGRESSIVE AND ADVERSELY IMPACT CLINICAL OUTCOMES. DESPITE THE HIGH CLINICAL SIGNIFICANCE, THERE ARE NO PROSPECTIVE STUDIES ON DEVELOPMENT, PROGRESSION AND PREDICTORS OF SARCOPENIA AND FRAILTY IN CIRRHOSIS. CO-MORBIDITIES ESPECIALLY INFECTION WITH HUMAN IMMUNE- DEFICIENCY VIRUS (HIV) PLACES PATIENTS WITH CIRRHOSIS AT HIGH RISK OF PROGRESSION OF FIBROSIS, DECOMPENSATION, AND SARCOPENIA/FRAILTY SYNDROME. THE GUT MICROBIOME AND THEIR METABOLITES (XENOMETABOLITES) PLAY A MECHANISTIC ROLE IN HEPATIC INJURY AND COMPLICATIONS OF CIRRHOSIS INCLUDING HCC AND SARCOPENIA BUT THERE ARE VERY LIMITED PROSPECTIVE STUDIES IN HUMAN PATIENTS. MOST STUDIES ON THE PROGRESSION, LONG TERM COMPLICATIONS, IMPACT OF CO- MORBIDITIES AND OUTCOMES IN CIRRHOSIS ARE CROSS-SECTIONAL, HAVE SMALL NUMBER OF SUBJECTS, AND DO NOT TRANSLATE ADVANCES IN MECHANISTIC UNDERSTANDING OF DEVELOPMENT OF CIRRHOSIS OR ITS COMPLICATIONS INTO CLINICAL PRACTICE. THEREFORE, PROSPECTIVE STUDIES IN WELL CHARACTERIZED CIRRHOSIS ARE CRITICAL TO DEVELOP EFFECTIVE MANAGEMENT STRATEGIES AND IMPROVE OUTCOMES. THERE IS INCREASING INTEREST IN THE USE OF STATINS IN THE MANAGEMENT OF CIRRHOSIS DUE TO ANTI-INFLAMMATORY AND ANTIFIBROTIC EFFECTS THAT MAY PREVENT DECOMPENSATION AND HCC. THE CLEVELAND CLINIC HEALTH SYSTEM IS ONE OF THE LARGEST CLINICAL PROGRAMS WITH A LARGE POPULATION OF PATIENTS WITH CIRRHOSIS WHO ARE REFERRED FOR LONG-TERM MANAGEMENT INCLUDING LIVER TRANSPLANTATION, BECAUSE OF OUR EXPERTISE IN INNOVATIVE APPROACHES TO PATIENT CARE INCLUDING TELEVISITS AND APPLICATIONS OF DIGITAL HEALTH INCORPORATED INTO INTEGRATED ELECTRONIC MEDICAL RECORDS. IN RESPONSE TO THE RFA PAR DK-20-003, WE PROPOSE TO BE A PART OF A LIVER CIRRHOSIS NETWORK TO ESTABLISH A LONGITUDINAL COHORT OF PATIENTS WITH CIRRHOSIS, PRIMARILY ALCOHOL RELATED AND NON-ALCOHOLIC FATTY LIVER DISEASE WITH CO-MORBIDITIES INCLUDING HIV INFECTION. WE WILL DEVELOP A DATABASE OF WELL CHARACTERIZED PATIENTS AND A BIOREPOSITORY FROM THESE PATIENTS TO ADVANCE OUR MECHANISTIC UNDERSTANDING OF PROGRESSION OF CIRRHOSIS, DEVELOPMENT OF COMPLICATIONS AND IDENTIFY NOVEL BIOMARKERS AND THERAPIES TO IMPROVE CLINICAL OUTCOMES. WE WILL ALSO CONDUCT A PROSPECTIVE RANDOMIZED CLINICAL TRIAL USING SIMVASTATIN/PLACEBO IN WELL CHARACTERIZED PATIENTS WITH CIRRHOSIS AS PART OF THE NETWORK TO DETERMINE CLINICAL RESPONSES AND SAFETY.

BIOLOGICAL DETERMINANTS OF COLORECTAL CANCER OUTCOMES IN LATINOS OF DIVERSE ANCESTRAL ORIGINS

CHRONIC ANTIBODY-MEDIATED REJECTION OF KIDNEY ALLOGRAFTS - ABSTRACT ANTIBODY-MEDIATED MECHANISMS LEADING TO ACUTE AND CHRONIC RENAL ALLOGRAFT INJURY AND LOSS REMAIN POORLY UNDERSTOOD. INVESTIGATION INTO THESE MECHANISMS IS HAMPERED BY THE LACK OF APPROPRIATE ANIMAL MODELS TO STUDY THE DEVELOPMENT OF ALLOGRAFT INJURY AS THE DONOR-SPECIFIC ANTIBODY (DSA) RESPONSE IS INITIATED AND PROGRESSES. WE HAVE DEVELOPED A NOVEL MODEL OF ANTIBODY-MEDIATED REJECTION (ABMR) OF KIDNEY ALLOGRAFTS IN CCR5-/- RECIPIENTS WHERE DSA ELICITED IN RESPONSE TO COMPLETE MHC-MISMATCHED RENAL ALLOGRAFTS ARE >50-FOLD HIGHER THAN THE TITERS ELICITED IN WILD-TYPE RECIPIENTS. THE ALLOGRAFTS ARE ACUTELY REJECTED BY THE DSA IN CCR5-/- RECIPIENTS BETWEEN DAYS 17 AND 22 WITH FEATURES THAT ARE VIRTUALLY IDENTICAL TO THOSE DURING ACUTE ABMR OF CLINICAL KIDNEY GRAFTS, INCLUDING IDENTICAL HISTOPATHOLOGY AND GENE EXPRESSION SIGNATURES INDICATING NK CELL ACTIVATION. WE HAVE RECENTLY DEMONSTRATED NK CELL ACTIVATION WITHIN THE KIDNEY ALLOGRAFTS THAT IS REQUIRED FOR ACUTE ABMR. IN THE ABSENCE OF NK CELL ACTIVATION, HOWEVER, THE HIGH DSA TITERS INDUCED IN CCR5-/- KIDNEY ALLOGRAFT RECIPIENTS ARE INCAPABLE OF MEDIATING ACUTE ABMR BUT SLOWLY INDUCE DEVELOPMENT OF TUBULAR FIBROSIS AND CHRONIC GLOMERULAR INJURY THAT LEADS TO EVENTUAL GRAFT FAILURE SIMILAR TO THAT OBSERVED DURING LATE FAILURE OF CLINICAL KIDNEY TRANSPLANTS. OUR PRELIMINARY RESULTS FURTHER INDICATE MARKED CHANGES IN THE PHENOTYPE AND FUNCTIONAL TRANSCRIPTOME OF GRAFT INFILTRATING MONOCYTES AND MACROPHAGES DURING CHRONIC VS. ACUTE ABMR. DEVELOPMENT OF THE CHRONIC ABMR IS ALSO ACCOMPANIED BY THE APPEARANCE OF ANTIBODIES TO SEVERAL AUTOANTIGENS THAT IS NOT OBSERVED DURING ACUTE AMR. THESE RESULTS SUGGEST THE GENERATION OF ALTERED MYELOID CELLS AND AUTOANTIBODIES AS KEY MECHANISMS UNDERLYING THE DEVELOPMENT OF THIS CHRONIC PATHOLOGY AND HAVE LED US TO HYPOTHESIZE THAT DSA BINDING TO KIDNEY ALLOGRAFT ENDOTHELIUM IN THE ABSENCE OF NK CELL ACTIVATION STIMULATES PRODUCTION OF MYELOID CELL RECRUITMENT AND DIFFERENTIATION FACTORS THAT SKEW THEIR FUNCTION TO PROMOTE AUTOANTIBODY PRODUCTION AND DEVELOPMENT OF CHRONIC ANTIBODY-MEDIATED GRAFT INJURY. THIS HYPOTHESIS WILL BE TESTED IN THREE SPECIFIC AIMS. IN SPECIFIC AIM 1 WE WILL TEST MECHANISMS OF DSA-INDUCED KIDNEY ALLOGRAFT ENDOTHELIAL PRODUCTION OF FACTORS DIRECTING MYELOID CELL RECRUITMENT AND FUNCTION DURING DEVELOPMENT OF CHRONIC ABMR. IN SPECIFIC AIM 2 WE WILL TEST THE ROLE AND FUNCTIONS OF GRAFT INFILTRATING MYELOID CELLS THAT PROMOTE DEVELOPMENT OF ANTIBODY-MEDIATED ALLOGRAFT INJURY. IN SPECIFIC AIM 3 WE WILL TEST THE PRODUCTION AND ROLE OF AUTO-ANTIBODIES IN THE DEVELOPMENT OF CHRONIC ABMR. THE PROPOSED EXPERIMENTS WILL UTILIZE NOVEL AND CLINICALLY RELEVANT MODELS OF KIDNEY ALLOGRAFT CHRONIC ABMR TO DIRECTLY IDENTIFY THE INFLAMMATORY COMPONENTS AND THEIR MECHANISMS MEDIATING CHRONIC INJURY OF THE KIDNEY GRAFTS LATE AFTER TRANSPLANT. WE ANTICIPATE THAT OUR STUDIES WILL CONTINUE TO REVEAL NOVEL MECHANISMS CRITICAL TO THE DEVELOPMENT OF KIDNEY GRAFT CHRONIC ABMR AND TO IDENTIFY NEW THERAPEUTIC TARGETS TO INHIBIT OR ATTENUATE THIS PATHOLOGY AND IMPROVE KIDNEY GRAFT FUNCTION AND SURVIVAL.

FORWARD GENETIC ANALYSIS OF BILIARY SYSTEM FORMATION

ELUCIDATING GENETICS OF RESPONSE TO IMMUNE CHECKPOINT BLOCKADE IN LUNG CANCER

IMPROVING SPINAL CORD INJURY REHABILITATION INTERVENTIONS BY RETRAINING THE BRAIN

A GUT-RESTRICTED SMALL MOLECULE THERAPEUTIC FOR A NOVEL TARGET TO TREAT INFLAMMATORY BOWEL DISEASE

IMPROVING PHENOTYPIC CLASSIFICATION AND PREDICTION OF TREATMENT OUTCOMES IN PATIENTS WITH NON-ISCHEMIC CARDIOMYOPATHY AND FUNCTIONAL MITRAL REGURGITATION - PROJECT SUMMARY FUNCTIONAL MITRAL REGURGITATION (FMR) PORTENDS A BLEAK PROGNOSIS AND IS A COMMON CONSEQUENCE OF ISCHEMIC AND NON-ISCHEMIC CARDIOMYOPATHY (ICM, NICM), WHERE ADVERSE ANNULAR AND LEFT VENTRICULAR (LV) REMODELING AND/OR INFARCTION ALTERS MITRAL VALVE (MV) FUNCTION. PRIOR STUDIES DEMONSTRATE SIGNIFICANT INCREASES IN MORTALITY RISK AS SEVERITY OF FMR INCREASES; MORTALITY RATES RANGE FROM 15-40% AT 1 YEAR. FURTHERMORE, AS THE PREVALENCE OF HEART FAILURE (HF) IS RISING, FMR IS PROJECTED TO DOUBLE FROM OVER 2 MILLION PATIENTS IN 2000 TO OVER 4 MILLION PATIENTS IN THE UNITED STATES BY 2030. DEFINING FMR SEVERITY, OPTIMAL TIMING OF INTERVENTION, AND MOST APPROPRIATE METHOD FOR INTERVENTION REMAIN CONTROVERSIAL. RECENTLY, MITRA-FR AND COAPT TRIALS DEMONSTRATED CONTRASTING SURVIVAL BENEFIT WITH PERCUTANEOUS MV REPAIR, DEMONSTRATING THE IMPORTANCE AND NEED FOR MORE OPTIMAL SELECTION CRITERIA. CURRENTLY, THE PATIENT SELECTION CRITERIA FOR MITRACLIP THERAPY ARE SOLELY BASED ON MV ANATOMY AND CONTROVERSIAL ECHOCARDIOGRAPHIC CRITERIA FOR FMR SEVERITY. CARDIAC MAGNETIC RESONANCE (CMR) PROVIDES AN EXCITING OPPORTUNITY TO ADDRESS NUMEROUS UNMET NEEDS REGARDING CHARACTERIZING FMR AND THE NEED FOR MORE OPTIMAL SELECTION CRITERIA FOR IMPROVING OUTCOMES. SUPERIOR ACCURACY AND REPRODUCIBILITY FOR QUANTIFICATION OF LV SIZE AND FUNCTION, AND GOLD STANDARD TISSUE CHARACTERIZATION, POSITIONS CMR AS THE IDEAL IMAGING MODALITY FOR COMPREHENSIVELY CHARACTERIZING FMR AND THE UNDERLYING MYOPATHIC PROCESSES THAT SIGNIFICANTLY IMPACT RESPONSE TO FMR THERAPIES. THE GOAL OF THE CURRENT RESEARCH IS TO DEVELOP PERSONALIZED RISK PREDICTION FOR FMR PATIENTS THROUGH EXPLAINABLE UNSUPERVISED PHENOMAPPING ENRICHED WITH ADVANCED CMR IMAGING BIOMARKERS, AND TO DETERMINE THE CMR PREDICTORS OF REVERSE REMODELING FOLLOWING MODERN THERAPIES FOR FMR.

HDL STRUCTURE, FUNCTION AND DYSFUNCTION

BIOENERGETIC AND EPIGENETIC REPROGRAMMING BY OBESITY IN SEPSIS

COMPLEMENT REGULATES MACROPHAGE AND PLATELET FUNCTION IN KIDNEY TRANSPLANTS - ABSTRACT THE POTENTIAL FOR COMPLEMENT ACTIVATION DURING ORGAN RECOVERY, ISCHEMIA REPERFUSION AND ANTIBODY-MEDIATED REJECTION (AMR) IS WELL-RECOGNIZED. AS A RESULT, NUMEROUS THERAPEUTIC INHIBITORS OF COMPLEMENT HAVE BEEN DEVELOPED AND TESTED IN THE TREATMENT OF AMR. INHIBITORS OF THE TERMINAL COMPLEMENT COMPONENT C5, AND MORE RECENTLY, C1 THE FIRST COMPONENT OF THE CLASSICAL PATHWAY HAVE BEEN TESTED MOST EXTENSIVELY. EXTENSIVE EVIDENCE INDICATES C1Q FUNCTIONS AS A PATTERN RECOGNITION RECEPTOR (PRR) THAT BINDS APOPTOTIC CELLS AND MEDIATES A NON- INFLAMMATORY CLEARANCE BY MACROPHAGES. IN FACT, C1Q DEFICIENT PATIENTS AND MICE DO NOT CLEAR APOPTOTIC CELLS EFFICIENTLY AND DEVELOP FLORID AUTOIMMUNITY. REMARKABLY LITTLE IS KNOWN ABOUT THE EFFECTS OF C1Q IN TRANSPLANTATION. HOWEVER, IN RECENT CLINICAL TRIALS, INJURY CAUSED BY DELAYED GRAFT FUNCTION HAS BEEN DIMINISHED BY TREATMENT WITH C1 INHIBITOR (C1INH), A SERINE PROTEASE INHIBITOR THAT TERMINATES COMPLEMENT ACTIVATION, BUT LEAVES C1Q INTACT. THESE RESULTS INVITE THE OBVIOUS QUESTION: DOES C1INH WORK BECAUSE IT TRUNCATES THE COMPLEMENT CASCADE AND DECREASES PRODUCTION OF DOWNSTREAM INFLAMMATORY MEDIATORS OR DOES C1INH WORK BECAUSE IT LEAVES C1Q INTACT TO MODULATE MACROPHAGES AND CELLS THAT EXPRESS C1Q RECEPTORS? OF COURSE, THESE ARE NOT MUTUALLY EXCLUSIVE. MORE IS KNOWN ABOUT THE FUNCTIONS OF C5A IN ANTIBODY INDUCED INFLAMMATION. C5A IS A POTENT CHEMOATTRACTANT AND ACTIVATOR OF NEUTROPHILS AND MACROPHAGES. HOWEVER, PLATELETS ALSO EXPRESS C5AR. WE HAVE DEMONSTRATED THAT PLATELETS ACCUMULATE WITHIN MINUTES AFTER ANTIBODY BINDS AND ACTIVATES COMPLEMENT ON GRAFT ENDOTHELIUM. WE PROPOSE THE HYPOTHESIS THAT C1Q DOWN MODULATES WHEREAS C5A UPREGULATES INFLAMMATORY RESPONSES IN TRANSPLANTS; THEREFORE PRESERVING C1Q FUNCTIONS AND INHIBITING C5A FUNCTIONS WILL DECREASE AMR. WE WILL TEST THIS HYPOTHESIS IN THE FOLLOWING 3 SPECIFIC AIMS: 1) TEST THE CAPACITY OF C1Q TO FUNCTION AS A PRR TO REMOVE POTENTIALLY IMMUNOGENIC EXTRACELLULAR VESICLES DURING REPERFUSION AFTER TRANSPLANTATION AND DECREASE THE INDUCTION OF ALLO- AND AUTOANTIBODY RESPONSES. 2) TEST THE EFFECTS OF C5A ON NEUTROPHIL AND MACROPHAGE FUNCTIONS IN ISCHEMIA- REPERFUSION AND CHRONIC AMR. 3) TEST THE EFFECTS OF C5A ON PLATELET FUNCTIONS IN ACUTE AND CHRONIC AMR. THESE SPECIFIC AIMS ENCOMPASS AND ENHANCE COMMON THREADS OF OUR PPG THAT FOCUSES ON MECHANISMS UNDERLYING AMR. THE GOAL OF THIS PROPOSAL IS TO PROVIDE THE BASIS FOR RATIONAL THERAPEUTIC ENHANCEMENT OF THE BENEFICIAL FUNCTIONS OF C1Q IN TRANSPLANTS AND MODULATION OF PROINFLAMMATORY EFFECTS OF C3A AND C5A.

CONTRALATERALLY CONTROLLED FES COMBINED WITH BRAIN STIMULATION FOR SEVERE UPPER LIMB HEMIPLEGIA

DIET, GUT MICROBIOTA, AND HEART FAILURE

FEASIBILITY AND ACCEPTABILITY OF TELEPHONE-DELIVERED MBCT FOR MIGRAINE AND DEPRESSION (FATE-MD) - PROJECT SUMMARY MIGRAINE IS A COMMON, PAINFUL AND HIGHLY DISABLING DISORDER. DEPRESSIVE SYMPTOMS ARE COMMON IN PEOPLE WITH MIGRAINE AND ARE ASSOCIATED WITH INCREASED SUFFERING. DESPITE THE SIGNIFICANT EMOTIONAL, SOCIAL, AND ECONOMIC BURDEN OF THIS COMORBIDITY, NO EXISTING TREATMENT ADEQUATELY ADDRESSES BOTH MIGRAINE DISABILITY AND ELEVATED DEPRESSIVE SYMPTOMS. DEVELOPMENT OF SCALABLE AND INTEGRATIVE TREATMENTS THAT ADDRESS BOTH MIGRAINE DISABILITY AND COMORBID DEPRESSIVE SYMPTOMS HAS BEEN IDENTIFIED AS A MAJOR GAP IN MIGRAINE TREATMENT. MINDFULNESS-BASED COGNITIVE THERAPY (MBCT) IS AN EVIDENCE-BASED AND STANDARDIZED TREATMENT THAT IS AN EXCELLENT CANDIDATE FOR REDUCING DISABILITY AND DEPRESSIVE SYMPTOMS IN PEOPLE WITH MIGRAINE. OUR PRELIMINARY DATA SHOW THAT MBCT REDUCED MIGRAINE-RELATED DISABILITY; HOWEVER, DURING EXIT INTERVIEWS, PARTICIPANTS NOTED THE BURDEN OF ATTENDING 8 WEEKLY IN-PERSON SESSIONS WAS A SUBSTANTIAL BARRIER TO ACCESSING CARE. OUR PRELIMINARY DATA SUGGEST AN ABBREVIATED AND TELEPHONE-DELIVERED MBCT (MBCT-T) PROTOCOL IS A FEASIBLE ALTERNATIVE TO IN-PERSON TREATMENT. HOWEVER, VIDEO- BASED DELIVERY (MBCT-V) MAY IMPROVE PATIENT ENGAGEMENT. THIS PROJECT AIMS TO PREPARE FOR A DEFINITIVE MULTI-SITE PHASE III TRIAL OF REMOTE-DELIVERED MBCT-T OR MBCT-V OR BOTH IN PATIENTS WITH MIGRAINE AND DEPRESSIVE SYMPTOMS. DURING STUDY 1, WE WILL USE GUIDELINES FROM THE NIH’S BEHAVIORAL CHANGE CONSORTIUM TO OPTIMIZE FIDELITY MBCT-T AND MBCT-V FOR PATIENTS WITH MIGRAINE (AS DEFINED BY THE INTERNATIONAL CLASSIFICATION OF HEADACHE DISORDERS – 3) AND DEPRESSIVE SYMPTOMS (AS DEFINED BY EMPIRICAL CUT-OFFS ON THE PATIENT HEALTH QUESTIONNAIRE – 9). PRODUCTS DEVELOPED DURING AIM 1 WILL BE: TRAINING PROTOCOLS, FACILITATOR TREATMENT MANUALS, FIDELITY CHECKLISTS, MBCT-T/V ADHERENCE AND COMPETENCE SCALES, FIDELITY CODING MANUALS. PRODUCTS WILL BE ITERATIVELY OPTIMIZED USING QUANTITATIVE AND QUALITATIVE FIDELITY METRICS ASSESSED AT THE PATIENT-LEVEL (COMPREHENSION) AND FACILITATOR-LEVEL (CORRECTIVE FEEDBACK, INTERVENTION DELIVERY) FOLLOWING THE DELIVERY OF ONE MBCT-T AND ONE MBCT-V GROUP LED BY A CERTIFIED MBCT FACILITATOR AT A SITE IN NEW YORK, NY. ALSO, DURING STUDY 1, A 22-HOUR TRAINING WILL BE CONDUCTED FOR FACILITATORS ACROSS TWO ADDITIONAL SITES IN THE BRONX, NY AND WINSTON-SALEM, NC. DURING STUDY 2, WE WILL CONDUCT A MULTI-SITE PILOT RCT (N=144) OF MBCT-T, MBCT-V, AND ENHANCED USUAL CARE (EUC). WE WILL EVALUATE FIDELITY, FEASIBILITY, AND ACCEPTABILITY OF EACH ARM ACROSS THREE SITES IN PATIENTS WITH MIGRAINE AND DEPRESSIVE SYMPTOMS. PRE- SPECIFIED DECISION RULES FOR FIDELITY, FEASIBILITY, ACCEPTABILITY, AND CLINICAL UTILITY WILL DETERMINE THE ARMS OF THE FUTURE PHASE III TRIAL EVALUATING REMOTE-DELIVERED MBCT FOR MIGRAINE DISABILITY AND DEPRESSIVE SYMPTOMS.

UNDERSTANDING AND USING VARIATION IN SOURCE MATERIALS FOR MSC FABRICATION

CONTRIBUTION OF MYELOID-DERIVED SUPPRESSOR CELLS TO NEURO-INFLAMMATORY ALTERATIONS AND DISEASE PROGRESSION IN GLIOBLASTOMA - ABSTRACT: GLIOBLASTOMA (GBM), THE MOST COMMON PRIMARY BRAIN TUMOR, REMAINS UNIFORMLY LETHAL DUE TO MANY FACTORS, INCLUDING A POTENTLY IMMUNE-SUPPRESSIVE MICROENVIRONMENT. WHILE ATTEMPTS TO ALTER IMMUNE ACTIVATION HAVE BEEN SUCCESSFUL IN OTHER ADVANCED CANCERS, A SERIES OF DIVERSE STRATEGIES HAS YET TO MARKEDLY INCREASE GBM PATIENT SURVIVAL. THESE RESULTS DEMONSTRATE A KEY CLINICAL BARRIER TO SUCCESS AND UNDERSCORE THE NEED TO BETTER UNDERSTAND THE IMMUNE-SUPPRESSIVE GBM MICROENVIRONMENT, WHICH IS PART OF A UNIQUE NEUROIMMUNE SYSTEM. CENTRAL TO IMMUNE SUPPRESSION IN GBM IS THE PRESENCE OF MYELOID-DERIVED SUPPRESSOR CELLS (MDSCS), AN IMMATURE LINEAGE COMPRISED OF MONOCYTIC (M) AND GRANULOCYTIC (G) SUBSETS THAT POTENTLY SUPPRESSES CYTOTOXIC IMMUNE RESPONSE. INTERROGATING THE FUNCTION OF MDSCS IN GBM HAS BEEN A MAJOR FOCUS OF OUR LABORATORY. USING AN INTEGRATED APPROACH, WE HAVE SHOWN THAT MDSCS ASSOCIATE WITH POOR GBM PROGNOSIS, DRIVE CANCER STEM CELL FUNCTION, AND INTERACT WITH THE TUMOR THROUGH MULTIPLE SIGNALING NETWORKS THAT CAN BE NEUTRALIZED TO INCREASE IMMUNE ACTIVATION. WE HAVE ALSO INTERROGATED MDSC SUBSETS TO REVEAL DIFFERENCES IN LOCALIZATION AND FUNCTION IN A SEX-SPECIFIC MANNER AND IDENTIFIED MDSC SUBSET SIGNALING PROGRAMS THAT CAN BE ALTERED TO INCREASE IMMUNE ACTIVATION AND DECREASE GBM GROWTH. WHILE OUR WORK HAS IMPLICATED MDSCS AS BIOMARKERS AND DRIVERS OF GBM PROGRESSION AND IDENTIFIED THEM AS NEXT-GENERATION THERAPEUTIC TARGETS, THERE ARE SEVERAL KNOWLEDGE GAPS THAT REMAIN, AND ADDRESSING THEM IS THE FOCUS OF THIS APPLICATION: IT REMAINS UNCLEAR HOW MDSCS ORIGINATE AND THE EXTENT OF THEIR PLASTICITY; IT IS UNCLEAR HOW MDSC LINEAGE COMMITMENT IS INFORMED BY CELL-INTRINSIC PROGRAMS AND IS ALTERED AS A RESULT OF INTERACTION WITH UNIQUE NEURAL MICROENVIRONMENTS, MICROBIAL INTERACTIONS, AND SIGNALING PROGRAMS; AND THE EFFICACY OF TARGETING MDSC SUBSETS IN COMBINATION WITH IMMUNE ACTIVATING STRATEGIES HAS YET TO BE DETERMINED. THE OVERARCHING HYPOTHESIS OF THIS APPLICATION IS THAT MDSC SUBSET LINEAGE COMMITMENT IS DRIVEN THOUGH THE INTEGRATION OF CELL-INTRINSIC (INCLUDING SEX-SPECIFIC GENETIC AND EPIGENETIC PROGRAMS) AND CELL- EXTRINSIC (INCLUDING SYSTEMIC FACTORS FROM THE GUT-BRAIN AXIS) INTERACTIONS THAT CAN BE LEVERAGED FOR THE DEVELOPMENT OF MORE EFFECTIVE ANTI-GBM THERAPIES. THROUGH THIS R35 MECHANISM THAT ALLOWS FOR LONGER- TERM/FLEXIBLE FUNDING TO DEVELOP PARALLEL AREAS WITH SYNERGISTIC POTENTIAL, WE WILL TEST DISTINCT ASPECTS OF THIS HYPOTHESIS THOUGH THREE COMPLEMENTARY BUT INTEGRATED FOCUS AREAS: (1) THE CELLULAR AND MOLECULAR BASIS FOR MDSC LINEAGE COMMITMENT AND PLASTICITY, (2) THE RESPONSE OF MDSCS TO MICROENVIRONMENTAL CUES, AND (3) PRE- CLINICAL MDSC TARGETING IN COMBINATION WITH IMMUNE ACTIVATING THERAPIES. THESE STUDIES HAVE IMMEDIATE IMPLICATIONS FOR GBM AND OTHER NEUROLOGICAL DISORDERS AND ESTABLISH A PLATFORM FOR UNDERSTANDING IMMUNE RESPONSES IN OTHER NEUROLOGICAL DISORDERS BY PROVIDING UNIQUE INSIGHTS INTO NEURAL/IMMUNE INTERACTIONS MEDIATED VIA MDSCS, AS WELL AS BY ASSESSING BRAIN-PENETRANT IMMUNE-ALTERING THERAPEUTIC STRATEGIES.

RECOVERY OF BLADDER REFLEXES AND NERVE REGENERATION AFTER SPINAL CORD INJURY

ACUTE ANTIBODY MEDIATED KIDNEY ALLOGRAFT REJECTION - ABSTRACT ANTIBODY-MEDIATED LYMPHOCYTE DEPLETION IS A COMMON STRATEGY TO ELIMINATE DONOR-REACTIVE T CELLS IN TRANSPLANT RECIPIENTS. HOWEVER, MEMORY T CELLS ARE MORE RESISTANT TO DEPLETION AND HAVE BEEN ASSOCIATED WITH ACUTE REJECTION EPISODES IN TRANSPLANT RECIPIENTS TREATED WITH POLYCLONAL RABBIT ANTI-THYMOCYTE GLOBULIN (ATG) OR ANTI- CD52 MAB. UNDERSTANDING THE MECHANISMS AND COMPOSITION OF T CELLS RECONSTITUTED IN LYMPHOPENIC TRANSPLANT RECIPIENTS IS THUS CRITICAL FOR THE RATIONAL USE OF LYMPHOABLATIVE THERAPIES AND FOR IMPROVING THEIR GRAFT-PROLONGING EFFICACY. THE ULTIMATE GOAL OF OUR STUDIES IS TO DEVELOP APPROACHES THAT MINIMIZE HOMEOSTATIC EXPANSION AND SHIFT THE BALANCE TOWARDS THYMOPOIESIS, THUS AVOIDING OVER-IMMUNOSUPPRESSION. DURING THE PREVIOUS FUNDING CYCLE, WE USED A MURINE ATG ANALOG (MATG) IN A MOUSE HEART ALLOGRAFT MODEL TO ESTABLISH THAT HOMEOSTATIC RECONSTITUTION OF THE ENTIRE T CELL COMPARTMENT IS DRIVEN BY DEPLETION-RESISTANT MEMORY CD4+ T CELLS VIA B CELLS AND CD40/CD154 PATHWAY. WHILE COGNATE TCR-PMHC INTERACTIONS BETWEEN B CELLS AND T CELLS WERE DISPENSABLE, WE IDENTIFIED POSTTRANSPLANT INFLAMMATION AND B CELL-DERIVED CYTOKINES IL-1SS, IL-6 AND IL-27 AS KEY FACTORS FACILITATING HOMEOSTATIC T CELL RECOVERY. OUR PRELIMINARY DATA INDICATE THAT SIGNALING THROUGH PATTERN RECOGNITION RECEPTORS TLR4, TLR9 AND A MACROPHAGE-INDUCIBLE C-TYPE LECTIN (MINCLE, OR CLEC4E) IS REQUIRED TO INITIATE B CELL PRODUCTION OF PROINFLAMMATORY CYTOKINES. WE FURTHER IDENTIFIED INNATE-LIKE MARGINAL ZONE (MZ) B CELLS ACTING AS INITIAL SENSORS OF POSTTRANSPLANT INFLAMMATION IN LYMPHOPENIC RECIPIENTS. GENETIC DEFICIENCY OR SPECIFIC DEPLETION OF MZ B CELLS MARKEDLY DELAYS T CELL RECONSTITUTION IN MATG TREATED HEART ALLOGRAFT RECIPIENTS. WE HYPOTHESIZE THAT INFLAMMATION INDUCED BY TRANSPLANTATION AT THE TIME OF LYMPHOABLATION PROMOTES RAPID T CELL RECONSTITUTION. DAMPS RELEASED BY THE GRAFT ACTIVATE B CELLS TO SECRETE PROINFLAMMATORY CYTOKINES THAT FURTHER AMPLIFY B CELL ACTIVATION AND DIRECTLY ENHANCE T CELL PROLIFERATION. IN PARTICULAR, MZ B CELLS ACTIVATED VIA C-TYPE LECTIN RECEPTOR MINCLE AND TLRS ACT AS INITIAL SENSORS OF POSTTRANSPLANT INFLAMMATION FACILITATING PROINFLAMMATORY FUNCTIONS OF FOLLICULAR B CELLS. THEREFORE, THE HOMEOSTATIC RECOVERY OF MEMORY T CELLS AND ENSUING ALLOGRAFT REJECTION MAY BE DECREASED BY MINIMIZING DAMPS SIGNALING OR BY TARGETING MZ B CELL ACTIVATION AND FUNCTIONS. WE WILL TEST THIS HYPOTHESIS IN TWO SPECIFIC AIMS: AIM 1. TO TEST THE ROLE OF MZ B CELLS AS PRIMARY SENSORS OF GRAFT TISSUE INJURY IN LYMPHOPENIC RECIPIENTS. AIM 2. TO INVESTIGATE THE MECHANISMS BY WHICH C-TYPE LECTIN RECEPTOR MINCLE FACILITATES B CELL PROINFLAMMATORY FUNCTIONS AFTER MATG LYMPHOABLATION. THE PROPOSED STUDIES WILL MECHANISTICALLY DISSECT HOW INFLAMMATORY PATHWAYS TRIGGERED BY ALLOGRAFT ISCHEMIA/REPERFUSION INJURY DRIVE RAPID RECONSTITUTION OF DEPLETION-RESISTANT MEMORY T CELLS. BASED ON THESE INSIGHTS, WE WILL TEST THE EFFICACY OF SEVERAL CLINICALLY RELEVANT APPROACHES FOR INHIBITING RECOVERY OF PATHOGENIC DONOR-REACTIVE MEMORY T CELLS AND PROLONGING HEART ALLOGRAFT SURVIVAL IN ATG TREATED RECIPIENTS.

IMMUNE CELL ACTIVATION AND ASSOCIATED BLOOD BRAIN BARRIER CHANGES ACROSS DIFFERENT STAGES OF ALZHEIMER'S DISEASE - PROJECT SUMMARY/ABSTRACT AGE-RELATED DISEASES, SUCH AS ALZHEIMER'S DISEASE (AD), ARE DEFINING PUBLIC HEALTH CONCERNS OF THE 21ST CENTURY AND ARE THE LEADING CAUSE OF DISABILITY WORLDWIDE. A GROWING BODY OF EVIDENCE NOTES THAT CENTRAL NERVOUS SYSTEM IMMUNE CHANGES SUPERIMPOSED ON ONGOING CHRONIC NEURODEGENERATION MAY HAVE A MAJOR IMPACT ON DISEASE PROGRESSION IN AD AND OTHER NEURODEGENERATIVE CONDITIONS. THE ROLE OF THE PERIPHERAL IMMUNE SYSTEM IN AD, INCLUDING CELLULAR ELEMENTS OF BOTH THE INNATE AND ADAPTIVE IMMUNITY ARE STILL ENIGMATIC. IN THIS PROPOSAL, WE INVESTIGATE IMMUNE CELL IDENTITY AND ABUNDANCE IN THE PERIPHERY AND CEREBROSPINAL FLUID (CSF) AT SINGLE CELL RESOLUTION AMONG PRECLINICAL-AD, MCI-AD, AND AD-DEMENTIA STAGES AND AMONG NORMAL AGING CONTROLS. FURTHERMORE, A NOVEL ASPECT OF THIS STUDY WILL BE TO CORROBORATE THE SEVERITY OF BLOOD BRAIN BARRIER (BBB) CHANGES ASSOCIATED WITH SPECIFIC IMMUNE CELL PROFILES NOTED IN CSF AND THE PERIPHERY ACROSS AD STAGES AND ITS IMPACT ON LONGITUDINAL COGNITIVE DECLINE. OUR OVERALL GOAL IS TO CHARACTERIZE IMMUNE CELL IDENTITY AND ABUNDANCE IN THE PERIPHERY AND THE CNS AT SINGLE CELL RESOLUTION ACROSS DIFFERENT AD CLINICAL STAGES AND IN TANDEM, TO CLARIFY THE PERIPHERAL ANATOMICAL CONTEXT OF IMMUNE CELL ACTIVATION AND THE DEGREE OF BBB CHANGES, WITH THE GOAL OF UNDERSTANDING THE TEMPORAL COURSE OF IMMUNE RESPONSE IN RELATION TO INITIATION OF CLINICAL DECLINE AND SUBSEQUENT PHENOTYPIC HETEROGENEITY OF AD TRAJECTORIES. TOWARDS THIS, WE WILL INVESTIGATE IMMUNE CELL PHENOTYPES IN THE CSF AND PERIPHERAL BLOOD BY A POWERFUL SINGLE-CELL PROTEOMIC ANALYSIS TECHNIQUE, MASS CYTOMETRY, AND BBB CHANGES WILL BE DELINEATED BY DYNAMIC CONTRAST-ENHANCED MRI TECHNIQUES IN THE SAME SUBJECTS. DATA INTEGRATING PERIPHERAL AND CSF ADAPTIVE AND INNATE IMMUNE CELL ACTIVATION ALONG WITH BBB CHANGES ACROSS DIFFERENT STAGES OF AD WILL HELP DEVELOP A CLEAR RATIONALE FOR THE USE OF THESE MARKERS IN THE AMYLOIDTAU(NEURODEGENERATION) FRAMEWORK. ADDITIONALLY, THESE INSIGHTS WILL HELP FUTURE THERAPEUTIC TARGETING OF SPECIFIC IMMUNE AND BBB CHANGES AT THE MOST EFFECTIVE CLINICAL STAGE OF DISEASE. THE RESULTS FROM THE STUDY WILL ALSO ENABLE THE IDENTIFICATION OF NOVEL MECHANISMS THAT REGULATE IMMUNE CELL HOMEOSTASIS IN THE PERIPHERY AND THE CNS, AND THE STATE OF BBB, PROVIDING POTENTIAL MEANS TO MANIPULATE THESE IMMUNE CELLS FOR THERAPEUTIC PURPOSES IN THE FUTURE.

DEVELOPMENT OF A MINIATURIZED, PEDIATRIC CONTINUOUS-FLOW TOTAL ARTIFICIAL HEART WITH A SINGLE MOVING PART

MECHANISMS OF ALLOANTIBODY PRODUCTION FOLLOWING RENAL TRANSPLANTATION - ABSTRACT ACUTE AND CHRONIC ANTIBODY-MEDIATED REJECTION (AMR) IS A SERIOUS THREAT TO THE SURVIVAL AND FUNCTION OF TRANSPLANTED ORGANS. THE CURRENT OPTIONS FOR AMR PREVENTION AND TREATMENT ARE LIMITED BY THE INCOMPLETE UNDERSTANDING OF THE MECHANISMS UNDERLYING DONOR SPECIFIC ALLOANTIBODY (DSA) GENERATION AND PATHOGENIC FUNCTIONS. WHEREAS THE PRODUCTION OF HIGH AFFINITY ISOTYPE-SWITCHED DSA IS TYPICALLY ASSOCIATED WITH GERMINAL CENTER FORMATION BY FOLLICULAR B CELLS, THE CONTRIBUTION OF MARGINAL ZONE (MZ) B CELLS TO ANTI-DONOR RESPONSES FOLLOWING TRANSPLANTATION HAS NOT BEEN PREVIOUSLY ADDRESSED. OUR PRELIMINARY STUDIES IDENTIFY MZ B CELLS AS IMPORTANT PLAYERS IN ORCHESTRATING DSA RESPONSES AND WARRANT DETAILED INVESTIGATION OF THIS B CELL SUBSET WITH AN ULTIMATE OBJECTIVE OF REDUCING HUMORAL ALLOIMMUNITY IN TRANSPLANT RECIPIENTS. PROLONGED COLD ISCHEMIA STORAGE (CIS) OF DONOR ALLOGRAFTS AND ENSUING ISCHEMIA/REPERFUSION INJURY (IRI) REMAIN AMONG LEADING RISK FACTORS FOR POOR TRANSPLANT OUTCOME. USING A MOUSE MODEL OF KIDNEY TRANSPLANTATION IN WHICH ALLOGRAFTS ARE SUBJECTED TO 6 H CIS, WE FOUND THAT POSTTRANSPLANT INFLAMMATION SPECIFICALLY AUGMENTS GENERATION OF CLASS II-REACTIVE DSA THAT MEDIATE ALLOGRAFT GLOMERULAR INJURY. THESE FINDINGS ARE HIGHLY RELEVANT TO CLINICAL STUDIES REVEALING CORRELATIONS BETWEEN LONGER COLD ISCHEMIA TIME, ANTI-CLASS II DSA AND LATE AMR IN RENAL TRANSPLANT PATIENTS. HOWEVER, THE MECHANISMS BY WHICH POSTTRANSPLANT INFLAMMATION AFFECTS GENERATION OF PATHOGENIC CLASS II DSA AND THE VERY SOURCE OF DONOR CLASS II ANTIGENS FOR B CELL ACTIVATION ARE POORLY DEFINED. BASED ON OUR PRELIMINARY DATA, WE HYPOTHESIZE THAT IRI AMPLIFIES CLASS II DSA PRODUCTION THROUGH THE FOLLOWING STEPS: 1) IRI UP-REGULATES MHC CLASS II EXPRESSION ON DONOR ENDOTHELIAL CELLS (EC) AND EC RELEASE OF CLASS II CONTAINING EXTRACELLULAR VESICLES (EEVS); 2) SPLEEN MZ B CELLS RAPIDLY ACQUIRE CIRCULATING EEVS, PRODUCE EARLY DSA AND FACILITATE FURTHER DSA PRODUCTION BY FO B CELLS; AND, 3) IN ADDITION TO DONOR ALLOANTIGENS, MZ B CELL ACTIVATION IS INITIATED AND ENHANCED BY DAMPS CARRIED BY GRAFT-DERIVED EVS AS WELL AS BY SYSTEMIC EFFECTS OF IRI. THEREFORE, TARGETING MZ B CELL TRAFFICKING, ACTIVATION AND FUNCTIONS WILL INHIBIT GENERATION OF PATHOGENIC CLASS II DSA AND IMPROVE OUTCOME OF RENAL ALLOGRAFTS SUBJECTED TO PROLONGED CIS. WE WILL TEST THIS HYPOTHESIS IN THREE SPECIFIC AIMS: AIM 1. TO TEST WHETHER ISCHEMIA/REPERFUSION INJURY (IRI) AUGMENTS CLASS II DSA BY ENHANCING ENDOTHELIAL EXTRACELLULAR VESICLES (EEV) GENERATION. AIM 2. TO TEST THE ROLE OF MZ B CELLS IN DSA PRODUCTION FOLLOWING RENAL TRANSPLANTATION. AIM 3. TO INVESTIGATE THE CONTRIBUTION OF MZ B CELLS IN THE GENERATION OF PATHOGENIC CLASS II DSA AFTER PROLONGED COLD ISCHEMIA STORAGE OF RENAL ALLOGRAFTS. THE PROPOSED STUDIES WILL FILL SEVERAL GAPS IN CURRENT KNOWLEDGE OF HUMORAL ALLOIMMUNE RESPONSES TO VASCULARIZED ORGAN TRANSPLANTS AND IDENTIFY POTENTIAL TARGETS OF THERAPEUTIC INTERVENTION TO INHIBIT ANTIBODY- MEDIATED REJECTION.

TARGETING TET DNA DIOXYGENASES AS THERAPEUTIC PRINCIPLE IN MYELOID NEOPLASMS - PROJECT SUMMARY/ABSTRACT TEN ELEVEN TRANSLOCATION (TET1, TET2 AND TET3) ARE A-KETOGLUTARATE (AKG) AND FE2+ DEPENDENT DNA- DIOXYGENASES THAT IS A KEY REGULATOR OF EPIGENETIC LANDSCAPE. THESE ENZYMES PROGRESSIVELY OXIDIZE 5- METHYLCYTOSINE TO 5-HYDROXYMETHYLCYTOSINE AND FURTHER TO 5-FORMYLCYTOSINE AND 5-CARBOXYLCYTOSINE IN DNA CULMINATING INTO DNA DEMETHYLATION ESSENTIAL FOR EFFICIENT TRANSCRIPTION OVER LARGE TIME SCALES. LOSS-OF-FUNCTION TET2 MUTATIONS (TET2MT), IS ONE OF THE MOST FREQUENT PATHOGENIC LESION IN MDS AND RELATED HEMATOLOGIC MALIGNANCIES IN HUMANS. TET2MT ARE FOUND IN ALL DISEASE STAGES AND LEVELS OF AGGRESSIVENESS. IN ADDITION, RECENT STUDIES HAVE DEMONSTRATED THAT SOMATIC TET2MT ARE VERY FREQUENTLY FOUND IN “HEALTHY” ELDERLY. THE PRESENCE OF TET2MT IN CHIP IMPLIES THAT IT IS AN EARLY EVENT IN THE CREATION OF HEMATOLOGIC DISORDERS. IT ALSO SUPPORTS MURINE STUDY CONCLUSIONS THAT TET2MT CAUSE EXPANSIONS OF HEMATOPOIETIC STEM AND PROGENITOR CELLS (HSPC). EARLY EVENTS IN THE EVOLUTION OF DEFECTIVE CLONES ARE RATIONAL TARGETS FOR PREVENTATIVE STRATEGIES SINCE CLONES IN SUCH STAGES ARE LIKELY TO BE DEPENDENT ON THESE EVENTS. THEIR PRESENCE IN ALL CELLS CAN, HOWEVER, ALSO BE EXPLOITED IN LATE STAGES OF THE DISEASE. FOR EXAMPLE, THE COMBINED LOSS OF TET1 AND TE2 IN HEMATOPOIETIC CELLS IN MURINE MODELS EXTENDS LIFE SUBSTANTIALLY RELATIVE TO TET2 LOSS ALONE. IN ADDITION, THE PROOF OF PRINCIPLE IS ALSO DERIVED FROM OBSERVATION IN PATIENTS WITH IDH1/2MT WHEREBY, THESE NEOMORPHIC MUTATIONS LEAD TO THE PRODUCTION OF A WEAK TET INHIBITOR, 2-HYDROXYGLUTYRATE (2-HG), KNOWN INHIBITOR OF DIOXYGENASES. OUR OBSERVATIONS THAT IDH1/2MT ARE MUTUALLY EXCLUSIVE WITH TET2MT, STRONGLY SUPPORTS OUR HYPOTHESES THAT THE TET INHIBITOR, 2-HG, PREVENTS EVOLUTION OF TET2MT CLONES. THIS OBSERVATION WAS FURTHER SUBSTANTIATED IN A CELLULAR MODEL OF MYELOID MALIGNANT CELLS AND SUPPORTS THAT SYNTHETIC LETHALITY CAN BE ACHIEVED THROUGH ELIMINATION OF REMAINING TET-ACTIVITY ESSENTIAL FOR EFFICIENT TRANSCRIPTION FOR PROLIFERATION IN TET-DIOXYGENASE DEFICIENT TET2MT CLONES. OUR OVERARCHING HYPOTHESIS IS THAT RATIONALLY DESIGNED AND SYNTHESIZED SMALL MOLECULES TETI76 CAN BE UTILIZED TO IMPEDE COMPENSATORY TET DIOXYGENASE ACTIVITY ORIGINATING FROM TET3 AND TET1, TO CAUSE EITHER SYNTHETIC LETHALITY OR LINEAGE REDIRECTION IN CASES WITH TET2MT INACTIVATION. OUR GOAL IS TO DEVELOP A NOVEL THERAPEUTIC APPROACH FOR TET2MT MDS BY EVALUATING THE POTENTIAL USE OF TETI76 AS TARGETED TREATMENTS IN PRECLINICAL MODELS. MORE SPECIFICALLY, WE AIM TO: I) STUDY THE MECHANISTIC CONSEQUENCES OF TET INHIBITION IN NORMAL AND MALIGNANT HEMATOPOIESIS IN VITRO. II) ESTABLISH THE EFFECTS OF TETI COMPOUNDS IN VITRO USING HUMAN AND MURINE CELLULAR MODELS AND III) CHARACTERIZE TETI PREVENTATIVE AND THERAPEUTIC EFFICACY AS WELL AS TOLERABILITY IN PRE-CLINICAL MURINE MODELS. OUR PROPOSAL, IF SUCCESSFUL, WILL LEAD TO A NOVEL CLASS OF THERAPEUTIC AGENTS FOR TET2MT ASSOCIATED HEMATOPOIETIC DISORDERS, AND PERHAPS ALSO OTHER DIOXYGENASE MUTATIONS. THIS WORK COULD ALSO HAVE IMPLICATIONS FOR CANCERS MUTATED IN HISTONE DEMETHYLASE KDM6A, WHICH IS SIMILAR TO TET2 AND FREQUENTLY MUTATED IN BLADDER CANCER.

MULTI-OMIC BIOMARKERS FOR NEUROPATHIC PAIN SECONDARY TO CHEMOTHERAPY - PROJECT SUMMARY TAXANES ARE AMONG THE MOST EFFICACIOUS CHEMOTHERAPEUTIC AGENTS AND ARE FREQUENTLY USED IN THE TREATMENT OF EARLY STAGE AND METASTATIC BREAST CANCER, BUT ARE KNOWN TO PRODUCE A PAIN CONDITION KNOWN AS CHEMOTHERAPY- INDUCED PERIPHERAL NEUROPATHIC PAIN (CIPNP). CIPNP IS ONE OF THE PRIMARY DOSE-LIMITING TOXICITIES OF TAXANE ADMINISTRATION. NO DIAGNOSTIC TOOL EXISTS TO IDENTIFY PATIENTS THAT WILL DEVELOP CIPNP IN RESPONSE TO TAXANE THERAPY. TO ADDRESS THIS UNMET CLINICAL NEED, THIS PROPOSAL SEEKS TO DEVELOP BIOMARKER SIGNATURES ASSOCIATED WITH TAXANE-INDUCED NEUROPATHIC PAIN TO 1) IDENTIFY PATIENTS AT RISK FOR DEVELOPING DEBILITATING TAXANE NEUROPATHIC PAIN BEFORE CHEMOTHERAPY IS INITIATED AND 2) TO IDENTIFY PATIENTS ON TREATMENT THAT ARE AT RISK OF DEVELOPING NEUROPATHIC PAIN AND NEED DOSE ADJUSTMENTS TO PREVENT THE ONSET OF SYMPTOMS. OUR LONG-TERM GOAL IS TO IDENTIFY BIOMARKER SIGNATURES THAT CAN IDENTIFY PATIENTS AT HIGH RISK OF DEVELOPING TAXANE-INDUCED CIPNP TO ENABLE PERSONALIZED DOSE ADJUSTMENTS TO MINIMIZE CIPNP AND OPTIMIZE THERAPEUTIC OUTCOMES. DURING THE R61 PHASE, WE WILL PERFORM GENOME-WIDE, EPIGENOME-WIDE, MIRNOME-WIDE, AND UNTARGETED METABOLOME-WIDE ASSOCIATIONS OF THE 11-POINT PAIN-INTENSITY NUMERICAL RATING SCALE (NRS), CHEMOTHERAPY-INDUCED PERIPHERAL NEUROPATHY 20- ITEM (CIPN20), THE PAIN CATASTROPHIZING SCALE (PCS), AND PATIENT REPORTED OUTCOME MEASUREMENT SYSTEM (PROMIS) SCORES FOR PHYSICAL FUNCTION, ANXIETY, DEPRESSION, AND PAIN INTERFERENCE TO IDENTIFY NOVEL BIOMARKERS ASSOCIATED WITH IDENTIFIABLE PHENOTYPES (AIM 1A). NEXT, WE WILL TEST PANELS OF CANDIDATE MRNA AND CYTOKINE BIOMARKERS FOR THEIR ASSOCIATION WITH NRS, CIPN20, PCS, AND PROMIS SCORES (AIM 1B). AN ADDED STRENGTH OF THIS STUDY IS THAT BY COLLECTING SAMPLES BEFORE, DURING, AND AFTER TAXANE TREATMENT WE WILL BE ABLE TO COMPARE THOSE PATIENTS WHO DEVELOP CIPNP TO THOSE THAT REMAIN PAIN FREE, PROVIDING A BETTER UNDERSTANDING OF THE MOLECULAR MECHANISM PREDISPOSING A PATIENT TO DEVELOPING NEUROPATHIC PAIN. R61 MILESTONES INCLUDE DEVELOPING BIOMARKER SIGNATURES THAT IDENTIFY PATIENTS AT RISK OF DEVELOPING NEUROPATHIC PAIN THAT COMPROMISE QUALITY OF LIFE (I) BEFORE TAXANE THERAPY IS STARTED AND (II) OVER THE TIME COURSE OF TAXANE THERAPY. ONE RATIONALE IS THAT SOME PATIENTS WILL HAVE A DETECTABLE BIOMARKER PROFILE SUGGESTING A HIGH LIKELIHOOD OF LONG TERM (I.E., GREATER THAN 1 YEAR) NEUROPATHIC PAIN AND A NEED TO EITHER AVOID OR REDUCE TAXANE DOSES. A SECOND RATIONALE IS THAT SOME PATIENTS MAY NOT HAVE A BIOMARKER SIGNATURE INDICATING AN ADVERSE RESPONSE TO TAXANE THERAPY AT BASELINE, BUT DEVELOP ONE THROUGH THE COURSE OF TAXANE THERAPY. THIS BIOMARKER SIGNATURE WILL BE USED TO DETECT THESE SUSCEPTIBLE PATIENTS EARLY AND PERSONALIZE THEIR TAXANE THERAPY TO MINIMIZE CIPNP. IN THE R33 PHASE (AIM 2), WE WILL USE MACHINE LEARNING TO DEVELOP AND VALIDATE ALGORITHMS USING BIOMARKER SIGNATURES FROM ASSOCIATIONS WITH PHENOTYPIC CHANGES FROM AIM 1 (A AND B) TO PREDICT INDIVIDUALS AT RISK OF DEVELOPING CIPNP AND TO IDENTIFY PATIENTS WHO ARE AT RISK OF DEVELOPING AN ADVERSE PROFILE AS TAXANE THERAPY IS ADMINISTERED.

CRYOPEN: AN INNOVATIVE TREATMENT FOR CERVICAL PRECANCER IN LOW-RESOURCE SETTING

RADIOMIC APPROACHES TO IMPROVE TARGETING FOR ATRIAL FIBRILLATION CATHETER ABLATION - PROJECT SUMMARY ALTHOUGH ABLATION TO ISOLATE PULMONARY VEIN (PV) TRIGGERS HAS REVOLUTIONIZED ATRIAL FIBRILLATION (AF) MANAGEMENT, PERFORMING EFFECTIVE AF ABLATION REMAINS CHALLENGING. THE PROCEDURE REMAINS LIMITED BY TARGETING OF ILL-DEFINED SUBSTRATES, A 2-6% RISK OF MAJOR COMPLICATIONS AND LIMITED SUCCESS (SINGLE PROCEDURE 5-YEAR SUCCESS AS LOW AS 17-56%; 63-81% AFTER THE LAST ABLATION). A MAJOR RECOMMENDATION OF A RECENT NHLBI-SPONSORED REPORT ON THE RESEARCH NEEDS AND PRIORITIES FOR AF CATHETER ABLATION WAS TO STUDY HOW CARDIAC STRUCTURE AFFECTS AF ABLATION SUCCESS. THERE IS A CLEAR UNMET NEED FOR NON-INVASIVE IMAGING TOOLS TO AID IN IMPROVED PATIENT SELECTION, ANATOMIC TARGETING AND PERSONALIZATION OF ABLATION OR MEDICAL THERAPIES. OUR TEAM HAS DEVELOPED NOVEL COMPUTATIONAL IMAGING (RADIOMICS) METHODS TO ANALYZE CARDIAC COMPUTED TOMOGRAPHY (CT) SCANS THAT WERE SHOWN TO PREDICT THE RISK OF RECURRENT AF POST-ABLATION (AUC=0.84, N=167). THESE APPROACHES INCLUDED NOVEL MORPHOLOGIC, FRACTAL AND ATLAS BASED FEATURES THAT TEASED OUT DIFFERENCES BETWEEN PVS AND THE LEFT ATRIAL APPENDAGE (LAA), SOLELY FROM ANALYSES OF CT SCANS. WE PROPOSE TO BUILD UPON OUR PRELIMINARY DATA USING RADIOMIC (COMPUTER EXTRACTED) FEATURES FROM RADIOGRAPHIC IMAGES TO USE SUPERVISED AND UNSUPERVISED MACHINE LEARNING METHODS THAT CAN ANALYZE DIGITIZED RADIOGRAPHIC AND ELECTRO-ANATOMIC IMAGES FROM THE LEFT ATRIUM (LA) AND PVS IN OVER 2000 PATIENTS FROM TWO LARGE AF ABLATION CENTERS (CLEVELAND CLINIC, VANDERBILT). OUR PROJECT WILL FOCUS ON TACKLING THE FOLLOWING MAIN OBJECTIVES: 1) IDENTIFY, EVALUATE AND VALIDATE RADIOMIC FEATURES AND IMAGING-CLINICAL NOMOGRAMS PREDICTIVE OF RECURRENT AF AFTER ABLATION; 2) IDENTIFY AND VALIDATE REGIONAL RADIOMIC SITES PREDICTIVE OF POST-ABLATION AF RECURRENCE WITH THE GOAL OF IDENTIFYING PERSONALIZED TARGETS FOR PATIENTS UNDERGOING AF ABLATION; AND 3) IDENTIFY BIOLOGICAL CORRELATES OF RADIOMIC FEATURES TO UNDERSTAND THE ARRHYTHMOGENIC MECHANISMS UNDERLYING ANATOMIC SUSCEPTIBILITY TO RECURRENT AF, USING GENOMIC ANALYSES. OUR 3 AIMS WILL TEST THE FOLLOWING HYPOTHESES: 1) RADIOGRAPHIC IMAGING CAN DETECT ANATOMIC FEATURES THAT PREDICT AF RECURRENCE AFTER ABLATION; 2) REGIONAL RADIOMIC FEATURES CAN PREDICT SITES THAT CAN BE CONSIDERED FOR ADDITIONAL ABLATION; AND 3) RADIOMIC MORPHOLOGIC FEATURES ARE CORRELATED WITH ELECTROANATOMIC FEATURES AND GENOMIC VARIANTS ASSOCIATED WITH AF SUSCEPTIBILITY. TOOLS DEVELOPED WILL ENABLE INTEGRATION OF RADIOGRAPHIC AND CLINICAL DATA THAT MAY LEAD TO IMPROVED PATIENT SELECTION, ANATOMIC TARGETING AND PERSONALIZATION OF ABLATION OR MEDICAL THERAPIES. SUCCESSFUL PROJECT COMPLETION WILL YIELD A NOVEL ARTIFICIAL INTELLIGENCE-BASED IMAGING PLATFORM THAT CAN BE TESTED FOR PERSONALIZED TARGETING OF AF ABLATION, AS WELL AS INSIGHTS INTO THE BIOLOGIC BASIS OF AF.

ROLES OF SOX8 AND SOX9 IN ADULT ARTICULAR CARTILAGE

THE ROLE OF HIPPOCAMPAL NEUROGENESIS IN ALCOHOL WITHDRAWAL SEIZURE AND COGNITION

ALCOHOLIC HEPATITIS CLINICAL AND TRANSLATIONAL NETWORK - LATE PHASE CLINICAL TRIALS AND OBSERVATIONAL STUDIES (COLLABORATIVE U01)

SINGLE VISIT CLINICAL VALIDATION OF SCREENFIRE, A LOW-COST HPV TEST: EFFICACY AND COST EFFECTIVENESS (SCALE) - PROJECT SUMMARY/ABSTRACT IN 2018, THE WORLD HEALTH ORGANIZATION (WHO) CALLED FOR ACTION TOWARDS ACHIEVING THE GLOBAL ELIMINATION OF CERVICAL CANCER. A STRATEGY FOR ACHIEVING THIS GOAL WAS RATIFIED BY MEMBER STATES IN AUGUST 2020. THE WHO PLAN CALLS FOR AN AGGRESSIVE APPROACH OF VACCINATION, SCREENING, AND TREATMENT OF THE HUMAN PAPILLOMAVIRUS (HPV), THE SINGLE CAUSE OF CERVICAL CANCER. IN LOW- AND MIDDLE-INCOME COUNTRIES (LMIC), WHICH BEAR 90% OF THE INCIDENCE AND MORTALITY OF CERVICAL CANCER GLOBALLY, IT IS ESTIMATED THAT THESE GOALS WILL NOT BE REACHED UNTIL 2120 – A CENTURY FROM NOW. ONE WAY TO MARKEDLY SHORTEN THIS TIMELINE IS THROUGH HPV SELF-SAMPLING AND IMMEDIATE TREATMENT FOR THOSE WHO SCREEN POSITIVE. IN ORDER TO SCALE-UP HPV THERE MUST BE A LOW-COST, EFFECTIVE TEST THAT IS EASY TO USE IN THE FIELD. AN OPTIMAL FIELD TEST WOULD ALLOW WOMEN TO COLLECT THEIR OWN SAMPLES WITHOUT COMPROMISING EFFICACY. THE ONLY WHO PREQUALIFIED TEST SPECIFICALLY DEVELOPED FOR LOW RESOURCE SETTINGS IS CAREHPV, WHICH HAS RELATIVELY GOOD SENSITIVITY WHEN A SAMPLE IS TAKEN BY A CLINICIAN, BUT WHICH PERFORMS POORLY WHEN SELF-SAMPLED. THERE IS AN URGENT NEED FOR EVALUATION OF A LOW-COST SELF- SAMPLING TEST THAT IS EFFECTIVE AND CAN BE EASILY IMPLEMENTED IN LMIC. AMPFIRE® (ATILA BIOSYSTEMS, CA), A NEW LOW-COST HPV TEST, HAS DEMONSTRATED HIGH SENSITIVITY IN SELF-COLLECTED SAMPLES AND MAY PERFORM BETTER THAN PROVIDER-COLLECTED CAREHPV. FURTHERMORE, THE CURRENT PROTOCOL FOR CAREHPV REQUIRES A 2-DAY VISIT BECAUSE THE MACHINES ARE IN A CENTRAL LOCATION AND REQUIRE 3-4 HOURS FOR PROCESSING. AMPFIRE® CAN BE RUN FROM LOCAL LOCATIONS USING A BATTERY-OPERATED PCR MACHINE. SINCE THESE TESTS PROCESS IN APPROXIMATELY 1 HOUR, A SINGLE- VISIT APPROACH IS FEASIBLE, WHICH IS MORE EFFICIENT FOR THE HEALTH SYSTEM AND MORE CONVENIENT FOR PATIENTS. THE PURPOSE OF THIS STUDY IS TO COMPARE THE EFFICACY OF AMPFIRE® VERSUS CAREHPV FOR THE DETECTION OF HIGH GRADE PRECANCER (CERVICAL INTRAEPITHELIAL NEOPLASIA, GRADE 2 OR HIGHER, OR CIN2+) AND TO COMPARE THE EFFICACY AND FEASIBILITY OF EACH TEST THROUGH THE FOLLOWING AIMS: AIM 1: TO COMPARE SELF-COLLECTED AMPFIRE® TO STANDARD PROVIDER-COLLECTED CAREHPV FOR THE DETECTION OF HIGH GRADE CERVICAL PRECANCER. AIM 2: DEMONSTRATE THAT A SINGLE-VISIT APPROACH USING SELF-COLLECTED AMPFIRE® WILL RESULT IN LOWER LOSS TO FOLLOW-UP THAN THE CURRENT CONVENTIONAL TWO-VISIT APPROACH USING PROVIDER-COLLECTED CAREHPV. AIM 3: PERFORM A COST- EFFECTIVENESS ANALYSIS TO COMPARE EFFICACY, LOSS TO FOLLOW-UP AND GENERALIZABILITY OF SELF-SAMPLED AMPFIRE® HPV VERSUS PROVIDER-COLLECTED CAREHPV.

MECHANISMS OF METABOLIC DYSREGULATION IN PULMONARY HYPERTENSION

TICKBORNE SFTS VIRUS VACCINE DEVELOPMENT

MECHANISMS OF CANCER IMMUNOTHERAPY-ASSOCIATED THROMBOSIS - CANCER IMMUNOTHERAPY IS ONE OF THE MOST IMPORTANT ADVANCES IN CANCER TREATMENT IN DECADES, AND HAS RAPIDLY MOVED TO FRONT-LINE THERAPY FOR MANY CANCERS. THE MECHANISM OF CANCER IMMUNOTHERAPY IS TO DISABLE NORMAL IMMUNOREGULATORY PATHWAYS THROUGH ADMINISTRATION OF IMMUNE CHECKPOINT INHIBITORS (ICI), WHICH ARE MONOCLONAL ANTIBODIES DIRECTED TOWARD KEY IMMUNE REGULATORY PROTEINS INCLUDING PD-1, PD-L1 AND CTLA-4. DISABLING THESE PATHWAYS ENHANCES ANTI-TUMOR IMMUNITY. HOWEVER, SINCE THESE RESPONSES ARE NOT TUMOR-SPECIFIC, ICIS ARE ASSOCIATED WITH A VARIETY OF IMMUNE-RELATED ADVERSE EVENTS (IRAES). WE AND OTHERS HAVE RECENTLY REPORTED A HIGH INCIDENCE OF THROMBOSIS, WHICH MAY EXCEED 20%, IN PATIENTS TREATED WITH ICI; GIVEN THE INCREASING USE OF ICIS IN CANCER TREATMENT AND THE FREQUENCY OF CANCER DIAGNOSES, IT IS CLEAR THAT ICI-CANCER ASSOCIATED THROMBOSIS (ICI-CAT) HAS BECOME A MAJOR CLINICAL PROBLEM AND THAT BETTER UNDERSTANDING OF THIS DISORDER IS URGENTLY NEEDED. HOWEVER, THERE IS LITTLE INFORMATION AVAILABLE CONCERNING MECHANISMS OF ICI-CAT, AND THERE ARE NO PUBLISHED STUDIES ADDRESSING THIS ISSUE. WE HYPOTHESIZE THAT ICI-CAT IS AN IRAE RESULTING FROM ICI-INDUCED CELLULAR ACTIVATION AND PROTHROMBOTIC ACTIVITY IN THE SETTING OF UNDERLYING TUMOR-ASSOCIATED INFLAMMATION. OUR MURINE MODEL DEMONSTRATES THAT ICI-CAT REQUIRES THE PRESENCE OF AN UNDERLYING TUMOR, WITH MARKEDLY INCREASED EXPRESSION OF TUMOR CELL TISSUE FACTOR (TF) OCCURRING AFTER ICI TREATMENT. OUR MODEL ALSO SUPPORTS A ROLE FOR NEUTROPHIL EXTRACELLULAR TRAPS (NETS) AND PLATELET ACTIVATION IN ICI-CAT; PLATELET ACTIVATION IS ALSO SUGGESTED IN PATIENTS TREATED WITH ICI BY OUR DEMONSTRATION OF INCREASED LEVELS OF CIRCULATING PLATELET-NEUTROPHIL AGGREGATES. ON A CELLULAR LEVEL, ANTI-PD-1 ANTIBODIES STIMULATE NEUTROPHIL NET RELEASE AND PROTHROMBOTIC ACTIVITY, AND MAY ALSO ENHANCE PLATELET ACTIVATION IN THE PRESENCE OF SUBTHRESHOLD THROMBIN CONCENTRATIONS. IN THIS APPLICATION, WE PROPOSE TO ADVANCE OUR UNDERSTANDING OF ICI-CAT USING BOTH CELLULAR AND ANIMAL MODELS, AND TO EXTEND THESE STUDIES TO CLINICAL SAMPLES FROM PATIENTS BEFORE AND AFTER INITIATING TREATMENT WITH ICI. IN AIM 1, WE WILL DETERMINE THE EFFECT OF DIFFERENT ICI AND ICI COMBINATIONS ON THE DEVELOPMENT OF THROMBI IN TUMOR-BEARING MICE, AND ASSESS OUR MODEL USING OTHER TUMOR TYPES AND MOUSE STRAINS. WE WILL EXAMINE THROMBUS SIZE AND COMPOSITION, AND IDENTIFY CRITICAL CELL TYPES INVOLVED IN THROMBUS FORMATION BY DEPLETING MICE OF T CELLS, NEUTROPHILS, MONOCYTES OR PLATELETS. WE WILL ALSO FURTHER DEFINE THE CELLULAR MECHANISMS INVOLVED IN EXPRESSION OF PROTHROMBOTIC ACTIVITY USING ISOLATED LEUKOCYTES FROM NORMAL HUMAN DONORS, MIXED LEUKOCYTE POPULATIONS, AND ENDOTHELIAL CELLS, INCUBATED WITH CYTOKINES AND ICI. IN AIM 2, WE WILL EXTEND THESE STUDIES BY MEASURING MECHANISTIC MARKERS OF INFLAMMATION AND VASCULAR ACTIVATION IN PATIENT PLASMA BEFORE AND AFTER INITIATING ICI, AND BY COMPARING PROCOAGULANT GENE EXPRESSION IN MYELOID CELLS FROM PATIENTS TREATED WITH ICI WHO DID, OR DID NOT DEVELOP THROMBOSIS. TAKEN TOGETHER, THESE STUDIES SHOULD PROVIDE IMPORTANT NEW INFORMATION LEADING TO BETTER UNDERSTANDING OF MECHANISMS, AND POTENTIALLY NEW APPROACHES FOR PREVENTION AND TREATMENT OF ICI-CAT.

IMAGING ASSESSMENTS OF ARPKD KIDNEY DISEASE PROGRESSION

MODELING AND FORECASTING ATHEROSCLEROTIC RISK: A COMPLEX SYSTEMS APPROACH

ELUCIDATING AND TARGETING THE MOLECULAR FOUNDATIONS OF IDH MUTANT GLIOMA

DISCOVERING THE ROLE OF YES1 IN TRIPLE NEGATIVE BREAST CANCER - PROJECT SUMMARY/ABSTRACT SRC FAMILY KINASES (SFKS) ARE A GROUP OF 9 NON-RECEPTOR TYROSINE KINASES THAT MEDIATE THE EFFECTS OF MANY EXTRACELLULAR AND INTRACELLULAR SIGNALING PATHWAYS. WHILE THESE PROTEINS HAVE OFTEN BEEN CONSIDERED TO BE FUNCTIONALLY SIMILAR TO SRC, THE FOUNDING MEMBER OF THE FAMILY, IT IS NOW CLEAR THAT THEY HAVE NON-REDUNDANT AND UNIQUE ACTIVITIES. IN CANCER, THESE PROTEINS ARE DIFFERENTIALLY EXPRESSED AND THEIR ROLES CAN EVEN BE ANTAGONISTIC. THIS PROJECT LAUNCHES FROM OUR DISCOVERY THAT ONE OF THESE FAMILY MEMBERS, YES1, IS SELECTIVELY OVEREXPRESSED IN TRIPLE NEGATIVE BREAST CANCER (TNBC), IS ASSOCIATED WITH POOR OUTCOMES, AND IS NECESSARY FOR SUSTAINED GROWTH OF TNBC CELLS. TNBC IS A COLLECTION OF HIGHLY AGGRESSIVE DISEASES WITH LIMITED THERAPEUTIC OPTIONS PRIMARILY INVOLVING CYTOTOXIC CHEMOTHERAPY. WHILE MANY PATIENTS INITIALLY RESPOND TO THESE TREATMENTS, RESISTANCE IS COMMON, RESULTING IN POOR PATIENT OUTCOMES. THUS, IDENTIFYING VULNERABILITIES IN THIS GROUP OF DISEASES IS ESSENTIAL TO YIELD APPROACHES FOR IMPROVING SURVIVORSHIP. WE PROPOSE THAT YES1 IS ONE OF THESE VULNERABILITIES. IN PRELIMINARY DATA, WE SHOW THAT YES1 IS ESSENTIAL FOR MAINTAINING EXPRESSION OF EPIDERMAL GROWTH FACTOR RECEPTOR (EGFR) AS WELL AS ENSURING MITOTIC FIDELITY. EGFR IS A MAJOR DRIVER OF TNBC GROWTH. WHEN YES1 IS DEPLETED, EGFR IS DEGRADED AND ITS SIGNALING IS LOST. IN ADDITION, WE REPORT THAT YES1 SILENCING CAUSES SEVERAL NUCLEAR DEFECTS INCLUDING MICRO-, MULTI-, AND DYSMORPHIC NUCLEI INDICATING THAT YES1 IS ESSENTIAL FOR ACCURATE COMPLETION OF MITOSIS. WE HYPOTHESIZE THAT THESE TWO FUNCTIONS OF YES1 ARE ESSENTIAL FOR CELLULAR VIABILITY IN TNBC. MOREOVER, WE PROPOSE THAT YES1 MAY BE A USEFUL THERAPEUTIC TARGET TO IMPROVE THE EFFICACY OF DRUGS TARGETING EGFR AND MITOSIS. IN AIM 1, WE WILL DETERMINE HOW YES1 CONTROLS EGFR DEGRADATION AND ASSESS WHETHER YES1 OVEREXPRESSION UNDERLIES RESISTANCE TO EGFR INHIBITORS IN TNBC. AIM 2 WILL FOCUS ON IDENTIFYING SPECIFIC EVENTS IN MITOSIS THAT ARE IMPACTED BY YES1 AND THE MECHANISMS INVOLVED. IT WILL ALSO DISCERN WHETHER MODULATING YES1 CAN IMPACT RESPONSE TO TAXANES, MAINSTAY CHEMOTHERAPIES FOR TNBC. LASTLY, IN AIM 3, WE WILL DETERMINE IF EGFR AND YES1 PARTICIPATE IN A FEEDFORWARD LOOP THAT CONTROLS MITOSIS. THIS WILL INVOLVE INTERROGATING THE ROLE OF EGFR IN MEDIATING THE EFFECTS OF YES1 ON MITOSIS AND DETERMINING IF EGFR IS ALSO AN UPSTREAM REGULATOR OF YES1. MAJOR INNOVATIONS OF THIS PROJECT INCLUDE THE IDENTIFICATION OF YES1 AS A NEW ONCOGENIC DRIVER OF TNBC, THE DISCOVERY THAT YES1 AND EGFR MAY CONTROL THE GENOMIC COMPLEXITY THAT IS ASSOCIATED WITH THIS DISEASE, THE THERAPEUTIC ASSESSMENT OF A NOVEL INHIBITOR OF YES1 IN PDX MODELS OF TNBC, AND THE POTENTIAL FOR IMPROVING THE EFFICACY OF EGFR INHIBITORS AND TAXANES IN A DISEASE THAT REQUIRES NEW THERAPEUTIC APPROACHES TO IMPROVE PATIENT OUTCOMES.

INTRACELLULAR FUNCTIONS OF APOL1 IN THE KIDNEY - ABSTRACT CHRONIC KIDNEY DISEASE (CKD) IN AFRICAN AMERICANS IS ONE OF THE LARGEST RACIAL HEALTH DISPARITIES IN THE UNITED STATES. THE CAUSE FOR THE INCREASED RISK HAS BEEN ATTRIBUTED TO RECESSIVE INHERITANCE OF ALLELIC VARIANTS IN THE GENE FOR APOLIPOPROTEIN L1 (APOL1). THESE APOL1 VARIANTS, KNOWN AS G1 AND G2, DO NOT CAUSE CKD ON THEIR OWN, BUT CKD IS CAUSED BY A COMBINATION OF THE INHERITED GENETIC RISK PLUS EXPOSURE TO A TRIGGERING ENVIRONMENTAL STRESSOR (A GENE-ENVIRONMENT INTERACTION). DESPITE THE ASSOCIATION OF CKD RISK WITH APOL1 VARIANTS MORE THAN TEN YEARS AGO, THE BIOLOGICAL FUNCTION OF APOL1 IN THE KIDNEY AND THE MECHANISM OF PATHOGENESIS IN THE SETTING OF A DISEASE STRESSOR REMAIN UNCLEAR. OUR LONG-TERM GOAL IS TO UNDERSTAND THE GENETICS AND BIOCHEMICAL MECHANISM OF CKD IN AFRICAN AMERICANS CAUSED BY THESE APOL1 POLYMORPHISMS. TO ACCOMPLISH THIS GOAL, WE ARE STUDYING HIV-ASSOCIATED NEPHROPATHY (HIVAN), THE CKD MOST STRONGLY ASSOCIATED APOL1 VARIANTS, AND THE ONLY CKD WHERE THE ENVIRONMENTAL STRESSOR IS KNOWN (HIV INFECTION). HIVAN IS AN IDEAL DISEASE MODEL TO DISSECT BIOCHEMICAL PATHWAYS AND CELLULAR EVENTS INTERSECTED BY APOL1 FUNCTION, VIRAL INFECTION, AND CKD. OUR RECENT STUDIES HAVE DEMONSTRATED, FOR THE FIRST TIME, A FUNCTION FOR THE COMMON APOL1 ALLELE, KNOWN AS G0, IN PROVIDING PROTECTION AGAINST PODOCYTE LOSSES IN HIVAN. SINCE APOL1 RISK IS A RECESSIVELY INHERITED TRAIT, THIS SUGGESTS CKD MAY BE CAUSED, IN PART, BY A LOSS-OF-FUNCTION PROCESS (I.E. ABSENCE OF G0). IN NEW PRELIMINARY DATA, G0 APPEARS TO ASSOCIATE WITH TOLL-LIKE RECEPTORS (TLRS) IN INTRACELLULAR VESICLES CONTAINING HIV AND FACILITATE SIGNALING EVENTS INITIATED BY INTERFERON REGULATORY FACTOR (IRF)-3, AND THESE PROCESSES WERE ABSENT WITH THE APOL1 RISK VARIANTS. METHODS WILL USE ESTABLISHED IN VITRO HIV INFECTION OF HUMAN PODOCYTE CELL LINES, IN VIVO MODELS OF HIVAN AND BAC-APOL1 TRANSGENIC MICE THAT REPLICATE ENDOGENOUS HUMAN APOL1 EXPRESSION. AIMS WILL EXAMINE BOTH INITIAL RESPONSE TO HIV INFECTION IN PODOCYTES, AND LONG-TERM IN VIVO STUDIES OF INTERCROSSES BETWEEN THE HIVAN AND BAC-APOL1 TRANSGENIC MICE EVALUATING EFFECTS ON RENAL FUNCTION AND PATHOLOGY. THESE STUDIES ALSO WILL ESTABLISH TEMPORAL AND MAGNITUDE OF STRESSOR-INDUCED APOL1 EXPRESSION, THE MECHANISM OF THE ALTERED INNATE IMMUNE ACTIVATION THROUGH TLRS TO IRF-3/7 SIGNALING, AND EFFECT ON PODOCYTE PHENOTYPE (SURVIVAL AND CELL ADHESION). THE DOMINANCE OF THE G0 PROTECTIVE EFFECT OVER THE RISK VARIANT DYSFUNCTION WILL BE TESTED IN PODOCYTES AND MOUSE MODELS CO-EXPRESSING G0 AND THE RISK VARIANTS. THESE STUDIES SHOULD ADVANCE OUR UNDERSTANDING OF GAIN- VERSUS LOSS-OF-FUNCTION MECHANISM ASSOCIATED WITH THE RECESSIVE INHERITANCE OF APOL1 RISK ALLELES, AND THE NECESSITY OF INDUCED APOL1 EXPRESSION TO DRIVE STRESS RESPONSES. DETERMINING THE CONTRIBUTION OF G0 FUNCTION VERSUS RISK VARIANT DYSFUNCTION WILL HAVE IMPORTANT CLINICAL IMPACT ON FURTHER THERAPY DESIGN, AS IT WILL ESTABLISH WHETHER REPLACEMENT OF G0 OR SUPPRESSION OF THE RISK VARIANTS WOULD BE THE MORE EFFECTIVE STRATEGY.

REGULATION OF HOST INNATE IMMUNITY AGAINST VIRAL INFECTION - RIG-I AND MDA5 ARE KEY CYTOSOLIC RECEPTORS FOR SENSING VARIOUS RNA VIRUSES, INCLUDING CORONAVIRUSES, INFLUENZA VIRUS, AND FLAVIVIRUSES. UPON BINDING OF VIRAL RNA, RIG-I-LIKE RECEPTORS INITIATE SIGNAL TRANSDUCTION PATHWAYS THAT RESULT IN TYPE-I INTERFERON (IFN) INDUCTION TO SUPPRESS VIRAL REPLICATION AND THE SPREAD OF INFECTION. FOR THEIR EFFICIENT REPLICATION, HOWEVER, VIRUSES HAVE EVOLVED ELEGANT STRATEGIES TO EVADE THE IFN-MEDIATED HOST IMMUNE RESPONSE. WORK FROM MANY LABORATORIES HAS IDENTIFIED THE PRECISE MECHANISMS THAT LEAD TO ACTIVATION OF RIG-I, WHICH INCLUDE RNA BINDING, K63-LINKED UBIQUITINATION, OLIGOMERIZATION, AND THE RE-DISTRIBUTION OF RIG-I FROM THE CYTOPLASM TO MITOCHONDRIA. IN STRIKING CONTRAST, OUR KNOWLEDGE ABOUT THE MECHANISMS THAT GOVERN ACTIVATION OF THE RELATED SENSOR MDA5 REMAINS ELUSIVE. THE PROPOSED STUDY BUILDS ON A RECENT DISCOVERY BY THE GACK LABORATORY THAT COVALENT MODIFICATION OF MDA5 BY ISG15 ACTIVATES MDA5 SIGNALING BY PROMOTING ITS CARD-DEPENDENT OLIGOMERIZATION AND CYTOSOL-TO-MITOCHONDRIA TRANSLOCATION. MOLECULAR AND CELL BIOLOGICAL STUDIES IN ISG15-DEFICIENT HUMAN AND MOUSE CELLS, INCLUDING PRIMARY IMMUNE CELLS, DEMONSTRATED THAT ISG15 IS REQUIRED FOR AN EFFECTIVE ANTIVIRAL IFN RESPONSE MEDIATED BY MDA5 (BUT NOT RIG-I). OUR DATA ALSO SHOWED THAT MDA5 ISGYLATION IS REQUIRED FOR RESTRICTING THE INFECTION OF SARS-COV-2, ZIKA, DENGUE AND PICORNAVIRUSES. FURTHERMORE, OUR MOST RECENT WORK REVEALED THAT SARS-COV- 2 USES ITS PAPAIN-LIKE PROTEASE (PLPRO) TO ANTAGONIZE MDA5 BY ACTIVELY REMOVING ITS ISGYLATION. MOLECULAR, BIOCHEMICAL, AND CELL BIOLOGICAL APPROACHES COMBINED WITH GENE-TARGETING SCREENS WILL FOCUS ON DEFINING IN PRECISE DETAIL HOW ISGYLATION REGULATES MDA5-MEDIATED INNATE IMMUNITY, AND ALSO IDENTIFY THE E3 LIGASE THAT MEDIATES MDA5 ISGYLATION AND THEREBY ITS ACTIVATION. WE WILL ALSO DETERMINE THE RELEVANCE OF MDA5 ISGYLATION FOR EFFECTIVE VIRUS RESTRICTION USING MDA5 GENE-EDITED CELLS USING CRISPR-CAS9 TECHNOLOGY, AND A NOVEL MUTANT MDA5 KNOCK-IN MOUSE MODEL. THIS STUDY WILL ALSO GIVE DETAILED INSIGHT INTO THE MECHANISMS BY WHICH SARS-COV-2 AND OTHER CORONAVIRUSES COUNTERACT MDA5 ISGYLATION AND DETERMINE THE PHYSIOLOGICAL ROLE OF THIS NOVEL IMMUNE ESCAPE STRATEGY IN IFN ANTAGONISM, VIRUS REPLICATION AND PATHOGENESIS. OUR STUDIES WILL PROVIDE A MOLECULAR UNDERSTANDING OF THE MECHANISMS THAT LEAD TO ACTIVATION OF ANTIVIRAL INNATE IMMUNITY, AND ALSO DEFINE A NOVEL IMMUNE EVASION STRATEGY OF CORONAVIRUSES, WHICH MAY PROVIDE THE FOUNDATION FOR NEW THERAPEUTIC APPROACHES OR HELP DESIGN BETTER VACCINES.

NOVEL MECHANISM BASED TREATMENT TO IMPROVE TISSUE INJURY IN ALCOHOLIC HEPATITIS

PRIMARY IMMUNOPREVENTION OF TRIPLE-NEGATIVE BREAST CANCER

IMPROVING INTRACORTICAL CONTROL OF REACHING AFTER PARALYSIS

IMPACT OF DISPARITIES AFFECTING SURVIVAL AMONG ADVANCED HEART FAILURE PATIENTS AND TRANSPLANT RECIPIENTS

SYSTEMIC NEUROTROPIC VIRUS INFECTION EFFECTS ON GI DYSMOTILITY

A PLACE-BASED APPROACH TO GEOGRAPHIC DISPARITIES IN LUNG TRANSPLANT - PROJECT SUMMARY WHILE LUNG TRANSPLANT IS A LIFESAVING SURGERY FOR PATIENTS WITH FATAL LUNG DISEASES, THERE REMAINS A CRITICAL LIMITATION AS MEDIAN POST-TRANSPLANT SURVIVAL IS MERELY 5.5 YEARS WHICH IS ONLY HALF THE SURVIVAL EXPERIENCED BY OTHER ORGAN TRANSPLANT TYPES. UNFORTUNATELY, THIS LOW SURVIVAL IS FROM THE ACCELERATED DETERIORATION IN LUNG FUNCTION IN THE YEARS AFTER TRANSPLANT AND CANNOT BE FULLY EXPLAINED BY TRANSPLANT CENTER PRACTICES, DONOR FACTORS, OR RECIPIENT FACTORS. TROUBLINGLY, THERE IS ALSO DISPARATE SURVIVAL BY GEOGRAPHY WHICH VARIES UP TO 39% DEPENDING ON THE REGION OF THE U.S. WE HYPOTHESIZE THAT PLACE, DEFINED AS A SOCIAL AND ENVIRONMENTAL LOCATION WITH MEANING TO A PERSON, IMPACTS RECIPIENT HEALTH AND IN A MECHANISTIC HYPOTHESIS THESE PLACE-BASED FACTORS CAUSE SUBCLINICAL MICRO-INJURIES TO THE LUNGS WHICH ACCELERATE LUNG FUNCTION LOSS IN THE YEARS AFTER TRANSPLANT. THERE IS A GAP IN KNOWLEDGE ON HOW PLACE-BASED FACTORS EXACERBATE BOTH THE GEOGRAPHICALLY DISPARATE AND POOR SURVIVAL OF LUNG TRANSPLANT RECIPIENTS. IN THE ABSENCE OF THIS KNOWLEDGE, IT IS DIFFICULT TO PRECISELY RISK STRATIFY RECIPIENTS AND DEVELOP PATIENT AND POLICY LEVEL INTERVENTIONS. TO ADDRESS THIS GAP, WE LEVERAGE THE GEOGRAPHIC INFORMATION SCIENCES, DEFINED AS THE FRAMEWORK TO MEASURE, MAP, AND MODEL THE EFFECTS OF PLACE. THE SCOPE OF AIM 1 IS TO INFORM CLINICAL DECISIONS BY GEOCODING A MULTICENTER COHORT TO IDENTIFY INDIVIDUAL RECIPIENT LEVEL CENSUS TRACTS. THIS ENABLES LINKAGE TO HIGHLY GRANULAR FEDERAL DATASETS WITH A WIDE ARRAY OF SOCIAL AND ENVIRONMENTAL HEALTH MEASURES FOLLOWED BY THE APPLICATION OF MULTILEVEL MODELS AND ESTABLISHED SPATIAL CLUSTER DETECTION METHODS. THIS IS SIGNIFICANT AS THESE RESULTS WOULD ENABLE US TO PIVOT AWAY FROM AN EXISTING ONE-SIZE-FITS-ALL CLINICAL APPROACH BY SCREENING EARLIER FOR WORSENING LUNG FUNCTION OR TAILORING IMMUNOSUPPRESSION MEDICATIONS TO PREVENT LUNG FUNCTION LOSS FOR AT RISK PATIENTS. THE SCOPE OF AIM 2 IS TO INFORM POLICY THROUGH MAPPING DISPARITIES ACROSS THE U.S. AND IMPROVING THE ACCURACY OF ESTABLISHED CLINICAL PREDICTION MODELS. WE WILL MERGE THE SINGULAR NATIONAL TRANSPLANT REGISTRY WITH HIGHLY VALUED FEDERAL DATA MEASURING SOCIAL AND ENVIRONMENTAL HEALTH FACTORS, AND TEST THE INCLUSION OF PLACE-BASED FACTORS ON THE PERFORMANCE OF NOVEL MULTILEVEL MODELS AND ESTABLISHED CLINICAL COX MODELS. NEW MAPS AND MORE ACCURATE MODELS WOULD BE A SIGNIFICANT ADVANCE TOWARDS IDENTIFYING AND REDUCING GEOGRAPHIC DISPARITIES AND IMPROVING OUTCOMES THROUGH TARGETED RESOURCE ALLOCATION. OUR LONG-TERM GOAL IS TO IMPROVE THE SUBOPTIMAL LUNG TRANSPLANT SURVIVAL AND ENSURE THIS IMPROVEMENT IS ACHIEVED REGARDLESS OF PLACE. THIS PROJECT IS IMPACTFUL AS WE ARE THE FIRST IN LUNG TRANSPLANT TO LOOK BEYOND THE WALLS OF OUR HOSPITALS TO CREATE UNPRECEDENTED COMPREHENSIVE DATA TRANSLATABLE TOWARDS PATIENT AND POLICY LEVEL INTERVENTIONS. THIS PROPOSAL IS RESPONSIVE TO THE FINAL RULE MANDATE BY THE U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES THAT PATIENT GEOGRAPHY SHOULD HAVE A MINIMAL ROLE IN TRANSPLANT OUTCOMES, A MANDATE THE FIELD OF TRANSPLANTATION HAS MADE LITTLE PROGRESS TOWARDS OVER THE PAST TWO DECADES DUE, IN LARGE PART, TO GAPS IN KNOWLEDGE THAT OUR PROPOSAL AIMS TO FILL.

THE PURPOSE OF THE PROJECT IS TO PREVENT THE LEADING CAUSE OF POSTOPERATIVE MORTALITY AFTER HEART SURGERY LOW CARDIAC OUTPUT SYNDROME LCOS BY PROVIDING AT HOME ICU LEVEL MONITORING TO CHILDREN AND ADULTS WITH CONGENITAL HEART DISEASE (CHD). THIS WILL BE ACHIEVED THROUGH THE CREATION OF A BATTERY OPERATED FLEXIBLE ULTRATHIN QUAD MODAL WIRELESS DEVICE ETATTOO FOR DAILY ABSOLUTE CARDIAC OUTPUT (CO) MONITORING. THE CURRENT STATE OF THE ART MONITORING DEVICES RELY ON DOWNSTREAM METRICS (E.G. LACTIC ACIDOSIS) ARE INVASIVE REQUIRING PULMONARY ARTERIAL CATHETERIZATION OR ARE UNFIT FOR CHILDREN. ADDITIONAL STATE OF THE ART TOOLS FOR CARDIAC OUTPUT MONITORING ARE COSTLY TO OPERATE AND ARE LIMITED BY SINGLE TIME POINT MEASUREMENTS (E.G. MAGNETIC RESONANCE IMAGING MRI).ONE IN EVERY FOUR PATIENTS WITH CONGENITAL HEART DISEASE WILL REQUIRE SURGERY AT SOME POINT. LCOS IS THE LEADING CAUSE OF POSTOPERATIVE MORTALITY IN THOSE PATIENTS. THE PROPOSED PROGRAM IS CONSIDERED A CRITICAL TECHNOLOGY EFFORT BECAUSE IT AIMS TO NONINVASIVELY ESTIMATE CARDIAC OUTPUT IN CONGENITAL HEART DISEASE PATIENTS OF ALL AGES AND TO HELP PREVENT LCOS. IN ADDITION THE TECHNOLOGY CAN BE USED AT HOME AND IT CAN BE APPLIED TO OTHER HEART SENSING APPLICATIONS. CURRENT TECHNOLOGY AND APPROACHES TO NONINVASIVE CONTINUOUS CARDIAC OUTPUT MONITORING (NICCOM) ARE NOT IDEAL AND CAN PRESENT A HIGHER RISK PROFILE. CURRENTLY ECG PATCHES ARE USED TO MONITOR CHD PATIENTS INTERMITTENTLY EVERY YEAR AND THEY DO NOT MEASURE ALL RELEVANT INFORMATION. IMPLANTABLE SENSORS HAVE REVOLUTIONIZED LONGER TERM PATIENT MONITORING BUT THEY ARE NOT WELL STUDIED AND PRESENT INFECTION RISK.THE ADVANTAGE OF COLLABORATING WITH THE GOVERNMENT ON THIS ENDEAVOR IS THAT IF SUCCESSFUL IT CAN PROVIDE MEDICAL GRADE CLINICALLY BACKED AFFORDABLE ACCESSIBLE FLEXIBLE AND DISPOSABLE WEARABLE TECHNOLOGY TO AMERICANS. A SUCCESSFUL PROJECT WOULD RESULT IN STATE OF THE ART ALGORITHMS FOR BOTH NONINVASIVE CARDIAC OUTPUT MONITORING AND LCOS PREDICTIONS FOLLOWING SURGERY. ADDITIONALLY NONINVASIVE QUAD MODAL WEARABLE HARDWARE (ETATTOO) WOULD BE PRODUCED TO MEASURE PHYSIOLOGICAL PARAMETERS. THERE IS GREAT COMMERCIALIZATION AND MARKET POTENTIAL FOR THE ETATTOO WEARABLE MONITORING DEVICE (1) THE DEVICE COVERS BOTH PEDIATRIC AND ADULT MARKETS (2) THE AI ALGORITHMS CAN BE STANDALONE OR LICENSED TO OTHER COMPANIES AND (3) THE ETATTOO DEVICE HARDWARE CAN BE USED FOR APPLICATIONS OUTSIDE OF LCOS PREDICTION.

CHRONIC PHYSIOLOGIC AND BEHAVIOR CHANGES INDUCED BY NOVEL STN DBS PATTERNS FOR PD

LNCRNA REGULATION OF GLIOBLASTOMA PROGRESSION AND THERAPEUTIC RESISTANCE - PROJECT SUMMARY GLIOBLASTOMA (GBM) IS AN INCURABLE PRIMARY BRAIN TUMOR THAT IS CHARACTERIZED BY REGIONS OF HYPOXIA AND MARKED RESISTANCE TO RADIATION. GLIOMA STEM-LIKE CELLS (GSCS) ARE A HIGHLY MALIGNANT SUBPOPULATION OF CELLS THAT ARE HIGHLY RESISTANT TO STANDARD CYTOTOXIC TREATMENTS. GSCS HAVE A HIGH CAPACITY FOR SELF-RENEWAL AND ARE FREQUENTLY LOCATED IN HYPOXIC AREAS, MAKING THEM MORE EVEN DIFFICULT TO KILL WITH RADIATION. GSCS THEREFORE PLAY AN IMPORTANT ROLE IN DISEASE RECURRENCE. LONG NON-CODING RNAS (LNCRNAS) HAVE RECENTLY BEEN FOUND TO BE DYSREGULATED IN CANCER. LNCRNAS HAVE MULTIPLE FUNCTIONS INCLUDING REGULATION OF GENE EXPRESSION. WE HAVE FOUND THAT THE LNCRNA LUCAT1 IS AN IMPORTANT REGULATOR OF GSC RESPONSE TO HYPOXIA. LUCAT1 IS FREQUENTLY OVEREXPRESSED IN GBM AND IS ASSOCIATED WITH POOR PROGNOSIS IN THE AGGRESSIVE IDH WT SUBTYPE. OUR DATA SUPPORT THAT LUCAT1 IS INDUCED BY HYPOXIA AND FORMS A POSITIVE REGULATORY LOOP TO PROMOTE HIF1A SIGNALING. FUNCTIONALLY, OUR DATA SUPPORT THAT LUCAT1 HELPS TO MAINTAIN GSCS IN HYPOXIA AND PROMOTE TUMOR GROWTH. IN THIS STUDY, WE PROPOSE TO DETERMINE A MECHANISM BY WHICH LUCAT1 REGULATES HIF1 SIGNALING (AIM 1) AND ASSESS THE FUNCTION OF LUCAT1 IN GSC MAINTENANCE AND TUMOR PROGRESSION (AIM 2). THESE STUDIES WILL REVEAL A NEW AND IMPORTANT MECHANISM BY WHICH HYPOXIC INDUCTION OF LUCAT1 DRIVES GSC-MEDIATED TUMORIGENESIS. IF SUCCESSFUL, OUR FINDINGS WILL PROVIDE A NEW THERAPEUTIC APPROACH FOR TARGETING GSCS IN HYPOXIA TO IMPROVE GBM CONTROL. THIS TREATMENT STRATEGY MAY BE EXTENDED TO OTHER CANCERS INCLUDING SMOKING-ASSOCIATED LUNG CANCER AND RENAL CELL CANCER THAT EXPRESS HIGH LEVELS OF LUCAT1.

NEW MOUSE MODEL OF CISPLATIN-INDUCED AKI AND DEVELOPMENT OF PREVENTION THERAPY - SUMMARY CISPLATIN IS AMONG THE MOST WIDELY USED DRUGS AGAINST SOLID TUMORS, BUT IT CAN CAUSE SEVERE DAMAGE OF RENAL TUBULOINTERSTITIAL TISSUES, MANIFESTING ITSELF PATHOLOGICALLY IN THE FORM OF GLOMERULAR FILTRATION RATE REDUCTION, HYPE- RURICEMIA, HYPOKALEMIA, AND ACCELERATED SECONDARY GLOMERULOSCLEROSIS AND GLOMERULAR ISCHEMIA. THE END RE- SULT IS ACUTE KIDNEY INJURY (AKI). CISPLATIN INDUCES AKI, WHICH AFFLICTS 25% OF CISPLATIN-TREATED CANCER PATIENTS WORLDWIDE. WITH FEW EXCEPTIONS, THERE IS NO EFFECTIVE PREVENTIVE OR POST-EXPOSURE THERAPY FOR CISPLATIN-INDUCED AKI (C-AKI). OXIDATIVE STRESS AND MITOCHONDRIAL DAMAGE ARE DRIVERS OF AKI-ASSOCIATED PATHOLOGY; HOWEVER, THE MOLECULAR PATHWAYS THAT MEDIATE THESE EVENTS ARE POORLY DEFINED. USING MURINE C-AKI MODEL KIDNEY AND BI- OPSY SAMPLES FROM C-AKI PATIENTS, WE HAVE RECENTLY FOUND THAT BOTH OXIDATIVE STRESS AND MITOCHONDRIAL DAM- AGE ARE ASSOCIATED WITH UPREGULATION OF RENAL APURINIC/APYRIMIDINIC ENDONUCLEASE 2 (APE2). IN PRELIMINARY EX- PERIMENTS, WE DISCOVERED THAT CISPLATIN EXPOSURE IN VIVO INCREASES EXPRESSION OF APE2 IN MICE AND INHIBITS THE ACTION OF THE PROTEIN MYOSIN HEAVY CHAIN 9 (MYH9), A KEY REGULATORY MOLECULE OF KIDNEY FUNCTION. THESE ALTERA- TIONS TRANSLATE INTO IMPACT ON TUBULOINTERSTITIAL TISSUE. OUR OVERALL WORKING HYPOTHESIS IS THAT CISPLATIN ALTERS KIDNEY APE2 SIGNALING, LEADING TO INHIBITION OF MYH9 WITH CONCOMITANT TUBULOINTERSTITIAL DAMAGE. HERE WE PRO- POSE TO MORE PRECISELY ILLUMINATE THE TARGET ROLE OF APE2 IN AKI DEVELOPMENT, AND SPECIFICALLY HYPOTHESIZE (I) THAT HIGHLY EXPRESSED APE2 AFFECTS MTDNA INTEGRITY AND MYH9/ACTIN INTERMEDIATE MITOCHONDRIAL FISSION AND (II) THAT THIS IN TURN PROMOTES RENAL INJURY BY TRIGGERING CELL-DEATH PATHWAYS. TO ADDRESS THESE DUAL ISSUES, WE PROPOSE USING APE2 TRANSGENIC AND KNOCKOUT (KO) MOUSE MODELS TO PURSUE THE TWO SPECIFIC AIMS DETAILED BELOW. IN AIM 1, WE WILL VALIDATE A GENETICALLY ENGINEERED MURINE MODEL OF APE2 WITH DIRECT CLINICAL RELEVANCE TO C-AKI. WE WILL COMPREHENSIVELY DEFINE THE APE2 MOUSE PHENOTYPE AND COMPARE IT WITH THAT OF THE C-AKI MOUSE MODEL AND THE HUMAN C-AKI SIGNATURE WITH RESPECT TO PATHOPHYSIOLOGICAL FEATURES AT THE MOLECULAR LEVEL USING WHOLE GENOMIC, TRANSCRIPTIONAL, AND IMMUNOMIC APPROACHES. WE WILL DEFINE THE BINDING SITE OF APE2/MYH9 AND ITS FUNCTIONAL SIGNIFICANCE FOR FURTHER TRANSLATIONAL STUDIES. IN AIM 2, WE WILL CONDUCT PROOF-OF- CONCEPT STUDIES ON APE2 TARGETED THERAPY FOR PREVENTION OF C-AKI. ADDITIONALLY, WE WILL DEFINE A NOVEL TARGETED THERAPY TO PREVENT NEPHROTOXICITY BY DELIVERING APE2 GAPMER ANTISENSE OLIGONUCLEOTIDES OR SMALL-MOLECULE IN- HIBITORS TO SELECTIVELY SUPPRESS APE2 EXPRESSION IN PROXIMAL TUBULE CELLS IN APE2 TRANSGENIC AND C-AKI MICE. IMPACT: ACCOMPLISHING THESE SPECIFIC AIMS WILL BE THE FIRST STEP TOWARD CONSTRUCTING A TEMPORAL-MECHANISTIC MAP OF EVENTS THAT OCCUR UPON CISPLATIN EXPOSURE AND TRANSLATE INTO TUBULOINTERSTITIAL NEPHRITIS AND MAY ULTI- MATELY ENABLE DEVELOPMENT OF NOVEL TARGETED THERAPY FOR AKI DISEASE.

MECHANISMS OF KIDNEY DISEASES ASSOCIATED WITH APOL1 VARIATION - ABSTRACT SEVERE, PROGRESSIVE KIDNEY DISEASES (CKD) ARE MORE COMMON AMONG BLACK PATIENTS THAN IN OTHER POPULATIONS. THE EXCESS RISK FOR NON-DIABETIC CKD IN THIS POPULATION IS EXPLAINED, IN PART, BY THE PRESENCE OF GENETIC VARIANTS IN THE APOL1 GENE, WHICH ARE UNIQUE TO SOME AFRICAN ANCESTRAL POPULATIONS. THE APOL1 HIGH RISK (HR) GENOTYPE, DEFINED AS THE PRESENCE OF TWO RISK ALLELES (G1 OR G2), IS STRONGLY ASSOCIATED WITH SOME IDIOPATHIC PROTEINURIC PODOCYTOPATHIES, PRIMARILY FSGS. THE PATHOGENIC MECHANISMS RESPONSIBLE FOR THE GENETIC ASSOCIATION REMAIN POORLY UNDERSTOOD, ALTHOUGH OVER-EXPRESSION SYSTEMS SUGGEST KIDNEY DISEASES RESULT FROM HR VARIANT-DEPENDENT CYTOTOXICITY. HOWEVER PUBLISHED DATA SUGGEST THAT BOTH LOSS-OF PROTECTION AND GAIN OF TOXIC FUNCTION PATHWAYS MAY MEDIATE APOL-ASSOCIATED KIDNEY DISEASES. TO DEVELOP HYPOTHESES ABOUT MECHANISMS, WE CHARACTERIZED THE G0 AND G2 SPATIAL PROTEOMES USING PEROXIDASE-CATALYZED PROXIMITY LABELLING AND MASS SPECTROMETRY TO IDENTIFY CANDIDATE APOL1-REGULATED PATHWAYS IN HELA CELLS WITH REGULATED APOL1 EXPRESSION. APOL1 INDUCTION DID NOT CAUSE CELL DEATH AS ASSAYED BY A METHOD AGNOSTIC TO DEATH MECHANISM OR BY AN APOPTOSIS-SPECIFIC ASSAY. THE COMPOSITIONS OF THE G0 AND G2 SPATIAL PROTEOMES ARE MARKEDLY ENRICHED IN SECRETORY PATHWAY MEMBRANE AND LUMINAL PROTEINS BUT THEIR PROTEIN NEIGHBORHOODS DIVERGE IN THE GOLGI WHERE G0 AND HR-APOL1S ARE LOADED INTO A SUBSET OF RAB6+ VESICLES. THE G0 AND G2 SPATIAL PROTEOMES SUGGEST THAT CARGOS OF THESE VESICLES ARE APOL1-ISOFORM SPECIFIC, WITH G0 BUT NOT G2 BEING IN PROXIMITY OF CELL SURFACE PROTEINS. USING A N-HYDROXYSUCCINIMIDE BIOTIN ESTER ENRICHMENT METHOD, WE DEMONSTRATED G2 EXPRESSION ALTERED CELL SURFACE PROTEINS WITH FUNCTIONAL CONSEQUENCES. INTERFERENCE REFLECTANCE MICROSCOPY SUGGESTS G2 EXPRESSING BUT NOT G0 EXPRESSING CELLS FAIL TO MAINTAIN ADHESION. IMPORTANTLY, THESE PHENOTYPES ARE RECAPITULATED IN ISOGENIC IPSC-DERIVED PODOCYTES WITH G0, G1 AND G2 GENOTYPES. BASED ON THESE DATA, WE HYPOTHESIZE THAT APOL1 REGULATES PODOCYTE SURFACEOME TO MAINTAIN PODOCYTE ADHESION AND DIFFERENTIATED FUNCTIONS. G2 FAILS TO DO THIS RESULTING IN PODOCYTE DEPLETION AND PROGRESSIVE PODOCYTOPATHY. WE PROPOSE THE FOLLOWING EXPERIMENTS TO BETTER UNDERSTAND APOL1 REFERENCE AND KIDNEY RISK VARIANT FUNCTION: AIM 1: USE STABLE ISOTOPE LABELLING BY AMINO ACIDS IN CELL CULTURE (SILAC) AND MASS SPECTROMETRY TO CHARACTERIZE THE SURFACE PROTEOME OF PODOCYTES EXPRESSING APOL1-G0, G1 AND G2 VARIANTS IN THE PRESENCE AND ABSENCE OF INTERFERON-. AIM 2: CHARACTERIZE THE RAB6A+ POST-GOLGI VESICULAR COMPARTMENT USING BIOCHEMICAL AND MICROSCOPIC METHODS TO DEFINE THE DIFFERENTIAL TRAFFICKING OF RAB6A+ VESICLES AND THE ASSOCIATED CARGO THAT IS DEPENDENT UPON APOL1 GENOTYPE. AIM 3: PROVE THAT THE APOL1 ISOFORM-SPECIFIC CARGOS IN RAB6+ VESICLES REGULATE THE PODOCYTE SURFACEOME AND DEFINE THE APOL1–DEPENDENT MECHANISMS FOR THE PODOCYTE ADHESION DEFECT.

OPTIMIZATION OF CAMPARI FOR LARGE-SCALE, CELLULAR-RESOLUTION ACTIVITY RECORDING IN FREELY-MOVING MICE - PROJECT SUMMARY THE GOAL OF THIS PROPOSED RESEARCH IS TO OPTIMIZE A DUAL-USE CALCIUM ION SENSOR FOR RECORDING SINGLE-CELL ACTIVITY FROM THE ENTIRE DORSAL CORTEX OR HIPPOCAMPUS OF FREELY-MOVING MICE. IT IS WIDELY ACCEPTED THAT OPTICAL IMAGING WITH GENETICALLY-ENCODED FLUORESCENT CALCIUM SENSORS IS CURRENTLY THE ONLY METHOD TO OBTAIN MEASUREMENTS OF GENETICALLY-IDENTIFIED NEURONAL POPULATIONS WITH DENSE SAMPLING. OVER THE PAST SEVERAL YEARS, THE RECORDING CAPABILITIES OF TWO-PHOTON MICROSCOPES HAVE BEEN IMPROVED TO RECORD FROM MILLIMETER-SCALE TISSUE, AND NEW MINIATURIZED MICROSCOPES HAVE BEEN DEVELOPED TO RECORD FROM MOVING MICE. HOWEVER, MOST BEHAVIORS ARISE FROM COLLECTIVE INTERACTIONS BETWEEN NEURONS FROM MULTIPLE BRAIN AREAS, WHICH CANNOT BE SIMULTANEOUSLY MONITORED WITH THESE SYSTEMS. THEREFORE, THERE IS A CLEAR NEED TO DEVELOP A NEW APPROACH TO DIRECTLY MONITOR THE SYNCHRONIZED ACTIVITY OF DISTRIBUTED NEURAL CIRCUITS. CAMPARI IS A UNIQUE CALCIUM SENSOR THAT CAN DETECT ACTIVITY IN TWO CALCIUM-DEPENDENT WAYS: 1) PERMANENT COLOR CHANGE (GREEN TO RED) UPON ILLUMINATION WITH VIOLET LIGHT, A PROCESS KNOWN AS PHOTOCONVERSION, AND 2) DYNAMIC CHANGES IN FLUORESCENCE INTENSITY. OUR DATA SHOW THAT CAMPARI ALLOWS RECORDING OF BRAIN ACTIVITY FROM FREELY-BEHAVING MICE, WITHOUT USING MICROSCOPE OBJECTIVES OR IMPLANTED DEVICES. MOREOVER, NATURAL DEGRADATION OF THE RED CAMPARI PROTEIN ENABLES MULTIPLE LONGITUDINAL MEASUREMENTS. HOWEVER, PREVIOUS ATTEMPTS TO IMPROVE CAMPARI USING IN VITRO ASSAYS REDUCED SOME OF ITS IN VIVO PROPERTIES, WHICH RESULTED IN LOW DYNAMIC RECORDING SENSITIVITY AND A LOW PHOTOCONVERSION RATE THAT REQUIRES LONG ILLUMINATION TIMES TO ACCUMULATE A SUFFICIENT AMOUNT OF RED PROTEIN. THEREFORE, THE PROJECT GOAL IS TO OPTIMIZE CAMPARI TO ALLOW SENSITIVE RECORDING OF CELLULAR-RESOLUTION, CORTEX-WIDE ACTIVITY SNAPSHOTS IN FREELY-MOVING MICE, FOLLOWED BY SUBSEQUENT DYNAMIC RECORDING FROM THE SAME MOUSE USING TWO- AND THREE-PHOTON MICROSCOPY. TO OPTIMIZE CAMPARI’S PERFORMANCE, WE WILL COMBINE IN VITRO TESTING IN PURIFIED PROTEIN, HEK CELLS, AND NEURONS, AND THE MOST PREDICTIVE ASSAY: LARGE-SCALE IN VIVO SCREENING OF ~30-FOLD MORE CONSTRUCTS THAN PREVIOUS STUDIES. AIM 1 WILL FOCUS ON ENHANCING CAMPARI’S PHOTOCONVERSION EFFICIENCY TO FACILITATE LARGE-SCALE RECORDINGS IN FREELY-MOVING MICE. AIM 2 WILL FOCUS ON IMPROVING CAMPARI’S DYNAMIC RECORDING PROPERTIES AND SENSITIVITY. IN AIM 3, WE WILL COMBINE BENEFICIAL MUTATIONS FROM AIMS 1-2 TO GENERATE A NEW CAMPARI WITH OPTIMIZED PHOTOCONVERSION AND DYNAMIC RECORDING CAPABILITIES. OUR PROOF-OF-CONCEPT EXPERIMENTS WILL DEMONSTRATE MULTI- REGIONAL CORTICAL MAPPING DURING A BATTERY OF BEHAVIORAL AND COGNITIVE TESTS TO DETECT CELLULAR-RESOLUTION CHANGES IN CORTEX-WIDE ACTIVITY PATTERNS. THIS OPTIMIZED CAMPARI IS EXPECTED TO FACILITATE NEW HYPOTHESIS-DRIVEN STUDIES BY PROVIDING VOLUMETRIC, MULTI-REGIONAL BRAIN ACTIVITY DATA OF GENETICALLY-TARGETED NEURONS DURING COGNITIVE AND BEHAVIORAL TESTING OF FREELY-MOVING MICE, ENABLING STUDIES THAT INVOLVE BOTH HEAD-FIXATION AND FREE MOVEMENT IN THE SAME MICE, AND TO UTILIZE COMPLEMENTARY TECHNIQUES LIKE OPTOGENETIC STIMULATION AND SINGLE-CELL SEQUENCING METHODS TO ENABLE STUDYING THE PROPERTIES OF ACTIVE (RED-LABELED) CELLS DURING BEHAVIORAL STUDIES.

PLATELETS IN CANCER

MODULATION OF PROTECTIVE RAS-GPCR PATHWAYS IN AORTIC DISEASE

DNA REPAIR PATHWAYS CHOICE TO REGULATE THERAPEUTIC RESISTANCE IN CANCER CELLS

NEXT GENERATION DNMT1 DEPLETION THERAPY FOR LEUKEMIA

ELTOMBOPAG: NOVEL MODE OF ACTION ON NORMAL AND APLASTIC ANEMIA HEMATOPOIETIC STEM CELLS - SUMMARY IDIOPATHIC APLASTIC ANEMIA (AA) IS CHARACTERIZED BY IMMUNE-MEDIATED HEMATOPOIETIC STEM AND PROGENITOR CELLS (HSPCS) DESTRUCTION RESULTING IN DEFICIENCIES ACROSS ALL HEMATOPOIETIC LINEAGES AND BONE MARROW FAILURE. DESPITE OF THE THERAPEUTIC SUCCESSES OF IMMUNOSUPPRESSIVE THERAPIES (IST), APPROXIMATELY ONE THIRD OF PATIENTS REMAIN REFRACTORY AND MANY OF THE RESPONSES ARE INCOMPLETE. RECENTLY, A SYNTHETIC THROMBOPOIETIN RECEPTOR (TPOR) AGONIST, EPAG HAS BEEN SHOWN TO BE EFFECTIVE IN AA. IN ADDITION TO THE ANTICIPATED EFFECT ON PLATELET COUNTS, EPAG ALSO PRODUCED REMARKABLE TRI-LINEAGE HEMATOPOIESIS. THESE EFFECTS EXPANDED THE INDICATION SPECTRUM OF EPAG FROM IMMUNE THROMBOCYTOPENIC PURPURA TO AA, ESTABLISHING THIS DRUG AS AN ESSENTIAL HEMATOLOGIC THERAPEUTIC. RECENT STUDIES DEMONSTRATE THAT EPAG’S HEMATOPOIETIC ACTIVITY IS ALSO OBSERVED IN MURINE MODELS DESPITE THE LACK OF BINDING TO MURINE TPOR. WE CONFIRMED SIMILAR EFFECTS IN MURINE CELLS WHERE EPAG TREATMENT REMARKABLY EXPANDED HSCS WITHOUT ANY EFFECT ON THE TPOR SIGNALING. THESE OBSERVATIONS SUGGESTED THAT A SIGNIFICANT PART OF EPAG’S ACTIVITIES ARE TPOR INDEPENDENT, IN CONTRAST TO PEPTIDE TPO ANALOGS, E.G., ROMIPLOSTIN. THESE TPOR INDEPENDENT EFFECTS OF EPAG WERE HYPOTHESIZED TO BE DUE TO ITS IRON CHELATING PROPERTIES, BUT THE MOLECULAR MECHANISM AS TO HOW THIS IRON-BINDING COULD DRIVE THE HSPCS EXPANSION REMAINS SPECULATIVE. EPAG EFFECTS ON INTRACELLULAR IRON MAY AFFECT CERTAIN IRON-DEPENDENT EPIGENETIC PATHWAY/S THAT PROMOTE HSPCS SELF-REPLICATION. FOR INSTANCE, TET-DIOXYGENASES (TET1-3) ARE FE2+- AND A- KETOGLUTARATE (AKG) DEPENDENT DNA-DIOXYGENASES, WHICH MEDIATE CPG DEMETHYLATION OF PROMOTERS AND ENHANCERS IN HSPCS. CONSEQUENTLY, BY CHANGING GENE EXPRESSION PATTERNS, TETS CONTROL HSPCS EXPANSION AND DIFFERENTIATION. TET2 IS THE MOST ABUNDANT TET-DIOXYGENASE IN HSPCS, AND SOMATIC LOSS-OF-FUNCTION (LOF) MUTATIONS OF THIS GENE FREQUENTLY OCCUR IN MYELOID NEOPLASIA AND CLONAL HEMATOPOIESIS OF INDETERMINATE POTENTIAL (CHIP), A PRODROMAL CONDITION IN OTHERWISE HEALTHY ELDERLY INDIVIDUALS CHARACTERIZED BY HIGH PROGRESSION RATE TO A FRANK LEUKEMIA. OUR PROPOSAL IS DESIGNED TO DETERMINE THE EFFECTS OF EPAG ON NORMAL HEMATOPOIESIS MEDIATED BY ITS ABILITY TO INHIBIT TET-ACTIVITY. BASED ON OUR ANALYSIS OF CLINICAL DATA COUPLED WITH BIOCHEMICAL ANALYSIS WE HYPOTHESIZE THAT, EPAG-MEDIATED TET- INHIBITION IS RESPONSIBLE FOR TRI-LINEAGE RESPONSE IN AA VIA HSC EXPANSION AND ON THE BIOCHEMICAL LEVEL DECREASED 5HMC CONTENT LEADING TO HYPERMETHYLATION AS A RESULT OF DIRECT INHIBITION OF TET2. OUR CURRENT PROPOSAL WILL PROVIDE A PROOF OF CONCEPT FOR THE REVERSIBLE TRANSIENT TET-INHIBITION AS A BASIS FOR HSC EXPANSION THAT MAY RESTORE NORMAL HEMATOPOIESIS.

ROLE OF STRESS IN THE DEGRADATION OF INTESTINAL IGA AND INTESTINAL HEALTH

USING PATIENT-LEVEL DECISION MODELING TO IMPROVE USE OF TREATMENTS FOR ALCOHOL USE DISORDER - PROJECT SUMMARY ALCOHOL USE DISORDER (AUD), PREVIOUSLY ALCOHOL ABUSE AND DEPENDENCE, IS A SIGNIFICANT PUBLIC HEALTH PROBLEM. IN THE US, EXCESSIVE ALCOHOL USE IS RESPONSIBLE FOR 93,000 DEATHS AND 2.7 MILLION YEARS OF POTENTIAL LIFE LOST, AT A COST OF $250 BILLION ANNUALLY. IN 2012-13, THE 12-MONTH AND LIFE-TIME PREVALENCE OF AUD IN US ADULTS WAS 14% AND 29%, RESPECTIVELY, INCREASING BY MORE THAN 40% FROM A DECADE EARLIER. SINCE THE COVID-19 PANDEMIC, ALCOHOL CONSUMPTION HAS INCREASED EVEN MORE. EXCESSIVE ALCOHOL CONSUMPTION AFFECTS VARIOUS BODILY ORGANS, LEADING TO MANY HEALTH CONSEQUENCES. FORTUNATELY, AUD IS MORE TREATABLE THAN COMMONLY BELIEVED. COGNITIVE BEHAVIORAL THERAPY, BRIEF INTERVENTIONS, AND MOTIVATIONAL ENHANCEMENT THERAPY ARE THE MOST WIDELY STUDIED BEHAVIORAL INTERVENTIONS AND ARE SIMILARLY EFFICACIOUS, WHILE ALCOHOLICS ANONYMOUS (AA) IS THE MOST SOUGHT OUT MUTUAL SUPPORT GROUP. PHARMACOTHERAPY CAN BE COMBINED WITH BEHAVIORAL INTERVENTIONS TO INCREASE SUCCESS. HOWEVER, TREATMENTS FOR AUD ARE UNDERUTILIZED. THE MISCONCEPTION AMONG PATIENTS AND PHYSICIANS THAT TREATMENT IS INEFFECTIVE IS AN IMPORTANT OBSTACLE TO USE. ANOTHER IS THE COMMON VIEW THAT TREATMENT SUCCESS REQUIRES TOTAL ABSTINENCE. IN REALITY, RECOVERY FROM AUD CAN INCLUDE SOME HEAVY DRINKING, AND AUD TREATMENT CAN BE EFFECTIVE. EXPRESSING TREATMENT EFFECTIVENESS INTO LONG-TERM HEALTH BENEFITS COULD HELP CONVINCE PHYSICIANS TO OFFER TREATMENT AND ENCOURAGE PATIENTS TO USE IT. HOWEVER, RANDOMIZED CONTROLLED TRIALS (RCTS) ALONE CANNOT PROVIDE THIS INFORMATION BECAUSE THEY DO NOT MEASURE LONG-TERM HEALTH OUTCOMES, NOR DO THEY COMPARE ALL TREATMENTS HEAD-TO-HEAD, MAKING IT DIFFICULT TO CHOOSE AMONG THEM. SIMULATION MODELING OFFERS A COMPREHENSIVE APPROACH TO COMPARING TREATMENTS, BUT NONE OF EXISTING AUD MODELS WERE DESIGNED TO ASSESS ALL TREATMENTS. WE DESIGNED A DECISION AID (DA) DESCRIBING TREATMENT OPTIONS, BUT IT DOES NOT YET INCLUDE ESTIMATES OF LONG-TERM BENEFITS DUE TO LACK OF SUCH DATA AT THE TIME OF DEVELOPMENT. PROJECT OBJECTIVE: TO DEVELOP, VALIDATE, AND APPLY A COMPUTER SIMULATION MODEL TO INFORM POLICY MAKERS, PHYSICIANS, AND PATIENTS OF THE LIFETIME BENEFIT OF TREATMENTS FOR AUD. AIM 1: DEVELOP AND VALIDATE A MICROSIMULATION MODEL OF THE NATURAL HISTORY OF ALCOHOL-ASSOCIATED COMPLICATIONS ACCOUNTING FOR THE CHANGE IN DRINKING BEHAVIORS AND AUD STATUS OVER TIME. AIM 2: ASSESS THE COMPARATIVE EFFECTIVENESS OF AUD TREATMENTS. AIM 3: INCORPORATE PATIENT AND PROVIDER FEEDBACK TO ASSESS THE CLARITY, CONTENT, AND ACCEPTABILITY OF AUGMENTING AN EXISTING DA WITH COMPARATIVE EFFECTIVENESS DATA. IMPACT: OUR MODEL WILL OFFER A BETTER UNDERSTANDING OF THE EXPECTED BENEFIT OF AUD TREATMENTS AND PROVIDE AN INNOVATIVE APPROACH TO DECISION MODELING AND TREATMENT SELECTION. BY INCORPORATING LONG-TERM EFFECTIVENESS DATA INTO AN EXISTING DA, OUR STUDY WILL PROVIDE PHYSICIANS AND PATIENTS WITH VALUABLE EVIDENCE AND A TOOL TO SUPPORT SELECTION OF OPTIMAL TREATMENTS, THEREBY HELPING TO CLOSE THE TREATMENT GAP. ULTIMATELY, OUR PROJECT WILL ADDRESS NIAAA’S RESEARCH MISSIONS WHILE IMPROVING THE QUALITY OF LIFE FOR TENS OF MILLIONS OF US ADULTS SUFFERING FROM AUD.

ROLE OF TLR2 IN ANGIOGENESIS

FLAVIVIRUS NS3-MEDIATED INNATE IMMUNE ESCAPE

ALCOHOL AND INTESTINAL MICROVASCULAR ENDOTHELIUM-IMMUNE AXIS AND THE ROLE OF GUT DERIVED IMMUNE NUTRIENTS

ACTIVITY AND CONNECTIVITY OF HIPPOCAMPAL NEWBORN NEURONS UNDERLIE ALCOHOL WITHDRAWAL-ASSOCIATED SYNDROMES - ALCOHOL WITHDRAWAL (AW) AFTER CHRONIC ALCOHOL EXPOSURE (AE) PRODUCES A SERIES OF SYMPTOMS. AMONG THEM, GENERALIZED TONIC-CLONIC SEIZURES ARE THE MOST SEVERE AND DANGEROUS SYMPTOM. THE SEVERITY AND SUSCEPTIBILITY TO RELAPSE, AND PERPETUATION OF ALCOHOL ABUSE UNDERSCORE THE URGENT NEED TO UNDERSTAND MECHANISMS UNDERLYING ALCOHOL DEPENDENCE AND AW IN ORDER TO DEVELOP NEW THERAPEUTIC STRATEGIES TO INTERVENE AND TREAT AW-ASSOCIATED SYNDROMES SUCH AS SEIZURES. IN THIS APPLICATION, WE WILL TEST THE NOVEL HYPOTHESIS THAT ACTIVITY AND CONNECTIVITY OF HIPPOCAMPAL NEWBORN DENTATE GRANULE CELLS (DGCS) UNDERLIE AW-ASSOCIATED SEIZURES. DGCS ARE PRINCIPAL EXCITATORY NEURONS THAT ARE CONTINUOUSLY PRODUCED AND INTEGRATE INTO HIPPOCAMPAL NEURAL CIRCUITS, AND ALTERED HIPPOCAMPAL NEUROGENESIS HAS BEEN IMPLICATED IN SEIZURES. OUR PREVIOUS STUDIES HAVE REVEALED THE ESSENTIAL ROLES OF HIPPOCAMPAL NEWBORN DGCS IN THE EXPRESSIONS OF AW-ASSOCIATED SEIZURES. AE REDUCED SPINE FORMATION WHILE AW INCREASED SYNAPTIC CONNECTIVITY OF HIPPOCAMPAL NEWBORN DGCS. OUR RABIES VIRUS-MEDIATED RETROGRADE TRACING STUDY DISCOVERED ALTERED NEURONAL CONNECTIVITY OF HIPPOCAMPAL NEWBORN DGCS WITH BOTH EXCITATORY AND INHIBITORY NEURONS DURING AW SEIZURES. MOREOVER, OUR FUNCTIONAL STUDY WITH A DREADD (DESIGNER RECEPTORS EXCLUSIVELY ACTIVATED BY DESIGNER DRUGS) METHOD DEMONSTRATED THAT ACTIVITY OF HIPPOCAMPAL NEWBORN NEURONS PLAYS AN ESSENTIAL ROLE IN THE EXPRESSION OF AW-ASSOCIATED SEIZURES. THESE OBSERVATIONS PROVIDED THE THEORETICAL FOUNDATION FOR OUR HYPOTHESIS THAT ALTERED NEURONAL CONNECTIVITY AND ACTIVITY OF HIPPOCAMPAL NEWBORN DGCS DISRUPTS THE BALANCE OF EXCITATORY AND INHIBITORY (E/I) SIGNALS, ULTIMATELY LEADING TO AW-ASSOCIATED SEIZURES. THUS, THE CENTRAL GOAL OF THIS PROPOSAL IS TO USE NOVEL MAPPING METHODS, IMAGING TOOLS, AND CELLULAR AND MOLECULAR APPROACHES IN ORDER TO UNDERSTAND ACTIVITY AND CONNECTIVITY OF HIPPOCAMPAL NEURAL CIRCUITS THAT ARE RESPONSIBLE FOR AW-ASSOCIATED SEIZURES. IN AIM 1, WE WILL DETERMINE WHETHER AW ALTERS NEURONAL AND FUNCTIONAL CONNECTIVITY OF DGCS BY USING A RABIES VIRUS- AND WHEAT GERM AGGLUTININ (WGA)-MEDIATED RETROGRADE AND ANTEROGRADE TRACING METHODS, RESPECTIVELY. WE WILL ALSO USE MULTIPLE DREADDS AND ASSESS THE ESSENTIAL ROLE OF DE NOVO NEURAL CIRCUITS FORMED BETWEEN HIPPOCAMPAL NEWBORN DGCS AND INPUT NEURONS IN AW-ASSOCIATED SEIZURES. AIM 2, USING CA2+ IMAGING AND VARIOUS MAGNETIC RESONANCE IMAGING (MRI) MODALITIES THAT ALLOWS US FOR LONGITUDINAL STUDIES, WE WILL DETERMINE ACTIVITY AND CONNECTIVITY OF HIPPOCAMPAL AND GLOBAL NEURAL CIRCUITS UNDERLYING AW-ASSOCIATED SEIZURES. IN AIM 3, WE WILL IDENTIFY AND VALIDATE TRANSCRIPTOME THAT MAY UNDERLIE ALTERED SYNAPTIC AND NEURONAL CONNECTIVITY OF HIPPOCAMPAL NEWBORN DGCS IN RESPONSE TO AE AND AW. THE USAGE OF SINGLE CELL RNA SEQUENCING WILL ALLOW US TO NOT ONLY REGISTER DIFFERENTIALLY EXPRESSED GENES TO CELL TYPE-SPECIFIC MANNER, BUT DETERMINE TRANSCRIPTOMES THAT DISTINGUISH PATHOLOGICAL NEWBORN DGCS FROM NORMAL DGCS DURING AE AND AW. ALTOGETHER, OUR PROPOSAL WILL DISSECT THE MOLECULAR, CELLULAR, AND NEURAL CIRCUITRY MECHANISMS BY WHICH HIPPOCAMPAL NEWBORN DGCS UNDERLIE AW-ASSOCIATED SEIZURES.

DEEP BRAIN STIMULATION OF THE CEREBELLAR DENTATE NUCLEUS TO ENHANCE CHRONIC, POST-TRAUMATIC BRAIN INJURY REHABILITATION

NOVEL REGULATION OF BETA-ADRENERGIC RECEPTOR FUNCTION BY PHOSPHOINOSITIDE 3-KINASE - PROJECT SUMMARY/ABSTRACT THE CURRENT PROPOSAL IS FOCUSED ON DETERMINING PROTEIN PHOSPHATASE 2A (PP2A) REGULATION OF SS-ADRENERGIC RECEPTORS (SSARS) FUNCTION AS SSAR DYSFUNCTION IS A HALLMARK OF HEART FAILURE. HORMONES BINDING TO SSAR RESULTS IN PHOSPHORYLATION BY SSAR KINASES (GRKS) PROMOTING DESENSITIZATION AND ENDOCYTOSIS. SSAR UNDERGOES RESENSITIZATION BY PP2A-MEDIATED DEPHOSPHORYLATION IN THE ENDOSOMES. STUDIES ON REGULATION OF SSAR FUNCTION HAVE MAJORLY FOCUSED ON KINASES DUE TO THE BELIEF THAT PP2A REGULATION IS HOMEOSTATIC IN NATURE. UNEXPECTEDLY, WE IDENTIFIED THAT PI3K ACUTELY REGULATES PP2A BY PHOSPHORYLATING AN ENDOGENOUS INHIBITOR OF PP2A(I2PP2A). PHOSPHORYLATED I2PP2A BINDS TO PP2A INHIBITING ITS ACTIVITY THAT IMPAIRS SSAR RESENSITIZATION. STUDIES IN HUMAN HEART FAILURE SHOWED ACCUMULATION OF SS1 AND SS2ARS IN THE ENDOSOMES WITH REDUCTION IN SSAR-ASSOCIATED PHOSPHATASE ACTIVITY REFLECTING INHIBITION OF RESENSITIZATION. SUBJECTING MICE WITH CARDIOMYOCYTE-SPECIFIC EXPRESSION OF WILD TYPE I2PP2A (WT I2PP2A) TO TRANSVERSE AORTIC CONSTRICTION (TAC) RESULTED IN DILATION, WHILE EXPRESSION OF PHOSPHO-I2PP2A (PI2PP2A) MIMETIC THAT PERSISTENTLY INHIBITS SSAR RESENSITIZATION DID NOT SURVIVE PAST FOUR WEEKS TAC. IN CONTRAST, EXPRESSION OF DEPHOSPHO-I2PP2A (DE-PI2PP2A) MIMETIC THAT PRESERVES SSAR RESENSITIZATION SHOWED SIGNIFICANT AMELIORATION OF CARDIAC DYSFUNCTION POST-TAC REFLECTING A QUINTESSENTIAL ROLE FOR RESENSITIZATION IN CARDIAC REMODELING. SINCE MECHANISTIC UNDERPINNINGS OF PI2PP2A INTERACTION WITH PP2A IS NOT KNOWN, WE USED A COMBINATION OF COMPUTATIONAL AND EXPERIMENTAL APPROACHES TO SHOW THAT ISOPROTERENOL (ISO) STIMULATION OF SSARS LEADS TO PI3K-MEDIATED PHOSPHORYLATION OF I2PPA, PRIMING ITS HOMO-DIMERIZATION RESULTING IN ROBUST BINDING TO PP2A. DOCKING STUDIES FURTHER SHOWED THAT I2PP2A BINDS TO THE C-TERMINAL REGION OF PP2A (PP2A-CT). EXPRESSION OF PP2A-CT AS A DOMINANT NEGATIVE STRATEGY IN CELLS PRESERVED SSAR RESENSITIZATION, WHILE CARDIOMYOCYTE-SPECIFIC EXPRESSION OF PP2A-CT IN MICE RESULTED IN PRESERVATION OF CARDIAC FUNCTION FOLLOWING 2 WEEKS OF ISO ADMINISTRATION SUPPORTING THE PREMISE THAT TARGETING RESENSITIZATION MAY BE BENEFICIAL. BASED ON THESE FINDINGS, WE HYPOTHESIZE THAT RELIEVING PP2A INHIBITION FROM PI2PP2A PRESERVES SSAR RESENSITIZATION AND FUNCTION UNDERLYING BENEFICIAL CARDIAC REMODELING, COUNTER-INTUITIVE TO THE CURRENT OPTION OF BLOCKING G-PROTEIN COUPLING WITH SS-BLOCKERS IN HEART FAILURE. IN THIS REGARD, WE SURPRISINGLY OBSERVED THAT SSARS IN THE DE-PI2PP2A MICE SWITCH THEIR G-PROTEIN COUPLING FROM STIMULATORY GAS (CAMP GENERATING) TO THE INHIBITORY GAI UPON TAC. THIS SUGGESTS THAT SUSTAINING RESENSITIZATION ALLOWS THE SSARS TO FLEXIBLY SWITCH BETWEEN G-PROTEINS MAINTAINING CARDIAC FUNCTION DESPITE CARDIAC STRESS AND SYMPATHETIC OVERDRIVE INDICATING A, YET TO BE UNDERSTOOD FUNDAMENTAL MECHANISM OF SSAR REGULATION AND THEREFORE, PROPOSE THE FOLLOWING AIMS- 1) DETERMINE WHETHER TARGETING I2PP2A DIMERIZATION PRESERVES PP2A ACTIVITY AND RESTORES SSAR FUNCTION, 2) TEST WHETHER RELIEVING OF PP2A INHIBITION PRESERVES SS1 AND SS2AR RESENSITIZATION AND CARDIAC FUNCTION POST-STRESS, 3) TO INVESTIGATE HOW PRESERVATION OF SSAR FUNCTION IN CONDITIONS OF CARDIAC STRESS MEDIATES BENEFICIAL EFFECTS.

NOVEL CYTOSKELETAL MECHANISMS OF PATHOGENIC BACTERIA INTERACTIONS WITH INTESTINAL EPITHELIUM - ABSTRACT INTESTINAL EPITHELIAL CELLS (IEC) PROVIDE THE FIRST LINE OF DEFENSE AGAINST ENTERIC PATHOGENIC BACTERIA. HOWEVER BACTERIAL PATHOGENS DEVELOPED DIFFERENT STRATEGIES TO COLONIZE INTESTINAL EPITHELIUM CAUSING SEVERE GASTROINTESTINAL DISORDER. ONE STRATEGY, USED BY ‘ATTACHING AND EFFACING’ BACTERIA (AEB), SUCH AS ENTEROPATHOGENIC E. COLI (EPEC) AND ENTEROHEMORRHAGIC E. COLI (EHEC), INVOLVES BACTERIAL ADHESION TO IEC WITHOUT ENTERING THE HOST CELLS. THE OTHER STRATEGY USED BY ADHERENT INVASIVE E. COLI (AIEC) OR SALMONELLA RESULTS IN BACTERIAL INTERNALIZATION INTO IEC. THESE DIFFERENT COLONIZATION ROUTES BEAR A KEY MECHANISTIC SIMILARITY: THEY REQUIRE REARRANGEMENTS OF THE EPITHELIAL ACTIN CYTOSKELETON TO ENABLE BACTERIAL ATTACHMENT AND INVASION. ONE IMPORTANT BUT UNDERSTUDIED MECHANISM OF PATHOGEN-INDUCED CYTOSKELETAL REMODELING INVOLVES A MAJOR ACTIN CYTOSKELETON MOTOR, NON-MUSCLE MYOSIN II (NM-II). OUR PRELIMINARY DATA DEMONSTRATE A STRIKING DUALISM OF NM-II-DEPENDENT REGULATION OF BACTERIAL-IEC INTERACTIONS. PHARMACOLOGIC AND GENETIC INHIBITION OF NM-II INCREASES AEB ATTACHMENT TO IEC, BUT INHIBITS AIEC INVASION IN VITRO AND IN VIVO. THESE CONTRASTING ROLES OF NM-II IN IEC-BACTERIAL INTERACTIONS LIKELY REFLECT EXPLOITING TWO DIFFERENT NM II-DEPENDENT FUNCTIONS: ITS ACTIN FILAMENT CONTRACTILITY AND ACTIN FILAMENT CROSS-LINKING ACTIVITY, BY DIFFERENT PATHOGENS. THIS EXCITING DATA PROVIDES A SCIENTIFIC PREMISE FOR THE FOLLOWING INNOVATIVE HYPOTHESIS: NM-II PLAYS A DUAL ROLE IN REGULATING ENTERIC PATHOGEN INTERACTIONS WITH INTESTINAL EPITHELIUM BY INHIBITING AEB ATTACHMENT TO EPITHELIAL CELLS, BUT PROMOTING EPITHELIAL ENTRY OF INVADING PATHOGENIC BACTERIA. THIS HYPOTHESIS WILL BE TESTED IN THE FOLLOWING AIMS: (1) TO DETERMINE THE ROLES OF NM-II IN REGULATING INTESTINAL EPITHELIAL CELL INTERACTIONS WITH ATTACHING AND EFFACING BACTERIA; (2) TO DISSECT THE MECHANISMS OF NM-II-DRIVEN INTERNALIZATION OF INVADING BACTERIA INTO INTESTINAL EPITHELIAL CELLS. WE WILL STUDY EPEC, EHEC, AIEC AND SALMONELLA COLONIZATION OF MODEL IEC CELL LINES, ORGANOID-DERIVED IEC MONOLAYERS IN VITRO AND INTESTINAL COLONIZATION BY CITROBACTER RODENTIUM, AIEC AND SALMONELLA IN VIVO. NM-II ACTIVITY WILL BE MODULATED BY PHARMACOLOGIC AND GENETIC APPROACHES. THE GENETIC APPROACH WILL TARGET TWO MAJOR EPITHELIAL NM-II ISOFORMS, NM-IIA AND NM-IIC, BY SELECTIVELY DOWNREGULATING THEIR EXPRESSION IN HUMAN IEC USING CRISPR/CAS9 GENE EDITING AND RNA INTERFERENCE. IN VIVO NM-II FUNCTIONS WILL BE EXAMINED BY USING MICE WITH EITHER INTESTINAL EPITHELIAL SPECIFIC KNOCKOUT OF NM-IIA, OR TOTAL KNOCKOUT OF NM-IIC. TO DETERMINE WHICH NM-II ACTIVITY IS ESSENTIAL FOR BACTERIAL-IEC INTERACTIONS, WE WILL UTILIZE IEC CELLS AND TRANSGENIC MICE EXPRESSING NM-IIA MUTANTS DEFICIENT IN EITHER ACTIN FILAMENT CROSS-LINKING, OR FILAMENT CONTRACTION ACTIVITIES. SIGNIFICANCE: THE PROPOSAL WILL REVEAL NOVEL INSIGHTS INTO UNDERSTANDING HOW THE INTESTINAL EPITHELIUM CONTROLS PATHOGENIC BACTERIAL INFECTIONS AND WILL IDENTIFY NEW TARGETS FOR THERAPEUTIC INTERVENTIONS TO TREAT DISEASES CAUSED BY ENTERIC PATHOGENS.

INTESTINAL T CELLS AND MICROBIOTA AS THERAPEUTIC TARGETS IN AUTOIMMUNE UVEITIS - PROJECT SUMMARY CHRONIC AUTOIMMUNE UVEITIS REPRESENTS A MEDICAL MANAGEMENT CONUNDRUM, WHICH LEFT UNTREATED, IS A SIGNIFICANT CAUSE OF BLINDNESS IN THE US AND WORLDWIDE. YET DUE TO AN INCOMPLETE UNDERSTANDING OF ITS PATHOGENESIS, TREATMENT HAS OFTEN REQUIRED USE OF NON-SPECIFIC ANTI-INFLAMMATORY AGENTS THAT CAN HAVE UNTOWARD SIDE EFFECTS. EMERGING EVIDENCE LINKS THE INTESTINAL MICROBIOTA TO EXTRAINTESTINAL AUTOIMMUNE DISEASES LIKE UVEITIS, PROVIDING NEW CLUES INTO PATHOGENESIS, AND NOVEL AVENUES FOR THERAPEUTIC TARGETING. OUR GOAL IS TO INVESTIGATE HOW THE INTESTINAL MICROBIOTA CAN BE MANIPULATED TO RE-ESTABLISH INTESTINAL AND THUS SYSTEMIC IMMUNE HOMEOSTASIS THAT HAS GONE AWRY DURING THE COURSE OF AUTOIMMUNE UVEITIS. TO DO THIS, WE ARE USING A QUINTESSENTIAL T-CELL MEDIATED MODEL, EXPERIMENTAL AUTOIMMUNE UVEITIS (EAU). WE WILL UTILIZE TWO DIFFERENT INTERVENTIONS TO ALTER THE INTESTINAL MICROBIOTA, ANTIBIOTICS AND SHORT CHAIN FATTY ACID (SCFA) METABOLITES OF INTESTINAL BACTERIAL FERMENTATION OF DIETARY FIBER, IN EAU. ENHANCEMENT OF INTESTINAL TREGS (CELLS THAT USUALLY SUPPRESS THE IMMUNE SYSTEM) BY THESE MICROBIOTA-ALTERING INTERVENTIONS WILL BE TESTED BY ADOPTIVE TRANSFER OF TREGS AND IN VITRO TREG SUPPRESSION ASSAYS. THE IMPACT OF MICROBIOTA-ALTERING INTERVENTIONS ON INTESTINAL IMMUNE CELL MIGRATION TO PERIPHERAL LYMPHOID TISSUES AND THE EYE DURING EAU WILL ALSO BE INVESTIGATED. DIRECT MICROBIOTA EFFECTS ON ENHANCEMENT OF INTESTINAL TREG ABUNDANCE AND UVEITIS SEVERITY WILL BE TESTED BY FECAL MICROBIAL TRANSPLANTATION FROM ANTIBIOTIC OR SCFA-PRE-TREATED ANIMALS. THESE RESULTS ARE EXPECTED TO YIELD NOVEL INSIGHTS INTO AUTOIMMUNE UVEITIS PATHOGENESIS AS WELL AS LAY GROUNDWORK FOR NEW TREATMENT STRATEGIES TARGETING THE INTESTINAL MICROBIOTA TO RE-ESTABLISH IMMUNE HOMEOSTASIS.

REDUCING ANTIMICROBIAL OVERUSE THROUGH TARGETED THERAPY FOR PATIENTS WITH COMMUNITY-ACQUIRED PNEUMONIA - PROJECT SUMMARY/ABSTRACT COMMUNITY-ACQUIRED PNEUMONIA (CAP) IS A LEADING CAUSE OF HOSPITALIZATIONS AND INPATIENT MORBIDITY AND MORTALITY IN THE UNITED STATES. HOWEVER, THE ETIOLOGICAL DIAGNOSIS OF CAP IS CHALLENGING AS >75% OF CULTURES ARE NEGATIVE AND A CAUSATIVE PATHOGEN CANNOT BE IDENTIFIED. THE ATS/IDSA GUIDELINES RECOMMEND THAT EXTENDED SPECTRUM EMPIRIC ANTIMICROBIAL THERAPY BE LIMITED TO ADULT PATIENTS WITH RISK FACTORS FOR RESISTANT PATHOGENS. HOWEVER, IN THE FACE OF NEGATIVE CULTURES AND DIAGNOSTIC UNCERTAINTY, CLINICIANS ARE OFTEN UNCOMFORTABLE DE- ESCALATING THERAPY, BECAUSE NARROWING TREATMENT COULD RESULT IN INADEQUATE COVERAGE. THE PROLONGATION OF EMPIRIC TREATMENT HAMPERS ANTIMICROBIAL STEWARDSHIP EFFORTS AND ENCOURAGES THE DEVELOPMENT OF ANTIMICROBIAL RESISTANCE. OUR OVERALL GOAL IS TO IMPROVE ANTIMICROBIAL PRESCRIBING FOR PATIENTS WITH CAP BY EMPHASIZING PATHOGEN-DIRECTED THERAPY. AN ACCURATE PATHOGENIC DIAGNOSIS COULD CONTRIBUTE TO ANTIMICROBIAL STEWARDSHIP IN 2 WAYS: 1) BY ALLOWING FOR INITIAL NARROW-SPECTRUM THERAPY AND 2) BY PROVIDING CONFIDENCE WHEN DE-ESCALATING THERAPY FOLLOWING NEGATIVE CULTURES. RAPID DIAGNOSTIC ASSAYS HAVE THE POTENTIAL TO PROVIDE ACCURATE RESULTS WITHIN HOURS AND THEREBY REDUCE THE DURATION OF EXPOSURE TO EXTENDED SPECTRUM EMPIRIC THERAPY. MULTIPLE OBSERVATIONAL STUDIES HAVE DEMONSTRATED THAT USE OF MOLECULAR DIAGNOSTIC ASSAYS IS ASSOCIATED WITH FAVORABLE OUTCOMES INCLUDING A REDUCTION IN THE TOTAL DURATION OF ANTIMICROBIAL USE AND LENGTH OF STAY IN THE HOSPITAL. THE MOST RECENT ANTIMICROBIAL STEWARDSHIP IMPLEMENTATION GUIDELINES RECOMMEND THE USE OF RAPID VIRAL TESTING FOR RESPIRATORY PATHOGENS AS A MEANS TO REDUCE THE USE OF INAPPROPRIATE ANTIBIOTICS. HOWEVER, THESE RECOMMENDATIONS ARE BASED ON LOW QUALITY EVIDENCE AND IT IS UNKNOWN WHETHER MORE WIDESPREAD EARLY DIAGNOSTIC TESTING COULD REDUCE THE USE OF BROAD-SPECTRUM ANTIBIOTICS AND/OR PROMPT INITIATION OF ANTIVIRAL THERAPY. DE-ESCALATION FOLLOWING NEGATIVE BACTERIAL CULTURES IS ANOTHER ANTIMICROBIAL STEWARDSHIP TARGET. WHILE MOST DE-ESCALATION FOLLOWS IDENTIFICATION OF A SUSCEPTIBLE PATHOGEN, THE ATS/IDSA GUIDELINES ALSO RECOMMEND DE-ESCALATION AT 48 HOURS IF CULTURES ARE NEGATIVE. HOWEVER, THESE RECOMMENDATIONS ARE ALSO BASED ON OBSERVATIONAL STUDIES. WE PROPOSE A LARGE, MULTICENTER 2 X 2 FACTORIAL CLUSTER RANDOMIZED CONTROLLED TRIAL TO TEST BOTH APPROACHES TO REDUCING THE USE OF BROAD-SPECTRUM ANTIBIOTICS IN PATIENTS WITH CAP: A) ROUTINE USE OF RAPID DIAGNOSTIC TESTING AT THE TIME OF ADMISSION AND B) PHARMACIST-LED DE-ESCALATION AFTER 48 HOURS FOR CLINICALLY STABLE PATIENTS WITH NEGATIVE CULTURES. OUR STUDY WILL BE THE LARGEST RANDOMIZED TRIAL TO DETERMINE THE IMPACT OF RAPID DIAGNOSTIC TESTING ON ANTIMICROBIAL STEWARDSHIP AND PATIENT OUTCOMES. OUR RANDOMIZED TRIAL DESIGN WILL ALLOW US TO ESTABLISH CAUSALITY AND DETERMINE WHETHER BROAD SPECTRUM ANTIBIOTICS CAN BE SAFELY DE-ESCALATED IN STABLE PATIENTS. FINDINGS FROM OUR PROPOSED TRIALS WILL GENERATE IMPORTANT NEW KNOWLEDGE ABOUT PATHOGEN TARGETED THERAPY AND ANTIMICROBIAL DE-ESCALATION IN PATIENTS WITH CAP. SPECIFICALLY, KNOWLEDGE GAINED FROM THIS PROPOSAL WILL ALLOW PHYSICIANS TO LIMIT THE USE OF BROAD-SPECTRUM ANTIMICROBIALS AND INITIATE TARGETED THERAPY.

PLATELETS AS BIOSENSORS AND MEDIATORS OF AORTIC ANEURYSM GROWTH - PROJECT SUMMARY ASYMPTOMATIC ABDOMINAL AORTIC ANEURYSMS (AAA) CAN PROGRESS TO RUPTURE WITH AN OUT-OF-HOSPITAL MORTALITY OF 90%. ACCELERATED AAA GROWTH IS ASSOCIATED WITH PLATELET ACTIVATION AND PLATELET AGGREGATES (THROMBI) IN ANEURYSMAL SEGMENTS. ANTIPLATELET DRUGS MAY LIMIT AAA GROWTH AND RUPTURE RISK. A MECHANISTIC EXPLANATION FOR THESE OBSERVATIONS HAS NEVER BEEN ELUCIDATED. THIS PROJECT BUILDS UPON OUR DISCOVERY THAT PLATELETS FROM PATIENTS WITH AAA ARE HYPERACTIVATED THROUGH SELECTIVE SURFACE RECEPTORS AND THAT THE ANTIPLATELET DRUG ASPIRIN PARTIALLY INHIBITS AAA GROWTH AND RUPTURE. THIS SUGGESTS A NEW LINK BETWEEN PLATELET ACTIVATION AND ANEURYSM DEVELOPMENT, AND IMPLIES ASPIRIN MAY NOT BE THE BEST ANTIPLATELET DRUG TO SUPPRESS. ANEURYSM GROWTH. USING AN EX VIVO SYSTEM TO RECAPITULATE DISTURBED (TURBULENT) BLOOD FLOW IN ANEURYSMAL ARTERIES, LOCALIZATION OF AN OLFACTORY RECEPTOR ON THE SURFACE OF THE PLATELET MEMBRANE IS A NEW AND PROMISING TARGET TO SUPPRESSES AAA GROWTH. DISCOVERING THIS PATHWAY IN PLATELETS FROM PATIENTS WITH AAA EXPOSED TO TURBULENT BLOOD FLOW IS A CONCEPTUAL ADVANCEMENT IN OUR UNDERSTANDING OF HOW PLATELETS MECHANICALLY SENSE AND RESPOND TO THEIR EXTERNAL ENVIRONMENT. PLATELETS THEREFORE EMERGE AS CIRCULATING BIOSENSORS, RELEASING PROTEINS THAT ARE USEFUL BIOMARKERS FOR DISTINGUISHING FAST FROM SLOW-GROWING ANEURYSMS. THIS PROJECT OFFERS THE PROMISE OF THE FIRST MEDICAL THERAPY TO TREAT AAA.

KETAMINE TREATMENT OF YOUTH SUICIDE ATTEMPTERS FOR FAST REDUCTION OF SEVERE SUICIDE RISK AND FACILITATION OF LONG-TERM COLLABORATIVE CLINICAL ENGAGEMENT: A RANDOMIZED PLACEBO CONTROLLED TRIAL

DIGITAL TWIN NEIGHBORHOODS FOR RESEARCH ON PLACE-BASED HEALTH INEQUALITIES IN MID-LIFE - DIGITAL TWIN NEIGHBORHOODS FOR RESEARCH ON PLACE-BASED HEALTH INEQUALITIES IN MID-LIFE INCREASINGLY, IT IS RECOGNIZED THAT HIGH-VALUE THERAPIES, SCREENING TOOLS AND PREVENTIVE SERVICES HAVE CREATED OR INCREASED INEQUALITIES EXPERIENCED BY PERSONS FROM RACIAL AND ETHNIC MINORITY BACKGROUNDS, THOSE OF LOW SOCIOECONOMIC POSITION (SEP) AND OTHER VULNERABLE GROUPS. THIS PROJECT AIMS TO CHART A NEW COURSE FOR UNDERSTANDING PLACE-BASED POPULATION HEALTH STRATEGIES. A GROWING LITERATURE ON HEALTH AND PLACE HAS DEMONSTRATED THE OUTSIZE INFLUENCE OF SOCIAL AND NEIGHBORHOOD INDICATORS VERSUS TRADITIONAL CLINICAL MEASURES IN DRIVING INDIVIDUAL HEALTH OUTCOMES. THUS, OUR OVERALL OBJECTIVE IS TO EMPOWER COMMUNITY MEMBERS AND ORGANIZATIONS, LOCAL HEALTH SYSTEMS AND POPULATION HEALTH AND POLITICAL LEADERS TO USE EVIDENCE FROM PLACE- BASED RESEARCH TO INFORM, PRIORITIZE, EVALUATE AND IMPLEMENT HEALTH-PROMOTING STRATEGIES THAT CLOSE HEALTH DISPARITIES. THE CORNERSTONE INNOVATION OF OUR WORK IS THE DEVELOPMENT OF DIGITAL TWIN NEIGHBORHOODS WHICH WILL DRAMATICALLY EXPAND ACCESS TO DATA AND ALGORITHMS FOR UNDERSTANDING PLACED-BASED HEALTH AND SOCIAL INEQUALITIES. DIGITAL TWIN NEIGHBORHOODS (DTNS) ARE DIGITAL REPLICAS OF REAL COMMUNITIES, INCLUDING BIOLOGICAL, SOCIAL AND GEOGRAPHIC DATA AND ALGORITHMS IN A CLOUD COMPUTING ENVIRONMENT. IN THIS PROJECT, WE WILL I) ESTABLISH COMMUNITY- AND PRIVACY-FOCUSED PROCEDURES FOR CONSTRUCTING DIGITAL TWIN NEIGHBORHOODS WHICH INCORPORATE EHR DATA; II) EVALUATE THE EFFICACY OF A DTN APPROACH TO UNDERSTANDING MECHANISMS OF PLACE- BASED HEALTH INEQUITIES IN MID-LIFE ACROSS MULTIPLE HEALTH CONDITIONS AND GEOGRAPHIES; AND III) EXAMINE THE GENERALIZABILITY AND SCALABILITY OF THE DTN APPROACH FOR STUDYING PLACE-BASED MID-LIFE HEALTH INEQUALITIES. THE DEVELOPED OPEN SCIENCE DTN PLATFORM WILL MAKE THE COMBINATION OF MODELING CAPABILITIES AND PRIVACY PRESERVING FEATURES AVAILABLE TO MULTI-SECTOR INITIATIVES THAT ARE AIMED AT EVALUATING LOCAL HEALTH INEQUALITIES AND INFORMING STRATEGIC POPULATION HEALTH POLICY DECISIONS. THUS, THE RESULTS OF THIS WORK WILL I) PROVIDE A FRAMEWORK FOR MECHANISTIC UNDERSTANDING OF CLINICAL, SOCIAL AND ENVIRONMENTAL FORCES PRODUCING DISPARITIES IN LIFE EXPECTANCY, MULTI-MORBIDITY, AND THE ONSET AND MANAGEMENT OF CHRONIC DISEASE AND II) CATALYZE RESEARCHERS AND COMMUNITY AND HEALTH CARE INSTITUTIONS BOTH LOCALLY AND NATIONALLY TO IMPROVE EQUITY AND MEET THE NEEDS OF THE COMMUNITIES THEY SERVE.

CHILDHOOD-ONSET HYPOMYELINATING LEUKODYSTROPHY AND THE MULTI-TRNA SYNTHETASE COMPLEX - PROJECT SUMMARY/ABSTRACT CHILD-ONSET HYPOMYELINATING LEUKODYSTROPHY (HLD) IS A GENETICALLY HETEROGENEOUS GROUP OF NEURODEGENERATIVE DISEASES CHARACTERIZED BY REDUCED CEREBRAL MYELIN FORMATION. CLINICAL FEATURES INCLUDE COGNITIVE AS WELL AS MOTOR IMPAIRMENT APPEARING IN CHILDHOOD. THERE ARE NO CURATIVE TREATMENTS. OUR COLLABORATOR, DR. GRACE YOON, EVALUATED TWO SIBLINGS PRESENTING WITH SEVERE NEUROLOGICAL DEFICIT AND A SHARED PHENOTYPE CONSISTING OF GLOBAL DEVELOPMENTAL DELAY AND INTELLECTUAL DISABILITY, PRENATAL ONSET UNDERGROWTH AND MICROCEPHALY, ROTATORY NYSTAGMUS, ATAXIA AND PROGRESSIVE GAIT DISTURBANCE/SPASTICITY, AND HYPOMYELINATION ON BRAIN MRI. WHOLE EXOME SEQUENCING REVEALED HOMOZYGOSITY OF A C.4444C>A MUTATION IN THE EPRS1 GENE. THE MUTATION IS IN THE CODING SEQUENCE (CDS) OF THE GENE CAUSING A PRO-TO-THR POINT MUTATION AT AA POSITION 1482. THE EPRS1 GENE ENCODES THE BIFUNCTIONAL, GLUTAMYL-PROLYL TRNA SYNTHETASE (EPRS1) THAT RESIDES IN THE CYTOPLASMIC MULTI- TRNA SYNTHETASE COMPLEX (MSC). THE P1482T MUTATION IS LOCATED NEAR THE C-TERMINUS OF THE PROTEIN IN A REGION OF THE PRO SYNTHETASE OUTSIDE THE CATALYTIC OR ANTI-CODON RECOGNITION DOMAINS. THE SPECIFIC ACTIVITY OF RECOMBINANT P1482T MUTANT EPRS1 IS INDISTINGUISHABLE FROM WILD-TYPE. IN CONTRAST, EPRS1 PROTEIN EXPRESSION IN IMMORTALIZED LYMPHOBLASTOID CELL LINES (LCL) FROM AFFECTED SIBLINGS IS ABOUT 20% OF THAT IN UNAFFECTED CONTROLS. EPRS1 MRNA LEVELS ARE IDENTICAL IN SIBLINGS AND CONTROLS INDICATING POST-TRANSCRIPTIONAL REGULATION. OUR PRELIMINARY STUDIES SHOW A DUAL MECHANISM DETERMINING DIMINISHED EPRS1 LEVEL IN AFFECTED LCLS, NAMELY, DECREASED NUCLEAR EXPORT OF MUTANT MRNA, FOLLOWED BY DECREASED CYTOPLASMIC TRANSLATION. WE SHOW THAT DECREASED EPRS1 EXPRESSION CAUSES RELEASE OF OTHER MSC CONSTITUENTS. REMARKABLY, SUPPRESSION OF SEVERAL OTHER MSC COMPONENTS CAUSALLY IMPLICATED IN HLD ALSO INDUCE MSC CONSTITUENT RELEASE, SUGGESTING A COMMON ETIOLOGY OF AN ENTIRE CLASS OF HLDS. WE HYPOTHESIZE THAT INEFFICIENT NUCLEAR EXPORT AND TRANSLATION OF C.4444C>A EPRS1 MRNA REDUCES MSC-BOUND EPRS1, CAUSING RELEASE OF MSC CONSTITUENTS AND DRIVING THE HLD PHENOTYPE. LIKEWISE, DEFECTS IN GENES ENCODING OTHER MSC CONSTITUENTS SHARE A COMMON PATHWAY AND ETIOLOGY OF HLD. WE WILL TEST THIS HYPOTHESIS BY PURSUING THESE SPECIFIC AIMS: IN AIM 1 WE WILL ELUCIDATE MOLECULAR MECHANISMS UNDERLYING REDUCED EXPRESSION OF EPRS1P1482T VARIANT, FOCUSING ON THE ROLE OF M6A METHYLATION. AIM 2 DETERMINES THE HLD-RELATED PHENOTYPE OF OUR NEWLY GENERATED, GENETICALLY-MODIFIED EPRS1P1482T MICE. IN AIM 3 WE WILL INVESTIGATE EFFECT OF DEFICIENCY OF MSC CONSTITUENTS ON MSC INTEGRITY AND ADVERSE CONSEQUENCES IN MYELINATING OLIGODENDROCYTES. A CNS-SPECIFIC TRANSCRIPTOMIC ANALYSIS WILL BE DONE BY SINGLE-NUCLEUS RNA-SEQUENCING. COMPLETION OF THESE STUDIES WILL ELUCIDATE A UNIQUE MECHANISM OF GENE DYSREGULATION THAT INDUCES HLD, AND WILL PROVIDE A UNIQUE MOUSE MODEL OF HLD THAT WILL PERMIT DETAILED IN VIVO ANALYSIS OF THE CELLULAR DEFECTS IN HLD. IMPORTANTLY, THESE RESULTS SUGGEST THE POSSIBILITY OF SPECIFIC, RNA-BASED THERAPEUTIC INTERVENTION TO RESCUE TRANSLATION OF THE VARIANT EPRS1 MRNA AND RESCUE THE NEUROLOGIC DEFECT.

CEREBELLAR DEEP BRAIN STIMULATION TO ENHANCE CHRONIC POST-STROKE REHABILITATION

REDUCING ANTIMICROBIAL OVERUSE IN HCAP THROUGH PERSONALIZED ANTIMICROBIAL RECOMMENDATIONS

GENETIC MODIFIERS OF ATHEROSCLEROSIS AND MACROPHAGE PHENOTYPES - SUMMARY HEART DISEASE IS THE NUMBER ONE KILLER OF MEN AND WOMEN IN THE UNITED STATES. ALTHOUGH THE INCIDENCE OF CARDIOVASCULAR DISEASE DEATHS HAS DECLINED, IT STILL ACCOUNTS FOR ~1 OUT OF EVERY 3 DEATHS. CORONARY ARTERY DISEASE (CAD) DUE TO ATHEROSCLEROSIS WAS RESPONSIBLE FOR MOST OF THESE DEATHS. DESPITE INCREASED KNOWLEDGE ABOUT CAD RISK FACTORS AND THE AVAILABLY OF DRUGS TO TREAT THEM, THE CAD PROBLEM HAS NOT BEEN SOLVED. LARGE HUMAN GENOME WIDE ASSOCIATION STUDIES HAVE IDENTIFIED MANY COMMON GENETIC VARIANTS ASSOCIATED WITH CAD, BUT ONLY A SMALL FRACTION OF THE HERITABLE RISK HAS BEEN DISCOVERED. THIS PROPOSAL AIMS TO PERFORM MOUSE GENETIC AND GENOMIC STUDIES TO IDENTIFY ATHEROSCLEROSIS MODIFIER GENES AND GENETIC MODIFIERS OF MACROPHAGE FOAM CELL LIPID DROPLET METABOLISM AND INFLAMMATION, YIELDING INSIGHTS INTO THE MECHANISMS THAT REGULATE THESE PATHWAYS. THE FIRST AIM OF THE PROPOSED STUDIES INVOLVES IDENTIFYING THE RESPONSIBLE GENES AND GENETIC VARIATION THAT GIVE RISE TO IN VITRO MACROPHAGE PHENOTYPES USING SOPHISTICATED GENETIC AND GENE EDITING APPROACHES. THE SECOND AIM OF THE PROPOSED STUDIES IS TO IDENTIFY THE MOUSE ATHEROSCLEROSIS MODIFIER GENE IN A GENETIC LOCUS IDENTIFIED ON CHROMOSOME 2. THIS AIM WILL USE SOPHISTICATED GENE EDITING AS WELL AS A NEWER MOUSE MODEL OF ATHEROSCLEROSIS INDUCED BY TREATMENT WITH AN ANTISENSE OLIGONUCLEOTIDE TARGETING THE LOW DENSITY LIPOPROTEIN RECEPTOR. THESE FINDINGS MAY LEAD TO NOVEL DRUG TARGETS AND THERAPIES TO PREVENT OR TREAT CAD. THE RELEVANCE OF THE PROPOSED STUDIES IS THAT THEY ADDRESS A SIGNIFICANT HEALTH CONCERN, CORONARY ARTERY DISEASE, AND WILL YIELD INSIGHT INTO THE MECHANISMS THAT MODIFY ATHEROSCLEROSIS SUSCEPTIBILITY. THE DISCOVERY OF NOVEL PATHWAYS AND PROTEINS THAT REGULATE ATHEROSCLEROSIS AND FOAM CELL CHOLESTEROL METABOLISM AND INFLAMMATION OFFERS HOPE FOR NEW MODES OF RISK ASSESSMENT, PREVENTION, AND THERAPY.

ADVANCED IMAGING AND SIMULATION TOOLS FOR PERSONALIZED CORNEAL DISEASE ASSESSMENT AND SURGERY - SIMULATIONS BASED ON PATIENT-SPECIFIC INPUTS HAVE THE POTENTIAL TO DRIVE MAJOR ADVANCES IN PERSONALIZED MEDICINE. HOWEVER, IMPORTANT GAPS PERSIST THAT MUST BE ADDRESSED BEFORE SIMULATION-BASED ENGINEERING CAN BE FULLY LEVERAGED IN THE TREATMENT OF CORNEAL AND REFRACTIVE DISORDERS. THESE INCLUDE DEVELOPMENT OF CLINICAL TOOLS FOR BIOMECHANICAL CHARACTERIZATION, INTEGRATION OF MEASUREMENT AND SIMULATION SYSTEMS, AND MODEL VALIDATION AND VERIFICATION. IN THIS BIOENGINEERING RESEARCH GRANT, WE WILL ADDRESS EACH OF THESE CHALLENGES BY THE FOLLOWING SPECIFIC AIMS: AIM 1. DEVELOP OPTICAL COHERENCE ELASTOGRAPHY (OCE) FOR 3D CHARACTERIZATION OF HETEROGENOUS CORNEAL BIOMECHANICAL PROPERTIES. IN THIS DEVELOPMENTAL AIM, WE WILL DEVELOP AND OPTIMIZE A NEW SYSTEM CAPABLE OF 1) VOLUMETRIC REGIONAL SAMPLING; 2) TRUE 3D DISPLACEMENT TRACKING AND 3) SIMULTANEOUS IOP AND VISCOELASTIC PROPERTY MEASUREMENT. THE SYSTEM WILL BE USED TO ESTABLISH MORE SENSITIVE BIOMECHANICAL BIOMARKERS FOR KERATOCONUS (KC) AND TO INFORM INVERSE FE MODELS FOR 3D PROPERTY ESTIMATION. AIM 2. INTEGRATE 3D OCE AND INVERSE FE MODELING TO CHARACTERIZE AND COMPARE 3D CORNEAL BIOMECHANICAL PROPERTIES IN NORMAL, KC, AND SURGICALLY ALTERED STATES. TOMOGRAPHY AND 3D OCE-DERIVED MEASUREMENTS WILL BE USED TO ESTABLISH AND VALIDATE PATIENT-SPECIFIC FE MODELS. WE WILL CONDUCT HUMAN STUDIES TO TEST THE HYPOTHESIS THAT OCE-DERIVED BIOMARKERS WILL BETTER DISCRIMINATE MANIFEST KC AND SUBCLINICAL KC FROM NORMAL EYES THAN AVAILABLE TOMOGRAPHY AND AIR PUFF-DERIVED BIOMECHANICAL METRICS. WE WILL ALSO MEASURE SPATIAL BIOMECHANICAL CHANGES DURING A LONGITUDINAL STUDY OF KC PROGRESSION AND COMPARE DEPTH- DEPENDENT BIOMECHANICAL CHANGES IN LASIK, SMALL-INCISION LENTICULE EXTRACTION (SMILE), AND CORNEAL CROSSLINKING (CXL). THE LATTER COMPARISON WILL DIRECTLY TEST THE WIDELY PROMULGATED HYPOTHESIS THAT SMILE CAUSES LESS STROMAL WEAKENING THAN LASIK. AIM 3. DEVELOP AND EVALUATE PATIENT-SPECIFIC COMPUTATIONAL MODELS FOR PREDICTING INTERVENTIONAL OUTCOMES, KC PROGRESSION, AND POST-LASIK ECTASIA. WE WILL TEST THE HYPOTHESIS THAT MODELS POPULATED WITH SUBJECT-SPECIFIC GEOMETRY AND MATERIAL PROPERTY DATA ARE MORE ACCURATE PREDICTORS OF SURGICAL OUTCOMES METRICS, KC PROGRESSION RATE, AND POST-LASIK ECTASIA RISK THAN EXISTING METHODS. SUCCESSFUL COMPLETION OF THE AIMS IS EXPECTED TO LEAD TO THE DEVELOPMENT AND IMMEDIATE TRANSLATION OF A PERSONALIZED PRECISION-MEDICINE FRAMEWORK FOR LEVERAGING SUCH DATA FOR MORE EFFECTIVE DIAGNOSIS AND PERSONALIZED TREATMENT PLANNING IN KEY CLINICAL CONDITIONS.

STRUCTURAL ANALYSIS AND THERAPEUTIC NANOBODY DEVELOPMENT OF KSHV G-PROTEIN COUPLED RECEPTOR

A PHASE I ASSESSMENT OF MESENCHYMAL STEM CELLS FOR THE TREATMENT OF MULTIPLE SCLEROSIS

COMPREHENSIVE AUGMENTED REALITY TESTING (CART) PLATFORM FOR PARKINSON?S DISEASE - PROJECT SUMMARY/ABSTRACT PARKINSON’S DISEASE (PD) IS THE FASTEST GROWING NEUROLOGICAL DISEASE, OUTPACING EVEN ALZHEIMER’S. A GAP IN THE EFFECTIVE TREATMENT OF PD IS THE RELIANCE ON CAPACITY-BASED CLINICAL ASSESSMENTS, SUCH AS THE MDS-UPDRS III, TO GUIDE CLINICAL DECISION-MAKING. CLINICAL RATINGS, WHILE SIMPLE TO ADMINISTER, PROVIDE A POOR ESTIMATION OF PD PERFORMANCE OF INSTRUMENTAL ACTIVITIES OF DAILY LIVING (IADLS). DECLINES IN IADL PERFORMANCE ARE OF PARTICULAR IMPORTANCE, AS RECENT POPULATION-BASED COHORT STUDIES INDICATE THAT IADL DECLINE PRE-DATES CLINICAL DIAGNOSIS OF PD BY 5-7 YEARS AND PREDICT THE TRANSITION FROM INDEPENDENCE TO DEPENDENT CARE SETTINGS. IN ORDER TO ADVANCE PD TREATMENT AND DISCOVERY, OBJECTIVE MEASURES OF DISEASE SYMPTOMS DURING THE PERFORMANCE OF ‘REAL-WORLD’ IADL SCENARIOS MUST BE CREATED. AUGMENTED REALITY TECHNOLOGY, IN WHICH COMPUTER-GENERATED IMAGES ARE SUPERIMPOSED IN THE USER’S REAL-WORLD VIEW, ENABLE THE PRESENTATION OF DIGITAL SCENARIOS TO REPLICATE IADLS AND OBJECTIVELY QUANTIFY USER’S PERFORMANCE VIA INTEGRATED IMUS AND DEPTH CAMERA. THE PRIMARY AIM OF THIS PROJECT IS TO DEVELOP AN ACCURATE, VALID, AND RELIABLE AUGMENTED REALITY ASSESSMENT PLATFORM FOR THE QUANTIFICATION OF MOTOR AND NON-MOTOR PERFORMANCE OF PD UNDER CLINICAL AND IADL AUGMENTED REALITY ENVIRONMENTS. THE PROPOSED COMPREHENSIVE AUGMENTED REALITY TEST (CART) PLATFORM FOR PD WILL OBJECTIVELY QUANTIFY THE CARDINAL MOTOR SIGNS OF PD, AS WELL AS IADL PERFORMANCE, TO FACILITATE COMPREHENSIVE TREATMENT OF SYMPTOMS AND PRECISE TRACKING OF DISEASE PROGRESSION THAT CAN EVENTUALLY BE USED TO OPTIMIZE MEDICATION AND DEEP BRAIN STIMULATION PROGRAMMING. A MULTI-DISCIPLINARY TEAM OF EXPERTS IN THE AREAS OF SOFTWARE AND BIOMEDICAL ENGINEERING, NEUROLOGY, NEUROSURGERY, NEUROPSYCHOLOGY, PHYSICAL THERAPY AND STATISTICS WILL COLLABORATE TO DEVELOP THE CART PLATFORM AND DETERMINE ITS ACCURACY, VALIDITY AND RELIABILITY. PARTICULAR ATTENTION WILL BE SPENT ON QUANTIFYING THE INTERPLAY BETWEEN MOTOR AND COGNITIVE TASKS, AS DUAL-TASKS PERFORMANCE IS LINKED TO GAIT DYSFUNCTION AND FALLS AND HISTORICALLY HAS BEEN OVERLOOKED IN CLINICAL ASSESSMENTS. FOLLOWING THE DEVELOPMENT STAGE, A SERIES OF PSYCHOMETRIC STUDIES AND ANALYSES WILL BE CONDUCTED TO DETERMINE THE ACCURACY, VALIDITY AND RELIABILITY OF THE CART PLATFORM IN A COHORT OF 140 INDIVIDUALS WITH PD (N=35 IN EACH HOEHN AND YAHR STAGE I- IV) AND 35 AGE-MATCHED CONTROLS. THE CART PD PLATFORM HAS THE POTENTIAL TO TRANSFORM THE TREATMENT OF PD GLOBALLY BY CREATING AN EQUAL EMPHASIS ON CLINICAL SYMPTOMS AND IADL PERFORMANCE THROUGH THE USE OF AN AFFORDABLE CONSUMER-AVAILABLE ELECTRONIC DEVICE THAT IS SUITABLE FOR THE INTEGRATION INTO TREATMENT DELIVERY SYSTEMS.

HIGH RESOLUTION TRANSCRIPTOME AND GENE REGULATORY MAPPING OF HUMAN URETER AND BLADDER ACROSS THE LIFESPAN - ABSTRACT BENIGN UROLOGIC DISEASES OF THE LOWER URINARY TRACT SUCH AS URINARY TRACT INFECTIONS, BENIGN PROSTATIC HYPERPLASIA, VOIDING DYSFUNCTION, URINARY INCONTINENCE, INTERSTITIAL CYSTITIS, PAINFUL BLADDER SYNDROME, AND URETHRAL STRICTURES INCUR SIGNIFICANT HEALTH CARE BURDEN IN THE US. OUR LACK OF A COMPREHENSIVE MOLECULAR AND CELLULAR UNDERSTANDING OF THESE TISSUES IN THE NORMAL AND DISEASE STATES HAS DIRECTLY HINDERED THE DEVELOPMENT OF EFFECTIVE NOVEL THERAPIES FOR THESE CONDITIONS. WE ARE PROPOSING TO GENERATE HIGH-RESOLUTION TRANSCRIPTOME AND GENE REGULATORY DATA SET FOR HEALTHY HUMAN URETER AND BLADDER TISSUES ACROSS THE LIFESPAN IN BOTH SEXES. THE OVERALL GOAL IS TO CONSTRUCT A HIGHLY INTEGRATED CELLULAR AND MOLECULAR ANATOMICAL MAP OF THE URETER AND BLADDER IN HUMANS. THIS WILL ENABLE US TO COMPREHENSIVELY UNDERSTAND THE CELLULAR COMPOSITION AND LINEAGE RELATIONSHIPS OF THE LOWER URINARY TRACT SYSTEM. TOWARDS THIS AIM, WE HAVE ESTABLISHED A ROBUST TISSUE PROCUREMENT AND PROCESSING WORKFLOW. THIS ALLOWS US TO RECOVER THE LOWER URINARY TRACK EN BLOC FROM DECEASED ORGAN DONORS AND PROCESS THEM, WITH HIGH CELL VIABILITY AND TISSUE INTEGRITY, FOR MOLECULAR ASSAYS. WE PROPOSE TO COLLECT SCRNA-SEQ AND VISIUM SPATIAL TRANSCRIPTOMICS DATA ON ORGAN DONORS IN 3 AGE GROUPS THAT REFLECT DISTINCT PHYSIOLOGICAL STATES IN TERMS OF LOWER URINARY TRACT FUNCTIONS ACROSS THE HUMAN LIFE SPAN. FOR EACH DONOR, WE WILL ANALYZE FIVE SPECIFIED ANATOMIC LOCATIONS, LOWER URETER, DOME, URETERAL ORIFICE, BLADDER NECK, AND URETHRA, TO COMPREHENSIVELY CHARACTERIZE PARTS OF THE LOWER URINARY TRACT THAT HAVE UNIQUE TISSUES STRUCTURES AND FUNCTIONS. WE WILL USE THESE DATA TO UNDERSTAND THE LINEAGE RELATIONSHIPS BETWEEN THE DIFFERENT CELL TYPES AND VALIDATE NOVEL MARKERS FOR EACH NOVEL CELL TYPE BY IMMUNOSTAINING. FINALLY, WE WILL PERFORM SCATAC-SEQ ON THE MOST INFORMATIVE TISSUE LOCATIONS TO GAIN FURTHER INSIGHT INTO THE UNDERLYING GENE REGULATORY NETWORKS. TOGETHER THESE STUDIES WILL PROVIDE, FOR THE FIRST TIME, A COMPREHENSIVE ANALYSIS OF THE HUMAN BLADDER AND URETER THROUGHOUT LIFE. THESE MOLECULAR DATA WILL BECOME A VALUABLE RESOURCE TO THE RESEARCH COMMUNITY AND ULTIMATELY, SUPPORT EFFORTS IN ORGAN REPAIR AND REGENERATION.

KSHVMEDIATED REGULATION OF PROLINE METABOLISM

MODEL-BASED METHODS FOR SINGLE CELL CHROMATIN INTERACTOMIC DATA - PROJECT SUMMARY/ABSTRACT MILLIONS OF CIS-REGULATORY ELEMENTS (CRE) HAVE BEEN IDENTIFIED IN MAMMALIAN GENOMES, WHICH HARBOR LARGE PORTION OF GWAS VARIANTS ASSOCIATED WITH COMPLEX HUMAN DISEASES AND TRAITS. INTERPRETING THE REGULATORY TARGET GENES OF CRE AND GWAS VARIANTS REMAINS CHALLENGING, AS MAJORITY OF GENES ARE NOT MERELY REGULATED BY CRES IN CLOSE ONE-DIMENSIONAL (1D) VICINITY. INSTEAD, CRES CAN FORM DNA LOOPS AND REGULATE THE EXPRESSION OF GENE(S) FROM HUNDREDS OF KILOBASES (KB) AWAY. THUS, DEEP UNDERSTANDING OF CHROMATIN SPATIAL ORGANIZATION CAN SHED LIGHT ON GENE REGULATION AND DISEASE MECHANISMS. DURING THE LAST DECADE, CHROMATIN CONFORMATION CAPTURE (3C)-DERIVED TECHNOLOGIES (E.G., IN SITU HI-C, CAPTURE HI-C, CHIA-PET, PLAC-SEQ AND HICHIP) HAVE BEEN WIDELY USED TO PROVIDE A GENOME-WIDE VIEW OF CHROMATIN SPATIAL ORGANIZATION. HOWEVER, THESE TECHNOLOGIES ARE USUALLY APPLIED TO BULK TISSUE OR PURIFIED CELL LINES, AND CANNOT REVEAL CELL-TYPE-SPECIFIC CHROMATIN INTERACTOME WITHIN COMPLEX TISSUES. FORTUNATELY, HARNESSING THE POWER OF SINGLE CELL TECHNOLOGIES, SINGLE CELL HI-C (SCHI-C) AND SCHI-C-DERIVED MULTI-MODAL ASSAYS, INCLUDING SINGLE CELL METHYL-HIC AND SINGLE- NUCLEUS METHYL-3C, HAVE BEEN RAPIDLY ADVANCED TO STUDY CHROMATIN INTERACTOME AT SINGLE CELL RESOLUTION, PROVIDING POWERFUL TOOLS TO STUDY CHROMATIN SPATIAL ORGANIZATION IN COMPLEX TISSUES AND DISEASE RELEVANT CELL TYPES. WHILE GREAT STRIDES HAVE BEEN MADE IN SCHI-C EXPERIMENTAL TECHNOLOGIES, COMPUTATIONAL METHODS FOR ANALYZING SCHI-C DATA ARE LARGELY LAGGING BEHIND. THE METHODOLOGICAL GAPS FALL MAINLY IN THREE ASPECTS: (1) CURRENT METHODS ARE INEFFICIENT TO ENHANCE RESOLUTION FROM EXTREMELY SPARSE SCHI-C DATA. (2) FEW METHODS EXIST FOR REMOVING SYSTEMATIC BIASES OF SCHI-C DATA WITHIN EACH CELL, AND ADJUSTING FOR BATCH EFFECT ACROSS DIFFERENT CELLS. (3) NO METHOD IS AVAILABLE TO DETECT KB RESOLUTION CELL-TYPE-SPECIFIC CHROMATIN INTERACTIONS FROM SCHI-C DATA. TO FILL IN THESE GAPS, I PROPOSE MAJOR RESEARCH DIRECTIONS: (1) DEVELOP DEEP LEARNING-BASED METHODS TO IMPUTE SPARSE CHROMATIN CONTACTS IN EACH CELL, (2) DEVELOP NON-PARAMETRIC REGRESSION MODELS TO REMOVE SYSTEMATIC BIASES WITHIN EACH CELL, AND TO ADJUST BATCH EFFECTS ACROSS DIFFERENT CELLS, (3) DEVELOP A HYBRID APPROACH BASED ON BOTH GLOBAL AND LOCAL BACKGROUND MODELS TO IDENTIFY CELL-TYPE-SPECIFIC CHROMATIN INTERACTIONS, AND PREDICT PUTATIVE TARGET GENES OF GWAS VARIANTS ASSOCIATED WITH COMPLEX HUMAN DISEASES AND TRAITS, AND (4) DEVELOP STAND-ALONE, USER-FRIENDLY SOFTWARE PACKAGES TO ANALYZE SINGLE CELL CHROMATIN INTERACTOMIC DATA AND DISSEMINATE RESULTS. COMPLETION OF THE PROPOSED STUDY WILL PROVIDE ROBUST AND USER FRIENDLY COMPUTATIONAL METHODS THAT ALLOW US TO ANALYZE 3D GENOME ORGANIZATION AT SINGLE CELL RESOLUTION AND INTERPRET THEIR REGULATORY ROLE ON GENE EXPRESSION AND COMPLEX HUMAN DISEASES.

IRON DYSREGULATION AND NEUROPSYCHIATRIC COMPLICATIONS OF HIV ACROSS THE LIFESPAN: IMPACT OF BIOLOGIC FACTORS, ANTIRETROVIRAL THERAPY AND GENETICS - ABSTRACT PEOPLE WITH HIV EXPERIENCE A MUCH HIGHER PREVALENCE OF DEPRESSION AND NEUROCOGNITIVE IMPAIRMENT (NCI) THAN THE GENERAL POPULATION, AND THESE DISORDERS FREQUENTLY CO-OCCUR, REDUCING MEDICATION ADHERENCE AND QUALITY OF LIFE AND INCREASING MORTALITY AT LEAST TWO-FOLD. STUDIES WITH LARGE NUMBERS OF WOMEN AS WELL AS MEN ARE URGENTLY NEEDED TO ELUCIDATE THE NEUROBIOLOGIC MECHANISMS UNDERLYING THESE DISORDERS AS WELL AS RECENTLY IDENTIFIED SEX DIFFERENCES IN VULNERABILITY. MICROGLIA-MEDIATED NEURO-INFLAMMATION AND ALTERATIONS IN THE KYNURENINE PATHWAY (KP), WHICH IS ESSENTIAL FOR MAINTAINING BALANCED MONO-AMINE NEUROTRANSMITTER (E.G., DOPAMINE AND SEROTONIN) SYNTHESIS IN THE BRAIN, ARE IMPLICATED IN BOTH DISORDERS. IRON, DYSREGULATED BY HIV AND COMBINATION ANTIRETROVIRAL THERAPY (CART), IS INTIMATELY INVOLVED IN MICROGLIAL ACTIVATION, THE KP, AND NEUROTRANSMITTER METABOLISM. OUR PRELIMINARY STUDIES IN A SINGLE NEURO-HIV COHORT AND IN HIV(-) PERSONS INDICATE A SIGNIFICANT ROLE FOR DYSREGULATED IRON METABOLISM AND IRON-RELATED GENE NETWORKS IN DEPRESSION AND NCI, AS WELL AS SEX-SPECIFIC EFFECTS. THE PROPOSED STUDY LEVERAGES EXISTING CLINICAL, IRON-BIOMARKER, AND GENOME-WIDE DATASETS FROM THREE LARGE PROSPECTIVE HIV COHORT STUDIES TO POWERFULLY AND COMPREHENSIVELY INTERROGATE THE ROLE OF IRON IN DEPRESSION AND NCI IN BOTH SEXES. A COMBINATION OF SERUM AND CEREBROSPINAL-FLUID (CSF) BIOMARKER, IRON-CENTERED GENOME-WIDE ASSOCIATIONS, AND NETWORK BIOINFORMATICS APPROACHES WILL BE EMPLOYED. CENTRAL HYPOTHESES OF THIS PROPOSAL ARE THAT IRON DYSREGULATION, AND IRON-RELATED GENES/NETWORKS, ARE MAJOR CONTRIBUTORS TO DEPRESSION AND NCI IN PWH, WITH VARIABILITY IN OVERALL RISK EXPLAINED IN PART BY AGE, SEX, HIV, AND CART EFFECTS ON IRON. THE SPECIFIC AIMS ARE: 1) DETERMINE THE ROLE OF ALTERED IRON HOMEOSTASIS IN DEPRESSION AND NCI IN PWH ACROSS THE AGE SPECTRUM AND THE PROPORTION OF SUSCEPTIBILITY THAT IS ATTRIBUTABLE TO THE EFFECTS OF BIOLOGIC FACTORS, HIV, AND CART ON IRON; 2) IDENTIFY NOVEL IRON-DEPENDENT PATHWAYS, IRON-METABOLIC GENES, AND IRON-QUANTITATIVE TRAIT LOCI ASSOCIATED WITH DEPRESSION AND NCI IN WOMEN AND MEN WITH HIV, AND DETERMINE THE EXTENT TO WHICH THESE GENETIC EFFECTS ARE MEDIATED VIA IRON; 3) EXPLORE THE FUNCTIONAL IMPACT OF NETWORK-PRIORITIZED IRON-RELATED GENE KNOCKDOWN ON PRODUCTION OF KP METABOLITES, USING AN IN VITRO HIV(+/-) HUMAN MICROGLIA MODEL. IN ADDITION, AIM 1 WILL ASSOCIATE IRON INDICES IN A SUBSET OF INDIVIDUALS TO CSF MONO-AMINE METABOLITES. WE EXPECT THESE STUDIES TO ADVANCE UNDERSTANDING OF MECHANISMS LEADING TO DEPRESSION AND NCI IN PEOPLE WITH HIV, SUGGESTING NOVEL PRECISION- MEDICINE APPROACHES TO DISEASE MANAGEMENT AND POTENTIALLY IMPACTFUL, IRON- OR KP-MODULATING INTERVENTIONS.

ALZHEIMER'S DISEASE AND RELATED DEMENTIA-LIKE SEQUELAE OF SARS-COV-2 INFECTION: VIRUS-HOST INTERACTOME, NEUROPATHOBIOLOGY, AND DRUG REPURPOSING - PROJECT SUMMARY ABUNDANT CROSS-INTERDISCIPLINARY EVIDENCE INDICATES THAT THERE ARE MULTIPLE PATHOPHYSIOLOGICAL PROCESSES DRIVING DEVELOPMENT AND PROGRESSION OF ALZHEIMER’S DISEASE (AD) AND AD-RELATED DEMENTIAS (ADRD), INCLUDING NEUROINFLAMMATION AND MICROVASCULAR INJURY TO PATHOGENS, ESPECIALLY VIRUSES. WE ARE EXAMINING THESE ASPECTS OF AD/ADRD WITH RESPECT TO HUMAN CORONAVIRUS DISEASE 2019 (COVID-19), WHICH IS CAUSED BY THE SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS 2 (SARS-COV-2) WITH OVER 87 MILLION CASES IN THE UNITED STATES ALONE. NOTABLY, SUBSTANTIAL EVIDENCE INDICATES NEUROCOGNITIVE SEQUELAE OF COVID-19, WHICH ARE POISED TO ULTIMATELY LEAD TO A SURGE IN CASES OF AD/ADRD, AND OTHER FORMS OF NEUROCOGNITIVE IMPAIRMENT. PRELIMINARY EVIDENCE FROM OUR TEAM IDENTIFIED THAT SARS-COV-2 INFECTION CAUSED NEUROINFLAMMATION AND BRAIN MICROVASCULAR INJURY, TWO MAJOR RISK FACTORS FOR AD/ADRD. WE HAVE ALSO DEMONSTRATED THAT SYSTEMATIC CHARACTERIZATION OF HUMAN- AND VIRUS- HUMAN PROTEIN INTERACTOME MAPS CAN IDENTIFY NOVEL PATHOPHYSIOLOGICAL PATHWAYS AND DRUG TARGETS TO PROTECT THE SARS-COV-2-INFECTED BRAINS. WE THEREFORE POSIT THAT MULTIMODAL ANALYSES OF SARS-COV-2-HUMAN INTERACTOME MAPS IN PATIENT INDUCED PLURIPOTENT STEM CELL (IPSC)-DERIVED BRAIN MICROVASCULAR ENDOTHELIAL CELLS (BMECS) AND BRAIN SINGLE-NUCLEUS GENOMIC/EPIGENOMIC DATA FROM DE NOVO AD/ADRD-LIKE NEUROCOGNITIVE IMPAIRMENT IN COVID-19 PATIENTS (NEUROCOGN-COVID), WILL PROVIDE VALUABLE UNBIASED INSIGHTS INTO THE COMPLEX ETIOLOGY OF NEUROCOGNITIVE SEQUELAE OF SARS-COV-2 AT MOLECULAR, CELLULAR AND NETWORK LEVELS. THIS PROJECT WILL ELUCIDATE CRITICAL UNDERSTANDING OF BOTH BRAIN CELL TYPE-SPECIFIC VIRUS-HUMAN PROTEIN INTERACTOME-INHIBITORY TARGETS AND NEURO-IMMUNE GENE NETWORKS AND BRAIN MICROVASCULAR INJURY THAT MAY LEAD TO AD/ADRD AFTER VIRAL INFECTION. OUR IMMEDIATE GOAL IS TO BUILD A COMPREHENSIVE, BRAIN CELL TYPE-SPECIFIC VIRUS-HUMAN PROTEIN INTERACTOME MAP FOR IDENTIFYING MOLECULAR DRIVERS FOR NEUROCOGNITIVE SEQUELAE OF SARS-COV-2 INFECTION USING OUR HIGH-THROUGHPUT PROTEIN INTERACTOMICS PLATFORM. AIM 1 WILL INTERROGATE THE SARS-COV-2 VIRUS-HUMAN INTERACTOME TO IDENTIFY AND VALIDATE MOLECULAR DRIVERS OF ADRD-LIKE VIRAL MICROVASCULAR INJURY IN IPSC-DERIVED BMECS (AGE-, SEX-, APOE- MATCHED IPSC LINES). AIM 2 WILL INTERROGATE CELL TYPE-SPECIFIC NEUROIMMUNE AND BRAIN ENDOTHELIAL TRANSCRIPTIONAL NETWORKS TO IDENTIFY PATHOPHYSIOLOGY RELATED TO VIRUS-INDUCED NEURO-INFLAMMATION AND BRAIN MICROVASCULAR INJURY. WE WILL LEVERAGE SINGLE-NUCLEUS GENOMIC/EPIGENOMIC DATA GENERATED FROM BRAIN TISSUES OF DONORS WHO SUFFERED FROM NEUROCOGN-COVID, AD, MILD COGNITIVE IMPAIRMENT (MCI), AND AGE-, SEX-, APOE-MATCHED HEALTHY CONTROLS FROM THE CLEVELAND ALZHEIMER'S DISEASE RESEARCH CENTER (ADRC) AND NORTHWESTERN ADRC. AIM 3 WILL TEST THE HYPOTHESIS THAT POTENTIAL NEW OPPORTUNITIES FOR DRUG REPURPOSING IN NEUROCOGNITIVE SEQUELAE CAN BE IDENTIFIED THROUGH A COMBINATION OF LONGITUDINAL POPULATION-BASED VALIDATION AND FUNCTIONAL TESTING IN MOUSE MODELS. SUCCESSFUL COMPLETION OF THIS PROJECT WILL ELUCIDATE MECHANISTIC BIOMARKER FOR NEUROCOGNITIVE SEQUELAE OF SARS- COV-2 AND IDENTIFY NEW DRUG TARGETS AND TREATMENTS TO BE DIRECTLY TESTED IN CLINICAL TRIALS. 1

GLIOVASCULAR MECHANISMS OF BLOOD-BRAIN BARRIER DISRUPTION IN NEUROINFLAMMATORY DISEASE

CHARACTERIZE NEURONAL AND GLIAL CELL-SPECIFIC VULNERABILITY TO PROTEINOPATHIES IN ALZHEIMER'S DISEASE USING MULTIMODAL SINGLE-NUCLEI GENOMIC AND EPIGENOMIC APPROACHES - PROJECT SUMMARY ALZHEIMER’S DISEASE (AD) IS A DEVASTATING NEURODEGENERATIVE CONDITION THAT IT IS ESTIMATED TO AFFECT 16 MILLION AMERICANS BY 2050. AD IS MIXED PROTEINOPATHIES (E.G., AMYLOID-SS (ASS) AGGREGATION, TAU NEUROFIBRILLARY TANGLES, AND TDP-43 INCLUSIONS) AND SELECTIVELY AFFECT CERTAIN REGIONS OF THE BRAIN (E.G., NEOCORTEX AND HIPPOCAMPUS) WITH COMPLEX PATHOPHYSIOLOGY. AND CHARACTERIZATION OF SPECIFIC ALTHOUGH MULTIPLE STUDIES HAVE FOCUSED ON GENETIC FACTORS FOR AD, THE DELINEATION NEURONAL AND GLIAL CELL POPULATIONS WITH ENRICHED VULNERABILITY TO PROTEINOPATHY IN AD REMAINS UNKNOWN. OUR TEAM HAS DEMONSTRATED THAT SINGLE-CELL/NUCLEUS MULTI-OMICS (SNRNA-SEQ/SNATAC- SEQ) CAN BE USED TO INVESTIGATE BOTH “NORMAL” AND “PATHOLOGICAL” NEURONAL AND GLIAL SUBPOPULATIONS FROM HUMAN POST-MORTEM BRAINS AND WE HAVE ALSO ESTABLISHED A LARGE HUMAN BRAIN BIOBANK WITH DIVERSE PROTEINOPATHIES (INCLUDING ASS, TAU, TDP-43 AND OTHERS). MOREOVER, WE HAVE DEMONSTRATED HOW TARGETING PROTEINOPATHY-SPECIFIC NETWORKS, SUCH AS ACETYLATED TAU AND SYNERGISTIC PROTEINOPATHY NETWORKS SHARED BY TAU AND ASS, CAN IDENTIFY REPURPOSABLE TREATMENTS (E.G., SILDENAFIL AND DIFLUNISAL) FOR AD. OUR PRELIMINARY SNRNA-SEQ AND SNATACT-SEQ ANALYSES OF HUMAN POST-MORTEM CEREBELLUM REGIONS FROM 7 INDIVIDUALS (N=4 [AD] AND N=3 OTHER DEMENTIA CASES) HAVE REVEALED UNIQUE NEURONAL AND GLIAL CELL POPULATIONS AND GENES/NETWORKS WHEN COMPARING TO TRADITIONAL SNRNA-SEQ DATA FROM NEOCORTEX AND HIPPOCAMPUS. OUR INTEGRATIVE SNRNA-SEQ DATA ANALYSIS HAS ALSO IDENTIFIED DISEASE-RELEVANT MICROGLIAL SUBTYPES, INCLUDING MICROGLIA CONTAINING AMYLOID-B/PHOSPHOR-TAU, AS WELL AS MICROGLIA ENRICHED IN EXPRESSION OF PRO-INFLAMMATORY MARKERS, USING DEEP GENERATIVE MODELS. WE THEREFORE HYPOTHESIZE THAT COMPREHENSIVE CHARACTERIZATION OF HUMAN NEURONAL AND GLIAL CELL GENOMIC AND EPIGENOMIC SIGNATURES AND NETWORKS THAT ARE VULNERABLE TO PROTEINOPATHIES WILL HELP TO IDENTIFY NOVEL MECHANISTIC PATHWAYS AND DISEASE- MODIFYING TREATMENTS. IN AIM 1, WE WILL GENERATE COMPREHENSIVE MULTI-OME DATA OF HUMAN NEURONAL AND GLIAL CELLS VULNERABLE TO AD PROTEINOPATHIES. WE WILL USE A SAMPLE POOLING SNRNA-SEQ/SNATAC-SEQ TECHNOLOGY TO ANALYZE HUMAN POST-MORTEM NEOCORTEX, HIPPOCAMPUS, AND CEREBELLUM WITH VARYING DEGREES OF PROTEINOPATHY SEVERITY (AMYLOID-B, P-TAU AND TDP-43) AND AGE-, SEX- AND APOE-MATCHED COGNITIVE HEALTHY CONTROLS AVAILABLE FROM THE NORTHWESTERN ALZHEIMER’S DISEASE RESEARCH CENTER. IN AIM 2, WE WILL TEST THE HYPOTHESIS THAT NEURONAL AND/OR GLIAL CELL-SPECIFIC GENOMIC/EPIGENOMIC SIGNATURES AND NETWORKS IDENTIFY THE MOLECULAR MECHANISM(S) OF VULNERABILITY AND RESILIENCE IN AD. THESE MULTIMODAL DATA ANALYSES WILL INTEGRATE LARGE SNRNA-SEQ/SNATAC-SEQ PROFILES WITH EXISTING WHOLE GENOME-SEQUENCING DATA FROM THE ALZHEIMER’S DISEASE SEQUENCING PROJECT (ADSP). IN AIM 3, WE WILL TEST THE HYPOTHESIS THAT SELECTIVE CELLULAR VULNERABILITY LINKED TO GENES/NETWORKS CAN BE TARGETED VIA PHARMACOLOGIC TREATMENT TO SLOW PROGRESSION OF AD-LIKE DISEASE IN ANIMAL MODELS. SUCCESSFUL COMPLETION OF OUR PROJECT WILL IDENTIFY NEW TREATMENT OPPORTUNITIES THAT TARGET SPECIFIC NEURONAL AND/OR GLIAL CELL-SPECIFIC RISK/RESILIENCE GENES AND NETWORKS THAT CONFER VULNERABILITY TO PROTEINOPATHIES IN AD AND OTHER DEMENTIAS.

TARGETING ONCOGENIC DOPAMINE RECEPTOR SIGNALING IN GLIOBLASTOMA

UNRAVELING STRAIN-LEVEL VARIATION IN THE INDUCTION OF ANTI-INFLAMMATORY RESPONSES IN THE INTESTINE - PROJECT SUMMARY: INFLAMMATORY BOWEL DISEASE (IBD) IS AN INCREASINGLY PREVALENT DISEASE THAT CURRENTLY AFFECTS ~1.3% OF ADULTS IN THE US. IBD IS CHARACTERIZED BY CHRONIC INFLAMMATORY IMMUNE RESPONSES DIRECTED AGAINST THE GUT MICROBIOTA, AND SEVERELY IMPEDES THE HEALTH OF ITS SUFFERERS. CURRENT THERAPEUTIC APPROACHES INVOLVE NEUTRALIZATION OF PATHOGENIC INFLAMMATORY PATHWAYS. HOWEVER, MANY PATIENTS ARE NON-RESPONSIVE OR BECOME REFRACTORY TO TREATMENT, AND THE REQUIREMENT FOR SUSTAINED ADMINISTRATION OF THESE AGENTS CAN ENHANCE SUSCEPTIBILITY TO INFECTION. A MAJOR UNMET CLINICAL NEED ENTAILS DEVELOPMENT OF IMPROVED THERAPEUTIC REGIMENS THAT QUELL ONGOING INFLAMMATION WHILE SPARING PROTECTIVE IMMUNITY. STRATEGIES THAT SEEK TO RESTORE HOST IMMUNE- GUT MICROBIOTA HOMEOSTASIS THROUGH INTRODUCTION OF HEALTH-PROMOTING IMMUNOMODULATORY MICROBES (PROBIOTICS), REPRESENT AN ATTRACTIVE ALTERNATIVE TO BLOCKADE OF IMMUNE FUNCTION. TO DATE, THESE APPROACHES HAVE DEMON- STRATED LIMITED EFFICACY. OUR INCOMPLETE UNDERSTANDING OF THE MECHANISMS THROUGH WHICH MICROBES INDUCE ANTI- INFLAMMATORY RESPONSES, AND HOW TRANSPLANTED MICROBES SURVIVE THE HOSTILE ENVIRONMENT OF THE INFLAMED INTES- TINE TO ESTABLISH A NICHE HAVE SEVERELY HAMPERED THESE EFFORTS. AN APPROACH WHERE THE OPTIMAL FEATURES FROM DIFFERENT MICROBES ARE COMBINED, SO-CALLED DESIGNER PROBIOTICS, REPRESENTS AN IMPROVED TREATMENT STRATEGY. KNOWLEDGE GAP: THE IDENTITY OF THE BACTERIAL PATHWAYS THAT ACTIVELY PROMOTE INTESTINAL ANTI-INFLAMMATORY IM- MUNE RESPONSES AND ALLOW PROBIOTIC STRAINS TO COLONIZE THE INFLAMED INTESTINE HAVE REMAINED ENIGMATIC DUE TO MICROBIOTA COMPLEXITY AND DIFFICULTIES ASSOCIATED WITH THE GENETIC MANIPULATION OF GUT MICROBES. HYPOTHESIS: STRAIN-SPECIFIC DIFFERENCES IMPACT THE PROBIOTIC POTENTIAL OF GUT BACTERIAL SPECIES. PRELIMINARY STUDIES: THROUGH THE STUDY OF DISTINCT STRAINS OF THE GENETICALLY TRACTABLE GUT SYMBIONT BACTEROIDES THETAIOTAOMICRON, WE HAVE (I) IDENTIFIED EXTENSIVE STRAIN-LEVEL VARIATION IN THE ABILITY OF B. THETAIOTAOMICRON STRAINS TO INDUCE ACCUMULATION OF COLONIC TREGS IN MONOCOLONIZED GNOTOBIOTIC MICE, (II) REVEALED SIGNIFICANT STRAIN-LEVEL VARIATION IN THE BIOFILM- FORMING CAPACITY OF DIFFERENT STRAINS OF B. THETAIOTAOMICRON, AND (III) UNCOVERED THE EXISTENCE OF A NOVEL, B. THE- TAIOTAOMICRON-DERIVED, IMMUNOMODULATORY FACTOR THAT PROMOTES PRODUCTION OF THE ANTI-INFLAMMATORY CYTOKINE IL- 10. OUR SYSTEMS PROVIDE AN OPPORTUNITY TO LEVERAGE THE RELATEDNESS OF STRAINS WITHIN A SPECIES THAT IMPART DIF- FERENTIAL PHENOTYPES TO PROVIDE INSIGHT INTO PATHWAYS RELATED TO THE OPTIMAL FUNCTION OF PROBIOTICS. PROJECT OB- JECTIVE: TO LEVERAGE THE STRAIN-LEVEL VARIATION AND GENETIC TRACTABILITY OF B. THETAIOTAOMICRON TO DEFINE THE BACTE- RIAL GENES AND MOLECULES THAT MOST POTENTLY CONFER ANTI-INFLAMMATORY CAPACITY TO GUT MICROBES. IMPACT: RESULTS OF THESE STUDIES WILL ADVANCE EFFORTS TO DEVELOP DESIGNER PROBIOTIC THERAPEUTICS THAT PROVIDE DURABLE REMISSION FROM DISEASE FOR IBD PATIENTS. AIM 1: DEFINE THE MOLECULAR BASIS FOR STRAIN-LEVEL VARIATION IN BACTERIAL DRIVEN COLONIC TREG INDUCTION. AIM 2: DEFINE THE B. THETAIOTAOMICRON-DERIVED IMMUNOMODULATORY FACTOR(S) THAT LIMIT COLITIS. AIM 3: DEFINE THE GENETIC DETERMINANTS MEDIATING BACTERIAL STRAIN-LEVEL FITNESS IN THE INFLAMED INTESTINE.

INNATE IMMUNE RESPONSES TO TISSUE INFECTION BY INTESTINAL FUNGI INHIBIT WOUND REPAIR - ABSTRACT ALTERATIONS OF THE MYCOBIOME COMPOSITION THAT HAVE BEEN ASSOCIATED WITH CROHN’S DISEASE (CD) ARE CHALLENGING TO LINK TO DEFINING ELEMENTS OF PATHOPHYSIOLOGY OF THIS DISEASE SUCH AS POOR INJURY REPAIR. OUR PUBLISHED WORK SHOWS THAT USING CULTURE DEPENDENT AND INDEPENDENT METHODS, DEBARYOMYCES HANSENII, A YEAST BEST KNOWN FOR ITS ROLE AS AN ADDITIVE IN THE FOOD INDUSTRY, IS THE DOMINANT MICROBE FOUND TO NON-HEALED INTESTINAL WOUNDS OF SUBJECTS WITH CD IN TWO DIFFERENT MEDICAL CENTERS. D. HANSENII WAS NOT FOUND IN ADJACENT HEALTHY TISSUE. D. HANSENII STRAINS CULTURED FROM INFLAMED CD MUCOSAL TISSUE IMPAIRED COLONIC HEALING WHEN INTRODUCED INTO INJURED CONVENTIONALLY RAISED OR GNOTOBIOTIC MICE. D. HANSENII WAS RE-ISOLATED FROM INJURED AREAS OF THESE MICE, FULFILLING KOCH’S POSTULATES USING ANIMAL MODELS. MECHANISTICALLY, D. HANSENII IMPAIRED MUCOSAL HEALING VIA MACROPHAGE PRODUCTION OF TYPE 1 INTERFERON (IFN) (NOT TRADITIONAL PROINFLAMMATORY CYTOKINES SUCH AS TNF) THAT IN TURN LED TO OVERPRODUCTION OF THE CHEMOKINE CCL5 A DRIVER OF POOR REPAIR. WE HAVE NEW PRELIMINARY DATA THAT SHOWS D. HANSENII SPORES AND VEGETATIVE CELLS ARE BOTH IMPORTANT TO THE OVERALL PATHOLOGY OF POOR WOUND REPAIR IN THE INTESTINE. WE HYPOTHESIZE THAT IN DAMAGED INTESTINES, D. HANSENII SPORES ARE TAKEN UP BY F4/80+ INNATE IMMUNE CELLS IN THE WOUND BED WITHOUT ELICITING A TNF RESPONSE THEREBY ALLOWING FOR PERSISTENCE WITHIN THE MACROPHAGE. WE ALSO HYPOTHESIZE THAT THESE D. HANSENII SPORES GERMINATE INTO VEGETATIVE CELLS WITHIN THESE MACROPHAGES. THIS PROCESS STIMULATES TYPE I IFN PRODUCTION THROUGH TLR3 ENGAGEMENT AND THIS IMMUNE RESPONSE PREVENTS WOUND HEALING. OUR GOALS ARE TO DETERMINE HOW D. HANSENII SPORES EVADE MACROPHAGE KILLING (AIM 1) AND HOW GERMINATING/VEGETATIVE CELLS CAN STIMULATE TYPE I IFN (AIM 2). IN AIM 3, WE WILL ALSO DEVELOP TOOLS FOR THE DETECTION OF D. HANSENII SPORES AND VEGETATIVE CELLS IN MURINE MODELS OF INJURY AND WE WILL USE THESE TOOLS TO DETERMINE IN PRECLINICAL MODELS TO DETERMINE IF ANTIFUNGAL THERAPY CAN EFFECTIVELY ERADICATE CELLS IN DIFFERENT MORPHOLOGIES WITH THE GOAL OF RESTORING WOUND HEALING. OUR STUDIES WILL PROVIDE TOOLS NEW KNOWLEDGE TO UNDERSTAND WHY D. HANSENII BEHAVES AS A PATHOGEN IN CD, AND WE WILL USE THIS INFORMATION TO DEVELOP CONCEPTS FOR NEW DIAGNOSTICS AND THERAPIES FOR CD PATIENTS.

MANIPULATION OF HOST FACTORS THAT PROMOTE HCMV LATENCY - PROJECT SUMMARY/ABSTRACT – O’CONNOR, CHRISTINE M. HUMAN CYTOMEGALOVIRUS (HCMV) IS A WIDE-SPREAD PATHOGEN, INFECTING THE MAJORITY OF THE POPULATION IN THE UNITED STATES. THIS VIRUS POSES A SIGNIFICANT THREAT TO DEVELOPING FETUSES AS WELL AS TO CHILDREN AND ADULTS WHO LACK A COMPETENT IMMUNE SYSTEM, OFTEN CAUSING SEVERE DISEASE AND MORTALITY. ONCE INDIVIDUALS ACQUIRE AN HCMV INFECTION, THE VIRUS REMAINS WITH THE HOST FOR LIFE, IN A LATENT OR QUIESCENT STATE IN THE HEMATOPOIETIC COMPARTMENT. DURING TIMES OF SEVERE IMMUNOLOGICAL STRESS, THE VIRUS REACTIVATES TO ITS ACTIVE STATE, ALLOWING FOR DISSEMINATION AND SUBSEQUENT DISEASE. WITH THE EXCEPTION OF THE IMMUNO-NAÏVE AND SERO-NEGATIVE ORGAN TRANSPLANT RECIPIENTS, PRIMARY INFECTION WITH HCMV RARELY CAUSES DISEASE, BUT RATHER IT IS REACTIVATION THAT LEADS TO SIGNIFICANT COMPLICATIONS. THUS, TO PREVENT HCMV-ASSOCIATED DISEASE, WE MUST GAIN A COMPLETE UNDERSTANDING OF VIRAL LATENCY AND REACTIVATION. ONE OF A HANDFUL OF GENES ENCODED DURING LATENCY IS ONE OF THE FOUR VIRAL G PROTEIN-COUPLED RECEPTORS (GPCRS), US28. WE HAVE SHOWN PREVIOUSLY THAT US28 IS REQUIRED FOR THE ESTABLISHMENT AND MAINTENANCE LATENCY AND THAT US28-MEDIATED SIGNALING CONTRIBUTES TO THESE EFFECTS. WE HAVE ALSO FOUND THAT US28 MODULATES THE EXPRESSION OF SPECIFIC CELLULAR TARGETS THAT REGULATE THE MAJOR IMMEDIATE EARLY PROMOTER (MIEP), A MASTER REGULATOR IN THE LATENT-TO-LYTIC SWITCH. THEREFORE, WE HYPOTHESIZE THAT US28 MODULATES SPECIFIC HOST SIGNALING PATHWAYS TO REGULATE TRANSCRIPTIONAL SILENCING OF THE MIEP TO FACILITATE HCMV LATENCY. TO EXPLORE THIS HYPOTHESIS, WE WILL TAKE ADVANTAGE NOVEL APPROACHES COUPLED WITH OUR ARSENAL OF US28- SPECIFIC RECOMBINANTS, AS WELL AS BOTH IN VITRO AND EX VIVO LATENCY MODEL SYSTEMS. IN AIM 1, WE WILL DEFINE MECHANISMS UNDERLYING US28-MEDIATED ATTENUATION OF AP-1 TRANSCRIPTION FACTOR BINDING TO THE MIEP DURING LATENCY BY EXAMINING THE UPSTREAM SIGNALING PATHWAYS THAT REGULATE AP-1. IN AIM 2, WE WILL DETERMINE THE FACTORS THAT RECRUIT YY1 TO THE MIEP DURING LATENCY BY ASSESSING US28-REGULATED SIGNALING PATHWAYS AND FACTORS THAT PROMOTE THE BINDING OF THIS REPRESSIVE TRANSCRIPTION FACTOR. IN SUM, THE EXPERIMENTS PROPOSED HEREIN WILL LEAD TO A GREATER UNDERSTANDING OF US28’S BIOLOGICAL FUNCTIONS DURING LATENCY AND WILL LAY THE FOUNDATION FOR FUTURE STUDIES TO DEVELOP NOVEL THERAPEUTICS SPECIFICALLY TARGETING THE LATENT RESERVOIR OF HCMV INFECTION.

ADIPOKINES, AGING, AND ALZHEIMERS DISEASE

APOL1 STUDIES IN KIDNEY TRANSPLANTATION CONSORTIUM CLINICAL CENTERS (ASK-CCC)

SENSING ACTIVE MOVEMENT OF THE SELF: RECONSIDERING THE CELLULAR BASIS KINESTHESIA - ABSTRACT THE SENSE OF MOVEMENT (KINESTHESIA) PROVIDES AN INTEROCEPTIVE INTERNAL READOUT OF OUR PHYSICAL ACTIONS IN SPACE AND IS ESSENTIAL FOR OUR ABILITY TO MOVE FLUIDLY AND EFFECTIVELY THROUGH OUR ENVIRONMENT. DESPITE KINESTHESIA'S IMPORTANCE IN MOTOR FUNCTION AND SELF-REFERENCE, OUR UNDERSTANDING OF THIS SENSE IS PLAGUED BY GLARING KNOWLEDGE GAPS AND INCONSISTENCIES. MOVEMENT SENSATIONS ARE TRADITIONALLY BELIEVED TO BE A SPECIALIZED FUNCTION OF TYPE IA MUSCLE SPINDLE AFFERENTS. YET, THE APPARENT DISCONNECT BETWEEN THE PERIPHERAL CODING PROPERTIES OF THIS RECEPTOR AND THE SENSORY STIMULI KNOWN TO EVOKE A SENSE OF MOVEMENT HAVE RAISED QUESTIONS REGARDING THEIR PRIMARY ROLE IN KINESTHESIA. ALTHOUGH PROPRIOCEPTIVE INTERVENTIONS PROVIDE FUNCTIONAL MOTOR IMPROVEMENTS FOR MANY CONDITIONS SUCH AS STROKE, PARKINSON'S DISEASE, FOCAL DYSTONIA, PERIPHERAL NEUROPATHIES, AND MUSCULOSKELETAL INJURIES, THE LACK OF A CLEAR SCIENTIFIC FOUNDATION FOR KINESTHESIA IMPACTS OUR UNDERSTANDING OF SENSORY-MOTOR DEFICITS AND PREVENTS IMPORTANT BREAKTHROUGHS FROM TRANSLATING INTO CLINICAL SUCCESSES AND TARGETED INTERVENTION STRATEGIES. FROM OUR RECENT WORK WE HAVE MULTIPLE LINES OF EVIDENCE THAT SUGGEST THERE MAY BE SENSORY MUSCLE RECEPTORS, OUTSIDE OF THE TRADITIONAL MUSCLE SPINDLES AND GOLGI TENDON ORGANS THAT EXHIBIT FEATURES CONSISTENT WITH A KINESTHETIC SENSOR. FIRST, OUR PERIPHERAL ELECTROPHYSIOLOGICAL RECORDINGS IN RAT DEMONSTRATE A POPULATION OF FAST- CONDUCTING RAPIDLY-ADAPTING AFFERENTS, THAT ARE DISTINCT FROM MUSCLE SPINDLE AND GOLGI TENDON ORGAN AFFERENTS, YET ARE SELECTIVELY ACTIVATED IN THE FREQUENCY BANDWIDTH ASSOCIATED WITH KINESTHETIC ILLUSIONS. SECOND, OUR IMMUNOLOGICAL ANALYSES IN MOUSE SKELETAL MUSCLE REVEAL A NEW POPULATION OF LARGE CALIBER CALBINDIN28K+ AFFERENTS THAT DO NOT ASSOCIATE WITH MUSCLE SPINDLE OR GOLGI TENDON ORGAN RECEPTORS BUT INSTEAD TERMINATE IN FREE ENDINGS THAT SPREAD OUT ALONGSIDE EXTRAFUSAL MUSCLE FIBERS. IN A MOVEMENT-PERCEPTION STUDY WITH HUMAN NEURAL- MACHINE INTERFACE AMPUTEES, WE FOUND THAT VIBRATION-INDUCE ILLUSORY KINESTHETIC PERCEPTS WERE LINKED TO MUSCLE CONTRACTION NOT ELONGATION. THESE RESULTS WERE CORROBORATED IN A HUMAN STROKE MODEL WHERE WE AMPLIFIED KINESTHETIC PERCEPTION LINKED TO ACTIVE MUSCLE CONTRACTION WHICH RESULTED IN IMPROVED REACHING TRAJECTORIES. WITH THESE OBSERVATIONS WE HYPOTHESIZE THAT THERE ARE CANDIDATE MUSCLE SENSORY AFFERENTS, DISTINCT FROM TYPE IA AFFERENTS, WHICH SELECTIVELY RESPOND TO MUSCLE FIBER CONTRACTION. THE STUDIES IN THIS PROPOSAL WILL EXPLORE THE RELATIONSHIPS BETWEEN THE RESPONSE PROPERTIES AND PHYSICAL CHARACTERISTICS OF THESE CANDIDATE KINESTHETIC RECEPTORS AND THE TRADITIONALLY DEFINED MUSCLE SENSORY RECEPTORS USING GENETIC, HISTOLOGICAL, AND ELECTROPHYSIOLOGICAL APPROACHES. ADDITIONALLY, WE WILL EXAMINE THIS SYSTEMS FUNCTIONALITY WITH RESPECT TO CONTRACTILE FEATURES AND ITS ABILITY TO SERVE AS A STIMULUS FOR ACTIVE MOVEMENT SENSING. THE DISCOVERY AND EVALUATION OF THE CELLULAR BASIS OF KINESTHESIA WILL FUNDAMENTALLY TRANSFORM OUR UNDERSTANDING OF SENSORY-MOTOR CONTROL AND, BY EXTENSION, WILL IMPACT DESIGN STRATEGIES FOR ADVANCED NEURAL-MACHINE INTERFACE PROSTHETIC DEVICES FOR AMPUTEES, AS WELL AS OTHER DISORDERS WITH SENSORY-MOTOR DEFICIENCIES SUCH AS STROKE.

THE MAMMALIAN MULTI-TRNA SYNTHETASE COMPLEX - PROJECT SUMMARY/ABSTRACT MAMMALIAN CELLS CONTAIN A CYTOPLASMIC MULTI-TRNA SYNTHETASE COMPLEX (MSC) CONSISTING OF 8 AMINOACYL-TRNA SYNTHETASES (AARSS) AND 3 NON-SYNTHETASE PROTEINS. AARSS IN THE MSC FUNCTION AS “GENE DECODERS” DURING MRNA TRANSLATION, BUT ALSO EXHIBIT NON-CANONICAL FUNCTIONS OUTSIDE THE MSC. HOWEVER, THE ASSEMBLY, STRUCTURE, AND FUNCTION OF THE MSC ARE POORLY UNDERSTOOD. IMPORTANTLY, MUTATIONS IN GENES ENCODING 7/11 CONSTITUENTS CAUSE CENTRAL NERVOUS SYSTEM (CNS) DISORDERS – FIVE CAUSE HYPOMYELINATING LEUKODYSTROPHY (HLD), AND TWO OTHERS CAUSE PROGRESSIVE MICROCEPHALY. WE WILL UTILIZE STATE-OF-THE-ART MOLECULAR APPROACHES TO IMPROVE OUR UNDERSTANDING OF THE MSC, AND ITS POTENTIAL ROLE IN NEUROPATHOLOGY. OUR PROPOSED MULTIPLE-PI PROGRAM TAKES ADVANTAGE OF THE EXPERTISE OF TWO HIGHLY COLLABORATIVE PI'S – PAUL FOX (CONTACT PI), A MOLECULAR BIOLOGIST WITH LONG-TERM INTEREST IN TRNA SYNTHETASES AND THE MSC, AND VALENTIN GOGONEA (MULTIPLE PI), A PHYSICAL CHEMIST WITH EXPERTISE IN ANALYSIS AND MOLECULAR MODELING OF MULTI-PROTEIN COMPLEXES. WE WILL DETERMINE THE QUATERNARY STRUCTURE OF THE MSC BY CROSS-LINKING MASS SPECTROMETRY (XL-MS), A STATE-OF-THE-ART METHOD THAT FACILITATES ANALYSIS OF OTHERWISE INTRACTABLE COMPLEXES. TO DATE WE HAVE FOUND 19 INTER-PROTEIN CROSS-LINKS BETWEEN ALL 11 MSC CONSTITUENTS, AND 118 INTRA-PROTEIN CROSS-LINKS. WE HAVE GENERATED AN INITIAL MODEL OF THE MSC THAT WILL BE REFINED HERE BY XL-MS EXPERIMENTS WITH EXPANDED AMINO ACID SPECIFICITY, AND BY SIMPULL (SINGLE-MOLECULE PULLDOWN) COUPLED WITH SINGLE-MOLECULE FLUORESCENCE TO DETERMINE STOICHIOMETRY. IN ADDITION, WE WILL INVESTIGATE THE MECHANISM OF ASSEMBLY OF THE MSC. CONSTITUTIVE, MULTI-PROTEIN COMPLEXES ARE THOUGHT TO BE ASSEMBLED BY DOMAIN-SPECIFIC INTERACTIONS BETWEEN FULLY-FORMED, MATURE CONSTITUENTS (“POST-TRANSLATIONAL ASSEMBLY”). HOWEVER, ASSEMBLY OF SOME COMPLEXES UTILIZES A “CO-TRANSLATIONAL ASSEMBLY” MECHANISM IN WHICH A MATURE CONSTITUENT INTERACTS WITH THE NASCENT PEPTIDE OF A PARTNER CONSTITUENT AS IT EMERGES FROM THE RIBOSOME. IN PRELIMINARY DATA WE SHOW AT LEAST 10 PAIRS OF MSC CONSTITUENTS INTERACT CO-TRANSLATIONALLY. WE WILL APPLY THESE MECHANISTIC APPROACHES TO ELUCIDATE THE ROLE OF TWO MSC CONSTITUENTS IN CNS DISEASES – GENETIC DEFECTS IN QARS1 AND EPRS1 THAT CAUSE MICROCEPHALY AND HLD, RESPECTIVELY. OUR PRELIMINARY STUDIES INDICATE THAT CONSTITUENT MUTATION OR SUPPRESSION CAN LEAD TO EXTRA-MSC ACCUMULATION. OUR PRELIMINARY STUDIES HAVE LED US TO PROPOSE THE FOLLOWING HYPOTHESIS: THE MAMMALIAN MSC IS A COMPACT STRUCTURE ASSEMBLED IN PART BY AN ORDERLY SEQUENCE OF CO-TRANSLATIONAL INTERACTIONS, HOWEVER, MIS-ASSEMBLY OR MUTATION CAN INDUCE EXTRA-MSC ACCUMULATION OF CONSTITUENTS, WITH POTENTIALLY DELETERIOUS DOWNSTREAM CONSEQUENCES. WE WILL TEST THIS HYPOTHESIS BY (1) DETERMINING MSC QUATERNARY STRUCTURE AND COMPONENT STOICHIOMETRY, AND (2) DETERMINING THE ROLE OF CO-TRANSLATIONAL INTERACTIONS IN MSC FORMATION AND INTEGRITY. WE ANTICIPATE THAT ELUCIDATION OF THE STRUCTURE AND ASSEMBLY OF THE MSC WILL PROVIDE INSIGHTS INTO MECHANISMS BY WHICH MOLECULAR DEFECTS IN MSC CONSTITUENTS CAN CAUSE SEVERE PATHOLOGICAL DISTURBANCES, IN PARTICULAR, DEBILITATING DISORDERS OF THE CNS.

THE EFFECT OF NUCLEOSOMES ON THE EARLIEST STAGES OF RNA POLYMERASE II TRANSCRIPTION

AWARENESS AND ACCESS TO CARE FOR CHILDREN AND YOUTH WITH EPILEPSY

GUT MICROBIAL DIETARY PHENYLALANINE METABOLISM AND HEART FAILURE - PROJECT SUMMARY THIS APPLICATION IS ENTITLED “GUT MICROBIAL DIETARY PHENYLALANINE METABOLISM AND HEART FAILURE”. THERE IS RAPIDLY GROWING AWARENESS OF THE SIGNIFICANT ROLE OF DIET AND THE GUT MICROBIOME TO HUMAN HEALTH AND DISEASE. HERE WE EXAMINE HOW DIFFERENCES IN MACRONUTRIENT METABOLISM BETWEEN SUBJECTS AT RISK FOR HEART FAILURE ARE LINKED TO CHANGES IN GUT MICROBIAL COMMUNITY STRUCTURE AND FUNCTION, AND CONTRIBUTE TO DISEASE (HEART FAILURE) ETIOLOGY. SPECIFICALLY, HOW ALTERATIONS IN GUT MICROBIAL PHENYLALANINE METABOLISM IMPACTS PATIENT RISKS FOR DEVELOPMENT OF DISEASE, AND MULTIPLE PATHOPHYSIOLOGIC MECHANISMS LINKED TO HEART FAILURE RELEVANT PHENOTYPES. OUR STUDIES REVEAL HOW ALTERED MICROBIAL METABOLISM OF PHENYLALANINE, AN ESSENTIAL MACRONUTRIENT, ALTERS METABOLIC HOMEOSTASIS, EXAMINED NEW BIOMARKERS OF BOTH HEART FAILURE DEVELOPMENT RISK, AND HOW PROTEIN DIET / NUTRIENT EXPOSURE ALTERS SUSCEPTIBILITY TO DISEASE. THE STUDIES EXPLORE A META-ORGANISMAL PATHWAY (INVOLVING BOTH MICROBES, AND HOST ENZYMES) FOR A METABOLITE LINKED TO CARDIOVASCULAR DISEASE, WITH FOCUS ON HEART FAILURE. WE IDENTIFY IN PRELIMINARY STUDIES STRIKING CLINICAL ASSOCIATIONS BETWEEN MICROBE DEPENDENT METABOLITES DERIVED FROM DIETARY PHENYLALANINE, AND HEART FAILURE DEVELOPMENT. AND IN PRELIMINARY ANIMAL MODEL STUDIES, OBSERVE RECAPITULATION OF THESE CLINICAL OBSERVATIONS EXTENDING TO EVIDENCE OF A CAUSAL LINK BETWEEN GUT MICROBIAL METABOLITE GENERATION AND MULTIPLE HEART FAILURE RELATED PHENOTYPES. OUR STUDIES AIM TO EXPLORE WHETHER NUTRIENT INTAKE (DIETARY PROTEIN), VIA GUT MICROBE DEPENDENT CHANGES IN HOST METABOLISM (METABOLITE LEVELS), IS LINKED TO ADAPTIVE PHYSIOLOGICAL RESPONSES, AND WHEN IN EXCESS, MALADAPTIVE EFFECTS ON MYOCARDIAL FUNCTION AND REMODELING. THE STUDIES PROPOSED ARE MULTIDISCIPLINARY, RANGING FROM HUMAN CLINICAL OBSERVATIONAL STUDIES, TO MORE MECHANISTIC MICROBIAL ENGINEERING, CELLULAR, AND PRECLINICAL DISEASE MODEL INVESTIGATIONS. DEFINED MICROBIAL CONTRIBUTORS TO THE METAORGANISMAL PATHWAY ARE EXAMINED IN MODEL GNOTOBIOTIC SYSTEMS DESIGNED TO RECAPITULATE THE COMPLEX MICROBIAL COMMUNITY / HOST INTEGRATED NETWORK THAT COLLECTIVELY CONTRIBUTE TO THE METABOLIC AND PHYSIOLOGICAL EFFECTS OBSERVED IN VIVO. SUCCESSFUL COMPLETION OF THE PROPOSED STUDIES WILL FURTHER ADVANCE OUR KNOWLEDGE OF HOW DIETARY NUTRIENTS, THROUGH THE FILTER OF OUR GUT MICROBIAL COMMUNITY, CAN IMPACT HOST METABOLISM, PHYSIOLOGY AND DISEASE ETIOLOGY.

MECHANISMS OF ACTIVITY-DEPENDENT MYELINATION IN THE VISUAL SYSTEM

DARK GPCR SIGNALING UNDERLYING THE MICROBIOME-GUT-BRAIN AXIS FOR ALZHEIMER'S DISEASE AND RELATED DEMENTIA - PROJECT SUMMARY CUMULATIVE EVIDENCE INDICATES THE MICROBIOME-GUT-BRAIN AXIS PLAYS A CRUCIAL ROLE IN ALZHEIMER’S DISEASE (AD) AND SUPPORTS THE POTENTIAL OF MICROBIOME-TARGETED THERAPIES AS TREATMENTS FOR AD AND AD-RELATED DEMENTIA (ADRD). HOWEVER, THE PRECISE MECHANISM OF THE MICROBIOME-GUT-BRAIN AXIS AND THE IDENTITY OF ACTIONABLE GUT MICROBIAL BIOMARKERS UNDERLYING AD/ADRD PATHOGENESIS, DISEASE PROGRESSION, AND MODIFICATION REMIND UNDERSTUDIED. RECENT ADVANCES IN CHEMOGENOMIC TECHNOLOGIES HAVE DEMONSTRATED THAT G-PROTEIN-COUPLED RECEPTORS (GPCRS, THE LARGEST DRUGGABLE TARGET FAMILY IN THE HUMAN GENOME, AS DEFINED BY THE NIH-FUNDED ILLUMINATING THE DRUGGABLE GENOME PROGRAM) MEDIATE MUCH OF THE MICROBIOME-GUT-BRAIN AXIS, ESPECIALLY FOR GUT MICROBIOTA-DERIVED METABOLITES SUCH AS MEDIUM-CHAIN FATTY ACIDS (MCFAS). OUR PRELIMINARY EXPERIMENTS REVEAL STRONG SIGNIFICANT ASSOCIATIONS BETWEEN GUT-MICROBIOTA MCFA METABOLITES (E.G., 5-PHENYLVALERIC ACID) AND DARK GPCR SIGNALING (E.G., GPR84) IN AD USING MULTI-OMICS APPROACHES AND AN AD PATIENT-INDUCED PLURIPOTENT STEM CELLS (IPSC) MODEL. FURTHERMORE, WE IDENTIFIED TARGETING GUT MICROBIAL METABOLITE PATHWAYS IMPROVE COGNITIVE BEHAVIORS IN GERM-FREE MICE. WE POSIT THAT COMBINING AD PATIENT-INDUCED IPSC, CEREBRAL ORGANOIDS, AND GERM-FREE MOUSE MODELS, ALONG WITH MULTIMODAL ANALYSES OF PLASMA AND HIPPOCAMPUS GUT MICROBIAL METABOLOMICS DATA, WILL ENABLE IMPROVED MECHANISTIC UNDERSTANDING OF PRECISE PROTECTIVE MECHANISMS OF THE MICROBIOME-GUT-BRAIN AXIS IN AD/ADRD. OUR CENTRAL UNIFYING HYPOTHESIS IS THAT IDENTIFYING LIKELY MOLECULAR DRIVERS (E.G., GUT MICROBIAL METABOLITES) AND DRUGGABLE GPCR SIGNALING NETWORKS UNDERLYING THE MICROBIOME- GUT-BRAIN AXIS WILL ELICIT POTENTIAL PREVENTION AND TREATMENT STRATEGIES FOR AD. AIM 1 WILL TEST DARK GPR84 (A PUTATIVE MICROGLIAL GENE) AND ITS SIGNALING ACTIVATION UNDERLYING THE MICROBIOME-GUT-BRAIN AXIS OF MCFAS VIA FECAL MICROBIOTA TRANSPLANTATION (FMT) IN GERM-FREE MICE BY ASSESSING AD-RELATED COGNITIVE AND PATHOLOGICAL PHENOTYPES AND MECHANISMS. WE WILL EVALUATE DIFFERENTIAL GUT MICROBIAL COMMUNITIES, UNTARGETED AND TARGETED GUT METABOLOMICS ANALYSES OF PLASMA AND HIPPOCAMPUS IN GPR84-/-, 5XFAD, AND CROSS (5XFAD;GPR84-/-) GERM-FREE MICE DURING PRE-FMT AND POST-FMT. AIM 2 WILL SCREEN, TEST AND VALIDATE DARK GPCRS AND SIGNALING NETWORK PERTURBATION BY GUT MICROBIOTA-DERIVED MCFA METABOLITES USING AD PATIENT-DERIVED IPSC LINES IN CONJUNCTION WITH CEREBRAL ORGANOID MODELS. SPECIFICALLY, WE WILL EVALUATE PHYSICAL BINDING OF THE GUT MICROBIAL METABOLITE-GPCR INTERACTOME USING COMPLEMENTARY CALCIUM FLUX, CAMP GLOSENSOR, AND SS-ARRESTIN TANGO ASSAYS. AIM 3 WILL CONDUCT SUPERVISED ANALYSES OF GUT MICROBIAL METABOLITE BIOMARKER DISCOVERY FOR, AND PREDICTION MODELING OF, CLINICALLY RELEVANT AD PATHOLOGICAL FEATURES USING PATIENT PLASMA TARGETED AND UNTARGETED GUT MICROBIAL METABOLOMICS. IN SUMMARY, OUR MULTIDISCIPLINARY APPROACH COMPRISING GERM-FREE MICE, AD PATIENT- DERIVED IPSC, AND CEREBRAL ORGANOID MODELS, ALONG WITH HUMAN PLASMA GUT MICROBIAL METABOLOMICS, WILL IDENTIFY POTENTIAL MICROBIOME-TARGETED PREVENTION AND TREATMENT APPROACHES TO BE DIRECTLY TESTED IN PEOPLE WITH AD.

NUTRITION, TERMINAL NAD METABOLITES AND CARDIOVASCULAR DISEASE - SUMMARY IN THIS APPLICATION, WE USED UNTARGETED METABOLOMICS AS DISCOVERY PLATFORM TO STRUCTURALLY IDENTIFY AND CHARACTERIZE AN UNKNOWN METABOLITE WHOSE LEVELS IN PLASMA IN A DISCOVERY COHORT ARE STRONGLY ASSOCIATED WITH INCIDENT RISK FOR DEVELOPMENT OF CARDIOVASCULAR DISEASE (CVD), AND MAJOR ADVERSE CARDIOVASCULAR EVENTS (MACE = MYOCARDIAL INFARCTION (MI), STROKE OR DEATH). EXTENSIVE CHEMICAL STRUCTURAL ELUCIDATION EFFORTS REVEALED THE “ANALYTE” WAS IN FACT A PAIR OF RARELY STUDIED STRUCTURAL ISOMERS THAT SERVE AS TERMINAL METABOLITES OF NAD+ METABOLISM, NAMELY 2PY AND 4PY. VITAMIN B3 IS NIACIN (NICOTINIC ACID, AND NICOTINAMIDE), AND IS INTEGRALLY LINKED TO BOTH NAD+ AND CELLULAR ENERGY METABOLISM. THIS PROPOSAL INTEGRATES NUTRITION AND METABOLISM DATA IN BOTH HEALTH AND DISEASE. IT ALSO EXPLORES MECHANISMS OF MICRO-NUTRIENT PROCESSING (NAD+/NIACIN) AND ITS IMPACT ON METABOLISM, PATHOPHYSIOLOGICAL PATHWAYS AND DISEASE OUTCOMES. PRELIMINARY DATA INDICATE THE TERMINAL METABOLITE, 4PY, BUT NOT ITS STRUCTURAL ISOMER 2PY, IS CLINICALLY ASSOCIATED WITH CVD RISKS, AND PRELIMINARY CELLULAR AND DISEASE MODEL STUDIES SUGGEST 4PY DIRECTLY FOSTERS VASCULAR INFLAMMATION AND HEIGHTENED THROMBOSIS POTENTIAL. WE WILL USE A VARIETY OF APPROACHES TO EXPLORE THE METABOLISM, BIOLOGY AND (PATHO) PHYSIOLOGICAL LINKS BETWEEN 4PY AND CVD RELEVANT PHENOTYPES. OUR STUDIES ALSO SUGGEST THAT A SPECIFIC GENETIC VARIANT, COUPLED WITH DIETARY TRYPTOPHAN, CAN GIVE RISE TO BOTH ENHANCED 4PY LEVELS, AND VASCULAR ENDOTHELIAL CELL ACTIVATION, PRO-INFLAMMATORY GENE PROGRAMS, AND HEIGHTENED ATHEROSCLEROSIS AND THROMBOSIS POTENTIAL. THE PRESENT STUDIES HAVE SIGNIFICANT POTENTIAL PUBLIC HEALTH IMPORTANCE FOR MANY REASONS. THEY IDENTIFY A NEW POTENTIAL CONTRIBUTOR TO RESIDUAL CVD RISKS INDEPENDENT OF TRADITIONAL RISK FACTORS. THEY ALSO CALL FOR REASSESSMENT OF THE SAFETY OF CEREAL FORTIFICATION, A PRACTICE INSTITUTIONALIZED IN THE US SINCE WWII. FINALLY, THE PRESENT STUDIES MAY HELP EXPLAIN THE “NIACIN PARADOX” WHEREIN LARGE DOSES OF NIACIN USED TO LOWER LDL CHOLESTEROL, FAILED TO MATCH EXPECTATIONS IN CLINICAL TRIALS BASED ON THE DEGREE OF LDL LOWERING.

NOVEL OXIDATIVE PATHWAY TARGETS MICROGLIA IN ALZHEIMER'S DISEASE - ABSTRACT ALZHEIMER'S DISEASE IS THE MOST COMMON DEMENTIA CHARACTERIZED BY PROGRESSIVE COGNITIVE DECLINE. BY ESTIMATES, ALZHEIMER'S DISEASE AFFECTS MORE THAN ONE-THIRD OF THE POPULATION OVER 85, POSING A SUBSTANTIAL SOCIOECONOMIC BURDEN. THERE IS AN APPARENT LACK OF UNDERSTANDING OF THE MOLECULAR MECHANISMS OF ALZHEIMER'S DISEASE AND REPORTED PROBLEMS WITH ANIMAL MODELS. THESE OBSTACLES PREVENT THE DEVELOPMENT OF NEW THERAPEUTIC STRATEGIES, ESPECIALLY THEIR TRANSLATION TO HUMAN PATIENTS. THIS PROPOSAL IS BASED ON MOLECULAR AND CELLULAR EVIDENCE AND AN ANALYSIS OF HUMAN ALZHEIMER'S DISEASE CASES. IT BRIDGES SEVERAL ASPECTS OF ALZHEIMER'S DISEASE PATHOLOGY: DISCOVERY OF UNIQUE OXIDATIVE STRESS PATHWAY, NEW ASPECTS OF MICROGLIAL BIOLOGY AND DYSFUNCTION, AND BASIC MECHANISMS OF CELL ADHESION/PHAGOCYTOSIS IN THE CONTEXT OF DISEASE. WE WILL ANSWER HOW EXACTLY OXIDATION CAUSES MICROGLIAL DYSFUNCTION IN ALZHEIMER'S DISEASE AND HOW TO PREVENT/INHIBIT THEIR DETRIMENTAL CONSEQUENCES. BASED ON OUR PIONEERING PRELIMINARY STUDIES, WE PROPOSE THAT OXIDATIVE STRESS TARGETS IMPORTANT PROTEINS IN MICROGLIA RESPONSIBLE FOR PHAGOCYTOSIS, LEADING TO DEFICIENT PHAGOCYTOSIS AND MICROGLIA OVERACTIVATION IN ALZHEIMER'S DISEASE. THEREFORE, WE WILL FOCUS ON THE MECHANISMS OF THIS DAMAGE AND ITS PREVENTION IN ALZHEIMER'S DISEASE AND OTHER NEURODEGENERATIVE PATHOLOGIES. WE WILL DEVELOP A NEW INTERVENTION APPROACH (COMPOUNDS NEUTRALIZING OXIDATIVE STRESS) AND UNCOVER NEW SPECIFIC TARGETS FOR THE TREATMENT OF ALZHEIMER'S DISEASE.

REASSORTMENT OF BUNYAVIRUS IN TICKS AND ANIMAL MODELS - PROJECT SUMMARY/ABSTRACT LISTED IN THE WHO TOP NINE MOST DANGEROUS PATHOGENS, SFTSV HAS 12-30% FATALITY RATES, RAPIDLY SPREADS THROUGH TICK-TO-ANIMAL/HUMAN, HUMAN-TO-HUMAN AND ANIMAL-TO-HUMAN AND INDUCES IMMUNOPATHOGENIC DISEASE WITH A CHARACTERISTIC THROMBOCYTOPENIA. VIRUS REASSORTMENT IS A PROCESS OF GENETIC RECOMBINATION THAT IS EXCLUSIVE TO SEGMENTED RNA VIRUSES IN WHICH CO-INFECTION OF A CELL OF NATURAL HOST AND VECTOR WITH MULTIPLE VIRUSES MAY RESULT IN THE SHUFFLING OF GENE SEGMENTS TO GENERATE PROGENY VIRUSES WITH NOVEL GENOME COMBINATIONS. REASSORTMENT GREATLY AFFECTS VIRUS FITNESS AND DIRECTLY INFLUENCES ANTIGENIC VARIATION, CONFOUNDING AVAILABLE METHODS OF VIRUS CONTROL. IN THIS APPLICATION, WE WILL TEST THE HYPOTHESIS THAT TICK-MEDIATED NATURAL COURSE OF SFTSV INFECTION AND REASSORTMENT INTRODUCES PHENOTYPIC CHANGES OF FITNESS, TRANSMISSIBILITY, ANTIGENICITY, OR PATHOGENICITY INTO PROGENY REASSORTANTS. THE GOAL OF THIS STUDY IS TO DEMONSTRATE THE NATURAL COURSE OF SFTSV INFECTION AND REASSORTMENT FOR FITNESS, IMMUNOGENICITY, TRANSMISSIBILITY, AND PATHOGENESIS IN IN VITRO AND IN VIVO ANIMAL MODELS AND H. LONGICORNIS TICKS, ULTIMATELY BRIDGING BASIC RESEARCH TO CLINICAL APPLICATION.

SARCOPENIA IN CIRRHOSIS IS MEDIATED BY A HYPERAMMONEMIC STRESS RESPONSE

PROTECTIVE FACTORS IN DIABETIC KIDNEY DISEASE IN PATIENTS WITH TYPE 1 DIABETES

VITAMIN D AND PHOTODYNAMIC THERAPY FOR HUMAN SKIN CANCER (SCC AND BCC)

MACHINE AND DEEP LEARNING FOR FINDING MULTIMODAL IMAGING BIOMARKERS IN PRODROMAL AD - OUR PROPOSED STUDY FOCUSES ON DEVELOPING DEEP NEURAL NETWORKS AND SOPHISTICATED MULTIVARIATE ANALYSIS METHODS FOR STUDYING EPISODIC MEMORY ACTIVATIONS IN PRODROMAL AD SUBJECTS AND AGE-MATCHED NORMAL CONTROLS. WE ARE PARTICULARLY INTERESTED IN INVESTIGATING THE EFFECTS OF SPATIAL AND OBJECT PATTERN-SEPARATION IN SUBFIELDS OF THE HIPPOCAMPUS, NEARBY REGIONS OF THE MEDIAL TEMPORAL LOBE, AND FUNCTIONAL WHOLE-BRAIN CONNECTIONS. IN ORDER TO ACQUIRE A FULLER UNDERSTANDING OF THE UNDERLYING PHYSIOLOGICAL PROCESSES DRIVING AD PATHOLOGY, ACTIVATIONS IN HIPPOCAMPAL SUBFIELDS MUST BE INVESTIGATED IN FURTHER DEPTH AND WITH METHODOLOGIES THAT EXCEED THE CURRENT LIMITATIONS OF FMRI AT 3T. ACQUIRING DATA THAT WILL YIELD A MORE ACCURATE VIEW OF THESE HIPPOCAMPAL INTERACTIONS IS FAR MORE EASILY FACILITATED WITH 7T TECHNOLOGY, ALTHOUGH BARRIERS COMPLICATE SUCH A STUDY EVEN AT 7T. OUR PROPOSED STUDY CENTERS ON CIRCUMVENTING THESE BARRIERS, PARTICULARLY DATA CONTAMINATION FROM EXCESSIVE SYSTEM NOISE, HEAD-MOTION NOISE, AND PHYSIOLOGICAL NOISE WHICH IS PROPORTIONAL TO THE FIELD STRENGTH, TO DEVELOP METHODS THAT WILL ALLOW INVESTIGATORS TO WORK WITHIN THE PARAMETERS OF 7T AT ITS FULLEST CAPACITY TOWARD THE DEVELOPMENT OF MORE POWERFUL IMAGING BIOMARKERS FOR DIAGNOSING AD. TO INCREASE THE LIKELIHOOD OF THE SUCCESSFUL COMPLETION OF OUR STUDY, WE CONSIDER IT IMPERATIVE TO DEVELOP BETTER TASK FMRI DESIGNS AND IMAGING PROTOCOLS (AIM 1), AUTOMATIC SEGMENTATION METHODS (AIM 2), NOISE-REDUCTION METHODS (AIM 3), AND MULTIVARIATE ANALYSIS METHODS, SUCH AS NOVEL ALGORITHMS AND SOFTWARE TOOLS BASED ON CONSTRAINED CANONICAL CORRELATION ANALYSIS (CONSTRAINED CCA), KERNEL CCA, AND DEEP CCA AND RELEVANT GROUP-LEVEL ANALYSIS USING FUSION CCA, MULTISET CCA, AND MACHINE LEARNING AND DEEP LEARNING TECHNIQUES (AIM 4) FOR STUDYING MEMORY FUNCTION TO OBTAIN NOVEL IMAGING BIOMARKERS (AIM 5) TO IDENTIFY INDIVIDUALS AT RISK FOR AD. THIS STUDY WILL ENABLE THE CREATION OF CLEARER AND MORE DETAILED BRAIN ACTIVATION MAPS AND THUS PROMOTE THE DISCOVERY OF CURRENTLY UNKNOWN ASPECTS OF BRAIN FUNCTION IN PRODROMAL AD. THE SUCCESSFUL COMPLETION OF OUR OBJECTIVES COULD LEAD TO MORE EFFECTIVE DIAGNOSTIC TOOLS FOR AD, INCLUDING AN FMRI-BASED DIAGNOSTIC TEST FOR MEMORY IMPAIRMENT TO CHARACTERIZE ABNORMAL MEMORY FUNCTION IN PEOPLE AT RISK FOR AD. OUR ADVANCED METHODOLOGY, COMBINING 7T HIGH-RESOLUTION FMRI, AUTOMATIC SEGMENTATION, DATA DENOISING AND MULTIVARIATE ANALYSIS, WILL BE ESSENTIAL FOR DETECTING SUBTLE FUNCTIONAL CHANGES IN SUBFIELDS OF THE HIPPOCAMPUS AND ITS CONNECTIONS TO OTHER CORTICAL REGIONS. RESULTS FROM THIS STUDY ARE EXPECTED TO BROADLY IMPACT SCIENTIFIC UNDERSTANDING OF BRAIN FUNCTION BEYOND ONLY ENHANCING CURRENT UNDERSTANDING OF MEMORY FUNCTION IN AD. WE ANTICIPATE THAT THE METHODS DEVELOPED FROM FINDINGS ACQUIRED IN OUR PROPOSED STUDY WILL HAVE A FAR-REACHING INFLUENCE ON IMPROVING FMRI DATA QUALITY, ENABLE MORE ACCURATE DETECTION OF BRAIN ACTIVATION, OPEN A PATH TOWARD BETTER AUTOMATED AND INSTANTANEOUS HIPPOCAMPAL SUBFIELD SEGMENTATION FOR MANY OTHER MRI/FMRI APPLICATIONS OF NEURODEGENERATIVE DISEASES, AND CONTRIBUTE NEW AND VITAL DISCOVERIES TO THE FIELD OF NEUROSCIENCE IN GENERAL.

NOVEL ROLE FOR HOST IMMUNOSTIMULATORY RNA IN ANTIVIRAL IMMUNE DEFENSE - PROJECT SUMMARY WORK FROM MANY LABORATORIES HAS ESTABLISHED THAT DNA SENSORS – BOTH MEMBRANE-LOCALIZED TOLL-LIKE RECEPTORS (E.G. TLR9) AND INTRACELLULAR DNA SENSORS (E.G. CGAS) – SENSE HERPESVIRUS INFECTION. IN STRIKING CONTRAST, OUR KNOWLEDGE ABOUT THE PHYSIOLOGIC RELEVANCE OF CYTOPLASMIC RNA SENSORS OF THE RIG-I-LIKE RECEPTOR (RLR) FAMILY IN THE DETECTION OF HERPESVIRUSES, SUCH AS HERPES SIMPLEX VIRUS TYPE 1 (HSV-1), IS RUDIMENTARY. THE PROPOSED STUDY BUILDS ON A RECENT DISCOVERY BY THE GACK LABORATORY THAT RIG-I PLAYS A CRUCIAL ROLE IN THE DETECTION AND RESTRICTION OF HSV-1 INFECTION, WHERE RIG-I AND INTRACELLULAR DNA SENSORS ACT IN A TEMPORAL MANNER. FURTHERMORE, WE IDENTIFIED THAT DURING HSV-1 INFECTION THE HOST-DERIVED 5S RIBOSOMAL RNA PSEUDOGENE TRANSCRIPT 141 (RNA5SP141) ACTIVATES RIG-I AND ELICITS CYTOKINE-MEDIATED ANTIVIRAL INNATE IMMUNE DEFENSE. IN COLLABORATIVE WORK WE RECENTLY IDENTIFIED THAT ABLATED RNA5SP141 GENE EXPRESSION IS ASSOCIATED WITH SEVERE HSV-1 ENCEPHALITIS IN HUMANS. FURTHERMORE, WE DISCOVERED THAT RNA5SP141 IS EXPORTED FROM INFECTED CELLS VIA EXOSOMES AND TAKEN UP BY UNINFECTED CELLS, WHERE IT TRIGGERED RIG-I SIGNALING. USING A COORDINATED SERIES OF MOLECULAR, BIOCHEMICAL, AND CELL BIOLOGICAL APPROACHES COMBINED WITH CRISPR-GENE EDITING TECHNIQUES, WE WILL DEFINE THE ROLE OF RNA5SP141 GENE EXPRESSION IN THE ANTIVIRAL INNATE IMMUNE RESPONSE TO HSV-1 INFECTION AND IN HSV-1-ASSOCIATED DISEASE (AIM 1). FURTHERMORE, WE WILL ELUCIDATE THE MOLECULAR MECHANISM(S) UNDERLYING RNA5SP141 EXOSOMAL LOADING (AIM 2), AND ALSO DETERMINE THE PHYSIOLOGIC RELEVANCE OF EXOSOMAL DELIVERY OF RNA5SP141 IN HOST IMMUNITY TO HSV-1 INFECTION (AIM 3). OUR STUDIES WILL PROVIDE A MOLECULAR UNDERSTANDING OF INNATE IMMUNE DETECTION OF HERPESVIRUS INFECTION, WHICH MAY PROVIDE THE FOUNDATION FOR NEW THERAPEUTIC APPROACHES OR GUIDE THE DESIGN OF NOVEL ADJUVANTS.

HYPERTENSION AUGMENTED COVID-19 THROUGH RENIN-INDUCED INTERNALIZATION OF PLATELET-ACE2 / SARS-COV-2 COMPLEXES - HYPERTENSION, FOR UNKNOWN REASONS, IS A PRIMARY CO-MORBIDITY RISK FACTOR FOR POOR COVID-19 OUTCOMES. SARS- COV-2 BREACHES THE LUNG BLOOD-AIR BARRIER TO SPREAD AMONG ORGANS INDUCING ENDOTHELIAL CELL DYSFUNCTION AND MULTI-ORGAN THROMBOEMBOLISM. ACE2 IS THE HIGH AFFINITY RECEPTOR FOR SARS-COV-2 WITH TMPRSS2 CLEAVAGE THEN ENABLING FUSION. HOWEVER, ACE2 AND TMPRSS ARE NOT CO-EXPRESSED BY ALL SARS-COV-2 INFECTED ORGANS. WE DISCOVERED HUMAN PLATELETS EXPRESS ACE2 AND TMPRSS, BIND SARS-COV-2 SPIKE PROTEIN, AND INTERNALIZE ACE2-SPIKE PROTEIN COMPLEXES. SARS-COV-2 RNA ACCUMULATES WITHIN PLATELETS. PLATELETS ARE ACTIVATED IN ESSENTIAL HYPERTENSION, TRANSGENIC RENIN EXPRESSION STIMULATES FIBROSIS AND COAGULATION, AND INHIBITION OF COAGULATION BLOCKS RENIN FIBROSIS. THE DIRECT RENIN INHIBITOR ALISKIREN BLOCKS THROMBOSIS IN HYPERTENSIVE ANIMALS, SO RENIN INTERCALATES INTO COAGULATION TO INITIATE THROMBOEMBOLIC DISEASE. WE DISCOVERED CELLS RELEASING PRORENIN STIMULATE EXPLOSIVE PLATELET ACTIVATION, BUT IN A UNIQUE WAY; THE ONSET OF ACTIVATION WAS VERY DELAYED, AND ACTIVATION WAS ALWAYS MAXIMAL. MECHANISTICALLY, PRORENIN INTERACTED WITH QUIESCENT PLATELETS PROMOTING ESCAPE OF INTRACELLULAR PHOSPHATIDYLSERINE ONTO THE PLATELET SURFACE. PHOSPHATIDYLSERINE ORGANIZES TENASE AND PROTHROMBINASE COAGULATION COMPLEXES, FORMING FACTOR XA AND THEN THROMBIN THAT EXPLOSIVELY ACTIVATED THE PLATELET PAR1 THROMBIN RECEPTOR. ALISKIREN ABOLISHED PHOSPHATIDYLSERINE EXPRESSION, THROMBIN FORMATION, AND THROMBOSIS. THIS ESTABLISHES RENIN AS A NOVEL, DIRECT PLATELET AGONIST. PLATELETS DISPLAYING SURFACE PHOSPHATIDYLSERINE ARE RAPIDLY CLEARED BY ENGULFMENT BY ENDOTHELIAL CELLS AND PERIVASCULAR MACROPHAGES OF THE RETICULOENDOTHELIAL SYSTEM OF LIVER, LUNG, AND SPLEEN. WE POSTULATE HYPERTENSION AND RENIN EXPRESSION PROMOTES PHOSPHATIDYLSERINE DISPLAY ON PLATELETS, INITIATING COAGULATION AND PLATELET ACTIVATION, BUT ALSO PROMOTING RAPID PLATELET CLEARANCE. THIS, WE POSTULATE, INTERNALIZES SARS-COV-2 INTO CELLS THAT NEED NOT EXPRESS ACE2 OR TMPRSS2, THEMSELVES. AIM 1. TEST THE HYPOTHESIS THAT RENIN-ACTIVATED PLATELETS ARE ENTRY VECTORS FOR SARS-COV-2 INTO ENDOTHELIAL CELLS AND MACROPHAGES OF THE RETICULOENDOTHELIAL SYSTEM. AIM 2. TEST THE HYPOTHESIS RENIN-STIMULATED PLATELET TURNOVER IN VIVO INTRODUCES SARS-COV-2 PSEUDOTYPED LENTIVIRUS-PLATELET COMPLEXES INTO DIVERSE ORGANS. THIS PROJECT WILL ESTABLISH A FUNCTIONAL CONNECTION BETWEEN HYPERTENSION AND SARS-COV-2 INFECTION, IDENTIFY RENIN ACTIVATED PLATELETS AS NOVEL SARS-COV-2 ENTRY VECTORS, AND DEFINE A BASIS FOR ALTERED PLATELET CLEARANCE IN RENIN-CLAMPED HYPERTENSIVE MICE. THIS PROVIDES A TRANSLATIONAL BASIS FOR ALISKIREN USE TO NORMALIZE HYPERTENSION RISK IN COVID-19, ELUCIDATES NOVEL APPROACHES TO SUPPRESS SARS-COV-2 ORGAN INFECTION AND DAMAGE, AND ESTABLISH CIRCULATING PLATELET ACE2 EXPRESSION AS A MEASURE OF RISK FOR COVID-19 MULTI-ORGAN DAMAGE.

THE ROLE OF ACETOGENIC, METHANOGENIC, AND SULFATE-REDUCING BACTERIA IN OXALATE METABOLISM AND HYPEROXALURIA

THE ROLE OF TRIM23 IN AUTOPHAGY MEDIATED ANTIVIRAL DEFENSES

TWO NOVEL THREONINE PHOSPHORYLATIONS OF STAT2 IMPACT INFLAMMATORY RESPONSES TO BACTERIAL INFECTION.

ENDOCYTIC TRAFFICKING OF ADCS IN GBM CANCER STEM-LIKE CELLS

NEURONAL PROTECTIVE NANOPARTICLES FOR TREATING ACUTE SCI

TRPV4-PI3K AXIS MEDIATES PULMONARY AND CARDIAC FIBROSIS

UNDERSTANDING ROLE OF CIRCADIAN DISRUPTION IN PATHOGENESIS OF MS - ABSTRACT DISEASE PROGRESSION IN MULTIPLE SCLEROSIS (MS) IS CONCEIVABLY DRIVEN BY MECHANISMS THAT CONTRIBUTE TO REMYELINATION FAILURE, AND IDENTIFICATION OF THESE MECHANISMS IS CRITICAL FOR DEVELOPING NOVEL THERAPIES. CURRENT THERAPIES FOR MS PATIENTS ARE INEFFECTIVE AT TREATING SECONDARY COMPLICATIONS THAT SIGNIFICANTLY IMPACT QUALITY OF LIFE, SUCH AS SLEEP AND FATIGUE. THE LONG-TERM GOAL OF THIS WORK IS TO DISCOVER NEW STRATEGIES TO FACILITATE REMYELINATION IN MS PATIENTS. WHILE CIRCADIAN AND/OR SLEEP DISRUPTIONS ARE COMMON IN MS PATIENTS, THEIR ROLES IN AFFECTING THE PROCESS OF REMYELINATION IN THE CONTEXT OF MS ARE UNKNOWN. THE OVERALL OBJECTIVE OF THIS STUDY THEREFORE IS TO UNDERSTAND THE MECHANISMS BY WHICH CIRCADIAN RHYTHM DISRUPTION MAY ALTER REMYELINATION AND DISEASE PROGRESSION IN MS. THE RATIONALE FOR THESE STUDIES STEMS FROM PRELIMINARY OBSERVATIONS THAT CIRCADIAN RHYTHM DISRUPTION (CRD) CORRELATES WITH NEUROLOGICAL DEFICITS AND LEADS TO REMYELINATION FAILURE IN ANIMAL MODELS. OUR PRELIMINARY DATA DEMONSTRATES THAT THE KEY CIRCADIAN REGULATOR, BMAL1, IS EXPRESSED BY OLIGODENDROCYTE LINEAGE CELLS IN MICE AND MS BRAINS AND IS CRITICAL FOR REMYELINATION. BASED UPON THESE OBSERVATIONS, OUR CENTRAL HYPOTHESIS IS THAT “BMAL1-MEDIATED CIRCADIAN DISRUPTION CONTRIBUTES TO REMYELINATION FAILURE AND PROGRESSION OF DISEASE COURSE IN MS”. THIS HYPOTHESIS WILL BE TESTED BY THREE SPECIFIC AIMS. SPECIFIC AIM 1 WILL ASSESS THE CONTRIBUTION OF CRD GENE POLYMORPHISMS IN MS PATIENTS WITH SLEEP DISTURBANCES AND MS BRAIN TISSUES TO CORRELATE MAGNETIC RESONANCE IMAGING AND PATHOLOGY OF THE MS BRAINS WITH GENETIC POLYMORPHISMS IN KEY CIRCADIAN GENES. AS HUMAN STUDIES ARE NOT AMENABLE TO MANIPULATION, WE PROPOSE TO TEST THE CORRELATIVE FINDINGS FROM THE HUMAN STUDIES USING ANIMAL MODELS OF SLEEP DEPRIVATION AND DEMYELINATION/REMYELINATION. SPECIFIC AIM 2 WILL ADDRESS WHETHER CHANGES IN CIRCADIAN RHYTHM LEAD TO REMYELINATION DEFECTS IN ANIMAL MODELS. SPECIFIC AIM 3 WILL INVESTIGATE THE CONSEQUENCES OF CELL-SPECIFIC LOSS AND GAIN OF BMAL1 ON CRD-MEDIATED REMYELINATION DEFECTS. THIS PROPOSAL IS CONCEPTUALLY INNOVATIVE BECAUSE WE INVESTIGATE A PREVIOUSLY-UNEXPLORED LINK BETWEEN CIRCADIAN DISRUPTION AND THE PROCESS OF REMYELINATION AND OLIGODENDROCYTE BIOLOGY. THE APPROACH IS TECHNICALLY INNOVATIVE BECAUSE WE PROPOSE TO USE MS PATIENT SAMPLES WITH SLEEP DISRUPTION, MS AUTOPSY TISSUES, AND ANIMAL MODELS WITH OLIGODENDROCYTE- AND OLIGODENDROCYTE PROGENITOR CELL-SPECIFIC DELETION AND OVEREXPRESSION OF THE KEY CIRCADIAN GENE, BMAL1. THE FUTURE OF MS THERAPEUTICS LIES IN THE IDENTIFICATION OF ADDITIONAL THERAPEUTIC TARGETS AND IN DEVELOPING MORE COMPREHENSIVE COMBINATORIAL STRATEGIES. THIS WORK WILL MAKE A SIGNIFICANT IMPACT ON THE FIELD OF MS RESEARCH BECAUSE IT WILL REVEAL THE LINK BETWEEN CIRCADIAN DISRUPTION AND MYELIN REPAIR FAILURE, AND IN THE FUTURE WILL GUIDE THE USE OF DRUGS DIRECTED TOWARDS RESETTING THE CIRCADIAN RHYTHM TO IMPROVE CLINICAL OUTCOME FOR MS PATIENTS.