Biomedical Research Institute

THE SOUTHWEST NATIONAL PRIMATE RESEARCH CENTER

SOUTHWEST NATIONAL PRIMATE RESEARCH CENTER

ESTABLISHMENT OF A SPF RHESUS MACAQUE COLONY

DIET AND GENOTOYPE IN PRIMATE ATHEROSCLEROSIS

OMICS FOR TB DISEASE PROGRESSION (OTB)

WISCONSIN CENTER OF EXCELLENCE IN GENOMICS SCIENCE

GENETICS OF ATHEROSCLEROSIS IN MEXICAN AMERICANS

PURE AND AUTHENTIC HIV-1 ENV IMMUNOGENS

OPTIMIZING HIV IMMUNOGEN-BCR INTERACTIONS FOR VACCINE DEVELOPMENT

EXPLORING THE IMPACT OF INFLAMMAGING ON IMMUNE FUNCTION DURING M. TB INFECTION

FURTHER EXPANSION OF THE SOUTHWEST NATIONAL PRIMATE RESEARCH CENTER SPECIFIC PATHOGEN FREE RHESUS MACAQUE RESOURCE - ABSTRACT THE SOUTHWEST NATIONAL PRIMATE RESEARCH CENTER (SNPRC) HOUSES A SPECIFIC PATHOGEN FREE (SPF) RHESUS MACAQUE COLONY OF INDIAN ORIGIN (IND RM), SUPPORTED BY THE NIH SPF RHESUS BREEDING PROGRAM (U42OD010442), AND A SMALLER SPF P51 SUPPORTED COLONY. WE CURRENTLY HOUSE ~1000 RHESUS MACAQUES, OF WHICH THE U42 COLONY OF APPROXIMATELY 800 ANIMALS SUPPORTS AIDS-RELATED RESEARCH BOTH AT SNPRC AND THROUGH SALES TO AIDS INVESTIGATORS AT OTHER INSTITUTIONS. HOWEVER, THERE IS A MAJOR NATIONAL SHORTAGE OF RESEARCH NONHUMAN PRIMATES (NHPS), AND PARTICULARLY OF SPF IND RM. THIS HAS SEVERELY IMPACTED OUR ABILITY TO SUPPORT THE NATIONAL AND INTERNATIONAL RESEARCH MISSION OF HIV/AIDS AS WELL AS IN THE AREA OF NON-AIDS/OTHER INFECTIOUS DISEASES, AIDS CO-INFECTIONS, MALARIA, TB, COVID-19, ETC. THE NIH HAS RECOGNIZED THIS NEED FOR EXPANSION OF SPF RHESUS PRODUCTION AND STRONGLY RECOMMENDS INCREASING IND RM BREEDING CAPACITY. THE SNPRC IS WELL POSITIONED TO EXPAND ITS SPF IND RM PRODUCTION AS A CENTER LOCATED IN A CLIMATE HOSPITABLE TO LARGELY OUTDOOR HOUSING, AT AN INSTITUTION WITH CAPACITY FOR EXPANSION. OUR HOST INSTITUTION, TEXAS BIOMED, RAISED FUNDS AND IS CURRENTLY BUILDING NEW NHP FACILITIES WHICH WILL HOUSE ~600 NHPS. TEXAS BIOMED/SNPRC HAS ALSO BEEN FUNDED BY THE NIH/ORIP TO EXPAND OUR PRODUCTION CAPACITY BY 30% OVER OUR EXISTING CAPACITY VIA OTHER MECHANISMS. FINALLY, THROUGH TEXAS BIOMED SUPPORT, WE HAVE PROCURED ~150 CONVENTIONAL FOUNDER/BREEDER IND RM. THIS FOUNDER COLONY HAS INCREASED THE NUMBER OF BREEDING PAIRS THE SNPRC MACAQUE COLONY MANAGEMENT TEAM CAN GENERATE, AND IT IS EXPECTED THAT IN THE UPCOMING BIRTHING SEASON, ~200 RM LIVE BIRTHS WILL OCCUR, DOUBLE THAT OF EACH OF THE LAST SEVERAL YEARS. AS SUCH THE SNPRC IND RM POPULATION CAN INCREASE TO 1500+ IN THE NEXT FIVE YEARS. FUNDED PARTIALLY BY THIS C06, WE SEEK TO CONSTRUCT AN ADDITIONAL ANIMAL HOUSING/BREEDING FACILITY ADJACENT THE TWO NEW HOLDING/BREEDING FACILITIES CURRENTLY UNDER CONSTRUCTION, AS PART OF THE OVERALL $45M LONG TERM ANIMAL CARE COMPLEX PROJECT. THIS WILL FURTHER ALLOW US TO HOUSE, BREED AND MAINTAIN AN ADDITIONAL SUPPLY OF ~300 IND RM FOR AIDS/EMERGING INFECTIOUS DISEASES RESEARCH.

MAPPING DRUG RESISTANCE GENES IN PLASMODIUM FALCIPARUM

THE IMPACT OF DIFFUSE MILD BRAIN INJURY ON CLINICAL OUTCOMES IN CHILDREN

PHASE III COBRE: MULTIMODAL IMAGING OF NEUROPSYCHIATRIC DISORDERS (MIND)

HOST-DIRECTED THERAPY TO AUGMENT ANTI-M. TUBERCULOSIS RESPONSES IN THE SETTING OF HIV CO-INFECTION AND TO STERILIZE THE TUBERCULOMA

LPS REGULATION OF MACROPHAGE FUNCTION

STRUCTURAL STUDIES OF THE AUTOPHAGIC UBIQUITIN-LIKE PROTEINS

MAPPING DRUG RESISTANCE GENES IN PLASMODIUM FALCIPARUM

GENOME PLASTICITY DURING ES CELL DIFFERENTIATION TO NEURAL LINEAGES

UNDERSTANDING CO-MORBIDITIES: COVID-19 IN INDIVIDUALS LIVING WITH HIV/AIDS - SUMMARY WHILE COVID-19 CONTINUES TO BE A HEALTH CHALLENGE, VERY LITTLE IS KNOWN ABOUT HOW COVID-19 AFFECTS PEOPLE LIVING WITH HIV (PLHIV). BASED ON THE MOST RECENT REPORTS ORIGINATING FROM CDC AND WHO, HOWEVER, IT APPEARS THAT PEOPLE WITH HIV MAY HAVE A 30% GREATER LIKELIHOOD OF DEVELOPING SEVERE COVID-19 DISEASE WHEN INFECTED WITH SARS-COV-2. WE WILL LEVERAGE THE ESTABLISHED RHESUS MACAQUE MODELS OF SARS-COV-2 INFECTION RESULTING IN COVID-19 AND SIV INFECTION TO CHARACTERIZE THE EFFECTS OF UNDERLYING SIV INFECTION ON THE MANIFESTATION OF BOTH ACUTE AND POST-ACUTE COVID-19 SEQUELAE. OUR GROUP WAS AMONGST FEW THAT ESTABLISHED THE RHESUS MACAQUE MODELS OF COVID-19 INFECTION EARLY ON DURING THE PANDEMIC. OUR MODEL HAS BEEN UTILIZED TO BOTH STUDY THE IMMUNOLOGICAL MECHANISMS OF PROTECTION FROM SARS-COV-2 INFECTION, AS WELL AS FOR ACCELERATED DEVELOPMENT OF VACCINE AND THERAPEUTICS AGAINST COVID-19. HERE WE PROPOSE TO COUPLE THIS MODEL WITH THE LONG-STANDING, HIGHLY VALIDATED, PATHOGENIC AIDS NHP MODEL IN SIV INFECTED RHESUS MACAQUES TO STUDY A CENTRAL HYPOTHESIS THAT UNDERLYING SIV INFECTION AND THE RESULTING IMMUNODEFICIENCY/IMMUNE ACTIVATION PROMOTES THE PROGRESSION OF A MORE SEVERE COVID-19 PRESENTATION DUE TO SARS-COV-2 INFECTION. AS COROLLARY, WE HYPOTHESIZE THAT ART DOES NOT COMPLETELY SUPPRESS THE ILL EFFECTS OF CHRONIC IMMUNE ACTIVATION DUE TO SIV, IT WILL NOT COMPLETELY PREVENT THE PROGRESSION OF SEVERE COVID-19 DUE TO SARS-COV-2 INFECTION IN THE MACAQUE MODEL. WE HAVE THE EXPERIENCE IN INFECTING RHESUS MACAQUES WITH SIV AND TREATING THESE ANIMALS WITH ART TO SUPPRESS VIRAL REPLICATION AND STUDY IMMUNE MECHANISMS. BY PROFILING THE DIFFERENCES IN DYNAMICS OF VIRAL TITERS, INDUCED TISSUE PATHOLOGY, AND UNDERLYING IMMUNOLOGICAL PERTURBATIONS, WE WILL PROVIDE DEFINITIVE KNOWLEDGE IN WHETHER SIV INFECTED RHESUS MACAQUES EXHIBIT HIGHER SUSCEPTIBILITY TO SEVERE COVID-19. FURTHERMORE, OUR STUDIES WILL ALSO BE ABLE TO HINT AT THE SPECIFIC MECHANISMS WHICH RESULT IN THIS SUSCEPTIBILITY. DELINEATING THESE COMORBID IMMUNOLOGICAL FACTORS DRIVING SUSCEPTIBILITY WILL ENABLE BETTER CLINICAL MONITORING AND INFORMED DECISIONS FOR PATIENT CARE. MECHANISTIC INSIGHTS DEVELOPED BY THIS STUDY IS ALSO IMPERATIVE FOR THE DEVELOPMENT OF HOST-DIRECTED IMMUNOTHERAPEUTIC INTERVENTIONS FOR COMBATING COVID-19 IN PLHIV.

DURABLE HIV VACCINE TARGETING MUCOSAL EPITHELIUM - ABSTRACT THE IMPRESSIVE AMOUNT OF DATA GENERATED BY EXPERIMENTAL HIV/SIV VACCINES HAS LED TO THE REALIZATION THAT PROTECTION WILL MOST LIKELY REQUIRES 2 LEVELS OF BARRIERS, THE INITIAL ONE AT THE MUCOSAL PORT OF ENTRY AND IF BREACHED, A SECOND SET OF SYSTEMIC DEFENSES. THE CAPACITY OF HUMORAL AND CELLULAR IMMUNE RESPONSES IN MUCOSAL TISSUES TO BLOCK OR CONTAIN REPLICATION AT THE INITIAL STAGE OF VIRUS TRANSMISSION MAY HAVE A PROFOUND IMPACT ON THE ABILITY OF A VACCINATED HOST TO RESIST INFECTION, EVEN IN THE CASE WHEN VIRUS PROGRESSES BEYOND THE PORT OF ENTRY, ALLOWING THE SYSTEMIC RESPONSE MORE TIME TO CONTROL OR ERADICATE THE INCOMING PATHOGEN. WE HYPOTHESIZED THAT THERE ARE TWO NECESSARY FEATURES FOR A SUCCESSFUL VACCINE: 1) A PROLONGED IF NOT LIFE-LONG STIMULATION OF THE IMMUNE SYSTEM WITH VIRAL ANTIGENS TO MAINTAIN “ALERT” IMMUNE RESPONSES; AND 2) A TARGETED IMMUNE RESPONSE AT THE SITE OF PRIMARY REPLICATION OF HIV. A VACCINE APPROACH THAT SIMULTANEOUSLY ADDRESSES THESE TWO ISSUES WOULD HAVE THE POTENTIAL TO ACHIEVE SOLID, LONG-TERM ACTIVE PROTECTION. TO FULFILL THESE REQUIREMENTS, WE HAVE DEVELOPED AN ORIGINAL STRATEGY TO SUCCESSFULLY DELIVER A VACCINE TO MUCOSAL SITES THAT PROVIDE ANTIGEN STIMULI AT RECURRENT INTERVALS AND ELICIT PROTECTIVE MUCOSAL IMMUNE RESPONSES. OUR STRATEGY LEVERAGES EPITHELIAL STEM CELLS AS PERMANENT BUT NON-EXPRESSING SOURCE OF VIRAL ANTIGEN WHILE THEIR DIFFERENTIATED OFFSPRING EXPRESS AND PRESENT ANTIGEN TO THE LOCAL IMMUNE SYSTEM, ALONG THE REPRODUCTIVE CYCLE. USING A SINGLE CYCLE SIV (SIVSC) APPROACH, WHICH HAS BEEN SHOWN TO BE SAFE COMPARED TO TRADITIONAL ATTENUATED VACCINES, WE HAVE CLONED THE SIVSC GENOME UNDER THE CONTROL OF THE INVOLUCRIN PROMOTER (PINV- SIVSC), A TERMINALLY DIFFERENTIATED KERATINOCYTE SPECIFIC PROMOTER. WHEN ADMINISTERED, THE VACCINE TARGETS AND TRANSDUCES BASAL EPITHELIAL STEM CELLS FROM VAGINAL TISSUES. THESE THEN PROLIFERATE AND DIFFERENTIATE INTO MATURE EPITHELIAL CELLS, TRIGGERING SIV ANTIGEN EXPRESSION VIA THE PROMOTER AND LEADING TO BOTH DIRECT AND CROSS PRIMING. FOR THIS PROJECT, WE PROPOSE: 1) TO CONFIRM AND FURTHER IMPROVE THE EFFICACY AND SAFETY PROFILE OF THE PINV-SIVSC VACCINE IN FEMALE MACAQUES; 2) TO VISUALIZE AND OPTIMIZE VACCINE DELIVERY, AND INVESTIGATE THE MECHANISMS OF ACTION UNDERLYING PROTECTION; AND, 3) USING OUR BEST OPTIMIZED VACCINE STRATEGY, DEMONSTRATE PROTECTION FROM VIRUS ACQUISITION AND/OR VIRAL REPLICATION IN VIVO AND DETERMINE THE CORRELATES OF PROTECTION OR CONTROL AGAINST REPEATED LOW-DOSE VAGINAL CHALLENGES WITH HETEROLOGOUS SIV.

PTPRD LIGANDS FOR STIMULANT AND OPIATE USE DISORDERS

ACCELERATED MOLECULAR PROBE PIPELINE

INTERDISCIPLINARY NEXGEN TB RESEARCH ADVANCEMENT CENTER (IN-TRAC) - TEXAS BIOMED IN-TRAC OVERALL ABSTRACT THE INTERDISCIPLINARY NEXGEN TB RESEARCH ADVANCEMENT CENTER (IN-TRAC) AT TEXAS BIOMEDICAL RESEARCH INSTITUTE (TEXAS BIOMED) WILL ATTRACT THE NEXT GENERATION OF DIVERSE RESEARCHERS TO THE TUBERCULOSIS (TB) RESEARCH FIELD, AND DEVELOP THEM INTO INDEPENDENT RESEARCHERS WITH MULTI-DISCIPLINARY SKILLS AND REAL-WORLD EXPERIENCE OF CLINICAL TB. THE PRODUCT OF IN-TRAC WILL BE RESEARCHERS THAT CHOOSE TO WORK ON SOME OF THE MOST CHALLENGING AND RELEVANT TRANSLATIONAL PROBLEMS IN TB, AND THAT WORK ACROSS ACADEMIC DISCIPLINES AND WITHIN A FRAMEWORK OF HIGHLY COLLABORATIVE RESEARCH. THIS WILL BE ACHIEVED THROUGH 6 INTERRELATED CORES. ADMINISTRATIVE CORE: THE CENTRAL HUB OF IN-TRAC MANAGEMENT AND OVERSIGHT, WITH RESPONSIBILITY FOR ADMINISTRATIVE AND SCIENTIFIC LEADERSHIP; COORDINATING COMMUNICATION BETWEEN CORES, RESEARCHERS AND NIH/NIAID PROGRAM STAFF AND PROVIDING AN ORGANIZATIONAL STRUCTURE TO OPTIMIZE MULTIDISCIPLINARY COLLABORATIONS AND INTERACTIONS BETWEEN THE CORES AND AMONG A DIVERSE COHORT OF IN-TRAC PARTICIPANTS. DEVELOPMENT CORE: OVERSIGHT OF ALL CAREER DEVELOPMENT PROGRAMMING AND COURSES, TAILORING THE RESEARCH AND CLINICAL EXPERIENCES, BOTH INTERN AND EXTERNSHIPS, TO EACH INDIVIDUAL IN-TRAC PARTICIPANT, AND MANAGEMENT OF THE IN-TRAC PILOT GRANT PROGRAM. BIOSAFETY & BIOCONTAINMENT CORE: EXTENDING THE REQUIRED BIOSAFETY TRAINING TO A PERSONALIZED CURRICULUM INCLUDING THEORETICAL AND HANDS-ON BIOCONTAINMENT TRAINING, UNDERSTANDING THE MECHANICAL AND REGULATORY REQUIREMENTS OF A BSL3/ABSL3, COACHING ON HOW TO DEVELOP BIOSAFETY PROTOCOLS, AND WORKING THROUGH RISK ASSESSMENTS FROM THE PERSPECTIVE OF A BIOSAFETY OFFICER. RESEARCH IMAGING CORE: PROVIDING THEORETICAL AND HANDS-ON TRAINING FROM SINGLE CELL IMAGING (CONFOCAL MICROSCOPY, CYTOMETRY, LIVE CELL IMAGING, SINGLE CELL RNASEQ) THROUGH TO WHOLE BODY IMAGING IN MICE (IVIS) AND NON-HUMAN PRIMATES (NHP) (PET/CT) WITHIN LARGE, FULLY OUTFITTED BSL3/ABSL3 FACILITIES. ANIMAL MODEL CORE: IMPLEMENTING A TRAINING PROGRAM TO INTRODUCE ALL IN-TRAC PARTICIPANTS TO THE REGULATORY REQUIREMENTS FOR WORKING WITH RODENTS AND NHPS, HOW TO DEVELOP A ROBUST EXPERIMENTAL PROTOCOL, HOW TO WRITE AN IACUC PROTOCOL, AND HANDS-ON ANIMAL HANDLING AND EXPERIMENTAL PROCEDURES. CLINICAL RESEARCH & PATIENT CARE CORE: INTRODUCING IN-TRAC PARTICIPANTS TO TB PATIENT CARE AT THE ONLY FREE-STANDING TB HOSPITAL IN THE US (TEXAS CENTER OF INFECTIOUS DISEASES) THAT MANAGES SOME OF THE MOST CHALLENGING CASES OF TB NATIONALLY. PARTNERING WITH THIS WILL BE A CLINICAL RESEARCH EXPERIENCE AT THE US- MEXICO BORDER, TO EXPERIENCE TB STUDIES IN UNDER-SERVED AND UNDER-RESOURCED COMMUNITIES.

DO EARLY MATERNAL ANTIBODIES FACILITATE ORAL TRANSMISSION OF HIV IN INFANTS?

A NOVEL LIVE NON-REPLICATING UNIVERSAL VACCINE FOR INFLUENZA

SIGH BASED ATTENUATED, EFFICACIOUS MTB VACCINES TO PROTECT AGAINST LETHAL TB

ROLE OF MICRORNAS IN B-CELL DYSFUNCTION IN HIV/SIV INFECTION

DNA REPAIR CAPACITY ASSAYS FOR LUNG DISEASE RISK ASSESSMENT

MOLECULAR ANALYSIS OF THE PFEMP1 BINDING ACTIVITY

IMMUNE CORRELATES OF PROTECTION FROM TB

HUMORAL CORRELATES OF PROTECTION AGAINST HIV

BRAIN MYELOID CELL-TARGETED MULTIPLEXED GENE EDITING FOR SIV/HIV ERADICATION - PROJECT SUMMARY THE LONG-LIVED MYELOID CELLS SUCH AS PERIVASCULAR MACROPHAGES AND MICROGLIA IN THE CENTRAL NERVOUS SYSTEM (CNS) PERSISTENTLY HARBOR HIV. THESE INFECTED CELLS COULD CONTRIBUTE TO THE SOURCE OF RESIDUAL VIREMIA DURING LONG-TERM ANTIRETROVIRAL THERAPY (ART) OR TO REBOUNDING VIRUS UPON ART CESSATION. IT IS UNDOUBTFULLY AND URGENTLY NEEDED TO DEVELOP NOVEL STRATEGIES TO SPECIFICALLY TARGET CNS MYELOID CELLS FOR HIV ERADICATION AND A CURE. BE- CAUSE THE EFFECTS OF THE WIDELY-STUDIED “SHOCK AND KILL” APPROACH COULD EXACERBATE NEUROINFLAMMATION, GENE THERAPY EMERGES AS THE OPTIMAL STRATEGY, PARTICULARLY THE ADVANCED CRISPR GENOME EDITING TECHNOLOGY. SIMIAN IMMUNODEFICIENCY VIRUS (SIV) INFECTION OF MACAQUES IS THE BEST AVAILABLE MODEL FOR TESTING NOVEL STRATEGIES PRIOR TO CLINICAL STUDIES. WE HAVE SHOWN THAT SIV INFECTION HAS A BROAD SPREAD IN THE CNS EVEN IN ANIMALS ON ART. BECAUSE THE VIRUS ENTERS THE BRAIN WITHIN A FEW DAYS AFTER INFECTION AND THE ESTABLISHMENT OF THE LATENT RESERVOIR OCCURS VERY EARLY, THE INITIATION OF ART SHOULD BE AS EARLY AS POSSIBLE. IN ADDITION, NUMEROUS STUDIES SUGGEST THAT CCR5/CCR2 PLAY A MAJOR ROLE IN HIV ENTRY AND NEUROINFLAMMATION. MOST IMPORTANTLY, WE HAVE USED AAV DELIVERY OF A CRISPR/CAS GENOME EDITOR TO ERADICATE HIV/SIV PROVIRUS IN MODELS OF HUMANIZED MICE AND NON- HUMAN PRIMATES. HOWEVER, THE LACK OF AAV SEROTYPES THAT ARE HIGHLY EFFECTIVE AND RELIABLE TO TRANSDUCE MYELOID CELLS IN THE CNS REMAINS A KEY CHALLENGE. THEREFORE, WE HYPOTHESIZE THAT THE MULTIPLE-TARGETING GENE EDITING SYSTEM ACROSS THE BLOOD BRAIN BARRIER (BBB) CAN REMOVE SIV PROVIRUS IN INFECTED MYELOID CELLS, PROTECT CELLS AGAINST NEW INFECTION, AND INHIBIT NEUROINFLAMMATION. TO TEST THIS HYPOTHESIS, WE WILL OPTIMIZE A NOVEL AAV SERO- TYPE WITH BBB PENETRATION AND MYELOID-SPECIFIC TRANSDUCTION (NAMELY AAV-BM) TO EFFECTIVELY DELIVER THE SMALLER CJCAS9 WITH MULTIPLEX SGRNAS (BMCJ4) SPECIFIC FOR 4 TARGET SITES (SIV LTR, GAG, AND HOST CCR5, CCR2) INTO THE ENTIRE CNS FOR IN VIVO HIV/SIV ERADICATION (AIM 1). WE WILL EVALUATE THE EFFICACY OF BMCJ4 EARLY TREATMENT IN PREVENTING BRAIN SIV INFECTION OR/AND EXCISING SIV PROVIRAL DNA FROM BRAIN MYELOID CELLS IN ACUTE SIV INFECTION WITH EARLY ART (AIM 2A). WE WILL ALSO DETERMINE THE THERAPEUTIC EFFECT OF BMCJ4 WITH ART AND THEN BOOST IT WITH AN ALTERNATIVE AAV-BM FOR ERADICATION OF PERSISTENT BRAIN SIV LATENT INFECTION (AIM 2B). WE EXPECT THAT EARLY OR LONG-TERM REPEATED BMCJ4 AAV GENE THERAPY WILL EFFECTIVELY ERADICATE ACUTE AND LATENTLY-INFECTED HIV PROVIRUS AND EXTENSIVELY MINIMIZE THE SIZE OF THE BRAIN VIRAL RESERVOIR TO ACHIEVE A STERILIZING OR FUNCTIONAL CURE OF HIV/AIDS, PARTICULARLY NEUROAIDS.

PRIMATE ANIMAL FACILITY

EPIGENETIC MECHANISMS UNDERLYING CANNABINOID MODULATION OF NEUROINFLAMMATION IN HIV/SIV INFECTION

INFANT IMMUNOPROPHYLAXIS AGAINST A PRIMATE LENTIVIRUS

INHALED DELIVERY OF VIDAZA FOR TARGETED EPIGENETIC LUNG CANCER THERAPY

IMPACT OF CONCURRENT HIV AND LATENT TB THERAPIES ON MTB-SPECIFIC IMMUNE FUNCTION

MACROPHAGE NUCLEAR RECEPTORS, METABOLISM AND IMMUNE EFFECTORS DURING HEALTH AND M. TUBERCULOSIS INFECTION

NIH-OWNED CHIMPANZEE RESEARCH RESOURCE AT THE SNPRC

MINDFULNESS FOR ALCOHOL ABUSING OFFENDERS: MECHANISMS AND OUTCOMES

PERTURBATION OF ANTIGEN-SPECIFIC T CELL RESPONSES IN LATENT TB/SIV CO-INFECTION

GENETIC BASIS OF PRAZIQUANTEL RESISTANCE

MALARIA VACCINE DEVELOPMENT: PROTECTING OUR TROOPS

RISK OF NEONATAL VACCINATION FOR HIV/SIV EXPOSED INFANTS

GENETICS OF BRAIN STRUCTURE AND FUNCTION

TRANS-COMPLEMENTING PAPILLIOMA VIRUS FOR AIDS VACCINE

MICROBIOME-MEDIATED THERAPIES FOR AGING AND HEALTHSPAN IN MARMOSETS

CNS MYELOID CELLS AS SIV RESERVOIRS: PERSISTENT INFECTION AND REBOUND

PREDICTING TUBERCULOSIS OUTCOMES USING GENOTYPIC AND BIOMARKER SIGNATURES

RATIONAL POLYTHERAPY IN THE TREATMENT OF CHOLINERGIC SEIZURES

GENETICS OF BONE STRUCTURE AND METABOLISM

EXTERNALIZING OUTCOMES IN HIGH RISK YOUTH

PREVENTING PREGNANCY MALARIA: MATERNAL-INFANT OUTCOMES

CVD IN AMERICAN INDIANS GENETICS CENTER

NEUROCOGNITIVE ABNORMALITIES IN STIMULANT ABUSE AMONG HIGH-RISK WOMEN - PROJECT ABSTRACT THERE CONTINUES TO BE GREAT INTEREST AND PUBLIC/HEALTH/RELEVANCE WITH REGARD TO UNDERSTANDING THE NEUROBIOLOGICAL SYSTEMS THAT UNDERLIE THE COMORBIDITY OF SUBSTANCE USE DISORDERS AND OTHER PSYCHIATRIC CONDITIONS. IN A PREVIOUS R01 AWARD, WE FOCUSED OUR EFFORTS UPON CHARACTERIZING THE NEURAL CIRCUITRY UNDERLYING MORAL DECISION MAKING IN INCARCERATED MEN WITH VARYING LEVELS OF TWO FREQUENTLY CO-OCCURRING CONDITIONS: STIMULANT ABUSE AND PSYCHOPATHY. HERE WE PROPOSE TO EXTEND THIS WORK TO INCARCERATED WOMEN, EXAMINE LONGITUDINAL OUTCOMES, AND APPLY STATEOF-THE-ART NETWORK ANALYSES FOR PREDICTIVE MODELS. STUDIES PUBLISHED BY OUR RESEARCH TEAM HAVE DEMONSTRATED SEX DIFFERENCES IN THE DEGREE AND EXPRESSION OF PSYCHOPATHIC TRAITS, PATTERNS OF STIMULANT ABUSE, AND MORAL DECISION-MAKING. HOWEVER, THE NEURAL CIRCUITRY THAT UNDERLIES THESE SEX DIFFERENCES IS NOT WELL UNDERSTOOD. WE HAVE ALSO IDENTIFIED SUBSTANTIAL SEX DIFFERENCES IN REGIONAL GRAY MATTER VOLUME AND DENSITY IN OUR EXTANT SAMPLES. COLLECTIVELY, SEX DIFFERENCES IN PATHOPHYSIOLOGY COULD HAVE SIGNIFICANT IMPLICATIONS FOR TREATMENT STRATEGIES AND DIFFERENTIAL BIOMARKERS OF TREATMENT PREDICTION AND OUTCOME IN MEN AND WOMEN. WE WILL IMPLEMENT THE RESEARCH STRATEGY WITH A LARGE INCARCERATED POPULATION BY DEPLOYING A UNIQUE MOBILE MRI SCANNER TO THE REGIONAL WOMEN’S PRISON. PARTICIPANTS WILL BE STRATIFIED BY THEIR LEVEL OF LIFETIME STIMULANT (COCAINE, AMPHETAMINE) USE SEVERITY AND PSYCHOPATHIC TRAITS (HIGH, MEDIUM, LOW) AND WILL UNDERGO ANATOMICAL AND FUNCTIONAL MRI SCANNING WHILE COMPLETING MULTI-MODAL (I.E., LINGUISTIC AND PICTURE) DECISION-MAKING TASKS. WE WILL ALSO EXAMINE FUNCTIONAL NETWORK AND DYNAMIC NETWORK CONNECTIVITY IN WOMEN USING A NEW MULTIBAND EPI PULSE SEQUENCE, AND COLLECT LONGITUDINAL OUTCOMES AFTER RELEASE TO THE COMMUNITY AND TEST BEHAVIORAL AND NEUROPREDICTIVE MODELS OF RELAPSE AND FUTURE ANTISOCIAL BEHAVIOR. THIS WORK IS EXPECTED TO GENERATE A LARGE, ROBUST DATASET THAT CHARACTERIZES THE OVERLAPPING AND UNIQUE ASPECTS OF NEURAL CIRCUITRY UNDERLYING STIMULANT USE AND PSYCHOPATHY IN FEMALES. THE PROPOSED RESEARCH IS IN LINE WITH RECENT PRIORITIES EMPHASIZED BY NIDA FOR PROJECTS AIMED AT EXAMINING GENDER DIFFERENCES, AND EFFECTS SPECIFIC TO FEMALES, TO IMPROVE OUR UNDERSTANDING OF THE NATURE AND ETIOLOGY OF DRUG ABUSE.

ESTABLISHMENT OF A SPF RHESUS MACAQUE COLONY

MATHEMATICAL MODELING OF MYCOBACTERIUM TUBERCULOSIS DISSEMINATION - RESEARCH SUMMARY TUBERCULOSIS (TB), A DISEASE CAUSED BY THE BACTERIA MYCOBACTERIUM TUBERCULOSIS (MTB), REMAINS A MAJOR INFEC- TIOUS DISEASE OF HUMANS IN THE WORLD. AFTER THE INITIAL LOCAL INFECTION OF ONE SITE IN THE LUNG MTB SOMEHOW DISSEM- INATES IN THE LUNG AND OFTEN SPREADS BEYOND THE LUNG. IN FACT, EXTRAPULMONARY TB IS A HALLMARK OF THE DISEASE IN YOUNG CHILDREN AND IMMUNOCOMPROMISED ADULTS THAT IS DIFCULT TO DIAGNOSE AND TREAT. OUR UNDERSTANDING OF MTB DISSEMINATION, BOTH WITHIN THE LUNG AND BEYOND, REMAINS LIMITED, HOWEVER. IN THIS PROPOSAL WE ASSEMBLED A TEAM OF SCIENTISTS WITH EXPERTISE IN COMPUTATIONAL BIOLOGY (GANUSOV, AITCHISON, DUFFY, LANGSTON) AND TB PATHOGENESIS (URDAHL, SHERMAN, BEHAR) TO PROVIDE QUANTITATIVE UNDERSTANDING OF MECHANISMS OF MTB DISSEMINATION THE LUNG AND SYSTEMICALLY. TO THIS END, WE WILL BE USING A NUMBER OF HIGHLY INNOVATIVE TECHNIQUES SUCH AS I) A NOVEL ANIMAL MODEL OF TB: INFECTION OF MICE WITH AN ULTRA LOW DOSE (ULD, 1-3 COLONY FORMING UNITS, CFU) OF MTB ALONG WITH A SET OF 50 BARCODED MTB STRAINS, II) AN MTB STRAIN H37RV-PBP10 WITH THE REPLICATION CLOCK PLASMID, ALLOWING TO ESTIMATE HOW QUICKLY BACTERIA ARE ELIMINATED IN VIVO, AND III) MRNA-BASED GENE SIGNATURES PREDICTING BACTERIAL NUMBERS IN MURINE LUNGS AND TB DISEASE PROGRESSION RISK IN HUMANS. WITH THREE COMPLEMENTARY SPECIC AIMS WE WILL PRO- VIDE DETAILED, QUANTITATIVE UNDERSTANDING OF FUNDAMENTAL PROCESSES OF HOW MTB DISSEMINATES FROM THE DEPOSITION IN LUNG ALVEOLI TO THE WHOLE LUNG AND SYSTEMICALLY. IN AIM 1 WE WILL DETERMINE THE PATHWAY OF MTB DISSEMINATION WITHIN THE LUNG USING A NOVEL MODEL OF ULD-INFECTED MICE THAT MIMICS BETTER HUMAN INFECTION THAN MANY OTHER ANIMAL MODELS. IN PARTICULAR, WE WILL DISCRIMINATE BETWEEN ALTERNATIVE HYPOTHESES OF MTB SPREAD IN THE LUNGS SUCH THE “BUBBLE MODEL” (IN WHICH MTB SPREADS LOCALLY BETWEEN LUNG LOBES) AND THE “RESEEDING MODEL” (IN WHICH MTB SPREADS HEMATOGENOUSLY TO DIFFERENT PARTS OF THE LUNG AFTER DISSEMINATING SYSTEMICALLY). IN AIM 2 WE WILL DETERMINE THE CONTRIBUTION OF DIFFERENT CELL POPULATIONS, INCLUDING MTB-SPECIC CD4 T CELL RESPONSE, TO KINETICS OF MTB DISSEMINATION SYSTEMICALLY IN MICE INFECTED WITH CONVENTIONAL DOSES (CD, 150 CFU) OF MTB. TO PARAMETERIZE BEST T MODELS WE WILL USE DATA FROM EXPERIMENTS WITH MTB H37RV CARRYING THE REPLICATION CLOCK PLASMID PBP10. FINALLY, IN AIM 3 WE WILL ATTEMPT TO IMPROVE ON OUR RECENTLY DERIVED MRNA-BASED GENE SIGNATURES PREDICTING CFU IN MURINE LUNGS USING CUTTING-EDGE GRAPH THEORY-BASED METHODS OF DATA DIMENSIONALITY REDUCTION. WE WILL ALSO PERFORM EXPERIMENTS AND DENE A NEW SIGNATURE PREDICTING DISSEMINATED TB IN MICE, AND TEST ITS ACCURACY USING DATA FROM MONKEYS AND HUMANS. TAKEN TOGETHER, BY COMBINING EXPERIMENTAL DATA FROM HIGHLY INNOVATIVE EXPERIMENTS INVOLVING NOVEL TECHNIQUES (ULTRA LOW DOSE INFECTIONS, BARCODED STRAINS, REPLICATION CLOCK PLASMID, MICROARRAY-BASED GENE SIGNATURES) WE WILL PROVIDE A QUANTITATIVE UNDERSTANDING OF HOW MTB DISSEMINATES IN THE LUNG AND SYSTEMICALLY IN THE BODY.

A LONGITUDINAL STUDY OF TRAUMATIC BRAIN INJURY IN A HIGH-RISK POPULATION - PROJECT SUMMARY/ABSTRACT TRAUMATIC BRAIN INJURY (TBI) IS A GLOBAL CONCERN THAT PRECIPITATES A DIVERSE SET OF HEALTH CONSEQUENCES. TBI IS A MAJOR CONCERN IN FORENSIC POPULATIONS -- AN ISSUE RECENTLY HIGHLIGHTED BY CALLS FOR RESEARCH BY NATIONAL PUBLIC HEALTH INSTITUTIONS [1]. INDEED, UPWARDS OF 60% OF INMATES HAVE A HISTORY OF TBI, A RATE SEVEN TIMES HIGHER THAN THE GENERAL POPULATION (8.5%) [2, 3]. TBI IS A CRIMINOGENIC RISK FACTOR [3, 4] AND EVEN MILD TBI INCREASES THE RATES OF RISK-TAKING BEHAVIOR, SUBSTANCE USE, AND FUTURE OFFENDING [2, 4]. OVER THE LAST DECADE OUR TEAM HAS BEEN CONTINUOUSLY FUNDED BY NIH TO COLLECT DETAILED CLINICAL AND NEUROIMAGING PROTOCOLS FROM OVER 4000 INCARCERATED MEN AND WOMEN. WE UTILIZE A UNIQUE PAIR OF TWIN MOBILE MRI UNITS THAT ARE DEPLOYED DIRECTLY ON THE SECURE GROUNDS OF FORENSIC FACILITIES, MAKING IT POSSIBLE TO CONDUCT THIS RESEARCH. OUR DATASET INCLUDES MULTIMODAL NEUROIMAGING PROTOCOLS, THOROUGH CLINICAL ASSESSMENT, NEUROPSYCHOLOGICAL EVALUATIONS, AND HISTORIES OF TBI. ALL OF OUR N>4000 OFFENDERS HAVE CONSENTED TO PARTICIPATE IN LONGITUDINAL FOLLOW-UP STUDIES. FOR THIS PROJECT, WE WILL COMMENCE A LONGITUDINAL STUDY ON N=300 INDIVIDUALS AGED 35+, EXAMINING VARIABLES THAT WILL DIFFERENTIATE TRAJECTORIES ASSOCIATED WITH COGNITIVE IMPAIRMENTS. WE WILL ALSO LEVERAGE ADVANCED NEUROIMAGING TOOLS DEVELOPED BY OUR TEAM TO PROVIDE BRAIN-BASED MEASURES OF COGNITIVE DECLINE AND IMAGING-BASED CLASSIFICATION TOOLS FOR IDENTIFYING BRAIN INJURIES AND COMORBID CONDITIONS (I.E., SUBSTANCE ABUSE, PSYCHOPATHY SCORES, DEPRESSION, ETC.). WE WILL ALSO EXAMINE HOW THE BRAIN INJURY PROFILES AND COMORBID CONDITIONS CHANGE OVER TIME. THE AVAILABILITY OF THESE UNIQUE RESOURCES TO EXAMINE THIS HIGHLY UNDERSERVED POPULATION MAKES THIS AN EXCITING PROJECT WITH THE POTENTIAL TO BREAK NEW GROUND IN OUR UNDERSTANDING OF INDIVIDUAL RISK NEEDS AND DIVERGENT OUTCOMES IN TBI AND NEURODEGENERATIVE DISEASE.

GENOMIC CONSEQUENCES OF SCHISTOSOME HYBRIDIZATION - HYBRIDIZATION BETWEEN PARASITE SPECIES HAS THE POTENTIAL TO TRANSFER BIOMEDICALLY IMPORTANT GENES ACROSS SPECIES BOUNDARIES WITH POTENTIAL IMPACT ON HOST SPECIFICITY, PATHOGENESIS AND DRUG RESISTANCE. IT IS WIDELY ASSUMED THAT THERE IS FREQUENT ONGOING HYBRIDIZATION BETWEEN THE LIVESTOCK PARASITE SCHISTOSOMA BOVIS AND THE HUMAN PARASITE S. HAEMATOBIUM IN WEST AFRICA: THIS HAS BECOME A POSTER CHILD FOR “ONE HEALTH” APPROACHES TO DISEASE MANAGEMENT. GENETIC CROSSES BETWEEN THESE SCHISTOSOME SPECIES CAN BE CONDUCTED IN THE LABORATORY, AND MULTIPLE PAPERS HAVE DESCRIBED “HYBRID” SCHISTOSOMES BETWEEN S. HAEMATOBIUM INFECTING HUMANS AND S. BOVIS INFECTING CATTLE. HOWEVER, A CENTRAL ISSUE WITH THESE FIELD STUDIES IS THAT SINGLE MITOCHONDRIAL AND RIBOSOMAL DNA MARKERS ARE USED TO CHARACTERIZE PARASITE LARVAE. WITH THIS LIMITED GENOMIC RESOLUTION IT IS UNCLEAR WHETHER HYBRIDIZATION OCCURS FREQUENTLY, WHETHER IT IS RARE AND ANCIENT, OR IF HYBRIDIZATION HAS NEVER OCCURRED AND THE DISCORDANCE RESULTS FROM ANCESTRAL LINEAGE SORTING. OUR PRELIMINARY DATA ARE CONSISTENT WITH RARE ANCIENT HYBRIDIZATION AND SUBSEQUENT INTROGRESSION, RATHER THAN WIDESPREAD, ONGOING HYBRIDIZATION. WE SEQUENCED EXOMES FROM MIRACIDIA COLLECTED FROM NIGER AND TANZANIA REVEALING (A) NO EVIDENCE FOR RECENT HYBRIDS, (B) THAT ALL S. HAEMATOBIUM FROM NIGER CARRY 5-8% OF S. BOVIS DNA IN THEIR GENOME (C) THE SIZE OF INTROGRESSED S. BOVIS FRAGMENTS INDICATED ANCIENT HYBRIDIZATION (100-600 GENERATIONS AGO) (D) THAT S. BOVIS DNA HAS RISEN TO HIGH FREQUENCY SOME REGIONS OF THE S. HAEMATOBIUM GENOME SUGGESTING ADAPTIVE INTROGRESSION. THE CENTRAL GOAL OF THIS APPLICATION IS TO USE GENOME SEQUENCING, POPULATION GENOMICS AND EXPERIMENTAL ANALYSES TO UNDERSTAND THE FREQUENCY AND GENOMIC CONSEQUENCES OF HYBRIDIZATION BETWEEN S. HAEMATOBIUM AND S. BOVIS. WE HAVE DEVELOPED METHODS FOR WHOLE GENOME SEQUENCING FROM SINGLE PARASITE LARVAE FROM FECAL SAMPLES OR SNAILS: IN AIM 1 WE WILL EXAMINE 395 GENOME SEQUENCES OF S. BOVIS AND S. HAEMATOBIUM FROM ARCHIVED PARASITE LARVAE OR ADULT WORMS FROM 14 COUNTRIES FROM ACROSS AFRICA AND FROM 10 STATES IN NIGERIA. WE WILL USE THESE DATA TO CRITICALLY EVALUATE: (A) EVIDENCE FOR RECENT (F1 OR F2) HYBRIDIZATION, (B) TO DETERMINE HOW MANY TIMES INTROGRESSION HAS OCCURRED; (C) IDENTIFY GENOME REGIONS THAT ARE ENRICHED OR DEPLETED IN S. BOVIS ALLELES; AND (D) TO DEFINE GEOGRAPHICAL REGIONS IN WHICH INTROGRESSION HAS OCCURRED. IN AIM 2 WE WILL STAGE EXPERIMENTAL GENETIC CROSSES BETWEEN S. BOVIS AND S. HAEMATOBIUM IN RODENTS TO DETERMINE GENOMIC AND PHENOTYPIC CONSEQUENCES OF HYBRIDIZATION. IN PARTICULAR, WE WILL DETERMINE GENOME REGIONS INVOLVED IN SNAIL PENETRATION OF MIRACIDIA LARVAE AND SKIN PENETRATION OF CERCARIAE TO DETERMINE THE IMPACT OF HYBRIDIZATION ON HOST SPECIFICITY. FINALLY, IN AIM 3 WE WILL EXAMINE BOTH ADULT WORMS AND EGGS RECOVERED FROM NATURAL SCHISTOSOME INFECTIONS OF WEST AFRICA RODENTS TO DETERMINE WHETHER RARE HYBRIDIZATION EVENTS MAY OCCUR. THE RESULTS WILL ADDRESS FUNDAMENTAL AND APPLIED QUESTIONS CONCERNING SPECIES BOUNDARIES, HYBRIDIZATION, HOST SPECIFICITY AND INTROGRESSION IN A BIOMEDICALLY IMPORTANT AND EXPERIMENTALLY TRACTABLE PARASITE SPECIES.

FUNCTIONAL CURE AND VIRUS ERADICATION BY EARLY HAART PLUS VACCINATION WITH LIVE ATTENUATED RUBELLA VIRUS VECTORS IN MACAQUE INFANTS AND NEONATES

GENETIC EPIDEMIOLOGY OF CHAGAS DISEASE PROGRESSION

IDENTIFICATION OF GENES INFLUENCING TOTAL ANTIOXIDANT STATUS

NIH DIRECTOR'S PIONEER AWARD

ANTIGEN PRESENTATION BY EPITHELIAL STEM CELLS TO PROMOTE LIFE LONG IMMUNITY

ATTENDING TO ALL CHILDREN: EXAMINING THE ROLE OF ALPHA OSCILLATIONS IN ATTENTION IN YOUNG CHILDREN WITH AND WITHOUT PRENATAL ALCOHOL EXPOSURE (ASCEND) - ABSTRACT ATTENTION DEFICITS CAUSE LIFE-LONG CHALLENGES AFFECTING ACADEMIC AND JOB PERFORMANCE, SOCIAL RELATIONSHIPS, AND LIFE SATISFACTION. ATTENTION PROBLEMS TYPICALLY BEGIN AT AN EARLY AGE AND CHILDREN WITH ATTENTION PROBLEMS STRUGGLE IN SCHOOL. IDEALLY, ATTENTION DEFICITS WOULD BE IDENTIFIED BEFORE A CHILD ENTERS SCHOOL. A PRIMARY LIMITATION IN THE FIELD IS OUR LIMITED KNOWLEDGE OF THE NEURAL SIGNATURES OF ATTENTION IN YOUNG CHILDREN. CHILDREN WITH PRENATAL ALCOHOL EXPOSURE (PAE) AND CHILDREN WITH A DIAGNOSIS OF FETAL ALCOHOL SPECTRUM DISORDER (FASD) RELATED TO PAE EXPERIENCE PERSISTENT ATTENTION DEFICITS, AND PAE IS PRESENT AT BIRTH. THEREFORE, STUDYING CHILDREN WITH AND WITHOUT PAE PROVIDES US WITH AN OPPORTUNITY TO EXAMINE NEURAL SIGNATURES OF ATTENTION AT A YOUNG AGE IN A GROUP OF CHILDREN AT HIGH RISK OF ATTENTION DEFICITS. CHILDREN RAISED IN LOWER SOCIOECONOMIC HOUSEHOLDS ARE ALSO AT GREATER RISK OF ATTENTION PROBLEMS, AND DUE TO HIGH RATES OF POVERTY IN NEW MEXICO, WE ARE WELL-POSITIONED TO EXAMINE ITS ROLE IN ATTENTION IN CHILDREN WITH AND WITHOUT PAE. RESEARCH IN ADULTS INDICATES THAT ALPHA OSCILLATIONS PLAY A KEY ROLE IN DIRECTING ATTENTION, BUT IT IS UNKNOWN HOW ALPHA OSCILLATIONS ARE RELATED TO ATTENTION EARLY IN DEVELOPMENT. ALPHA OSCILLATIONS ARE EASILY ELICITED DURING REST, MODIFIED DURING TASKS, AND MEASURABLE ACROSS DEVELOPMENT. THIS STUDY BUILDS ON (1) PRIOR RESEARCH DEMONSTRATING THALAMUS DRIVES SOME CORTICAL ALPHA OSCILLATIONS WHEREAS OTHERS ARE DRIVEN BY CORTICO-CORTICAL CONNECTIONS OR LOCAL NETWORK DYNAMICS AND (2) STUDIES IN PRECLINICAL PAE MODELS AND RESEARCH IN CHILDREN WITH PAE INDICATING DISRUPTED CORTICO- THALAMIC AND CORTICO-CORTICAL TRACTS WITH PAE. AIM 1 WILL ESTABLISH THE ROLE OF ALPHA OSCILLATIONS AND CORTICO- THALAMIC OR CORTICO-CORTICAL CONNECTIVITY IN ATTENTION BY MEASURING ALPHA OSCILLATIONS DURING REST AND TASK USING MAGNETOENCEPHALOGRAPHY (MEG) AND WHITE MATTER INTEGRITY (WMI) USING DIFFUSION TENSOR IMAGING (DTI) IN TYPICALLY DEVELOPING CHILDREN AGED 4-7 YEARS. AIM 2 WILL ASSESS ALTERATIONS IN ALPHA OSCILLATIONS AND WMI AND THEIR RELATION TO ATTENTION DEFICITS IN CHILDREN WITH PAE, RELATIVE TO TYPICALLY DEVELOPING CHILDREN. AIM 3 WILL EXAMINE THE DEVELOPMENTAL TRAJECTORY OF ALPHA OSCILLATIONS AND WMI AND THEIR ROLE IN ATTENTION BY FOLLOWING THE SAME CHILDREN LONGITUDINALLY UNTIL AGE SEVEN. WE HYPOTHESIZE THAT ALPHA OSCILLATIONS ARE DIRECTLY RELATED TO WMI AND ATTENTION IN TYPICALLY DEVELOPING CHILDREN. FURTHERMORE, ALPHA OSCILLATIONS WILL BE REDUCED IN CHILDREN WITH PAE, AND THESE REDUCTIONS WILL BE RELATED TO DISRUPTIONS IN WMI AND ATTENTION DEFICITS. WE EXPECT THESE EFFECTS TO BE MEDIATED BY POVERTY IN BOTH TYPICALLY DEVELOPING CHILDREN AND CHILDREN WITH PAE. WE WILL TEST OUR HYPOTHESES BY MEASURING ALPHA OSCILLATIONS USING MEG DURING 3 TASKS THAT ROBUSTLY MANIPULATE ALPHA OSCILLATIONS IN PARIETAL CORTEX, SOMATOMOTOR REGIONS AND THE FRONTO-PARIETAL NETWORK AND EXAMINING HOW THESE OSCILLATIONS RELATE TO WMI AND ATTENTION. THIS STUDY WILL PROVIDE FOUNDATIONAL KNOWLEDGE ABOUT THE ROLE OF ALPHA OSCILLATIONS IN ATTENTION IN YOUNG CHILDREN AND MAY INDICATE POTENTIAL BIOMARKERS TO IMPROVE EARLY DIAGNOSIS AND INTERVENTION FOR FASD TO INFORM OUR UNDERSTANDING OF ATTENTION DEFICITS BROADLY.

NEUROBEHAVIORAL DEFICITS IN HIV/HCV INFECTION PRE/POST ANTI-HCV THERAPY

RESEARCH TO IMPROVE AND STANDARDIZE MARMOSET NUTRITION AND DIETARY HUSBANDRY

OUTER MEMBRANE PROTEINS OF PATHOGENIC LEPTOSPIRA SPECIES

MODULATING INDOLEAMINES TO OPTIMIZE IMMUNITY IN THE SETTING OF MTB/HIV CO-INFECTION - PROJECT SUMMARY TB REMAINS THE LEADING CAUSE OF DEATH IN HIV-INFECTED PERSONS, WITH ONE IN FOUR DEATHS ATTRIBUTABLE TO TB. WHILE THE MAJORITY OF HEALTHY INDIVIDUALS INFECTED WITH MYCOBACTERIUM TUBERCULOSIS (MTB) CONTROL INFECTION, CO-INFECTION WITH HIV INCREASES THE RISK OF PROGRESSING TO TB DISEASE BY OVER 20 FOLD. ANTIRETROVIRAL THERAPY (ART) DECREASES THE INCIDENCE OF ATB AND REMAINS THE CORNERSTONE OF HIV CARE. HOWEVER, THE INCIDENCE OF TB IN HIV-CO-INFECTED INDIVIDUALS REMAINS FOUR-TO-SEVEN-FOLD HIGHER AFTER ART THAN IN HIV-UNINFECTED PEOPLE IN TB-ENDEMIC SETTINGS, REGARDLESS OF THE DURATION OF ART OR ATTAINMENT OF HIGH CD4+ T CELL COUNTS. WE HAVE DEVELOPED MACAQUE MODELS OF MTB/HIV CO-INFECTION WHICH UTILIZE ART. DEPENDING ON THE TIMING OF ART-INTERVENTION, SIV REPLICATION IN THE PERIPHERY AS WELL AS TISSUES IS EITHER EFFECTIVELY INHIBITED OR NOT, RECAPITULATING THE WHOLE SPECTRUM OF HUMAN TISSUE-SPECIFIC AND CLINICAL OUTCOMES. THIS ALLOWS FOR DETAILED LONGITUDINAL AND MECHANISTIC STUDIES THAT ARE NOT POSSIBLE IN HUMANS. OUR DATA SHOWS A CLEAR ROLE FOR INDOLE 2,3, DIOXYGENASE (IDO) IN BOTH THE INHIBITION OF EFFECTIVE IMMUNITY TO TB AS WELL AS IN ORCHESTRATING CHRONIC IMMUNE ACTIVATION IN SIV-INFECTED MACAQUES. BLOCKADE OF IDO PATHWAY IMPROVES THE OUTCOME OF TB AND HIV IN SINGLY INFECTED MACAQUES. HERE, WE WILL USE INHIBITION APPROACHES IN THE RM MODELS OF MTB/SIV/ART TO TEST THE HYPOTHESIS THAT THIS WILL IMPROVE ANTI-TB IMMUNE RESPONSES, INHIBIT HIV-INDUCED CHRONIC IMMUNE ACTIVATION, THUS ALLOWING THE IMMUNE SYSTEM TO BETTER CONTROL THE CO-INFECTION. OUR PROPOSED STUDIES WILL PROVIDE UNPRECEDENTED NOVEL INSIGHTS INTO THE MOLECULAR MECHANISMS THAT MEDIATE REACTIVATION OF TB IN THE SETTING OF HIV INFECTION, AND IDENTIFY PROTECTIVE IMMUNE MECHANISMS THAT WILL INFORM THE DEVELOPMENT OF NEW TREATMENT REGIMENS AND VACCINES FOR TB.

INTRARECTAL IMMUNIZATION FOR A BARRIER TO MUCOSAL HIV INFECTION - ABSTRACT THE IMPRESSIVE AMOUNT OF DATA GENERATED BY EXPERIMENTAL BUT MOSTLY UNSUCCESSFUL HIV/SIV VACCINES HAS LED TO THE REALIZATION THAT PROTECTION WILL MOST LIKELY REQUIRES 2 LEVELS OF BARRIERS, THE INITIAL ONE AT THE MUCOSAL PORT OF ENTRY AND IF BREACHED, A SECOND SET OF SYSTEMIC DEFENSES. THE CAPACITY OF HUMORAL AND CELLULAR IMMUNE RESPONSES IN MUCOSAL TISSUES TO BLOCK OR CONTAIN REPLICATION AT THE INITIAL STAGE OF VIRUS TRANSMISSION MAY HAVE A PROFOUND IMPACT ON THE ABILITY OF A VACCINATED HOST TO RESIST INFECTION, EVEN WHEN VIRUS PROGRESSES BEYOND THE PORT OF ENTRY, ALLOWING THE SYSTEMIC RESPONSE MORE TIME TO CONTROL OR ERADICATE THE INCOMING PATHOGEN. WE HYPOTHESIZED THAT THERE ARE TWO FEATURES NECESSARY FOR A SUCCESSFUL VACCINE: 1) A PROLONGED IF NOT LIFE-LONG STIMULATION OF THE IMMUNE SYSTEM WITH VIRAL ANTIGENS TO MAINTAIN “ALERT” IMMUNE RESPONSES; AND 2) A TARGETED IMMUNE RESPONSE AT THE SITE OF PRIMARY HIV REPLICATION. A VACCINE APPROACH THAT SIMULTANEOUSLY ADDRESSES THESE TWO ISSUES WOULD HAVE THE POTENTIAL TO ACHIEVE SOLID, LONG-TERM ACTIVE PROTECTION. TO FULFILL THESE REQUIREMENTS, WE HAVE DEVELOPED AN ORIGINAL STRATEGY THAT SUCCESSFULLY DELIVERS A VACCINE TO MUCOSAL SITES, PROVIDING ANTIGEN STIMULI AT RECURRENT INTERVALS AND ELICIT PROTECTIVE MUCOSAL AND SYSTEMIC IMMUNE RESPONSES. OUR STRATEGY LEVERAGES EPITHELIAL STEM CELLS AS PERMANENT BUT NON-EXPRESSING SOURCE OF VIRAL ANTIGEN WHILE THEIR SHORT-LIVED DIFFERENTIATED OFFSPRING EXPRESS AND PRESENT ANTIGEN TO THE LOCAL IMMUNE SYSTEM, ALONG THE MUCOSAL SURFACES OF VIRAL ENTRY. USING A SINGLE CYCLE REPLICATIVE-DEFICIENT SIV (SIVSC) APPROACH, WHICH HAS BEEN SHOWN TO BE SAFE COMPARED TO TRADITIONAL ATTENUATED VACCINES, WE HAVE CLONED THE SIVSC GENOME UNDER THE CONTROL OF THE INVOLUCRIN PROMOTER (PINV-SIVSC), A TERMINALLY DIFFERENTIATED KERATINOCYTE SPECIFIC PROMOTER. WHEN ADMINISTERED, THE VACCINE TARGETS AND TRANSDUCES BASAL EPITHELIAL STEM CELLS FROM COLORECTAL TISSUES. THESE CELLS THEN PROLIFERATE AND DIFFERENTIATE INTO MATURE EPITHELIAL CELLS, TRIGGERING SIV ANTIGEN EXPRESSION VIA THE PROMOTER AND LEADING TO BOTH DIRECT AND CROSS PRIMING. FOR THIS PROJECT, WE PROPOSE: 1) TO CONFIRM AND FURTHER IMPROVE THE EFFICACY AND SAFETY PROFILE OF THE PINV-SIVSC VACCINE VIA THE COLORECTAL ROUTE; 2) TO VISUALIZE AND OPTIMIZE VACCINE DELIVERY, AND INVESTIGATE THE MECHANISMS OF ACTION UNDERLYING PROTECTION; AND 3) USING OUR BEST OPTIMIZED VACCINE STRATEGY, DEMONSTRATE PROTECTION FROM VIRUS ACQUISITION AND/OR VIRAL REPLICATION IN VIVO AND DETERMINE THE CORRELATES OF PROTECTION OR CONTROL AGAINST REPEATED LOW-DOSE INTRARECTAL CHALLENGES WITH HETEROLOGOUS SIV.

MECHANISM AND COUNTERMEASURE OF FENTANYL-INDUCED SUDDEN DEATH - PROJECT SUMMARY OPIOIDS WERE INVOLVED IN 75,673 OVERDOSE DEATHS IN 2020 IN THE U.S. FENTANYL, AN OPIOID MU-RECEPTOR (MOR) AGONIST ACTING ON CENTRAL MORS PRODUCES ANALGESIA, BUT DEPRESSES VENTILATION THAT COULD BE LETHAL. RAPID INTRAVENOUS INJECTION OF OVERDOSE FENTANYL, PARTICULARLY IN ILLICIT USE, IS THE DEADLIEST. IT TRIGGERS AN APNEA LEADING TO SUDDEN DEATH WITHIN MINUTES IN ~1/3 OF ADULTS (MALE > FEMALE) WITH ~75% OF FENTANYL SEQUESTERED BY THE LUNGS AFTER INJECTION. NALOXONE IS THE PREFERABLE TREATMENT FOR POST-OVERDOSE, BUT NOT VIABLE FOR THE ILLICIT FNT USERS WHO ARE FOUND PULSELESS UPON THE ARRIVAL OF EMERGENCY MEDICAL SERVICES. MOREOVER, NALOXONE COUNTERACTS ANALGESIA WITH POTENTIALLY LIFE-THREATENING SIDE EFFECTS. CENTRAL APNEA, UPPER AIRWAY OBSTRUCTION, AND CHEST WALL RIGIDITY ARE ASSUMED TO BE ACCOUNTABLE FOR THE SUDDEN DEATH, BUT THE EXACT CAUSE OF THE DEATH AND THE EFFECTIVE COUNTERMEASURE REMAIN UNKNOWN, CONSTITUTING A CRITICAL AND UNMET THERAPEUTIC CLINICAL NEED. BRONCHOPULMONARY C-FIBERS (PCFS) ARE THE MAJOR SENSORY NERVE ENDINGS INNERVATING THE AIRWAYS AND LUNGS AND CRUCIAL IN THE CONTROL OF RESPIRATORY RHYTHM. PCF STIMULATION COULD ELICIT CENTRAL APNEA AND VOCAL CLOSURE LIKELY BY PROMOTING RELEASE OF GLUTAMATE OR SUBSTANCE P TO ACTIVATE PCF 2ND-ORDER NEURONS IN THE NUCLEUS TRACTUS SOLITARIUS VIA AMPA AND NEUROKININ-1 RECEPTOR. WE HAVE REPORTED THAT INTRAVENOUS BOLUS INJECTION OF A LOW DOSE FENTANYL TRIGGERS A PCF-MEDIATED BRIEF APNEA, CONSISTENT WITH PREVIOUSLY REPORTED EXCITATION OF DORSAL ROOT GANGLION C- NEURONS BY MOR AGONISTS VIA GS-CAMP PATHWAY. OUR PRELIMINARY DATA SHOW A TRIPLE APNEA (CENTRAL APNEA, VOCAL CLOSURE AND LARYNGEAL CONSTRICTION/COLLAPSE) COUPLED WITH CHEST WALL RIGIDITY, RESULTING IN DEATH WITHIN MINUTES IN ANESTHETIZED ADULT RATS AFTER OVERDOSE FENTANYL IS RAPIDLY INJECTED. IN THIS PROPOSAL, WE SEEK TO ESTABLISH THE FIRST ANIMAL MODEL OF FENTANYL-INDUCED SUDDEN DEATH FOLLOWING TRIPLE APNEA IN AWAKE ADULT RATS AND DETERMINE THE DEPENDENCY OF TRIPLE APNEA (DEATH) ON THE GENDER AND PERIPHERAL (ESPECIALLY VAGAL) MORS/Μ1RS (AIM 1). WE WILL FURTHER DEMONSTRATE THAT FENTANYL DIRECTLY STIMULATES PCFS TO EVOKE TRIPLE APNEA BY ACTING ON MOR/Μ1R AND GS- CAMP PATHWAY, THEREBY PROMOTE PCF RELEASE OF GLUTAMATE AND SP TO ACTIVATE THE 2ND-ORDER NEURONS (AIM 2). OUR PILOT STUDY HAS SHOWN THAT INTRAVENOUS INJECTION OF DERMORPHIN (A PERIPHERALLY ACTING MOR AGONIST) INDUCES MOR INTERNALIZATION IN VAGAL SENSORY NEURONS AND BLOCKS FENTANYL-INDUCED TRIPLE APNEA. THE SS-ARRESTIN-MEDIATED SIGNALING IS KNOWN TO BE RESPONSIBLE FOR MOR DESENSITIZATION/INTERNALIZATION. WE WILL DEFINE THAT DIAMORPHINE HAS LIMITED IMPACT ON BASELINE CARDIORESPIRATORY ACTIVITY, BUT INTERNALIZES PCFS' MORS/Μ1RS VIA ACTIVATION OF SS- ARRESTIN-SIGNALING TO PREVENTS THE TRIPLE APNEA (DEATH) WITH NO CHANGE IN ANALGESIA. THIS WILL PRESENT A POTENTIAL SAFER AND TRANSLATIONAL PRETREATMENT FOR OVERDOSE FENTANYL IN CLINICAL SETTINGS (AIM 3). A MULTIDISCIPLINARY APPROACH (ELECTROPHYSIOLOGY, IMMUNOHISTOCHEMISTRY, PHARMACOLOGY AND BIOCHEMISTRY) AT SYSTEMIC AND CELLULAR LEVELS WILL BE APPLIED TO ELUCIDATE THE MECHANISMS UNDERLYING NOT ONLY TRIPLE APNEA (DEATH) BUT ALSO THE LACK OF TRIPLE APNEA AFTER DERMORPHIN PRETREATMENT. THE EXPECTED DATA WILL, FOR THE FIRST TIME, MECHANISTICALLY REVEAL THE KEY ROLE OF PCFS IN GENERATING FENTANYL-INDUCED TRIPLE APNEA AND PROVIDE A NOVEL AND POTENTIAL COUNTERMEASURE OF DERMORPHIN PRETREATMENT TO PREVENT/DIMINISH THE SUDDEN DEATH.

PATHWAYS OF MATERNAL ANEMIA

GENETIC DETERMINANTS OF HUMAN TRANSCRIPTIONAL AGING

IMPROVING RAPID PHENOTYPIC DRUG SUSCEPTIBILITY TESTING FOR DRUG RESISTANT TUBERCULOSIS IN HIGH-BURDEN AREAS - ABSTRACT TUBERCULOSIS (TB), CAUSED BY MYCOBACTERIUM TUBERCULOSIS (M.TB), IS A LEADING INFECTIOUS DISEASE AND CAUSE OF DEATH WORLDWIDE. THE GROWING BURDEN OF DRUG-RESISTANT (DR)-TB IS COMPLICATING TB TREATMENT. EARLY DIAGNOSIS OF TB WITH DRUG SUSCEPTIBILITY TESTING (DST) IS CRITICAL FOR SUCCESSFUL TREATMENT AND IS THE FIRST PILLAR OF THE WORLD HEALTH ORGANIZATION’S (WHO) END TB STRATEGY. DST IS ACHIEVED VIA PHENOTYPIC OR GENOTYPIC METHODS. TRADITIONALLY, PHENOTYPIC DST IS PERFORMED ON SOLID (LÖWENSTEIN JENSEN) OR LIQUID MEDIA (MGIT) IN A TWO-STEP PROCESS: FIRST A CULTURE TO IDENTIFY M.TB GROWTH, AND THEN RE-CULTURE OF THE ISOLATE WITH THE DRUGS TO BE TESTED. IN ADDITION TO REQUIRING BIOSAFETY LEVEL II-PLUS LABS, THE DST PROCESS, IF AVAILABLE IN LOW-MIDDLE INCOME SETTINGS, CAN TAKE 42 TO ~6 MONTHS FROM SAMPLE COLLECTION TO NOTIFICATION OF RESULTS TO THE CLINICAL PROVIDER RESULTING IN TREATMENT DELAYS, CONTINUED TRANSMISSION, AND HIGHER MORTALITY. CONVERSELY, GENOTYPIC DST HAS MANY ADVANTAGES, INCLUDING A REDUCED TIME TO RESULT (< 2H FOR GENEXPERT) AND THE POSSIBILITY OF DEPLOYMENT TO AT OR NEAR POINT OF CARE (POC). HOWEVER, ITS WIDESPREAD USE IN HIGH TB BURDEN RESOURCE-LIMITED SETTINGS IS HINDERED BY THE NEED FOR REGULAR POWER SUPPLY AND IMPORTANTLY COST. THUS, NIH/NIAID IS REDIRECTING ATTENTION TO INNOVATIVE AND SIMPLE PHENOTYPIC DST SOLUTIONS TO BE DEPLOYED AT OR NEAR POC. THE GOAL OF THIS APPLICATION IS TO DEVELOP THE 1G TEST INTO THE 2G TEST, PROVIDING HIGHER FLEXIBILITY TO PERFORM DST FOR 1ST AND 2ND FRONTLINE DRUGS, INCLUDING DRUGS PRESCRIBED FOR DS- AND DR-TB REGIMENS SUCH RIPE (DS-TB ORAL REGIMEN COMPOSED OF RIF/INH/PZA/ETHAMBUTOL), HPMZ (DS-TB 4-MONTH SHORT COURSE ORAL DRUG REGIMEN COMPOSED OF INH/RIFAPENTINE/MFX/PZA) AND BPAL [MDR- AND PRE-XDR ORAL DRUG REGIMEN COMPOSED OF BEDAQUILINE (BDQ), PRETOMANID (PMD) AND LINEZOLID (LNZ)], AS WELL AS CLOFAZIMINE (CFZ) AND DELAMANID (DLM), OTHER WHO RECOMMENDED ORAL AGENTS FOR DR-TB. BECAUSE THE 2G TEST IS NON-PROPRIETARY, ITS COST IS EXPECTED TO BE EXTREMELY LOW (< $8) AND MAINLY DRIVEN BY THE COST OF DRUGS. FURTHER, FOR THE 1G TEST WE TESTED A SIMPLE STEP TO DIGEST/DECONTAMINATE SPUTA THAT DOES NOT REQUIRE EQUIPMENT, MEETING THE NEAR TO POC TEST DEFINITION. WE WILL OPTIMIZE THIS SPUTUM-PROCESSING PROTOCOL FOR USE WITH THE 2G TEST. WE PROPOSE TO: AIM 1) DEVELOP AND VALIDATE THE 2G TEST BY DEFINING THE STABILITY AND CRITICAL CONCENTRATION (CC) FOR NEW DRUGS AGAINST KNOWN DR-M.TB STRAINS, AND OPTIMIZE APPROPRIATE SPUTUM DIGESTION AND DECONTAMINATION PROTOCOLS FOR THIS TEST; AIM 2) DETERMINE THE AGREEMENT OF THE 2G TEST WITH CURRENT GOLD STANDARD METHODS FOR PHENOTYPIC DST FOR EACH OF THE 11 DRUGS, AND AIM 3) DETERMINE THE ACCURACY OF THE 2G TEST AGAINST REFERENCE PHENOTYPIC DST PROTOCOLS USING FRESHLY COLLECTED SPUTA IN FIELD SETTINGS AND ASSESS ITS USABILITY, ACCEPTABILITY, AND FEASIBILITY. WE EXPECT THAT THE NOVEL, SIMPLE, AFFORDABLE AND SUSTAINABLE 2G TEST WILL PROVIDE A SIGNIFICANT IMPROVEMENT WHEN COMPARED TO CURRENT PHENOTYPIC DST REFERENCE METHODS, ALLOWING RAPID AND TAILORED TREATMENT FOR DS-/DR-TB IN LOW- AND MIDDLE-INCOME COUNTRIES WITH HIGH TB BURDEN.

SENSOR TECHNOLOGY FOR RAPID MICROBIAL IDENTIFICATION AND SUSCEPTIBILITY TESTING

MICROBIOME-GUT-BRAIN DYSFUNCTION IN PRODROMAL AND SYMPTOMATIC LEWY BODY DISEASES - PROJECT SUMMARY/ABSTRACT COGNITIVE DECLINE IN LEWY BODY DEMENTIAS [PARKINSON’S DISEASE DEMENTIA (PDD) AND DEMENTIA WITH LEWY BODIES (DLB); A CATEGORY OF ALZHEIMER’S DISEASE AND RELATED DEMENTIAS (ADRD)], CAUSES SIGNIFICANT FUNCTIONAL IMPAIRMENT AND DOES NOT RESPOND WELL TO EXISTING TREATMENTS. PERHAPS THE GREATEST CHALLENGE FOR DEVELOPING EFFECTIVE TREATMENTS IS THE LACK OF A MECHANISTIC UNDERSTANDING OF THE KEY EVENTS DRIVING PATHOPHYSIOLOGY. INCREASING EVIDENCE SUGGESTS THAT A PERTURBED GUT MICROBIOME (DYSBIOSIS) DRIVEN BY A BLOOM IN SULFATE REDUCING BACTERIA AND INCREASED INTESTINAL PERMEABILITY MAY BE KEY MECHANISMS DRIVING DISEASE PATHOGENESIS IN LEWY BODY DISEASES. THERE IS A NEED TO EVALUATE WHETHER THESE FACTORS ARE PRESENT EARLY IN THE DISEASE COURSE AND ASSOCIATED WITH KNOWN DISEASE PROCESSES, SUCH AS MIDBRAIN DEGENERATION AND CLINICAL OUTCOMES. IDIOPATHIC REM SLEEP BEHAVIOR DISORDER (IRBD), IS ONE OF THE EARLIEST AND MOST SPECIFIC PRODROMAL INDICATORS OF LEWY BODY DISEASES AS UP TO 96.6% OF IRBD PATIENTS WILL PROGRESS TO AN ALPHA-SYNUCLEINOPATHY AT 14 YEARS FOLLOW UP. THIS POPULATION THEREFORE PROVIDES AN OPPORTUNITY TO CONDUCT A NOVEL EVALUATION OF THE PRESENCE OF A BLOOM IN SRB/DYSBIOSIS AND INCREASED INTESTINAL PERMEABILITY EARLY IN THE DISEASE AND WHETHER THESE MECHANISMS ARE ASSOCIATED WITH KNOWN DISEASE PROCESSES (INCREASES IN A-SYNUCLEIN, MIDBRAIN DEGENERATION AND CLINICAL OUTCOMES). THE PROPOSED PROJECT WILL CONDUCT A PROSPECTIVE, CROSS-SECTIONAL STUDY TO TEST THE HYPOTHESIS THAT ABNORMAL LACTULOSE BREATH TESTS WITH ELEVATED H2S CONCENTRATION (AS A MARKER OF DYSBIOSIS AND A BLOOM OF SRB IN THE GUT) ARE PRESENT AT PRODROMAL STAGES OF THE DISEASE AND ASSOCIATED WITH INCREASES IN MARKERS OF INCREASED INTESTINAL PERMEABILITY AND MICROBIAL TRANSLOCATION OF LIPOPOLYSACCHARIDE (TOTAL BACTERIAL 16S RRNA GENE AND LIPOPOLYSACCHARIDE BINDING PROTEIN; LBP), KNOWN DISEASE PROCESSES (PLASMA CONCENTRATION OF A-SYNUCLEIN, MRI BIOMARKERS OF INTEGRITY OF SNC AND LC) AND CLINICAL OUTCOMES. WE WILL RECRUIT PRODROMAL (IRBD), SYMPTOMATIC LEWY BODY DISEASE PATIENTS (PD AND DLB) AND HC TO EVALUATE OUR HYPOTHESES. CONSISTENT WITH SEVERAL OBJECTIVES OUTLINED IN PAR-22-211, THE COMPLETION OF THE PROPOSED PROJECT WILL ADVANCE OUR MECHANISTIC UNDERSTANDING OF THE EFFECTS OF A BOOM IN SRB/DYSBIOSIS, POTENTIALLY IDENTIFYING TARGETS FOR DISEASE MODIFYING TREATMENTS. SHOULD OUR RESULTS DETERMINE AN IMPORTANT ROLE OF GUT SRB BLOOM/DYSBIOSIS IN THE PATHOPHYSIOLOGY OF LEWY BODY DISEASES, THESE MEASUREMENTS COULD BE INCORPORATED INTO ROUTINE CLINICAL PRACTICE TO SCREEN FOR EMERGING PATHOGENIC PROCESSES. NOTABLY, MANY POTENTIAL TREATMENTS TARGETING THE MICROBIOME ARE READILY AVAILABLE AND SUBSEQUENT CLINICAL TRIALS CAN EVALUATE WHETHER INTERVENTIONS TARGETING A BLOOM IN SRB/DYSBIOSIS COULD BE USED TO INTERVENE BEFORE OVERT MOTOR AND COGNITIVE SYMPTOMS DEVELOP.

ONCOGENIC PATHWAY-INDUCED FRAGILE SITES: A NEW PARADIGM FOR UNDERSTANDING GENOME INSTABILITY IN CANCER - ABSTRACT ONE OF THE GRAND CHALLENGES IN CANCER RESEARCH IS THE VAST HETEROGENEITY IN RESPONSIVENESS TO TREATMENT FOR DIFFERENT CANCER TYPES. WE HAVE MADE GREAT STRIDES IN TREATING SOME CANCERS, WHILE THE PROGNOSIS FOR OTHERS REMAINS DISMAL. LARGE SCALE WHOLE GENOME SEQUENCE (WGS) ANALYSES HAVE IDENTIFIED “BREAKPOINT SIGNATURES” OF DIFFERENT CANCER TYPES THAT PRESUMABLY REFLECT HETEROGENEITY IN THEIR UNDERLYING DISRUPTED PATHWAYS BUT, DRUG- GABLE TARGETS HAVE EMERGED ONLY IN CASES WHERE THE BREAKS CREATE ONCOGENIC FUSION PROTEINS. THE ABILITY TO PREDICT DISRUPTED PATHWAYS USING WGS WOULD BE HIGHLY IMPACTFUL FOR CANCER DIAGNOSIS AND TREATMENT. CHROMO- SOME FRAGILE SITES (FSS) MANIFEST AS GAPS AND BREAKS IN METAPHASE CHROMOSOMES WHEN CULTURED CELLS EXPERI- ENCE REPLICATION STRESS. HOWEVER, MANY FSS ARE NOT CANCER-TYPE SPECIFIC AND MANY BREAKPOINT HOTSPOTS IN CANCER ARE NOT KNOWN FSS, WHICH HAS SEVERELY LIMITED THE IMPACT OF THE FS FIELD. SINCE CULTURED CELLS OFFER THE POTENTIAL TO PROSPECTIVELY DISSECT MECHANISMS INITIATING CHROMOSOME BREAKS AND TO TRACK THEIR EXPANSION INTO COMPLEX STRUCTURAL VARIATION, THERE IS A CRITICAL NEED TO MAKE THESE IN VITRO SYSTEMS MORE CANCER RELEVANT. OUR LONGTERM GOAL IS TO ESTABLISH A SUSTAINED RESEARCH STRATEGY THAT CAN PREDICT WHICH KNOWN PATHWAYS ARE DISRUPTED IN A GIVEN CANCER TYPE FROM THEIR BREAKPOINT PATTERNS. THE OVERALL OBJECTIVE OF THIS PROPOSAL IS TO REVEAL MECHANISMS BY WHICH PERTURBATION OF CANCER-RELEVANT CELLULAR PATHWAYS PRODUCE PATHWAY-SPECIFIC PATTERNS OF FSS IN CELL CULTURE AND DETERMINE WHETHER THOSE PATTERNS CAN BE IDENTIFIED IN CANCERS. OUR CENTRAL HYPOTHESIS IS THAT THE POWER OF IN VITRO SYSTEMS TO DISSECT MECHANISMS LEADING TO CANCER BREAKPOINT SIGNATURES WILL BECOME EVIDENT ONLY WHEN SHIFTED AWAY FROM CHEMICALLY-INDUCED FSS TOWARDS FSS INDUCED BY PERTURBING KNOWN CANCER-RELEVANT PATHWAYS. WE HAVE SHOWN THAT OVEREXPRESSION OF DIFFERENT ONCOGENES LEADS TO ONCOGENE-SPECIFIC SPECTRA OF FSS AND PRE- LIMINARY DATA SUGGEST SOME OF THE MECHANISMS BY WHICH THIS OCCURS. OUR RATIONALE IS THAT UNDERSTANDING CANCER- RELEVANT MECHANISMS THAT SPECIFY FSS WILL FILL THE GAP IN LINKING IN VITRO FSS TO CANCER BREAKPOINTS. THIS WOULD BE A MAJOR STEP TOWARD A STRATEGY TO PREDICT DISRUPTED PATHWAYS FROM CANCER WGS DATA, THEREBY SUGGESTING TREAT- MENTS FOR PREVIOUSLY INTRACTABLE CANCERS. AIM1 WILL USE HIGH THROUGHPUT/RESOLUTION REPLICATION AND OLIGOPAINTS ASSAYS TO IDENTIFY DOWNSTREAM MECHANISMS DISTINGUISHING WHICH OF MANY SITES OF ONCOGENE-INDUCED REPLICATION DELAY MANIFEST AS FSS. AIM2 WILL ELUCIDATE UPSTREAM MECHANISMS CAUSING REPLICATION DELAYS AND FS AT SPECIFIC SITES. AIM3 WILL MAP FSS AT UNPREDEDENTED RESOLUTION AND MINE TUMOR SEQUENCING DATABASES FOR SIGNATURES THAT MATCH THOSE OF ONCOGENE-SPECIFIC FSS. THIS CONTRIBUTION WILL BE SIGNIFICANT BECAUSE THE ABILITY TO IDENTIFY AFFECTED PATHWAYS SOLELY FROM BREAKPOINT SIGNATURES WOULD EXPOSE TUMOR-SPECIFIC VULNERABILITIES FOR PRECISION CANCER MEDICINE. THE PROPOSED RESEARCH IS INNOVATIVE BECAUSE IT WILL REJUVENATE THE IMPACT OF FS RESEARCH, CLOSING THE GAP BETWEEN IN VITRO SYSTEMS AND IN VIVO CANCERS WHILE LEVERAGING NOVEL TECHNOLOGIES TO MANIPULATE CELLS, MAP THEIR SIGNATURES OF GENOME INSTABILITY AND MATCH THEM TO CANCER BREAKPOINT SIGNATURES.

EQUIPMENT: MRI: TRACK 2 MEG SYSTEM WITH INTEGRATED HELIUM RECOVERY EQUIPMENT FOR COGNITIVE NEUROSCIENCE -THE MAJOR RESEARCH INSTRUMENTATION PROJECT SUPPORTS THE ACQUISITION OF A NEW MAGNETOENCEPHALOGRAPHY (MEG) INSTRUMENT, THE MEGIN TRIUS NEO WITH AN INTEGRATED HELIUM RECYCLER. MEG IS A FUNCTIONAL BRAIN IMAGING TECHNOLOGY THAT PROVIDES EXQUISITE SENSITIVITY TO THE TIMING OF BRAIN ACTIVITY AND IS A DIRECT MEASURE OF NEURONAL CURRENTS. THE TECHNOLOGY IS COMPLEMENTARY TO OTHER NEUROIMAGING TECHNIQUES, SUCH AS FMRI, WHICH HAS HIGH SPATIAL SENSITIVITY BUT LIMITED SENSITIVITY TO RAPID CHANGES IN BRAIN DYNAMICS. THE MEG INSTRUMENTATION IS CRITICAL FOR SUPPORTING COGNITIVE NEUROSCIENCE RESEARCH. SAMPLE PLANNED PROJECTS THAT USE THE INSTRUMENT INCLUDE NEW RESEARCH TO GAIN FURTHER INSIGHT INTO THE DEVELOPMENT OF BRAIN AND BEHAVIOR IN CHILDREN, EXAMINATION OF THE DYNAMICS OF MEMORY SYSTEMS, ELUCIDATION OF HOW BRAIN STIMULATION ENHANCES BRAIN FUNCTION AND BEHAVIOR, AND INVESTIGATION OF THE ROLE OF SLEEP FOR COGNITIVE AND BEHAVIORAL OUTCOMES ACROSS THE LIFESPAN. THE MEGIN TRIUS NEO IS A HIGH-CHANNEL DENSITY, COMMERCIALLY AVAILABLE MEG SYSTEM THAT PROVIDES HIGH SENSITIVITY TO WEAK SIGNALS ALLOWING FOR MEASUREMENT OF DEEP STRUCTURES AND LOW AMPLITUDE SIGNALS. THE EQUIPMENT ALSO DRAMATICALLY REDUCES USE OF HELIUM, A LIMITED RESOURCE. BY COMBINING MEG DATA WITH STRUCTURAL INFORMATION OBTAINED WITH MAGNETIC RESONANCE IMAGING, NEW KNOWLEDGE IS OBTAINED BY UNDERSTANDING HOW BRAIN FUNCTION AND STRUCTURE RELATE TO OUTCOMES ACROSS THE POPULATION. THE GOALS OF SOME RESEARCH SUPPORTED BY THIS INSTRUMENTATION IS TRANSLATIONAL AND AIMS TO INFORM UNDERSTANDING OF THE BRAIN TO IMPROVE HEALTH AND OUTCOMES FOR ALL. OTHER RESEARCH SUPPORTED BY THIS INSTRUMENTATION USES ARTIFICIAL INTELLIGENCE TO INTERPRET THESE COMPLEX DATASETS TO ENHANCE THE UNDERSTANDING OF THE LINKS BETWEEN BRAIN AND BEHAVIOR. OVERALL, THE INSTRUMENTATION WILL PROVIDE SIGNIFICANT RESEARCH AND TRAINING OPPORTUNITIES. THIS AWARD REFLECTS NSF'S STATUTORY MISSION AND HAS BEEN DEEMED WORTHY OF SUPPORT THROUGH EVALUATION USING THE FOUNDATION'S INTELLECTUAL MERIT AND BROADER IMPACTS REVIEW CRITERIA.- SUBAWARDS ARE NOT PLANNED FOR THIS AWARD.

GENETIC ANALYSIS OF CERCARIAL RELEASE IN SCHISTOSOMES

HIV ANTIBODIES AND NK CELL ADCC: NANOMETER-SCALE TRACKING OF IMMUNE SYNAPSE DYNAMICS. - SUMMARY STRUCTURAL STUDIES HAVE ESTABLISHED THAT NATURALLY ELICITED ANTIBODIES CAN BIND TO HIV ENVELOPE (ENV) OVER A WIDE RANGE OF EPITOPES AND ANGLES MEDIATED BY THEIR FAB ARM (I.E. “IMMUNE COMPLEX GEOMETRY”). HOW THIS AFFECTS ANTIBODY DEPENDENT CELLULAR CYTOTOXICITY (ADCC) ACTIVITY, ESPECIALLY BY NATURAL KILLER (NK) CELLS IS UNKNOWN. OUR KNOWLEDGE IS LIMITED BY A POOR UNDERSTANDING OF HOW ANTIBODY IMMUNE COMPLEXES ORCHESTRATE ANTIBODY RECEPTOR BASED SIGNALING (SPECIFICALLY FOR FC GAMMA RIIIA, FCRIIIA). THE LONG-TERM GOAL IS TO ACQUIRE A MORE DETAILED UNDERSTANDING OF HOW ANTIBODIES RECRUIT FC MEDIATED CELLULAR ACTIVITY AND TO DEVELOP NOVEL STRATEGIES TO ENGINEER ANTIBODIES, DRUGS, AND VACCINES THAT CAN RECRUIT SPECIFIC EFFECTOR FUNCTIONS WITH MAXIMAL POTENCY IN VIVO. THE OBJECTIVE OF THIS PROPOSAL IS TO DETERMINE HOW IMMUNE COMPLEX GEOMETRY IMPACTS NK CELL ADCC AGAINST HIV AND IGG RECEPTOR SPATIOTEMPORAL DYNAMICS WITHIN THE NK CELL IMMUNE SYNAPSE (NKIS). OUR CENTRAL HYPOTHESIS IS THAT ANTIBODY GEOMETRY WILL MODULATE FCRIIIA INTERACTION AND ADCC ACTIVITY IN RELATION TO IMMUNE COMPLEX GEOMETRY. THE RATIONALE FOR THIS WORK IS THAT CUTTING-EDGE MICROSCOPY OBSERVATIONS WILL PROVIDE NEW INSIGHT INTO ADCC FUNCTION WITH IMMEDIATE IMPACTS ON HIV ANTIBODY THERAPEUTIC DESIGN WITH APPLICATIONS TO A BROADER RANGE OF HUMAN DISEASES. OUR CENTRAL HYPOTHESIS WILL BE TESTED IN THREE SPECIFIC AIMS: 1) DETERMINE HOW IMMUNE COMPLEX GEOMETRY INFLUENCES FCGRIIIA INTERACTION DURING ADCC; 2) PERFORM SINGLE MOLECULE TRACKING OF FCRIIIA WITHIN THE NKIS DURING ADCC; 3) DETERMINE NANOMETER-SCALE LOCALIZATION OF FCRIIIA AND SIGNALING KINASES WITHIN THE NKIS DURING ADCC. WE WILL PURSUE THESE AIMS USING THE INNOVATIVE TECHNIQUE OF MINFLUX NANOSCOPY, A SUPER-RESOLUTION FLUORESCENCE MICROSCOPY TECHNIQUE THAT IS CAPABLE OF 1- TO 3-NM SPATIAL RESOLUTIONS IN BOTH 2- AND 3-DIMENSIONS AS WELL AS SUB-MILLISECOND TRACKING OF SINGLE MOLECULES IN LIVE CELLS. CAREFULLY MEASURED IN VITRO ADCC ACTIVITY AND FÖRSTER RESONANCE ENERGY TRANSFER (FRET) MEASUREMENTS WILL ALSO COMPLEMENT OUR MINFLUX OBSERVATIONS AND BROADEN THE INTERPRETATION OF OUR RESULTS. THESE STUDIES ARE SIGNIFICANT BECAUSE THEY WILL ESTABLISH A MOLECULAR BASIS FOR ANTIBODY EFFECTOR FUNCTION, ESPECIALLY IN RELATION TO NK CELL ADCC, THAT COULD IMPROVE THERAPEUTICS FOR HIV. THE TECHNIQUES ESTABLISHED IN THIS PROPOSAL WILL ALSO BE USEFUL FOR INTERROGATING ANTIBODY ADCC FUNCTION FOR OTHER VIRAL PATHOGENS. THE EXPECTED OUTCOME OF OUR STUDIES IS THE CHARACTERIZATION OF BIOPHYSICAL PRINCIPLES THAT ALTER NK CELL ADCC ACTIVITY AS WELL AS THE MOLECULAR MECHANICS THAT FORM THE BASIS FOR SUCH ACTIVITY. THESE FINDINGS WILL HAVE AN IMPORTANT IMPACT ON HUMAN HEALTH BY OFFERING A RATIONAL BASIS FOR DESIGNING IMPROVED ANTIBODY THERAPEUTICS FOR HIV, AS WELL AS OTHER VIRUSES AND DISEASES, AND WILL INCREASE OUR BASIC UNDERSTANDING OF NK CELL ADCC.

GENOME WIDE ASSESSMENT OF ESSENTIAL GENES IN TRYPANOSOMA BRUCEI

SYNERGISTIC EFFECTS OF SILICA EXPOSURE, VIRUS INFECTION AND GENETIC PREDISPOSITION IN SYSTEMIC AUTOIMMUNITY

HUMAN BRAIN AND SPINAL FLUID RESOURCE CENTER

THIS EDA INVESTMENT WILL LEVERAGE RECOVERY AND RESILIENCE IN SAN ANTONIO, TEXAS. TEXAS BIOMEDICAL RESEARCH INSTITUTE IS ADDRESSING THE LOCAL AND REGIONAL NEED FOR HIGH-VALUE LIFE SCIENCE JOBS BY CONSTRUCTING A FACILITY WHICH ENABLES EXPANDING PANDEMIC PREPAREDNESS CAPACITY FOR IDENTIFYING AND DEVELOPING NEW INFECTIOUS DISEASE DIAGNOSTICS, THERAPIES, AND CURES. THE FACILITY IS BEING DESIGNED TO COUNTER THE EFFECTS OF NATURAL DISASTERS AND EMPLOY REDUNDANCY AND DESIGN ELEMENTS THAT ENABLE OPERATION EVEN WHEN REGIONAL ELECTRICAL AND COMMUNICATIONS GRID GO DOWN. THE ECONOMIC IMPACT WILL DIRECTLY INCREASE THE AVAILABILITY OF HIGH PAYING JOBS WITHIN A FACILITY SPECIFICALLY DESIGNED FOR RESILIENCY TO EMERGING NATURAL DISASTER THREATS, INCLUDING WEATHER RELATED AND PANDEMICS. THIS PROJECT WAS MADE POSSIBLE BY THE REGIONAL PLANNING EFFORTS LED BY THE ALAMO AREA COUNCIL OF GOVERNMENTS. EDA FUNDS ALAMO AREA COUNCIL OF GOVERNMENTS TO BRING TOGETHER THE PUBLIC AND PRIVATE SECTORS TO CREATE AN ECONOMIC DEVELOPMENT ROADMAP TO STRENGTHEN THE REGIONAL ECONOMY, SUPPORT PRIVATE CAPITAL INVESTMENT AND CREATE JOBS.

IDENTIFICATION OF NOVEL MICRORNAS ASSOCIATED WITH BRAIN STRUCTURE AND FUNCTION

LARGE-SCALE METHYLATION PROFILING IN METABOLIC SYNDROME PHENOTYPES

DISCOVERING MORE JUVENILE MYOCLONIC EPILEPSY GENES BY A CONSORTIUM

THE PLASTID OF TOXOPLASMA GONDII

IDENTIFYING CHARACTERIZING AND INDUCING EFFECTIVE ANTI-HIV T CELL RESPONSES

MECHANISM AND EVOLUTION OF FILOVIRAL MONOCLONAL AFFINITY REAGENT SANDWICH ASSAYS

METHAMPHETAMINE AND HIV INTERACTIONS IN THE REGULATION OF GLIAL ACTIVATION

RESCUE OF BROADLY NEUTRALIZING MABS USING NATIVE TRIMER

MECHANISMS OF EFFICIENT CONTROL OF MYCOBACTERIUM TUBERCULOSIS IN THE LUNGS PRIOR TO GRANULOMA - PROJECT SUMMARY/ABSTRACT. NOVEL VACCINATION STRATEGIES ARE NECESSARY TO CONTAIN THE TB PANDEMIC, AS THE CURRENTLY LICENSED ANTI-TUBERCULAR VACCINE, BACILLE CALMETTE-GUERIN (BCG), HAS LIMITED AND VARIABLE EFFICACY. ATTENUATED, LIVE-REPLICATING MYCOBACTERIUM TUBERCULOSIS (MTB) EXPRESS THE FULL COMPLEMENT OF PROTECTIVE ANTIGENS NOT PRESENT IN BCG. AS A RESULT, THESE STRAINS ARE MOST LIKELY TO INDUCE LONG-LIVED IMMUNE RESPONSES AND GENERATE DURABLE PROTECTION. RHESUS MACAQUES VACCINATED WITH AN ISOGENIC MTB MUTANT IN THE ALLELE ENCODING THE STRESS-RESPONSE MASTER REGULAR SIGH (DSIGH) WERE PROTECTED FROM TB AFTER INFECTION WITH A LETHAL DOSE OF MTB, AND CHARACTERIZED BY THE PRESENCE OF INDUCIBLE BRONCHUS ASSOCIATED LYMPHOID TISSUE (IBALT) AND ROBUST T CELL RESPONSES IN THE LUNGS. PROTECTION BY DSIGH COULD BE REVERSED BY THE DEPLETION OF CD20+ B CELLS WHICH ABLATES IBALT. PROTECTION WITH DSIGH WAS VALIDATED IN CYNOMOLGUS MACAQUES, A SECOND NHP SPECIES USED AS A MODEL FOR TB VACCINATION. PRELIMINARY DATA PRESENTED IN THIS PROPOSAL INDICATES THAT PROTECTION ELICITED BY DSIGH IS BASED ON THE GENERATION OF VERY EARLY, POTENT, INNATE IMMUNE RESPONSES IN THE LUNG THAT DRIVE RIGOROUS IMMUNE DYNAMICS AND INTERACTIONS. WE PROPOSE TO USE CUTTING-EDGE TECHNIQUES THAT OUR GROUP HAS OPTIMIZED, SUCH AS SINGLE CELL RNA SEQUENCING IN AIRWAYS AND LUNGS, PET/CT SCANS OF WHOLE ANIMALS AND SINGLE CELL IMAGING, TO FULLY UNDERSTAND ELITE LUNG RESPONSES GENERATED BY DSIGH COMPARED TO MTB. OUR PROPOSED WORK WILL NOT ONLY PROVIDE IN-DEPTH KNOWLEDGE OF IMMUNE RESPONSES GENERATED BY A POTENTIAL HUMAN INTERVENTION FOR TB, BUT ALSO IDENTIFY MECHANISMS BY WHICH MTB INFECTION CAN BE STERILIZED PRIOR TO THE FORMATION OF THE GRANULOMA.

MYELOID-DERIVED SUPPRESSOR CELLS (MDSC) SUPPRESS INFANT IMMUNE RESPONSES

FATTY LIVER DISEASE AND ITS DETERMINANTS IN AN AMERICAN INDIAN POPULATION: THE STRONG HEART STUDY

IMMUNE PROFILE AND NETWORK ANALYSIS OF MALARIA INFECTION AND VACCINATION

DUODENAL MUCOSAL DEFENSE MECHANISMS

THE PROTECTIVE ROLE OF MICROGLIA IN PREVENTING HYPOXIC DISRUPTION OF BLOOD-BRAIN BARRIER INTEGRITY AND VCID - PROJECT SUMMARY/ABSTRACT DEMENTIA IS A MAJOR HEALTH PROBLEM IN THE UNITED STATES. VASCULAR CONTRIBUTIONS TO COGNITIVE IMPAIRMENT AND DEMENTIA (VCID) IS THE SECOND LEADING CAUSE OF DEMENTIA BEHIND ALZHEIMER’S DISEASE (AD) BUT DESPITE THE MASSIVE IMPACT OF VCID IN THE EXPANDING ELDERLY POPULATION, ITS PATHOGENESIS IS STILL ONLY POORLY UNDERSTOOD. THE CONSENSUS IS THAT IN THE AGING BRAIN, PARTICULARLY ON A BACKGROUND OF HYPERTENSION, BLOOD VESSELS UNDERGO DEGENERATIVE CHANGES, RESULTING IN LOSS OF BLOOD-BRAIN BARRIER (BBB) INTEGRITY AND INCREASED VASCULAR RESISTANCE, WHICH TOGETHER, LEAD TO CEREBRAL HYPOPERFUSION, NEURONAL DAMAGE AND COGNITIVE DECLINE. RECENTLY, WE DESCRIBED A NOVEL ROLE FOR MICROGLIA IN THE MAINTENANCE OF VASCULAR INTEGRITY. WE DEMONSTRATED THAT CHRONIC MILD HYPOXIA (CMH; 8% O2) INDUCES TRANSIENT VASCULAR LEAK IN SPINAL CORD BLOOD VESSELS IN YOUNG (10 WEEKS OLD) MICE, THAT IS ASSOCIATED WITH MICROGLIAL ACTIVATION AND CLUSTERING AROUND LEAKY BLOOD VESSELS. INTERESTINGLY, MICROGLIAL DEPLETION PROFOUNDLY INCREASED VASCULAR LEAK AND THIS WAS ASSOCIATED WITH ASTROCYTE-VASCULAR UNCOUPLING AND LOSS OF VASCULAR TIGHT JUNCTION PROTEINS, SUGGESTING THAT MICROGLIA PLAY AN IMPORTANT PROTECTIVE ROLE IN MAINTAINING VASCULAR INTEGRITY IN THE SPINAL CORD. WE HAVE SINCE FOUND THAT CMH ALSO TRIGGERS VASCULAR LEAK IN THE BRAIN AND THAT MICROGLIAL DEPLETION EXACERBATES THIS LEAK. STRIKINGLY, IN AGED (20 MONTHS OLD) MICE, THE EXTENT OF HYPOXIC-INDUCED CEREBROVASCULAR DISRUPTION IS GREATLY ENHANCED, AS SHOWN BY INCREASED VASCULAR LEAK AND THE EMERGENCE OF MICROHEMORRHAGES, THOUGH THE IMPACT OF MICROGLIAL DEPLETION IN AGED MICE HAS YET TO BE ADDRESSED. TOGETHER, OUR DATA SUGGESTS THAT MICROGLIA PLAY AN IMPORTANT VASCULOPROTECTIVE ROLE IN YOUNG MICE, BUT THIS MECHANISM MAY BE LESS EFFECTIVE IN THE AGED BRAIN. TAKEN WITH THE OBSERVATION THAT AGING INDUCES THE APPEARANCE OF A “PRIMED”, PRO-INFLAMMATORY, DESTRUCTIVE MICROGLIAL PHENOTYPE, WE HYPOTHESIZE THAT: (I) MILD HYPOXIA TRIGGERS VASCULAR LEAK AND MICROHEMORRHAGE IN THE BRAIN, RESULTING IN NEURONAL DAMAGE AND COGNITIVE DECLINE, (II) VASCULAR DISRUPTION IS WORSE IN THE AGED AND THE HYPERTENSIVE, (III) MICROGLIA PLAY AN IMPORTANT VASCULOPROTECTIVE ROLE IN STABILIZING THE BBB, BUT THIS DECLINES WITH AGE, AND (IV) REPOPULATING THE AGED BRAIN WITH YOUNG MICROGLIA OR ATTENUATION OF MICROGLIAL ACTIVATION STATE, COULD STABILIZE THE BBB AND REDUCE COGNITIVE IMPAIRMENT. TO INVESTIGATE THESE HYPOTHESES, WE PROPOSE THREE SPECIFIC AIMS: (1) CHARACTERIZE HYPOXIA-INDUCED VASCULAR LEAK IN THE BRAIN AND DEFINE HOW THIS IS INFLUENCED BY AGE, GENDER, SEVERITY OF HYPOXIA, HYPERTENSION AND BRAIN REGION, (2) DEFINE THE CONTRIBUTION OF MICROGLIA IN PREVENTING HYPOXIA-INDUCED CEREBROVASCULAR LEAK IN YOUNG AND AGED MICE, AND (3) DEMONSTRATE THAT HYPOXIA-INDUCED BBB DISRUPTION AND COGNITIVE IMPAIRMENT ARE REDUCED BY REPOPULATING THE AGED BRAIN WITH “YOUNG” MICROGLIA OR BY ATTENUATING MICROGLIAL ACTIVATION STATE. THESE STUDIES WILL PROVIDE IMPORTANT INSIGHT INTO THE LINK BETWEEN HYPOXIC EXPOSURE, BBB DISRUPTION, NEURONAL DAMAGE AND COGNITIVE DECLINE, AND INFORM ON THE THERAPEUTIC POTENTIAL OF MANIPULATING MICROGLIAL BEHAVIOR IN THE AGED BRAIN TO RESTORE VASCULOPROTECTIVE FUNCTION.

QUANTITATIVE TRAIT LOCUS MAPPING IN HUMAN PEDIGREES

NEW INFECTION-RELATED PROTEINS OF PLASMODIUM SPOROZOITES AND LIVER STAGES

THE INNATE IMMUNE RESPONSE IN THE MARMOSET MODEL OF GBV-B INFECTIONS: A SURROGATE

COGNITIVE SEQUELAE OF CEREBROVASCULAR AND GUT DYSFUNCTION IN POST-ACUTE COVID-19 SYNDROME. - ABSTRACT: APPROXIMATELY ONE THIRD OF NON-HOSPITALIZED CORONAVIRUS DISEASE OF 2019 (COVID-19) PATIENTS REPORT CHRONIC SYMPTOMS AFTER RECOVERING FROM THE ACUTE STAGE OF SEVERE ACUTE RESPIRATORY SYNDROME CORONAVIRUS 2 (SARS-COV-2) INFECTION. SOME OF THE MOST PERSISTENT AND COMMON COMPLAINTS OF THIS POST-ACUTE COVID-19 SYNDROME (PACS) ARE COGNITIVE IN NATURE, DESCRIBED SUBJECTIVELY AS “BRAIN FOG” AND ALSO OBJECTIVELY MEASURED AS DEFICITS IN EXECUTIVE FUNCTION, WORKING MEMORY, ATTENTION, PROCESSING SPEED. THE MECHANISMS OF THESE CHRONIC COGNITIVE SEQUELAE ARE CURRENTLY NOT UNDERSTOOD. MOST STUDIES TO-DATE HAVE FOCUSED ON DIRECT SARS-COV-2 INFECTION OF THE BRAIN; HOWEVER, WHILE DIRECT VIRAL BRAIN INFECTION IS PLAUSIBLE IN ACUTE CASES OF SEVERE AND FATAL COVID-19, IT IS OF INTEREST TO EXAMINE INDIRECT MECHANISMS OF CHRONIC COGNITIVE DYSFUNCTION THAT FOLLOW MILD AND ASYMPTOMATIC DISEASE CASES. SARS-COV-2 INFLICTS DAMAGE TO CEREBRAL BLOOD VESSELS AND THE INTESTINAL WALL BY BINDING TO ANGIOTENSIN-CONVERTING ENZYME 2 (ACE2) RECEPTORS AND ALSO BY PRODUCING HIGH LEVELS OF SYSTEMIC CYTOKINES, COMPROMISING THE BRAIN’S NEUROVASCULAR UNIT AND DEGRADING THE INTESTINAL BARRIER, POTENTIALLY INCREASING THE PERMEABILITY OF BOTH TO HARMFUL SUBSTANCES. SUCH SUBSTANCES ARE HYPOTHESIZED TO BE PRODUCED BY PATHOGENIC MICROBIOTA IN THE GUT THAT, GIVEN THE PROFOUND EFFECTS COVID-19 HAS ON THE GASTROINTESTINAL SYSTEM, MAY FLOURISH VIA INTESTINAL DYSBIOSIS. COVID-19 MAY THEREFORE CREATE A SCENARIO IN WHICH NEUROTOXIC AND NEUROINFLAMMATORY SUBSTANCES READILY PROLIFERATE FROM THE GUT LUMEN AND ENCOUNTER A WEAKENED NEUROVASCULAR UNIT, GAINING ACCESS TO THE BRAIN AND SUBSEQUENTLY PRODUCING COGNITIVE DEFICITS. WE INTEND TO EXAMINE SUCH EFFECTS OF SARS-COV-2 IN PACS PATIENTS LONGITUDINALLY OVER THE COURSE OF 3 STUDY VISITS (BASELINE, 4 MONTHS, AND 8 MONTHS). THE IMPAIRMENTS OF CEREBROVASCULAR FUNCTION AND INTESTINAL BARRIER, AS WELL AS THEIR EFFECTS ON COGNITIVE SYMPTOMOLOGY, WILL BE EXAMINED IN 80 FORMER COVID-19 PATIENTS WHO RECOVERED FROM NON-HOSPITALIZED ACUTE PHASES OF COVID-19, YET REPORT PERSISTENT COGNITIVE SYMPTOMS (PACS+). THESE PATIENTS WILL BE COMPARED WITH 80 FORMER SIMILAR COVID-19 PATIENTS WITHOUT SUCH SYMPTOMS (PACS-). FORTY HEALTHY CONTROL PARTICIPANTS WILL ALSO BE RECRUITED TO ESTABLISH GENERAL NEUROVASCULAR, INTESTINAL, AND COGNITIVE EFFECTS OF COVID-19 HISTORY. CEREBROVASCULAR FUNCTION WILL BE QUANTIFIED VIA INNOVATIVE FUNCTIONAL MAGNETIC RESONANCE IMAGING OF CEREBROVASCULAR REACTIVITY (CVR) TO RESPIRATION OF CO2 GAS, WHILE THE INTESTINAL BARRIER WILL BE ASSESSED VIA CONCENTRATIONS OF INTESTINAL WALL BIOMARKERS IN BLOOD PLASMA SUCH AS FATTY ACID-BINDING PROTEIN 2 (FABP-2) AND ZONULIN. GUT DYSBIOSIS WILL BE ESTABLISHED VIA LACTULOSE BREATH TESTING, AND LEVELS OF SUBSEQUENTLY PRODUCED AND SYSTEMICALLY RELEASED LIPOPOLYSACCHARIDE (LPS), PEPTIDOGLYCAN (PGN) AND PRO-INFLAMMATORY CYTOKINES WILL ALSO BE QUANTIFIED. IMPAIRMENTS IN THE NEUROVASCULAR UNIT AND INTESTINAL BARRIER IN THE CONTEXT OF GUT DYSBIOSIS ARE EXPECTED TO BE ASSOCIATED WITH GREATER COGNITIVE DEFICITS IN PACS+ PATIENTS. THIS WORK MAY REVEAL IMMEDIATE RECOURSES FOR RESOLVING PACS COGNITIVE EFFECTS VIA EXISTING TREATMENTS FOR VASCULAR DYSFUNCTION AND GUT HEALTH.

DOPAMINE SYSTEM AS REPORTER OF HIV STATUS AND INFLAMMATION IN METH ABUSERS

FACTORS INFLUENCING ORAL TRANSMISSION OF SIV

MAPPING THE 3D ARCHITECTURE OF NATIVE HUMAN REPLISOMES

A VASCULO-PROTECTIVE ROLE FOR BETA4 INTEGRIN IN NEUROINFLAMMATION

HIGH PERFORMANCE COMPUTING SYSTEM FOR HUMAN GENOMICS

A NOVEL PLASMINOGEN RECEPTOR IN BREAST CANCER

IDENTIFICATION OF PRE-ECLAMPSIA SUSCEPTIBILITY GENES

TROPONIN AND MYOSIN IN REGULATION OF MUSCLE CONTRACTION AND HEART DISEASE

STRONG HEART FAMILY STUDY

SIGNIFICANT EXPANSION OF THE SNPRC SPECIFIC PATHOGEN FREE RHESUS MACAQUE COLONY FOR AIDS RESEARCH - THE SOUTHWEST NATIONAL PRIMATE RESEARCH CENTER (SNPRC) SPF RHESUS COLONY, SUPPORTED BY THE NIH SPF RHESUS BREEDING PROGRAM (U42 OD010442), IS PRESENTLY AROUND 900 ANIMALS AND SUPPORTS AIDS-RELATED RESEARCH BOTH AT SNPRC AND THROUGH SALES TO AIDS INVESTIGATORS AT OTHER INSTITUTIONS. THE SNPRC IS WELL POSITIONED TO MAINTAIN AND EXPAND SPF RHESUS MACAQUE PRODUCTION AS A CENTER LOCATED IN A CLIMATE HOSPITABLE TO LARGELY OUTDOOR HOUSING AT AN INSTITUTION WITH CAPACITY FOR EXPANSION. THE NIH HAS RECENTLY RECOGNIZED THE NEED FOR EXPANSION OF SPF RHESUS PRODUCTION. SPECIFICALLY, THE ORIP NONHUMAN PRIMATE EVALUATION AND ANALYSIS COMPLETED IN DECEMBER, 2018, PROPOSES PROVIDING “NIH RESOURCES TO EXPAND EXISTING COLONIES OF RHESUS MACAQUES, INCLUDING SPF AND ENHANCED SPF COLONIES, BY 10-25% IN ORDER TO MEET GROWING DEMAND” AS ONE OF EIGHT KEY RECOMMENDATIONS STEMMING FROM THE EXPERT PANEL DISCUSSIONS (HTTPS://ORIP.NIH.GOV/ABOUT- ORIP/RESEARCH-HIGHLIGHTS/NONHUMAN-PRIMATE-EVALUATION-AND-ANALYSIS-PART-2-REPORT-EXPERT-PANEL). THIS APPLICATION PROPOSES RENOVATION AND NEW CONSTRUCTION IN ONE OF TWO SPF RHESUS HOUSING COMPLEXES THAT WILL SIGNIFICANTLY EXPAND OUR CAPACITY TO PRODUCE SPF RHESUS MACAQUES FOR USE IN AIDS RESEARCH AND WILL ENHANCE OUR ABILITY TO MANAGE THE POPULATION FOR RESEARCH USE IN THE MOST EFFICIENT AND HUMANE FASHION. THE PROPOSED RENOVATION WILL INCREASE OUR CAPACITY TO PRODUCE SPF RHESUS MACAQUES BY 41%; PROVIDE APPROPRIATELY DESIGNED SPACE TO ALLOW FOR EXAMINATION AND SAMPLING OF ANIMALS DESTINED FOR POTENTIAL STUDY OR SALE WHILE NOT COMPLETELY REMOVING THEM FROM THEIR ORIGINAL SOCIAL SETTING; MINIMIZE THE NEED TO MOVE ANIMALS AWAY FROM THE BUILDING COMPLEX BY PROVIDING A MODERN CLINIC AND PROCESSING ROOMS IN ASSOCIATION WITH THE HOUSING COMPLEX; IMPROVE THE CARE OF ANIMALS DURING THE HOLDING AND RECOVERY PHASE OF PROCESSING BY EXPANDING THE NUMBER OF ANIMALS PROCESSING HOLDING BAYS. HIGHLIGHTS OF THE PROPOSED RENOVATION INCLUDE IMPROVING CAGE WALL AND CEILING MATERIALS BOTH WITHIN THE SHELTERED OR INDOOR AREAS AND THE OUTDOOR AREAS IN ORDER TO MAKE THE CAGES SUITABLE FOR HOUSING RHESUS MACAQUES, PROVIDING FULLY ENCLOSED “CONDITIONED” HOLDING SPACE FOR EXAMS AND SAMPLING, AND MAKING SUSTAINABLE AND ENVIRONMENTALLY-SENSITIVE FACILITY IMPROVEMENTS THAT WILL LAST AT LEAST THE NEXT 20 YEARS. OUR PROPOSAL WILL ENSURE THAT EXISTING ANIMALS ARE ADEQUATELY HOUSED AND CARED FOR DURING THE RENOVATION.

INTEGRATIVE GENOMICS OF VANIN GENE EXPRESSION IN RELATION TO CVD RISK

IMPACT OF HIV ON THE HUMAN ALVEOLAR ENVIRONMENT DRIVINGTHE EARLY EVENTS OF MYCOBACTERIUM TUBERCULOSIS INFECTION - ABSTRACT TUBERCULOSIS (TB), CAUSED BY MYCOBACTERIUM TUBERCULOSIS (M.TB), REACHES THE LUNG ALVEOLI WHERE IT IS IN CONTACT WITH THE ALVEOLAR MUCOSA. THIS MUCOSAL SURFACE IS LINED BY ALVEOLAR EPITHELIAL CELLS (ATS) AND COMPOSED OF A MONOLAYER FORMED BY SURFACTANT LIPIDS AND A HYPOPHASE, CALLED ALVEOLAR LINING FLUID (OR ALF), CONTAINING SOLUBLE INNATE COMPONENTS AND ENZYMES. M.TB IS EXPOSED TO ALF FOR AN UNDETERMINED PERIOD OF TIME BEFORE AND DURING ITS ENGULFMENT BY HOST CELLS, PRIMARILY ALVEOLAR MACROPHAGES (AMS) BUT ALSO ATS, PROVIDING A SHIELD FOR M.TB AND PORTAL FOR DISSEMINATION. M.TB INFECTION DRIVES INFLAMMATION, WHICH EVENTUALLY ATTRACTS NEUTROPHILS, MONOCYTES, EOSINOPHILS AND OTHER INNATE CELLS ENTERING THE ALVEOLI FROM THE PERIPHERY. FOLLOWING PRIMARY INFECTION AND DISSEMINATION, TISSUE GRANULOMAS BEGIN TO DEVELOP. THIS PROPOSAL IS IN RESPONSE TO THE SPECIFIC RFA, “ANALYZING EARLY EVENTS IN TB AND TB/HIV INFECTION FOR INTERVENTIONAL TARGETS”. THESE EARLIEST INTERACTIONS, I.E., HOW THE LUNG ENVIRONMENT, SPECIFICALLY ALF AND ALVEOLAR HOST CELLS INTERACT WITH M.TB, ARE POORLY UNDERSTOOD, YET CRITICAL TO IMPACTING ERADICATION OR PROGRESSION OF M.TB INFECTION. OUR RESEARCH PROGRAM IS FOCUSED ON THESE EARLIEST EVENTS FOR M.TB IN THE ALVEOLI. OUR DATA SUPPORT THE FINDING THAT M.TB’S INTERACTION WITH ALF FROM HIV-INFECTED INDIVIDUALS (AS WELL AS ELDERLY INDIVIDUALS), FUNDAMENTALLY REMODELS THE BACTERIAL SURFACE AND ITS METABOLISM THEREBY AFFECTING HOST CELL ENTRY, TRAFFICKING AND HOST RESPONSE, CULMINATING IN ENHANCED HOST SUSCEPTIBILITY TO INFECTION. THUS, OUR CENTRAL HYPOTHESIS IS THAT THE FIRST INTERACTIONS OF M.TB WITH SOLUBLE HUMAN ALF COMPONENTS SHAPE ITS CELL SURFACE AND METABOLIC STATUS, IMPACTING ITS SUBSEQUENT INTERACTIONS WITH AMS AND ATS, THE TWO MAJOR RESIDENT ALVEOLAR CELL POPULATIONS, AND LEADING TO CONTROL OR SPREAD OF M.TB INFECTION. TO ADDRESS OUR HYPOTHESIS, WE WILL SYSTEMATICALLY INTEGRATE OUR UNIQUE IN VITRO AND IN VIVO MODELS STARTING WITH SINGLE CELL INTERACTIONS AND MOVING TO AN INNOVATIVE LUNG-ON-CHIP INFECTION MODEL WITH TIME-LAPSE IMAGING TO REVEAL THE DYNAMICS OF HOST- M.TB INTERACTIONS AT THE AIR-LIQUID INTERFACE WITH SPATIOTEMPORAL RESOLUTION, AND AN IN VIVO EXPERIMENTAL APPROACH TO ASSESS EARLY DYNAMIC INTERACTIONS IN THE ALVEOLI USING THE RHESUS MACAQUE (RM) MODEL. WE WILL ALSO DELINEATE THE EFFECTS OF HIV INFECTION ON ALVEOLAR COMPOSITION & FUNCTION, ADDRESSING HOW HIV’S EFFECTS DRIVE M.TB FASTER REPLICATION AND DISSEMINATION WITHIN THE ALVEOLI. THE SPECIFIC AIMS ARE TO: 1) DETERMINE HOW M.TB EXPOSURE TO PEOPLE LIVING WITH HIV ALF (HIV-ALF) VS. CONTROL ALF GENERATES EARLY-STAGE M.TB METABOLIC ADAPTATIONS THAT LEAD TO HOST CELL IMMUNE DYSREGULATION; 2) DETERMINE HOW THE COMBINED EFFECT OF EXPOSURE OF ALVEOLAR CELLS AND M.TB TO HIV-ALF (VS. CONTROL ALF) CAUSES DYSFUNCTIONAL ALVEOLAR IMMUNITY THAT ACCELERATES M.TB GROWTH AND DISSEMINATION USING A LUNG-ON-CHIP (LOC) MODEL; AND 3) DETERMINE HOW INFECTION OF BAL-ACQUIRED AMS BY HIV OR CONTROL ALF-EXPOSED M.TB INSTILLED IN THE AIRWAYS ALTERS THE ALVEOLAR CELLULAR IMMUNE RESPONSE USING THE NHP MODEL. RESULTS FROM THIS PROPOSAL WILL MOVE SCIENCE FORWARD BY GUIDING THE SCIENTIFIC COMMUNITY ON THE DEVELOPMENT OF EFFECTIVE IMMUNE-BASED EARLY INTERVENTIONS, SPECIFICALLY FOR PEOPLE LIVING WITH HIV.

ROLE OF CELLULAR LONG NON-CODING RNAS IN HIV REPLICATION AND DISEASE OUTCOME - SUMMARY THE MAJORITY OF THE HUMAN TRANSCRIPTOME CONSISTS OF LONG NON-CODING RNAS (LNCRNAS), WHICH REGULATE THE EXPRESSION AND FUNCTION OF PROTEIN-CODING GENES, IMMUNE CELL DEVELOPMENT, DIFFERENTIATION, AND RESPONSE TO PATHOGENS. INFECTIONS INDUCE GLOBAL CHANGES IN LNCRNA EXPRESSION. THE EXPRESSION OF LNCRNAS IN HIV-INFECTED CELLS AND HOW HOST LNCRNAS IMPACT THE REPLICATION AND PERSISTENCE OF HIV INFECTION ARE UNKNOWN. A SPONTANEOUS FUNCTIONAL CURE OF HIV-1 OCCURS IN 0.3-0.5% OF ALL HIV PATIENTS. THESE PATIENTS, TERMED ELITE CONTROLLERS (EC), MAINTAIN UNDETECTABLE LEVELS OF HIV IN THE ABSENCE OF TREATMENT, MAINTAIN STABLE CD4+ T CELL COUNTS, AND ARE LESS LIKELY TO TRANSMIT HIV. THE ECS EXEMPLIFY SPONTANEOUS CONTROL OF HIV, AND IDENTIFYING DEFENSE MECHANISMS IN THESE PATIENTS HOLDS PROMISE FOR FINDING A FUNCTIONAL CURE FOR HIV INFECTION. PREVIOUS STUDIES FOCUSED ON PROTEIN- CODING GENES. HOWEVER, EFFECTIVE HIV CONTROL IS LIKELY TO INVOLVE COMPLEX GENE NETWORKS, INCLUDING LNCRNAS. WE FOUND SEVERAL LNCRNAS SIGNIFICANTLY SUPPRESSED IN EC VS. HAART-TREATED CHRONIC HIV-PATIENTS AND HEALTHY UNINFECTED CONTROLS. THE FUNCTIONAL IMPACT OF LNCRNAS SUPPRESSED IN EC (SIEC) ON HIV OUTCOMES IS UNKNOWN. WE HAVE OBSERVED SIGNIFICANT GLOBAL CHANGES IN CELLULAR LNCRNA EXPRESSION IN HIV-INFECTED CD4+T CELLS COMPARED TO UNINFECTED CELLS IN OUR PRELIMINARY STUDIES. CELLULAR FUNCTIONS OF MOST OF THE HIV DEREGULATED (HIDE) LNCRNAS ARE YET TO BE DETERMINED. WE EMPLOYED A CRISPR/RFXCAS13D (CASRX)-SILENCING SCREEN TO DETERMINE THE FUNCTIONAL IMPACT OF SIEC AND HIDE LNCRNAS ON HIV REPLICATION. OUR PRELIMINARY DATA SHOWED THAT SILENCING OF SEVERAL SIEC AND HIDE LNCRNAS SIGNIFICANTLY REGULATED HIV REPLICATION. BASED ON THESE PRELIMINARY DATA, WE HYPOTHESIZE THAT HOST LNCRNA EXPRESSION IN HIV-INFECTED CELLS ORCHESTRATES NATURAL RESISTANCE AND DISEASE OUTCOME IN HIV INFECTION. WE WILL DETERMINE HOW THE SUPPRESSION OF SPECIFIC LNCRNAS IN ECS IS PROTECTIVE AND ELUCIDATE MOLECULAR MECHANISMS OF THEIR FUNCTION (AIM 1). WE WILL ALSO IDENTIFY HIV DEREGULATED (HIDE) LNCRNAS MODULATING VIRAL REPLICATION AND INVESTIGATE THE MECHANISM(S) UNDERLYING LNCRNA-MEDIATED REGULATION (AIM 2). WE WILL PURSUE THESE AIMS USING INNOVATIVE COMBINATIONS OF MOLECULAR AND BIOCHEMICAL TECHNIQUES, SUCH AS CRISPR/CASRX-SILENCING, TRANSCRIPTOMICS, RNA ANTISENSE PURIFICATION, AND MASS-SPECTROMETRY IN CELL LINE MODELS AS WELL AS PRIMARY CD4+ T CELLS. THE PROPOSED RESEARCH IS SIGNIFICANT BECAUSE IT WILL IDENTIFY CELLULAR LNCRNAS THAT INFLUENCE SPONTANEOUS CONTROL OF HIV AND DISEASE OUTCOMES, DELIVER UNPRECEDENTED INSIGHT INTO LNCRNA-MEDIATED REGULATION OF HIV REPLICATION, LAY THE GROUNDWORK FOR THE DEVELOPMENT OF NEW APPROACHES TO INTERVENTION.

REGULATION OF NORMAL AND DISEASED CARDIAC CA2+ CHANNELS

MECHANISM AND COUNTERMEASURE OF CARFENTANIL-INDUCED RESPIRATORY DISORDER AND DEATH - PROJECT SUMMARY OPIOIDS WERE INVOLVED IN 75,673 OVERDOSE DEATHS IN 2020 IN THE U.S, IN WHICH FENTANYL, A SYNTHETIC OPIOID, IS THE DEADLIEST DUE TO INDUCTION OF VENTILATORY DEPRESSION. CARFENTANIL (CFN) IS ANOTHER SYNTHETIC OPIOID ACTING ON OPIOID MU-RECEPTORS (MORS) WITH A POTENCY 100 TIMES THAT OF FENTANYL. THOUGH NO INTENDING FOR THERAPEUTIC USE, AEROSOLIZED CFN HAS BEEN APPLIED AS A LETHAL AGENT (BIOWEAPON) TO CAUSE RESPIRATORY DISORDER (RD) AND DEATH IN HUMANS. NALOXONE INCREASES SURVIVAL RATE OF OVERDOSE OPIOIDS IN EARLY REPORTS; HOWEVER, DEATHS HAVE BEEN INCREASING AFTER SYNTHETIC OPIOID ERA DUE TO UNSUCCESSFUL RESUSCITATION ATTEMPTS IN CLINICAL SETTINGS. IT ALSO FAILS TO REVERSE AEROSOLIZED CFN-INDUCED RD/DEATH IN ANIMALS. TO DATE, THE CHARACTERISTICS, MECHANISMS, AND EFFECTIVE COUNTERMEASURE OF ACUTE CFN EXPOSURE-INDUCED RD/DEATH ARE STILL UNCLEAR. OVERDOSE FENTANYL VIA INTRAVENOUS (IV) PERFUSION PRODUCES RD INCLUDING GRADUAL VENTILATORY DEPRESSION (DVE) AND THEN APNEAS/GASPING BEFORE DEATH. THE RD IS BELIEVED TO RESULT FROM INHIBITING THE CAROTID BODY O2-RECEPTOR AND CENTRAL CO2-CHEMORECEPTORS, ESPECIALLY THOSE IN THE RETROTRAPEZOID AND PARAFACIAL NUCLEUS (RTN/PFN) AND THE RESPIRATORY NEURONS IN THE PRE-BÖTZINGER COMPLEX (PBC, THE RESPIRATORY RHYTHM-GENERATOR). INTERESTINGLY, WE RECENTLY REPORTED THAT IV BOLUS INJECTION OF FENTANYL INDUCED AN IMMEDIATE APNEA SOLELY MEDIATED BY BRONCHOPULMONARY C-FIBERS (PCFS) IN ANESTHETIZED RATS. OUR PILOT STUDY FURTHER SHOWS THAT THE APNEA IS ASSOCIATED WITH CLOSURE OF THE LARYNX, CONSTRICTION OF CHEST EXPIRATORY MUSCLES, AND SILENCE OF INSPIRATORY MUSCLES (I.E., OBSTRUCTIVE AND CENTRAL APNEA WITH CHEST WALL RIGIDITY), WHICH IS LETHAL IF OVERDOSED. MOREOVER, AEROSOLIZED FENTANYL (100 MG/M3 FOR 10 MIN) INITIALLY TRIGGERS THE SIMILAR APNEA FOLLOWED BY DVE AND GASPING, LEADING TO DEATH ~10 MIN POST EXPOSURE IN ANESTHETIZED RATS. IN THIS PROPOSAL, WE, THEREFORE, WILL CHARACTERIZE THE CARDIORESPIRATORY (RESPIRATORY MUSCLES’) RESPONSES TO ACUTE CFN EXPOSURE IN AWAKE RATS AND DETERMINE THE CAUSAL ROLES OF PERIPHERAL AND CENTRAL MORS (AIM 1). SUBSEQUENTLY, WE WILL MECHANISTICALLY DEMONSTRATE THAT CFN EXPOSURE STIMULATES PCFS TO TRIGGER THE INITIAL APNEAS AND THEN INHIBITS THE CAROTID BODY, RTN/PFN CO2-SENSITIVE AND PBC RESPIRATORY NEURONS TO PRODUCE DVE AND GASPING (APNEAS) BY ACTING ON LOCAL MORS (AIM 2). DOXAPRAM IS A POTENT RESPIRATORY STIMULANT TO THE CAROTID BODY, WHILE CX717 CAN ENHANCE RESPIRATORY FREQUENCY AND RESPIRATORY DRIVE VIA ACTING ON THE PBC AND STRENGTHEN CENTRAL CO2-CHEMORECEPTION. BOTH AGENTS ARE CAPABLE OF REVERSING OPIOID-INDUCED RD IN ANIMALS AND HUMANS. THIS, ALONG WITH MOR INTERNALIZATION IN VAGAL SENSORY NEURONS AND BLOCKADE OF IV INJECTION OF FENTANYL-INDUCED APNEA BY DERMORPHIN PRETREATMENT (A PERIPHERALLY ACTING MOR AGONIST, IV) IN OUR PILOT STUDY, ALLOWS US TO DEFINE IF A COCKTAIL PRETREATMENT OF DERMORPHIN, DOXAPRAM, AND CX717 IS A COUNTERMEASURE FOR CFN-INDUCED RD/DEATH (AIM 3). A MULTIDISCIPLINARY APPROACH (ELECTROPHYSIOLOGY, IMMUNOHISTOCHEMISTRY, AND PHARMACOLOGY) AT SYSTEMIC AND CELLULAR LEVELS WILL BE EMPLOYED TO ELUCIDATE THE MECHANISMS UNDERLYING ACUTE CFN EXPOSURE-INDUCED RD/DEATH AND EVALUATE THE EFFICACY OF THE COCKTAIL PRETREATMENT. THE EXPECTED DATA WILL, FOR THE FIRST TIME, MECHANISTICALLY REVEAL THE KEY ROLES OF PCFS, THE CAROTID BODY, RTN/PFN AND PBC IN GENERATING THE CFN-INDUCED RD AND PROVIDE A NOVEL AND POTENTIAL COUNTERMEASURE TO PREVENT/DIMINISH THE RD/DEATH.

ASSESSING TOXICANT PROPERTIES AND HEALTH EFFECTS OF CIGARILLO AND HOOKAH TOBACCO AEROSOLS IN RATS

BI-DIRECTIONAL INTERACTIONS WITH TUMOR MICROENVIRONMENT ENHANCE BRCA1-IRIS OVEREXPRESSING TNBC TUMOR CELLS AGGRESSIVENESS

OLEATE-RESPONSIVE GENE REGULATORY NETWORKS GOVERNING PEROXISOME PROLIFERATION

PROTECTIVE IMMUNITY INDUCED BY P. YOELII GENETICALLY ATTENUATED VACCINES

ROLE OF A NOVEL INTERFERON RESPONSIVE T CELL SUBSET IN ALLERGY AND ASTHMA - DISEASE ASSOCIATED WITH ALLERGIES SUCH AS ASTHMA ARE A RISING HEALTH PROBLEM WITH NO CURRENT CURATIVE SOLUTIONS. CD4+ HELPER T CELLS (TH) THAT RESPOND TO COMMON ALLERGENS PLAY AN IMPORTANT ROLE IN DRIVING AIRWAY INFLAMMATION IN ASTHMA. TO BETTER UNDERSTAND THE DIVERSITY OF T CELL SUBSETS IN ALLERGY AND ASTHMA, WE ANALYZED THE SINGLE-CELL TRANSCRIPTOME OF ~50,000 HOUSE DUST MITE (HDM) ALLERGEN-REACTIVE TH CELLS FROM ASTHMATICS AND NON- ASTHMATICS, WITH AND WITHOUT HDM ALLERGY. FROM OUR ANALYSIS, BESIDES CANONICAL CLUSTERS OF CELLS SUCH AS TH2, TH17, AND TH1, WE IDENTIFIED A NOVEL SUBSET OF ALLERGEN-REACTIVE TH CELLS CHARACTERIZED BY AN IFN RESPONSIVE GENE SIGNATURE THAT WE CALLED THIFNR CELLS (SEUMOIS ET AL. SCIENCE IMMUNOLOGY, 2020). PROPORTIONS OF THIFNR CELLS WERE SIGNIFICANTLY INCREASED IN NONALLERGIC INDIVIDUALS COMPARED TO ALLERGIC PATIENTS, SUGGESTING AN ALLERGEN-SPECIFIC HOST SPECIFIC RESPONSE EVEN IN NON-ALLERGIC INDIVIDUALS. MOREOVER, THE EXCLUSIVE PRESENCE OF THE ALLERGEN-REACTIVE TH2 CELLS IN THE ALLERGIC PATIENTS SUGGESTS A PROTECTIVE ROLE (ANTI-TH2 RESPONSE) OF THE THIFNR CELLS IN THE NON-ALLERGIC PATIENTS WITH EXPOSURE TO ALLERGEN. THIS POTENTIAL PROTECTIVE ROLE WAS REINFORCED BY OUR IN VITRO STUDIES SHOWING THAT TNF-RELATED APOPTOSIS- INDUCING LIGAND (TRAIL) PRODUCED BY THIFNR CELLS DIRECTLY INHIBITS T CELL ACTIVATION TRIGGERED BY TCR ENGAGEMENT. IN FOLLOW-UP STUDIES, WE FOUND THIFNR CELLS AMONG VIRAL-REACTIVE TH CELLS DIRECTED TOWARDS FLU OR SARS-COV2, SUGGESTING A BROADER ROLE OF THOSE CELLS IN IMMUNE RESPONSES. ALSO, WE FOUND THIFNR CELLS AS A STABLE TH SUBSET IN A LARGE COHORT OF HEALTHY INDIVIDUALS. BECAUSE OF THE RECENT DISCOVERY OF THIFNR CELLS, VERY LITTLE IS KNOWN ABOUT THEIR ORIGINS, DIFFERENTIATION, PHENOTYPE, AND FUNCTION. WE HYPOTHESIZE THAT THESE THIFNR HDM-REACTIVE T CELLS COULD PLAY A ROLE THROUGH TRAIL ENGAGEMENT IN DAMPENING TH2 INFLAMMATION IN ALLERGY AND ASTHMA. IN AIM 1, WE WILL UTILIZE INTERFERON-STIMULATED RESPONSE ELEMENT (ISRE) REPORTER MICE AND T CELL-SPECIFIC INTERFERON RECEPTOR 1 (IFNAR1) KNOCKOUT MICE TO DETERMINE THE IMPORTANCE OF THIFNR CELLS IN CONTROLLING ALLERGIC AIRWAY INFLAMMATION IN ASTHMA MODELS. IN AIM 2, WE WILL PERFORM SINGLE-CELL ATAC-SEQ PROFILING TO IDENTIFY TRANSCRIPTION FACTORS THAT MAY BE INVOLVED ESTABLISHING AND MAINTAINING THE EPIGENETIC STATE OF THIFNR CELLS. FINALLY, WE WILL TEST FUNCTIONALLY THOSE TF BY USING SHRNA KNOCKDOWN EXPERIMENTS. OVERALL, STUDIES IN THIS PROGRAM WILL IMPROVE OUR UNDERSTANDING OF HOW THIFNR CELLS ARE GENERATED IN VIVO AND HOW THEY INTERACT WITH OTHER CD4+ T CELLS SUBSETS LIKE TH2 CELLS TO CURTAIL ALLERGIC AIRWAY INFLAMMATION IN ASTHMA MODELS.

SINGLE CELL GENOMICS FOR MALARIA PARASITES

USE OF FOCUSED ULTRASOUND STIMULATED BLOOD BRAIN BARRIER OPENING FOR CNS HIV/SIV RESERVOIR REDUCTION - ABSTRACT THE ADVENT OF COMBINATION ANTI-RETROVIRAL THERAPY (CART) HAS PROLONGED THE LIFESPANS OF PEOPLE LIVING WITH HIV. HOWEVER, UPON CESSATION OF ART TREATMENT THE VIRUS QUICKLY REBOUNDS. ONE OF THE LIKELY CAUSES FOR THIS REBOUND IS THE RAPID SEEDING AND ESTABLISHMENT OF VIRAL RESERVOIRS IN MULTIPLE TISSUE COMPARTMENTS INCLUDING THE CENTRAL NERVOUS SYSTEM (CNS). THE CNS IS A SANCTUARY SITE FOR HIV DUE TO THE PRESENCE OF BARRIERS SUCH AS THE BLOOD BRAIN BARRIER (BBB) AND CHOROID PLEXUS. THESE STRUCTURES SIGNIFICANTLY LIMIT THE CNS DELIVERY OF ART DRUGS WHICH PREVENTS CONTROL OF VIRAL REPLICATION AND THE ASSOCIATED NEGATIVE EFFECTS ON COGNITION. THEREFORE, NOVEL STRATEGIES NEED TO BE DEVELOPED FOR BOTH TREATMENT AND VIRAL RESERVOIR CLEARANCE IN THE BRAIN. WE RECENTLY OPTIMIZED A NOVEL TECHNIQUE CALLED FOCUSED ULTRASOUND INDUCED BLOOD BRAIN BARRIER OPENING (FUSS-BBBO) TO OVERCOME BBB PERMEABILITY CONSTRAINTS. FUS-BBBO HAS BEEN SAFELY AND SUCCESSFULLY USED TO DELIVER ANTICANCER CHEMOTHERAPY DRUGS DIRECTLY INTO BRAIN TUMORS IN HUMAN PATIENTS AND TARGET LARGE BRAIN REGIONS IN RODENTS AND NONHUMAN PRIMATES FOR GENE DELIVERY. FUS-BBBO USES LOW-INTENSITY ULTRASOUND WITH SYSTEMICALLY ADMINISTERED MICROBUBBLES, WHICH OSCILLATE IN BLOOD VESSELS AT THE ULTRASOUND FOCUS, RESULTING IN SAFE, LOCALIZED, TEMPORARY (6- 24 H), AND REVERSIBLE OPENING OF THE BBB. FUS-BBBO HAS BEEN SUCCESSFULLY USED TO DELIVER PROTEINS, SMALL MOLECULES, AND VIRAL VECTORS WITHOUT INFLICTING DETECTABLE TISSUE DAMAGE IN BOTH RODENTS AND NONHUMAN PRIMATES. THEREFORE, IN THE PRESENT STUDY, WE WILL USE FUS-BBBO TO DELIVER LONG-ACTING CART (CABOTEGRAVIR AND ISLATRAVIR) DRUGS IN CONJUNCTION WITH AN ANTI-INFLAMMATORY DELTA-9-TETRAHYDROCANNABINOL TREATMENT TO TWO SPECIFIC AREAS OF THE BRAIN (BASAL GANGLIA AND HIPPOCAMPUS) KNOWN TO BE TARGETED BY HIV/SIV FOR BETTER CONTROL OF VIRAL REPLICATION AND PERSISTENCE. IN ADDITION, WE WILL USE ADENO-ASSOCIATED VECTOR PARTICLES TO DELIVER A GENE THERAPEUTIC BLOCKING SIV INFECTION (ANTI-SIV GLYCOSYLPHOSPHATIDYLINOSITOL-ANCHORED SINGLE CHAIN VARIABLE FRAGMENTS (SCFVS) TO BRAIN RESIDENT MACROPHAGES AND MICROGLIA FOR DURABLE SUPPRESSION OF VIRAL REPLICATION IN THE BRAIN FOLLOWING CART INTERRUPTION. THESE NEW TECHNOLOGIES WILL BE TESTED IN THE SIV-INFECTED RHESUS MACAQUE MODEL THAT SHOWS CONSISTENT CNS INFECTION, AND NEUROINFLAMMATION UNDER SUPPRESSIVE CART. WHILE BOTH CABOTEGRAVIR AND ISLATRAVIR HAVE BEEN INDEPENDENTLY DEMONSTRATED TO REDUCE PLASMA VIREMIA IN THE HUMAN AND RHESUS MACAQUES, OUR STUDIES WILL FOR THE FIRST TIME DIRECTLY TEST THE EFFICACY OF THESE DRUGS ADMINISTERED USING SUBDERMAL NANOFLUIDIC IMPLANTS ON HIV/SIV REPLICATION IN BRAIN RESIDENT PERIVASCULAR MACROPHAGES AND MICROGLIA. THE PROPOSED STUDIES WILL HELP ESTABLISH THE FUS-BBBO METHOD TO INCREASE CART DRUG DELIVERY TO THE BRAIN FOR REDUCTION OF NEURO- HIV/SIV INFECTION AND ITS ADVERSE EFFECTS ON BRAIN FUNCTION. FINALLY, THESE STUDIES WILL ALSO HELP EVALUATE THE SAFETY AND EFFICACY OF FUS-BBBO DELIVERY OF LONG-ACTING CARTS AND NOVEL AAV-DELIVERED ANTI-HIV/SIV INHIBITORS TO THE BRAIN.

M. TUBERCULOSIS METABOLITES TO ACTIVATE HUMAN MUCOSAL-ASSOCIATED INVARIANT T CELLS - MYCOBACTERIUM TUBERCULOSIS (MTB) INFECTS OVER A QUARTER OF THE GLOBAL POPULATION AND REMAINS A SIGNIFICANT HEALTH THREAT CAUSING MILLIONS OF DEATHS ANNUALLY. MULTIDRUG RESISTANCE OF MTB LEADS TO A HIGHER RISK OF FAILED TREATMENT AND DEATH. THIS HIGH TUBERCULOSIS BURDEN WORLDWIDE DEMANDS THE DISCOVERY OF NOVEL CELLULAR AND MOLECULAR TARGETS FOR DEVELOPING EFFICACIOUS PROTECTIVE STRATEGIES. IT IS KNOWN THAT MUCOSAL-ASSOCIATED INVARIANT T (MAIT) CELLS RESPOND TO NON-PEPTIDIC BACTERIAL METABOLITES AND FUNCTION AS INNATE-LIKE SENSORS TO ELICIT RAPID IMMUNE RESPONSES AGAINST MTB INFECTIONS. MAIT CELL ACTIVATION IN MTB INFECTION REQUIRES THE RECOGNITION OF MTB METABOLITE ANTIGENS PRESENTED BY A MONOMORPHIC ANTIGEN-PRESENTING MOLECULE IN AN INDIVIDUAL-UNRESTRICTED MANNER, SIMILAR TO THE BINDING OF PATHOGEN-ASSOCIATED MOLECULAR PATTERNS TO INNATE RECEPTORS. ACTIVATED MAIT CELLS ARE EXPECTED TO INDUCE RAPID ANTI-MTB MAIT CELL RESPONSES AT EARLY OR CHRONIC TUBERCULOSIS INFECTIONS. ALTHOUGH RECENT STUDIES PROVIDED STRONG EVIDENCE SUPPORTING THE PROTECTIVE ROLE OF MAIT CELLS AGAINST TUBERCULOSIS IN MICE AND HUMANS, THE MODEL ANTIGEN FROM E. COLI INDUCED PARTIAL PROTECTION IN MICE AND PRIMATES, TOGETHER WITH SIDE EFFECTS IN SOME OTHER PRIMATE SUBJECTS. THIS SUBOPTIMAL PROTECTIVE MAIT CELL RESPONSE INDUCED BY THE E.COLI ANTIGEN AGAINST MTB INFECTION IS LIKELY BECAUSE MTB PROVIDES DIFFERENT ANTIGENS TO ACTIVATE AND RECOGNIZE MAIT CELLS OR THE POTENTIAL TOXIC EFFECT OF THE E. COLI COMPOUND. INDEED, MAIT CELLS RESPOND DIFFERENTLY TO VARIOUS PATHOGENS, AND THE CURRENT CRITICAL UNKNOWN IS WHICH MTB ANTIGENS STIMULATE ANTI-MTB MAIT CELL RESPONSES. BASED ON OUR VALIDATED FUNCTIONAL METABOLOMICS PLATFORM, WE WILL APPLY THESE CHEMICAL BIOLOGY APPROACHES TO TEST THE CENTRAL HYPOTHESIS THAT MTB METABOLITES STIMULATE HUMAN MAIT CELL RESPONSE AGAINST MTB INFECTIONS WITH TWO AIMS. IN AIM 1, WE WILL USE OUR PURIFIED AND PRELIMINARILY IDENTIFIED MTB AGONISTS TO INDUCE PROTECTIVE ANTI-MTB RESPONSES OF POLYCLONAL AND MONOCLONAL HUMAN MAIT CELLS IN COMPARISON WITH THE E. COLI ANTIGEN. MTB AGONISTS WILL STIMULATE MAIT CELLS FROM HEALTHY DONORS, TUBERCULOSIS PATIENTS, AND LUNG TISSUES. THE PROTECTION OF MAIT CELL RESPONSES WILL BE MAINLY MEASURED BY KILLING MTB-INFECTED CELLS AND INHIBITING MTB GROWTH. IN AIM 2, WE WILL DETERMINE THE CHEMICAL STRUCTURES OF MTB AGONISTS USING FUNCTIONAL METABOLOMICS TO STIMULATE ANTI-MTB MAIT CELL RESPONSES. WE HAVE OBTAINED MAIT-STIMULATORY FRACTIONS USING HIGH-PRESSURE LIQUID CHROMATOGRAPHY AND IDENTIFIED CANDIDATE MTB AGONISTS THAT ACTIVATED MAIT CELLS. OUR MASS SPECTROMETRY-BASED FUNCTIONAL METABOLOMICS WILL FURTHER DEFINE THE STRUCTURES OF MTB AGONISTS FROM ACTIVE CHEMICAL FRACTIONS. RESULTED IN NOVEL MTB METABOLITES WILL BE EITHER CHEMICALLY SYNTHESIZED OR PURIFIED FOR MAIT CELL ACTIVATION AND PROTECTION AGAINST MTB INFECTIONS. UPON SUCCESSFUL COMPLETION, WE WILL ELUCIDATE THE STRUCTURES AND FUNCTIONS OF MTB METABOLITES TO INDUCE A PROTECTIVE MAIT CELL RESPONSE AGAINST TUBERCULOSIS INFECTIONS. ULTIMATELY, MTB ANTIGENS CAN BE APPLIED TO UNDERSTAND MAIT ACTIVATION MECHANISMS IN TUBERCULOSIS DISEASE AND DEVELOP NOVEL ANTI-MYCOBACTERIAL STRATEGIES FOR FIGHTING TUBERCULOSIS IN HUMANS.

RECOGNITION OF BACTERIAL FLAGELLIN BY MACROPHAGES

VACCINE EFFICACY OF MODIFIED HIV ENVELOPES

MULTIMODAL PREDICTION OF BEHAVIORAL AND MENTAL HEALTH OUTCOMES IN INCARCERATED WOMEN - PROJECT SUMMARY/ABSTRACT RESEARCH IN PSYCHOLOGY AND NEUROSCIENCE HAS HELPED DEEPEN OUR UNDERSTANDING OF RELATIONSHIPS BETWEEN MENTAL HEALTH ISSUES, SOCIAL-ENVIRONMENTAL FACTORS, AND MALADAPTIVE BEHAVIORS CONTRIBUTING TO RISK FOR INCARCERATION. SUCH RESEARCH HOLDS SIGNIFICANT ECONOMIC IMPLICATIONS, AS THE SOCIETAL COST OF CRIME IN THE UNITED STATES IS A STAGGERING $3.2 TRILLION ANNUALLY, A SUM THAT EXCEEDS ANNUAL EXPENDITURES ACROSS ALL NATIONAL HEALTH CARE. WHILE MOST OF THESE COSTS ARE DUE TO MEN, INCARCERATED WOMEN REPRESENT THE SINGLE FASTEST GROWING SEGMENT OF THE CRIMINAL LEGAL POPULATION OVER RECENT DECADES. RELATIVE TO MEN, WOMEN IN THE LEGAL SYSTEM DISPLAY HIGHER RATES OF MENTAL HEALTH PROBLEMS, INCLUDING WITH COMORBID SUBSTANCE USE, THOSE OF WHICH RESULT IN A MULTITUDE OF NEGATIVE OUTCOMES THAT IMPACT WOMEN AND THEIR OVERALL PSYCHOSOCIAL FUNCTIONING, AS WELL AS SOCIAL NETWORKS AND PUBLIC HEALTH SYSTEMS, BROADLY. DESPITE THIS, THERE HAS BEEN LITTLE MODERN RESEARCH DEDICATED TO FEMALE FORENSIC POPULATIONS. INDEED, WE NEED PROSPECTIVE WORK IN FORENSIC SAMPLES OF WOMEN AIMED AT UNDERSTANDING ADAPTIVE AND MALADAPTIVE CHANGES IN MENTAL HEALTH FUNCTIONING OVER TIME, INCLUDING RISK AND PROTECTIVE FACTORS THAT INFLUENCE THESE CHANGES. THIS TYPE OF WORK IS FAR REACHING, AS IT CAN BE TRANSLATED TO TREATMENT OPTIONS TO PROVIDE BETTER LONG-RANGE OUTCOMES FOR THIS POPULATION. IN PRIOR NIH-SUPPORTED WORK, WE HAVE EXAMINED PSYCHOSOCIAL AND BRAIN FUNCTIONING IN 588 INCARCERATED WOMEN AND FEMALE YOUTH. HERE WE PROPOSE TO UTILIZE THIS SAMPLE TO CREATE A UNIQUE LONGITUDINAL STUDY IN ORDER TO DEVELOP ALGORITHMS THAT HELP PREDICT MENTAL HEALTH FUNCTIONING AND RELATED OUTCOMES (E.G., RECIDIVISM, SUBSTANCE USE RELAPSE). OUR ALGORITHMS WILL UTILIZE THE LATEST IN MACHINE LEARNING TECHNIQUES AND WILL INCLUDE MULTIMODAL (PSYCHOSOCIAL, FORENSIC, AND NEUROIMAGING) VARIABLES. UTILIZING THESE METHODS, WE INTEND TO IDENTIFY SPECIFIC BRAIN MEASURES, IN COMBINATION WITH PSYCHOSOCIAL AND FORENSIC VARIABLES, THAT ACCOMPANY LONG-RANGE OUTCOMES OF RESILIENCE (DESISTANCE) AND RISK (POOR GLOBAL MENTAL HEALTH, RECIDIVISM, SUBSTANCE USE RELAPSE). THE TRANSLATIONAL VALUE OF THIS WORK WILL BE TO CLARIFY INFORMATIVE PATTERNS OF DATA THAT MAY INDICATE POSITIVE OUTCOMES. FURTHERMORE, NEURAL MEASURES INDICATIVE OF SPECIFIC VULNERABILITY WILL BE IDENTIFIED AS TARGETS FOR TREATMENT AND NOVEL INTERVENTION STRATEGIES. BY IDENTIFYING VULNERABILITIES AND THE CHANGES THAT ACCOMPANY POSITIVE OUTCOMES, WE WILL BE CLOSER TO UNDERSTANDING WAYS TO IMPROVE OUTCOMES FOR VULNERABLE WOMEN.

GENETIC ANALYSIS OF HOST SPECIFICITY IN SCHISTOSOMA MANSONI

MALARIA CLINICAL TRIALS CENTER

TRYPANOSOME DRUG DEVELOPMENT CONSORTIUM

CONTROL OF MUSCLE PROTEIN SYNTHESIS DURING MYOGENESIS

EXPRESSION-BASED EMPIRICAL CANDIDATE GENES INFLUENCING BODY MASS INDEX

VIRAL AND HOST DETERMINANTS OF DONOR-SPECIFIC ANTI-HIV/SIV IMMUNITY MEDIATED BY APOBEC3-ENZYME - PROJECT SUMMARY HUMAN APOBEC3 ENZYMES, PARTICULARLY APOBEC3D, APOBEC3F, APOBEC3G, AND STABLE HAPLOTYPES OF APOBEC3H, CAN INDUCE G>A MUTATIONS IN THE HIV-1 GENOME. OFTEN, THESE MUTATIONS, PARTICULARLY THOSE INFLICTED BY APOBEC3G, WHICH IS A POTENT ANTI-HIV ENZYME, GENERATE STOP CODONS AND LEAD TO HIV-1 INACTIVATION. STUDIES HAVE SHOWN THAT APOBEC3 ENZYMES CAN ALSO INDUCE SUB-LETHAL LEVELS OF MUTATIONS THAT LEAD TO HIV-1 DIVERSIFICATION AND DRUG RESISTANCE. MUTATIONS INFLICTED BY DIFFERENT VARIANTS OF APOBEC3 ENZYMES VARY SUBSTANTIALLY IN TERMS OF BOTH THEIR EXTENT AND SEQUENCE CONTEXT. FOR EXAMPLE, APOBEC3H HAPLOTYPE II OFTEN INDUCES MANY G>A MUTATIONS. BY CONTRAST, APOBEC3H HAPLOTYPE I INDUCES LITTLE OR NO CHANGES IN THE HIV-1 GENOME. ANALYSIS OF ALL REPORTED HIV-1 SEQUENCES FROM ~37,000 PATIENTS SHOW THAT THE EXTENT AND PATTERN OF VIRAL MUTAGENESIS BY APOBEC3 ENZYMES ARE HIGHLY DONOR-SPECIFIC. IT IS HYPOTHESIZED THAT VARIATIONS IN BOTH APOBEC3 AND HIV-1 GENES ARE RESPONSIBLE FOR THE OBSERVED DIFFERENCES IN VIRAL MUTATION PROFILES. THE PRELIMINARY DATA SUGGEST THESE VARIATIONS CREATE A COMPLEX CASCADE OF PATIENT-SPECIFIC INTERACTIONS BETWEEN HIV- 1 AND APOBEC3 ENZYMES UNDERLYING HIV-1 HEALTH DISPARITIES INCLUDING RESPONSES TO ANTIRETROVIRAL TREATMENTS. THIS PROPOSAL AIMS TO INTEGRATE DIVERSE VIRAL AND HOST DATASETS AND USE A COMBINATION OF COMPUTATIONAL AND EXPERIMENTAL TECHNIQUES TO IDENTIFY THE MOLECULAR DETERMINATES OF DIFFERENTIAL VIRAL MUTAGENESIS, AND ELUCIDATE THEIR ROLES IN PATIENT-SPECIFIC RESPONSES TO ANTIRETROVIRAL THERAPIES.

NOVEL HIV 1 ENVELOPE IMMUNOGENS DERIVED FROM BROADLY NEUTRALIZING PLASMAS

T. BRUCEI MITOCHONDRIAL PROTEOME

ADVANCING INNOVATIVE NEXT_GENERATION HETEROLOGOUS VACCINES AGAINST TUBERCULOSIS - PROJECT SUMMARY/ABSTRACT THROUGHOUT MODERN HISTORY, TUBERCULOSIS (TB) HAS KILLED MORE PEOPLE THAN ANY OTHER INFECTIOUS DISEASE TO DATE (ESTIMATED > 2 BILLION PEOPLE OVER THE PAST 350 YEARS) AND TB CONTINUES TO KILL 4,000 PATIENTS EACH DAY. IN 2019 ALONE, THE WORLD HEALTH ORGANIZATION ESTIMATED THAT ~1.4 MILLION PEOPLE DIED OF TB AND 8-9 MILLION PATIENTS WERE NEWLY DIAGNOSED. THE ONLY APPROVED VACCINE, M. BOVIS BACILLE CALMETTE-GUERIN (BCG) HAS HAD MANY SUCCESSES BUT ITS PROTECTION IS VARIABLE AND BCG-VACCINATED TB PATIENTS STILL TRANSMIT M.TB. ACROSS THE GLOBE, AN EQUILIBRIUM OF TRANSMISSION AND DISEASE EXISTS SUCH THAT ONE NEW CASE OF PULMONARY TB ARISES FROM EACH EXISTING TB PATIENT; THUS, EFFORTS MUST BE REINVIGORATED TO DRIVE TB RATES LOWER. NEW STRATEGIES ARE NEEDED TO COMBAT TB INCLUDING DISCOVERY AND ADVANCEMENT OF NEW VACCINES TO CONTROL THE PATHOGEN MYCOBACTERIUM TUBERCULOSIS (M.TB). WITH THAT END GOAL IN MIND, WE ANSWER THE CALL FOR RFA AI-21-007: INNOVATION FOR TUBERCULOSIS VACCINE DISCOVERY (ITVD), FORMING PARTNERSHIPS BETWEEN PIS WITH VACCINE DEVELOPMENT EXPERTISE, AND EXPERTISE USING ANIMAL MODELS OF TB. WE PROPOSE THREE NOVEL MEANS TO ADVANCE TB VACCINES. IN THE R61 PHASE, WE DEVELOP AND IDENTIFY THE BEST PERFORMING NEW VACCINE CANDIDATES. SPECIFICALLY, WE (I) COMBINE THE ID93 PROTEIN ANTIGEN WITH NEW ADJUVANTS DESIGNED TO MAXIMIZE MUCOSAL IMMUNE RESPONSES AND DURABILITY; (II) CAPITALIZE ON NOVEL RNA PLATFORM TO CREATE VACCINES EXPRESSING ID93 AND RELATED M.TB ANTIGENS TO INDUCE RAPID AND DURABLE IMMUNITY; (III) OPTIMIZE HETEROLOGOUS PROTEIN/RNA PRIME-BOOST CANDIDATES FOR STRONG AND DURABLE MUCOSAL, HUMORAL, AND CELLULAR ANTI-M.TB IMMUNITY, AND SELECT THE FINAL CANDIDATES TO MOVE INTO CHALLENGE STUDIES. TO MAXIMIZE EARLY-STAGE DEVELOPMENT, RIGOR, AND REPRODUCIBILITY, WE PERFORM IMMUNOGENICITY STUDIES IN A SELECTED PANEL OF COLLABORATIVE CROSS (CC) INBRED STRAINS, REPRESENTING KNOWN DIFFERENTIAL SUSCEPTIBILITY TO M.TB INFECTION. IN THE R33 PHASE, I.E., M.TB CHALLENGE STUDIES, WE EXPLOIT THE DIVERSITY OUTBRED (DO) MOUSE POPULATION FOR ITS OUTSTANDING REPRESENTATION OF GENOTYPIC AND PHENOTYPIC DIVERSITY EQUIVALENT TO HUMANS, A MAJOR HURDLE IN TB VACCINE DEVELOPMENT EFFORTS TO DATE. WE WILL ALSO TEST THE FINAL VACCINE CANDIDATES IN GUINEA PIGS, THE CLASSIC PRECLINICAL MODEL FOR TB VACCINES TO ENSURE SUCCESS IN 2 ANIMAL MODELS.

BRAIN INJURY TREATMENT BY MODULATION OF HEMODYNAMICS WITH BLOOD SOLUBLE DRAG REDUCING MOLECULES

MATCHING TREATMENTS TO COGNITIVE DEFICITS IN OFFENDERS WITH SUBSTANCE USE DISORDERS - PROJECT SUMMARY/ABSTRACT PRIOR RESEARCH HAS IDENTIFIED TWO SUBTYPES OF ANTISOCIAL OFFENDERS, TYPIFIED BY DISTINCT DYSFUNCTIONAL COGNITIVE- EMOTION INTERACTIONS UNDERMINING SELF-REGULATION. ONE SUBTYPE IS CHARACTERIZED BY AN ATTENTION-BASED ABNORMALITY, WHICH IMPAIRS ADAPTIVE PROCESSING OF CONTEXTUAL INFORMATION, TANGENTIAL TO ONE’S PRIMARY FOCUS. THIS ABNORMALITY IS TYPIFIED BY OFFENDERS WITH HIGH LEVELS OF CALLOUS/UNEMOTIONAL TRAITS. A SECOND SUBTYPE IS CHARACTERIZED BY HYPER-REACTIONS TO PERSONALLY RELEVANT CUES, INTERFERING WITH EXECUTIVE FUNCTIONS THAT ARE OTHERWISE NEEDED TO REGULATE BEHAVIOR. THIS ABNORMALITY IS PREVALENT IN THOSE WITH HIGH EXTERNALIZING CHARACTERISTICS. OUR TEAM HAS DEVELOPED COGNITIVE SKILLS TRAINING THAT ADDRESSES EACH OF THESE SPECIFIC DEFICITS IN DISTINCT WAYS. THESE DISTINCT INTERVENTIONS ARE INTENDED TO ADDRESS MECHANISM-SPECIFIC COGNITIVE EMOTIONAL DYSFUNCTION, AS OPPOSED TO TREATING DISTINCT DYSFUNCTIONS WITH A UNIFORM APPROACH. UNDER AN R21, OUR PILOT WORK HAS SHOWN EVIDENCE OF IMPROVING OUTCOMES IN OFFENDERS BY MATCHING SPECIFIC DEFICITS WITH FOCUSED SKILLS TRAINING. HERE WE SEEK TO CONDUCT A WELL-POWERED, RANDOMIZED CLINICAL TRIAL TO DEMONSTRATE RELIABILITY AND GENERALIZABILITY OF THESE EFFECTS. WE EXPECT TO VERIFY PRIOR FINDINGS OF IMPROVED OUTCOMES BY MATCHING INDIVIDUAL DEFICITS WITH THE TARGETED INTERVENTIONS. WE WILL EXAMINE EFFECTS OF TREATMENT ON SPECIFIC COGNITIVE SKILLS (ASSESSED BY LABORATORY- BASED TESTS) AND REAL-WORLD BEHAVIORAL OUTCOMES INCLUDING SUBSTANCE USE BEHAVIORS, INSTITUTIONAL ADJUSTMENT, AND RECIDIVISM FOLLOWING RELEASE FROM PRISON. FURTHER, WE WILL EXAMINE ADVANCED NEUROIMAGING MEASURES TO ASSESS BRAIN CHANGES WITH TREATMENT. THIS WILL AID IN SPECIFYING POTENTIALLY DIFFERENT MECHANISMS OF TREATMENT SUCCESS IN EACH RESPECTIVE GROUP. THIS PROJECT ADDRESSES THE ONGOING NEED TO IMPROVE AND VALIDATE FOCUSED INTERVENTIONS THAT TARGET SPECIFIC DEFICITS, REPLACING LESS-EFFECTIVE ONE-SIZE-FITS-ALL APPROACHES.

MOLECULAR PHYSIOLOGY OF RESPIRATORY MUSCLES

GENETICS OF GALLBLADDER DISEASE IN MEXICAN AMERICANS

COPY NUMBER VARIATION IN MALARIA PARASITES

MOLECULAR EFFECTS OF CANNABINOIDS ON THE BLOOD BRAIN BARRIER IN HIV-INFECTED BRAIN - MOLECULAR EFFECTS OF CANNABINOIDS ON THE BLOOD BRAIN BARRIER IN HIV-INFECTED BRAIN INFECTION WITH THE HUMAN IMMUNODEFICIENCY VIRUS (HIV) CAUSES SIGNIFICANT DISEASE MORBIDITY IN THE BRAIN, LARGELY DUE TO HIV-ASSOCIATED BLOOD-BRAIN BARRIER (BBB) DISRUPTION, INFLAMMATION, AND PERSISTENCE OF HIV-INFECTED CNS TARGET CELLS SUCH AS MICROGLIA AND MACROPHAGES, IN SPITE OF ANTIRETROVIRAL THERAPIES (ART). CANNABIS IS A COMMONLY USED DRUG BY PEOPLE WITH HIV (PWH). RECENTLY, WE MADE THE IMPORTANT OBSERVATION THAT THE IMPACT OF CANNABIS ON BBB INTEGRITY MARKERS IS CONTEXT-DEPENDENT, WITH SIGNS OF BBB DISRUPTION IN NON-HIV SUBJECTS, BUT PARADOXICALLY IMPROVING BBB INTEGRITY IN PWH. THIS OBSERVATION MAKES IT A HIGH PRIORITY TO UNDERSTAND THE MECHANISTIC BASIS OF THIS DICHOTAMOUS INFLUENCE OF CANNABIS ON THE BBB. CANNABIS AND HIV INTERACT IN MULTIFACTORIAL WAYS THAT ARE STILL POORLY UNDERSTOOD. IMPORTANTLY, HIV BRAIN TARGET CELLS MICROGLIA/MACROPHAGES EXPRESS THE CANNABINOID RECEPTOR 2 (CB2R), WHILE ENDOTHELIAL CELLS OF THE BLOOD BRAIN BARRIER (BBB) EXPRESS BOTH CB1R AND CB2R AND OTHER CANNABINOID RECEPTORS SUCH AS GPR55. IT IS ALSO UNKNOWN HOW CANNABIS INFLUENCES HIV-ASSOCIATED BRAIN INFLAMMATION OR THE ACTIVE/LATENT STATUS OF HIV IN INFECTED MICROGLIA. BY PROTECTING VASCULAR INTEGRITY, IT FOLLOWS THAT CANNABIS WILL NOT ONLY AFFECT THE SELECT TRANSPORT OF SUBSTRATES INTO THE CNS, IT WILL ALSO DIRECTLY INFLUENCE THE INFILTRATION OF INFLAMMATORY CELLS AND THE PASSAGE OF ART OR OTHER TREATMENTS TO THE BRAIN. THE GOAL OF THIS APPLICATION IS TO TEST THE HYPOTHESIS THAT CANNABINOIDS POSITIVELY IMPACT BBB INTEGRITY IN THE HIV-INFECTED BRAIN ENVIRONMENT, LEADING TO REDUCED INFLAMMATORY CELL INFILTRATION AND ULTIMATELY PROTECTION FROM HIV-ASSOCIATED NEUROCOGNITIVE DISORDERS. OUR ANALYSIS OF TWO HUMAN BRAIN MICROVASCULAR ENDOTHELIAL CELL (HBMEC) LINES (HCMEC/D3 AND HBMEC/CI18) WITH OPPOSING VASCULAR INTEGRITY RESPONSES TO CANNABINOIDS IN THE CONTEXT OF HIV SUGGEST A MOLECULAR MECHANISM THAT CAN EXPLAIN THE CONTEXT-DEPENDENT EFFECTS OF CANNABIS IN HUMAN BBB PROPERTIES. BASED ON THIS PRELIMINARY DATA, OUR HYPOTHESIS POSITS THAT BENEFICIAL EFFECTS OF CANNABINOIDS IN THE CONTEXT OF HIV OCCUR VIA CB2R, WHILE CANNABINOID SIGNALING VIA GPR55 MAY BE DAMAGING. IN THE PROPOSED STUDIES WE WILL MODEL THE HUMAN BBB IN VITRO USING MULTICELLULAR SYSTEMS CONTAINING THE TWO HBMEC LINES IN PARALLEL, ALONG WITH PERICYTES, ASTROCYTES AND MICROGLIA, TO REPLICATE HIV-INDUCED VASCULAR PHENOTYPES CHARACTERIZED BY LOSS OF TIGHT JUNCTION PROTEINS AND INCREASED PERMEABILITY TO FLUORESCENT LABELLED- DEXTRAN. IMPLICATIONS TO INFECTION IN THE BRAIN WILL BE VALIDATED IN VIVO USING THE ECOHIV MOUSE MODEL, WHICH DEVELOPS BBB DISORDERS. SUCCESSFUL COMPLETION OF THESE EXPERIMENTS WILL DEFINE THE MOLECULAR MECHANISMS THAT UNDERLIE THE DICHOTOMOUS INFLUENCE OF CANNABIS ON THE BBB IN THE CONTEXT OF HIV, FORMING THE BASIS OF NEW APPROACHES AIMED AT OPTIMIZING BBB INTEGRITY.

GENE NETWORKS FOR DIFFERENTIAL RISK OF KIDNEY DAMAGE BY LONG-TERM DIABETES

ANTIBODY CYTOKINE FUSION PROTEINS AGAINST CRYPTOCOCCUS NEOFORMANS

A PROSPECTIVE OBSERVATIONAL STUDY ON THERAPEUTIC AND ADVERSE EFFECTS OF MEDICAL CANNABIS FOR CHRONIC TRAUMATIC BRAIN INJURY

MITOCHONDRIAL FUNCTION IN BLOODSTREAM TRYPANOSOMA BRUCEI

GASTRIC BIOLOGY OF HELICOBACTER PYLORI

IDENTIFICATION OF BROAD SPECTRUM TARGETS FOR THERAPEUTIC INTERVENTION AGAINST CRIMEAN CONGO, EBOLA AND LASSA HEMORRHAGIC FEVER VIRUS INFECTION

ANTI-INFLAMMATORY SIGNALS AND NEURODEGENERATION - ABSTRACT NEUROIMMUNE SIGNALS REGULATE NEURONAL FUNCTION AND SURVIVAL. WE HAVE OBTAINED MULTIPLE PIECES OF EVIDENCE INDICATING THAT ACTIVATION OF THE HETERODIMERIC INTERLEUKIN-13 RECEPTOR ALPHA 1/INTERLEUKIN-4 RECEPTOR ALPHA (IL- 13RΑ1/IL-4RΑ) AFFECTS THE VIABILITY OF MIDBRAIN DOPAMINERGIC (DA) NEURONS. WE FOUND THAT IN THE BRAIN, IL- 13RΑ1/IL-4RΑ IS PREFERENTIALLY EXPRESSED ON THE NEURONS OF THE SUBSTANTIA NIGRA PARS COMPACTA (SNC) THAT ARE LOST IN PARKINSON’S DISEASE (PD). WE ALSO SHOWED THAT INTERLEUKIN 13 (IL-13), PRODUCED DURING NEUROINFLAMMATION BY MICROGLIA AND NEURONS, CAN MODULATE THE ACTIVITY OF DOPAMINERGIC CELLS AND INCREASE THEIR SUSCEPTIBILITY TO OXIDATIVE DAMAGE. THUS, HAVING ESTABLISHED THAT ACTIVATION OF IL-13RΑ1 SIGNALING CAN AFFECT THE SURVIVAL OF DOPAMINERGIC NEURONS DURING NEUROINFLAMMATION, IN THE PRESENT APPLICATION WE WISH TO DETERMINE THE MOLECULAR AND CELLULAR MECHANISMS BY WHICH THIS OCCURS. SPECIFICALLY, WE WISH TO TEST THE HYPOTHESIS THAT IL-13 AND NEURONAL IL-13RΑ1 CAUSE DAMAGE BY STIMULATING A REGULATED CELL DEATH PATHWAY CALLED FERROPTOSIS. WE ALSO WISH TO DETERMINE TO WHAT EXTENT IL-13 AND IL-13RΑ1 CONTRIBUTE TO NEURODEGENERATION IN A MOUSE MODEL OF ALPHA- SYNUCLEINOPATHY (Α-SYN), A HALLMARK TRAIT OF PD THAT IS ASSOCIATED WITH NEUROINFLAMMATION AND OXIDATIVE DAMAGE. THIS WILL HELP US DETERMINE WHETHER TARGETING IL-13RΑ1 SIGNALING MIGHT BE A VIABLE APPROACH TO SLOW NEURODEGENERATION IN HUMANS AFFECTED BY AN Α-SYNUCLEINOPATHY SUCH AS PD. THE ABILITY OF RUXOLITINIB, AN FDA- APPROVED DRUG THAT INHIBITS IL-13RΑ1 SIGNALING, AND THAT OF THE NOVEL FERROPTOSIS INHIBITOR CMS121 TO REDUCE IL- 13-MEDIATED DAMAGE IN VIVO WILL ALSO BE TESTED. FINALLY, WE PROPOSE IN VIVO EXPERIMENTS TO TEST THE HYPOTHESIS THAT A RARE GENETIC VARIANT OF IL-13 FOUND IN INDIVIDUALS DIAGNOSED WITH EARLY-ONSET PD CAN CONTRIBUTE TO MORE RAPID LOSS OF DOPAMINERGIC NEURONS IN A MOUSE WITH THE HOMOLOGUE OF THIS MUTATION. IF SUCCESSFUL, THESE EXPERIMENTS WILL PROVIDE STRONG SUPPORT FOR THE HYPOTHESIS THAT IL-13 AND IL-13RΑ1 ARE NOVEL TARGETS FOR PREVENTING PD OR SLOWING ITS PROGRESSION, AT LEAST IN A SUB-SET OF PD PATIENTS.

DEVELOPMENT OF LIVE-ATTENUATED VIRUS VACCINE PLATFORM AGAINST HEMORRHAGIC FEVER CAUSING ARENAVIRUSES

TB VACCINE DEVELOPMENT IN NONHUMAN PRIMATE MODEL

MOUSE MODEL OF CHRONIC VIRAL HEPATITIS

IDENTIFYING RESPONSE DEFINING MECHANISMS FOR BIOLOGICAL THERAPIES IN SEVEREASTHMA (INSIGHTS ); ASSESSING THE ROLE OF CD4-CTLS - PROJECT SUMMARY SEVERE ASTHMA, CHARACTERIZED BY PERSISTENT AIRWAY INFLAMMATION AND CORTICOSTEROID RESISTANCE, AFFLICTS MILLIONS OF AMERICANS WITH LIMITED THERAPEUTIC OPTIONS. CLINICIANS CURRENTLY RELY ON TYPE 2 IMMUNE BIOMARKERS FOR PATIENT CLASSIFICATION (T2HIGH VS. T2LOW) AND TREATMENT DECISIONS. HOWEVER, REAL-WORLD STUDIES REVEALED THE OVER- REPRESENTATION OF THE T2HIGH ENDOTYPE IN SEVERE ASTHMA AND THOSE TREATED WITH ANTI-T2 BIOLOGIC THERAPIES EXHIBIT CONTRASTING RESPONSES, CHALLENGING THE EFFICACY OF THIS T2 DICHOTOMY AND HIGHLIGHTING THE NEED TO EXPLORE ALTERNATIVE PATHOPHYSIOLOGICAL DRIVERS. OUR SINGLE-CELL TRANSCRIPTOME ANALYSIS OF T CELLS FROM INFLAMED AIRWAYS IN SEVERE ASTHMATICS IDENTIFIED A NOVEL SUBSET OF PATHOGENIC AIRWAY TISSUE RESIDENT CYTOTOXIC CD4+ T CELLS (CD4-CTLS). DESPITE BIOLOGIC TREATMENT, CD4- CTLS REMAINED ACTIVATED, PRODUCING INFLAMMATORY MOLECULES. CELL PROPORTIONS ANALYSIS LINKED CD4-CTLS TO ASTHMA SEVERITY AND LUNG FUNCTION IMPAIRMENT, PARTICULARLY IN MALES WITH LATE-ONSET ASTHMA. THE STUDY STRONGLY SUPPORTS CD4-CTLS' SIGNIFICANT CONTRIBUTION TO AIRWAY INFLAMMATION, POTENTIALLY HINDERING T2-BIOLOGIC RESPONSE. LED BY A MULTIDISCIPLINARY TEAM OF EXPERTS IN CLINICAL ASTHMA, AND IMMUNO-GENOMICS, DRS. KURUKULAARATCHY AND SEUMOIS, OUR INSIGHTS STUDY AIMS TO ASSESS THE IMPACT OF CD4-CTLS ON SEVERE ASTHMA PATHOGENESIS AND T2- BIOLOGICS RESPONSE AT A LARGER SCALE. AIM 1: WE WILL ANALYZE AIRWAY CD4-CTLS FROM BLOOD AND SPUTUM SAMPLES COLLECTED FROM 160 SEVERE ASTHMATIC PATIENTS BEFORE, AND 12- AND 24- MONTHS AFTER INITIATING T2-BIOLOGIC THERAPY. WE WILL EVALUATE THE IMPACT OF CD4- CTLS ON T2-BIOLOGICS RESPONSE THROUGH EXTENSIVE CLINICAL ANALYSIS. SAMPLES WILL BE COLLECTED BY THREE CLINICAL CENTERS [UNIVERSITY OF SOUTHAMPTON, UK; UNIVERSITY OF MICHIGAN, ANN ARBOR, US, AND UNIVERSITY OF CALIFORNIA, SAN DIEGO, US] ESTABLISHING THE CLINICAL SIGNIFICANCE OF CD4-CTL AS A NEW PLAYER IN ASTHMA PATHOGENESIS. AIM 2: WITH THE PLANNED COLLECTION OF SAMPLES, WE WILL GENERATE UP TO 1 MILLION SINGLE-CD4-CTL CELL TRANSCRIPTOMES ALLOWING US TO COMPREHENSIVELY DECIPHER THE PATHOGENIC CELLULAR AND MOLECULAR FEATURES OF CD4- CTLS IN ASTHMA SEVERITY AND RESPONSE TO T2-BIOLOGICS, AS WELL AS INVESTIGATE THEIR FUNCTION THROUGH TCR- REACTIVITY AND INTERACTIONS WITH STRUCTURAL CELLS ANALYSIS. IN SUMMARY, THIS UNIQUE TRANSLATIONAL PROPOSAL AIMS TO UNVEIL MECHANISMS CAUSING A LACK OR LOSS OF RESPONSE TO T2-BIOLOGICS IN SEVERE ASTHMA, FOCUSING ON CD4-CTLS. FINDINGS COULD ESTABLISH CD4-CTLS AS A BETTER PREDICTOR FOR PATIENT STRATIFICATION, LEADING TO NOVEL THERAPIES FOR UNMET PATIENT NEEDS.

THE PROTECTIVE ROLE OF MICROGLIA IN PREVENTING HYPOXIC DISRUPTION OF BLOOD-BRAIN BARRIER INTEGRITY AND VCID - PROJECT SUMMARY/ABSTRACT DEMENTIA IS A MAJOR HEALTH PROBLEM IN THE UNITED STATES. VASCULAR CONTRIBUTIONS TO COGNITIVE IMPAIRMENT AND DEMENTIA (VCID) IS THE SECOND LEADING CAUSE OF DEMENTIA BEHIND ALZHEIMER’S DISEASE (AD) BUT DESPITE THE MASSIVE IMPACT OF VCID IN THE EXPANDING ELDERLY POPULATION, ITS PATHOGENESIS IS STILL ONLY POORLY UNDERSTOOD. THE CONSENSUS IS THAT IN THE AGING BRAIN, PARTICULARLY ON A BACKGROUND OF HYPERTENSION, BLOOD VESSELS UNDERGO DEGENERATIVE CHANGES, RESULTING IN LOSS OF BLOOD-BRAIN BARRIER (BBB) INTEGRITY AND INCREASED VASCULAR RESISTANCE, WHICH TOGETHER, LEAD TO CEREBRAL HYPOPERFUSION, NEURONAL DAMAGE AND COGNITIVE DECLINE. RECENTLY, WE DESCRIBED A NOVEL ROLE FOR MICROGLIA IN THE MAINTENANCE OF VASCULAR INTEGRITY. WE DEMONSTRATED THAT CHRONIC MILD HYPOXIA (CMH; 8% O2) INDUCES TRANSIENT VASCULAR LEAK IN SPINAL CORD BLOOD VESSELS IN YOUNG (10 WEEKS OLD) MICE, THAT IS ASSOCIATED WITH MICROGLIAL ACTIVATION AND CLUSTERING AROUND LEAKY BLOOD VESSELS. INTERESTINGLY, MICROGLIAL DEPLETION PROFOUNDLY INCREASED VASCULAR LEAK AND THIS WAS ASSOCIATED WITH ASTROCYTE-VASCULAR UNCOUPLING AND LOSS OF VASCULAR TIGHT JUNCTION PROTEINS, SUGGESTING THAT MICROGLIA PLAY AN IMPORTANT PROTECTIVE ROLE IN MAINTAINING VASCULAR INTEGRITY IN THE SPINAL CORD. WE HAVE SINCE FOUND THAT CMH ALSO TRIGGERS VASCULAR LEAK IN THE BRAIN AND THAT MICROGLIAL DEPLETION EXACERBATES THIS LEAK. STRIKINGLY, IN AGED (20 MONTHS OLD) MICE, THE EXTENT OF HYPOXIC-INDUCED CEREBROVASCULAR DISRUPTION IS GREATLY ENHANCED, AS SHOWN BY INCREASED VASCULAR LEAK AND THE EMERGENCE OF MICROHEMORRHAGES, THOUGH THE IMPACT OF MICROGLIAL DEPLETION IN AGED MICE HAS YET TO BE ADDRESSED. TOGETHER, OUR DATA SUGGESTS THAT MICROGLIA PLAY AN IMPORTANT VASCULOPROTECTIVE ROLE IN YOUNG MICE, BUT THIS MECHANISM MAY BE LESS EFFECTIVE IN THE AGED BRAIN. TAKEN WITH THE OBSERVATION THAT AGING INDUCES THE APPEARANCE OF A “PRIMED”, PRO-INFLAMMATORY, DESTRUCTIVE MICROGLIAL PHENOTYPE, WE HYPOTHESIZE THAT: (I) MILD HYPOXIA TRIGGERS VASCULAR LEAK AND MICROHEMORRHAGE IN THE BRAIN, RESULTING IN NEURONAL DAMAGE AND COGNITIVE DECLINE, (II) VASCULAR DISRUPTION IS WORSE IN THE AGED AND THE HYPERTENSIVE, (III) MICROGLIA PLAY AN IMPORTANT VASCULOPROTECTIVE ROLE IN STABILIZING THE BBB, BUT THIS DECLINES WITH AGE, AND (IV) REPOPULATING THE AGED BRAIN WITH YOUNG MICROGLIA OR ATTENUATION OF MICROGLIAL ACTIVATION STATE, COULD STABILIZE THE BBB AND REDUCE COGNITIVE IMPAIRMENT. TO INVESTIGATE THESE HYPOTHESES, WE PROPOSE THREE SPECIFIC AIMS: (1) CHARACTERIZE HYPOXIA-INDUCED VASCULAR LEAK IN THE BRAIN AND DEFINE HOW THIS IS INFLUENCED BY AGE, GENDER, SEVERITY OF HYPOXIA, HYPERTENSION AND BRAIN REGION, (2) DEFINE THE CONTRIBUTION OF MICROGLIA IN PREVENTING HYPOXIA-INDUCED CEREBROVASCULAR LEAK IN YOUNG AND AGED MICE, AND (3) DEMONSTRATE THAT HYPOXIA-INDUCED BBB DISRUPTION AND COGNITIVE IMPAIRMENT ARE REDUCED BY REPOPULATING THE AGED BRAIN WITH “YOUNG” MICROGLIA OR BY ATTENUATING MICROGLIAL ACTIVATION STATE. THESE STUDIES WILL PROVIDE IMPORTANT INSIGHT INTO THE LINK BETWEEN HYPOXIC EXPOSURE, BBB DISRUPTION, NEURONAL DAMAGE AND COGNITIVE DECLINE, AND INFORM ON THE THERAPEUTIC POTENTIAL OF MANIPULATING MICROGLIAL BEHAVIOR IN THE AGED BRAIN TO RESTORE VASCULOPROTECTIVE FUNCTION.

LYSOSOMAL DAMAGE, IMPAIRED AUTOPHAGY, AND ALCOHOLIC PANCREATITIS

GENETIC ANALYSIS OF IDIOPATHIC THROMBOSIS

CHOLECYSTOKININ IS A PHYSIOLOGICAL REGULATOR OF INTERMEAL INTERVAL

PROTEIN PHOSPHORYLATION IN TRYPANOSOMES

IDENTIFICATION OF REGULATORY VARIANTS IN NOVEL CANDIDATE GENES FOR DIABETES

CHARACTERIZATION OF MARMOSETS AS A GEROSCIENCE MODEL BY THE SAN ANTONIO MAP

DEVELOPMENT OF ASSAYS FOR HTS TO IDENTIFY INHIBITORS OF A NEW PPI INVOLVED IN CANCER METASTASIS

REGULATION OF C-MYC TRANSLATION BY HNRNP A1: ROLE IN MULTIPLE MYELOMA TUMOR RESPO

TRANSCRIPTION OF PROTEIN-CODING GENES IN LEISHMANIA

A GENOME SCAN FOR SUSCEPTIBILITY TO HELMINTHIC INFECTION

GENETICS OF CORONARY ARTERY DISEASE IN ALASKA NATIVES (*

EVALUATING THE THERAPEUTIC POTENTIAL OF HYPOXIA MIMETICS IN INFLAMMATORY DEMYELINATING DISEASE - MULTIPLE SCLEROSIS (MS) IS A CHRONIC INFLAMMATORY DISEASE THAT RESULTS IN DEMYELINATION AND DEGENERATION OF AXONS IN THE CENTRAL NERVOUS SYSTEM (CNS). DISRUPTION OF THE BLOOD-BRAIN BARRIER (BBB) OCCURS AT AN EARLY STAGE OF MS AND IN THE ANIMAL MODEL EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE) AND IS CENTRAL TO THE INITIATION AND MAINTENANCE OF MS PATHOGENESIS BY PERMITTING LEUKOCYTE INFILTRATION INTO THE CNS. RECENTLY, WE MADE THE IMPORTANT DISCOVERY THAT WHEN APPLIED TO PRE-EXISTING EAE DISEASE, CHRONIC MILD HYPOXIA (CMH; 10% O2) MARKEDLY ACCELERATES NEUROLOGICAL RECOVERY, LEADING TO LONG-TERM STABLE REDUCTIONS IN DISABILITY SCORE. HISTOLOGICALLY, THIS PROTECTION CORRELATED WITH REDUCED LEVELS OF VASCULAR DISRUPTION AND LEUKOCYTE ACCUMULATION AND FASTER REMYELINATION. MECHANISTICALLY, CMH PROMOTED A VIGOROUS ANGIOGENIC RESPONSE AND INCREASED ENDOTHELIAL EXPRESSION OF TIGHT JUNCTION PROTEINS WHILE REDUCING VCAM-1 EXPRESSION, KEY INDICATORS OF ENHANCED VASCULAR INTEGRITY. IN ADDITION, CMH GREATLY ENHANCED APOPTOTIC REMOVAL OF INFILTRATED MONOCYTES DURING DISEASE REMISSION. HAVING RECENTLY FOUND THAT HYPOXIA MIMETICS (HMS), DRUGS THAT STIMULATE HYPOXIA SIGNALING PATHWAYS, ALSO ACCELERATE NEUROLOGICAL RECOVERY FROM EAE, WE NOW PLAN TO EXTEND THESE STUDIES IN TWO WAYS. FIRST, BY DEFINING A HM TREATMENT PROTOCOL THAT OPTIMIZES RECOVERY FROM EAE. SECOND, DEFINE MECHANISTICALLY HOW HYPOXIA INDUCIBLE FACTOR (HIF)-MEDIATED VASCULAR REMODELING AND APOPTOSIS OF INFILTRATED MONOCYTES CONTRIBUTE TO THIS PROTECTION. AS MS ONSET TYPICALLY OCCURS IN YOUNG TO MIDDLE-AGED PATIENTS, WE WILL STUDY THESE EVENTS BOTH IN YOUNG (10 WEEKS) AND MATURE (8 MONTHS) MICE. THE GOAL OF THIS PROPOSAL IS TO TEST THE HYPOTHESIS THAT HMS PROMOTE RECOVERY FROM INFLAMMATORY DEMYELINATING DISEASE BY ENHANCING VASCULAR INTEGRITY AND SUPPRESSING NEUROINFLAMMATION VIA HIF-MEDIATED SIGNALING. OUR HYPOTHESIS WILL BE TESTED IN THREE SPECIFIC AIMS: (1) DEFINE A HM PROTOCOL THAT OPTIMIZES RECOVERY FROM EAE, (2) EVALUATE THE CONTRIBUTION OF VASCULAR REMODELING TO THE HYPOXIA PROTECTIVE RESPONSE IN EAE, AND (3) EVALUATE THE CONTRIBUTION OF MONOCYTE APOPTOSIS TO THE HYPOXIA PROTECTIVE RESPONSE IN EAE. BUILDING ON OUR FUNDAMENTAL OBSERVATION THAT CMH ACCELERATES RECOVERY FROM EAE AND THAT THE HM DRUG FG-4592 EXERTS SIMILAR PROTECTION, BOTH IN THE RELAPSING-REMITTING AND CHRONIC PROGRESSIVE EAE MODELS, WE NOW PLAN TO EVALUATE THE TRANSLATIONAL POTENTIAL OF THESE FINDINGS BY (I) DEFINING AN OPTIMAL FG-4592 TREATMENT PROTOCOL AND (II) DEFINE MECHANISTICALLY HOW HIF-MEDIATED VASCULAR REMODELING AND APOPTOSIS OF INFILTRATED MONOCYTES CONTRIBUTE TO THIS PROTECTION. SUCCESSFUL COMPLETION OF THESE STUDIES WILL FURTHER OUR GOAL OF DEVELOPING THIS APPROACH AS A NOVEL TREATMENT FOR MS.

KUPFFER CELLS IN LIVER IMMUNOPATHOLOGY

SENSITIVITY MULTIPLE MYELOMA CELLS TO MTOR INHIBITORS

(PQD6)THE ROLE OF IMMUNE HOMEOSTASIS IN PROTECTION FROM CANCER CACHEXIA

AIDS RELATED ORAL CANDIDIASIS: DRUGS, STEROLS, AND FUNGAL CELLS

DEMONSTRATING THE EFFICACY OF GROUP PROLONGED EXPOSURE TREATMENT OF PTSD IN OEF/OIF/OND MALE VETERANS

FMRI RESTING STATE NETWORKS IN CHRONIC PAIN

NORADRENERGIC A7 NEURONS AND UPPER AIRWAY MOTOR CONTROL

IMMUNE RESPONSE TO P. VIVAX IN DUFFY (-) HUMANS

METHAMPHETAMINE, HIV INTEGRATION AND LATENCY IN THE BRAIN - METHAMPHETAMINE, HIV INTEGRATION AND LATENCY IN THE BRAIN METHAMPHETAMINE (METH), A STIMULANT DRUG USED BY PEOPLE WITH HIV (PWH PATHWAYS IN ADDICTION, AGGRAVATING EFFECTS OF HIV IN THE BRAIN, WHERE RESERVOIR CELLS BEARING HIV INTEGRATED PROVIRUS CHALLENGE CURE STRATEGIES AND CONTRIBUTE TO PERPETUATING NEUROLOGICAL CONSEQUENCES, DESPITE ), INFLUENCES INFLAMMATION AND ANTIRETROVIRAL TREATMENTS (ART). NEUROTRANSMITTERS INVOLVED IN ADDICTION SUCH AS DOPAMINE (DA) MODULATE HIV TARGETS THAT EXPRESS DA RECEPTORS, INCLUDING THE MICROGLIA DIVERSE POPULATION. CHROMATIN ORGANIZATION IN HIV-1 INTEGRATION REPRESENTS AN IMPORTANT PREREQUISITE FOR UNDERSTANDING INFECTION AND LATENCY. HOWEVER, THERE IS A CRITICAL GAP IN UNDERSTANDING RELATIONSHIPS BETWEEN CHROMATIN ORGANIZATION AND PROVIRAL INTEGRATION ASSOCIATED WITH HIV-1 PERSISTENCE IN MICROGLIA CELLULAR RESERVOIRS, AND PARTICULARLY IN THE BEST EXPERIMENTAL MODEL, THE SIV- RHESUS MACAQUE. MOREOVER, IT IS UNKNOWN HOW METH MODIFIES THESE RELATIONSHIPS. WE HYPOTHESIZE THAT METH IMPACTS CHROMATIN ENHANCING VIRAL INTEGRATION SUSCEPTIBILITIES IN HIV/SIV BRAIN TARGET CELLS, AND THAT EFFECTS VARY BY BRAIN REGION THAT DIFFER IN DOPAMINERGIC PROJECTIONS. CRITICAL TO THIS GOAL IS THE PROGRESSION OF MODELS REFLECTED IN PHASES, FIRST IN VITRO USING HUMAN MICROGLIA CELL LINES AND IPSC-DERIVED, WHERE THE CHARACTERISTICS OF HIV INTEGRATION HAVE BEEN DEFINED, AND EFFECTS OF METH, OR DA, CAN BE CONTROLLED. THIS IS FOLLOWED BY MODELS EX VIVO AND IN VIVO IN THE SIV-RHESUS MACAQUES, TO REPLICATE HIV, CHRONIC METH AND ART-SUPPRESSION. IN THE R61 PHASE, WE WILL CONCENTRATE ON WHETHER CHROMATIN ACCESSIBILITY AND THE ARCHITECTURAL PROTEIN CTCF, WHICH INTERACTS WITH LEDGF/P75 TO TETHER THE HIV-1 INTEGRASE, MEDIATE INSERTIONS INTO GENOMIC BOUNDARIES OF TOPOLOGICALLY ASSOCIATED DOMAINS (TADS), MODIFIED BY METH, OR DA. WE WILL ASSESS THE IMPACT OF INTEGRATION PATTERNS ON THE INTEGRITY OF PROVIRAL GENOMES TO GAIN INSIGHTS ABOUT THE REAL SIZE OF THE FUNCTIONAL VIRAL RESERVOIR. AIM 1) DELINEATES RELATIONSHIPS BETWEEN EPIGENOMIC CHANGES IN MICROGLIA UPON METH AND PROVIRAL GENOMES INSERTED INTO THE PRE- ESTABLISHED REGIONS WITH MARKED PRESENCE OF CTCF AND H3K36ME3 (TAD BOUNDARIES), AIM 2) CONNECTS EPIGENETIC PROFILES AND HIV-1 INSERTION PATTERNS WITH THE CELLULAR AND VIRAL TRANSCRIPTION PROFILES, WHILE CONFIRMING THESE RULES IN THE SIV SYSTEM, AND AIM 3) DETERMINES THE CONTRIBUTION OF DA. GO-NO-GO: IF EPIGENOMIC CHANGES CAUSED BY METH OR DA ARE LINKED TO INTEGRATION SITES, WITH SIMILAR RULES IN ALL MODELS, THE R33 PHASE WILL MOVE TO THE IN VIVO SIV MODEL, TO TEST EFFECTS OF METH CHRONIC USE ON EPIGENOMIC VULNERABILITIES TO SIV INTEGRATION IN MICROGLIA, IN RELATION TO SUBSET HETEROGENEITY IN MESOLIMBIC AREAS WHERE DA PROJECTIONS ARE ABUNDANT COMPARED TO CONTROL REGIONS OF THE BRAIN. AIM 4) TESTS WHETHER SIV INTEGRATION SITE AVAILABILITY FOLLOWS RULES DICTATED BY CHROMATIN STATES, ACCESSIBILITY AND CTCF BINDING THAT CAN BE USED TO PREDICT SUSCEPTIBILITIES IN THE CONTEXT OF METH. AIM 5) TESTS WHETHER INTEGRATION SUSCEPTIBILITIES AND PATTERNS VARY IN BRAIN AREAS THAT DIFFER IN DOPAMINERGIC PROJECTIONS, AND HETEROGENEOUS MICROGLIA POPULATIONS. THIS PROJECT ENABLES NOVEL STUDIES ON MICROGLIA CHROMATIN AND SIV INTEGRATION DYNAMICS IN BRAIN AREAS THAT DIFFER BY FUNCTION, CELLULAR DENSITY, AND NEUROTRANSMITTERS.

CHARACTERIZATION OF A MENDELIAN FORM OF PSYCHOSIS IN A POPULATION ISOLATE

ROLE OF CO-CHAPERONE, BAG3, IN MUSCLE DEGENERATION UNDER PHYSIOLOGICAL STRESS

DIFFERENTIAL REGULATION OF CARDIOMYOCYTE SURVIVAL AND HYPERTROPHY

NICOTINE RECEPTOR DENSITY & DOPAMINE SYSTEM FUNCTION

TARGETED CRISPR/CAS EDITING TO ELIMINATE SIV CNS RESERVOIR IN METHAMPHETAMINE-EXPOSED RHESUS MACAQUES ON ART - SUMMARY IN THE ONGOING BATTLE AGAINST HIV, CURRENT ANTIRETROVIRAL THERAPIES (ART) FALL SHORT OF PROVIDING A CURE, PRIMARILY DUE TO LATENT RESERVOIRS IN THE BODY, PARTICULARLY IN THE CENTRAL NERVOUS SYSTEM (CNS). MYELOID CELLS, INCLUDING MICROGLIA AND MACROPHAGES (MM), SERVE AS KEY CELLULAR RESERVOIRS IN THE CNS, PERPETUATING THE INFECTION BY RELEASING TOXIC VIRAL PROTEINS AND INFLAMMATORY MEDIATORS. THIS PERSISTENT INFECTION IS FURTHER COMPLICATED BY SUBSTANCE USE DISORDERS (SUD), WITH METHAMPHETAMINE (METH) USE DISORDER (MUD) BEING NOTABLY PREVALENT. METH EXACERBATES THE SITUATION BY DISRUPTING THE BLOOD-BRAIN BARRIER (BBB), ACTIVATING MICROGLIA, AND TRIGGERING THE NLRP3 INFLAMMASOME, WHICH LEADS TO INCREASED NEUROINFLAMMATION AND NEURONAL DAMAGE. RECENT ADVANCEMENTS IN CRISPR/CAS GENOME EDITING OFFER A GLIMMER OF HOPE. THIS TECHNOLOGY HAS SHOWN PROMISE IN PRECLINICAL STUDIES FOR ERADICATING HIV PROVIRUS FROM VARIOUS TISSUES, INCLUDING THE BRAIN. THE PROPOSED RESEARCH AIMS TO HARNESS THIS POTENTIAL BY USING A NOVEL AAV SEROTYPE (R2MAC) CAPABLE OF PENETRATING THE BBB AND TARGETING MM. THIS SEROTYPE WILL DELIVER MULTIPLEX SSO7D-MEDIATED ENOSCAS12F (SOS12F) EDITORS TO ELIMINATE HIV/SIV PROVIRUS AND THE NLRP3 INFLAMMASOME. THE RESEARCH WILL BE CONDUCTED USING A RHESUS MACAQUE MODEL OF SHIV INFECTION ON ART, WHICH CLOSELY MIMICS HUMAN CONDITIONS. THE STUDY WILL LEVERAGE THE COMBINED EXPERTISE OF DR. LING AND DR. HU IN NONHUMAN PRIMATE MODELS, METH NEUROTOXICITY, MM INFLAMMASOME, CRISPR/CAS HIV ERADICATION, AND TARGETED GENE THERAPY. THE PROPOSAL INCLUDES TWO SPECIFIC AIMS: FIRST, TO DETERMINE THE EFFICIENCY AND SPECIFICITY OF THE R2MAC-MEDIATED MULTIPLEX SOS12F GENE THERAPY IN ELIMINATING SHIV PROVIRUS AND INFLAMMASOME IN SHIV-INFECTED MM; AND SECOND, TO EVALUATE THE THERAPEUTIC EFFICACY OF THIS APPROACH AT ANALYTICAL ART INTERRUPTION (ATI) USING THE SHIV+ART+ MODEL, BOTH WITH AND WITHOUT METH ADMINISTRATION. THIS INNOVATIVE APPROACH, UTILIZING THE R2MAC SEROTYPE AND MULTIPLEX EDITING, HOLDS THE POTENTIAL TO SIGNIFICANTLY ADVANCE OUR UNDERSTANDING AND TREATMENT OF NEUROHIV AND MUD, OFFERING NEW HOPE FOR THOSE AFFECTED BY THESE CHALLENGING CONDITIONS.

THE ROLE OF ARSS IN GASTRIC INFECTION BY HELICOBACTER PYLORI

MOLECULAR MECHANISM OF FSHD PATHOLOGY

MIF: A PRO-INFLAMMATORY CYTOKINE AS A NOVEL TARGET TO REDUCE BLADDER INFLAMMATION

TROJAN HORSE GENE THERAPY OF INCLUSION BODY DISEASE

A NOVEL ADVANCED RESUSCITATION FLUID FOR A SWINE POLYTRAUMA MODEL (TRAUMATIC BRAIN INJURY AND HEMORRHAGIC SHOCK)

1/5 - GENETICS OF TRANSCRIPTIONAL ENDOPHENOTYPES FOR SCHIZOPHRENIA

BIOQUEST ACADEMY: CREATING AN INNOVATIVE IMMERSION PROGRAM FOR TEENS

SPINAL GLIA ACTIVATION IN CHRONIC STRESS-INDUCED VISCERAL HYPERALGESIA

IT'S CONTAGIOUS! PROMOTING THE BIOMEDICAL WORKFORCE PIPELINE THROUGH INFECTIOUS DISEASES - OUR COUNTRY’S BIOMEDICAL WORKFORCE NEEDS MORE PROFESSIONALS. IN PARTICULAR THERE IS A GRAVE NEED TO PROMOTE DIVERSITY WITHIN THIS POPULATION. INTERVENTIONS, FROM ELEMENTARY TO COLLEGE, TO ADDRESS THIS NEED AIM TO EDUCATE STUDENTS ABOUT OPPORTUNITIES IN BIOMEDICINE. HOWEVER, IT’S IMPORTANT TO NOTE THE EDUCATORS OF OUR FUTURE BIOMEDICAL WORKFORCE ARE OFTEN NOT INCLUDED IN THESE INTERVENTIONS. THE LACK OF ENGAGEMENT OF TEACHERS RESULTS IN A GROWING GAP IN KNOWLEDGE ABOUT THE BIOMEDICAL WORKFORCE PIPELINE AND CONTEMPORARY BIOMEDICAL RESEARCH TOPICS WHICH MUST BE REMEDIED. INNOVATIVE INTERVENTIONS ARE NEEDED TO EXPAND STUDENT EXPOSURE TO BIOMEDICAL CAREERS AS WELL AS ENHANCE TEACHER PROFESSIONAL DEVELOPMENT IN BIOMEDICAL RESEARCH AREAS. FURTHER, WHEN TEACHERS ARE PROVIDED THE TOOLS TO APPLY DATA LITERACY TO THEIR TEACHING PRACTICES, THEY ARE THEN EMPOWERED TO MAXIMALLY ENGAGE THEIR STUDENTS IN THESE NOVEL INTERVENTIONS. THE PROPOSED SEPA PROGRAM, “IT’S CONTAGIOUS! PROMOTING THE BIOMEDICAL WORKFORCE PIPELINE THROUGH INFECTIOUS DISEASES”, WILL FEATURE THE CURRENT SCIENCE ISSUE OF INFECTIOUS DISEASES. USING DATA FROM INFECTIOUS DISEASE RESEARCH AND INTERACTIONS WITH BIOMEDICAL RESEARCHERS THE TEACHERS AS RESEARCHERS (TAR) PROGRAM WILL (AIM 1), ENGAGE SECONDARY TEACHERS TO CREATE AND INTEGRATE CLASSROOM TO CAREER CONNECTIONS (C2C2) ACTIVITIES AND SUPPLEMENTAL NARRATIVES INTO THEIR CLASSROOM INSTRUCTION (AIM 2). THE TAR PROGRAM WILL PROVIDE DATA LITERACY PROFESSIONAL DEVELOPMENT FOR TEACHERS AS THEY WILL COLLECT DATA FROM THE C2C2 ACTIVITIES. TAR COHORTS WILL PARTICIPATE IN A COMMUNITY OF PRACTICE (COP) (AIM 3) WHERE THEY WILL APPLY DATA LITERACY SKILLS TO EVALUATE C2C2 ACTIVITIES ASSESSING STUDENT KNOWLEDGE OF INFECTIOUS DISEASES AND APPLY DATA OUTCOMES TO INFORM TEACHING PRACTICES. WITHIN THE COP TEACHERS WILL ENGAGE IN DISCOURSE WITH COP COLLEAGUES AS THEY EVALUATE STUDENT DATA AND PRESENT DATA OUTCOMES. PARTICIPANTS WILL REFLECT UPON HOW DATA OUTCOMES FROM STUDENT ACTIVITIES INFORM DECISIONS REGARDING THEIR OWN TEACHING PRACTICES GALVANIZING DATA LITERACY SKILLS THAT TEACHERS WILL APPLY THROUGHOUT THEIR CAREER. THROUGH THESE AIMS, “IT’S CONTAGIOUS! PROMOTING THE BIOMEDICAL WORKFORCE PIPELINE THROUGH INFECTIOUS DISEASES” WILL SUPPORT TEACHERS IN THEIR MISSION AS THEY INSPIRE THE NEXT GENERATION OF BIOMEDICAL SCIENTISTS.

RECEPTOR TRAFFICKING IN ENTRY OF MURINE LEUKEMIA VIRUSES

LUMINEX TECHNOLOGY FOR THE QUANTIFICATION OF CYTOKINES IN NON-HUMAN PRIMATES

THE METABOLIC SYNDROME IN MEXICAN AMERICAN CHILDREN

GENETICS OF INFECTION AND ITS RELATIONSHIP WITH CVD RISK

ZIKA VIRUS COUNTERMEASURES: PRE-CLINCIAL PREGNANCY MODELS TO ASSESS PROTECTIVE EFFICACY AGAINST PLACENTAL DAMAGE AND FETAL DEMISE

MACROPHAGE MIGRATION INHIBITORY FACTOR MEDIATES BLADDER PAIN

STABILIZING HIV-1 TRIMERS BY LINKING GP120 SUBUNITS

EVALUATING THE SAFETY, IMMUNOGENICITY AND EFFICACY OF A ROBUST ATTENUATED MTB VACCINE IN THE SETTING OF HIV CO-INFECTION - PROJECT SUMMARY/ABSTRACT. NOVEL VACCINATION STRATEGIES ARE NECESSARY TO CONTAIN THE TB PANDEMIC, AS THE CURRENTLY LICENSED ANTI-TUBERCULAR VACCINE, BACILLE CALMETTE-GUERIN (BCG), HAS LIMITED AND VARIABLE EFFICACY. ATTENUATED, LIVE-REPLICATING MYCOBACTERIUM TUBERCULOSIS (MTB) EXPRESS THE FULL COMPLEMENT OF PROTECTIVE ANTIGENS ABSENT IN BCG. AS A RESULT, THESE STRAINS ARE MOST LIKELY TO INDUCE LONG-LIVED IMMUNE RESPONSES TOWARDS A WIDER ANTIGENIC REPERTOIRE AND COULD GENERATE DURABLE PROTECTION. RHESUS MACAQUES VACCINATED WITH AN ISOGENIC MTB MUTANT IN THE ALLELE ENCODING THE STRESS-RESPONSE MASTER REGULAR SIGH (SIGH) WERE PROTECTED FROM TB AFTER INFECTION WITH A LETHAL DOSE OF MTB AND CHARACTERIZED BY THE PRESENCE OF INDUCIBLE BRONCHUS ASSOCIATED LYMPHOID TISSUE (IBALT) IN THE LUNGS. PROTECTION BY SIGH COULD BE REVERSED BY THE DEPLETION OF IBALT. WE HAVE NOW VALIDATED THE PROTECTION AGAINST TB BY MUCOSAL VACCINATION WITH SIGH IN CYNOMOLGUS MACAQUES, A SECOND NHP SPECIES. OUR RESULTS SHOW THAT SIGH VACCINATION PROTECTS CYNOMOLGUS MACAQUES FROM LETHAL TB BY INDUCING T CELL RESPONSES, T-B CELL COOPERATION AND IBALT IN THE LUNGS. THIS LEADS TO THE INDUCTION OF STRONG IFNG RESPONSES, WHICH INHIBITS TYPE I IFN SIGNALING, CONDITIONS MACROPHAGES TOWARDS AN IFNG-, RATHER THAN A TYPE I IFN-RESPONSIVE PHENOTYPE, RESULTING IN ALMOST COMPLETE PROTECTION IN THE AIRWAYS AND LUNG GRANULOMAS. IT IS IMPORTANT HOWEVER, TO UNDERSTAND THE SAFETY, IMMUNOGENICITY AND EFFICACY OF ATTENUATED MTB BASED VACCINE CANDIDATES, AS I) CONCERNS REMAIN ABOUT WHETHER THE CANDIDATE WILL BE SAFER THAN BCG IN THE IMMUNOCOMPROMISED POPULATION AND II) WHETHER MTB/HIV CO-INFECTED INDIVIDUALS WILL BE ABLE TO MOUNT PROTECTIVE T CELL-BASED RESPONSES TO THIS VACCINE, RESULTING IN PROTECTION. OUR CURRENT PROPOSAL ADDRESSES THIS BY DIRECTLY TESTING THE HYPOTHESIS IN THE RHESUS MACAQUE MODEL OF MTB/HIV CO-INFECTION. ADDITIONALLY, WE PROPOSE TO DEVELOP THE SIGH VACCINE PLATFORM BY TESTING THE IMMUNOGENICITY AND EFFICACY OF TWO UNMARKED DOUBLE/TRIPLE KNOCK OUT MUTANTS INCLUDING SIGH WHICH WE HAVE GENERATED. WE CONTEND THAT AT THE END OF THIS PROPOSAL, WE WILL BE ABLE TO MOVE SIGH BASED DKO/TKO STRAINS INTO HUMAN CLINICAL DEVELOPMENT AS ANTI-TB VACCINES.

APPLYING NEUROMODULATION TO BOOST THE EFFICACY OF MOTIVATIONAL INTERVIEWING - THE USE OF CANNABIS AND ALCOHOL IS ASSOCIATED WITH MYRIAD HEALTH CONSEQUENCES INCLUDING MOTOR VEHICLE CRASHES, MYOCARDIAL INFARCTION, AND CANCER RISK. FOR EXAMPLE, ALCOHOL ACCOUNTS FOR NEARLY A THIRD OF HEAD AND NECK CANCERS. DESPITE THIS, FEW ADULTS WITH A SUBSTANCE USE DISORDER (SUD) RECEIVE ANY FORM OF TREATMENT. WHEN ADULTS WITH AN SUD DO RECEIVE BEHAVIORAL TREATMENT, THEY TYPICALLY RECEIVE ONLY A SINGLE SESSION. GIVEN THIS, IT IS ESSENTIAL TO MAKE THAT ONE SESSION COUNT. THERE IS A CLEAR NEED FOR ADJUNCTS TO TREATMENT FOR PEOPLE WHO HAVE AN SUD. ONE POTENTIAL WAY TO BOOST THE EFFICACY OF TREATMENT IS NON-INVASIVE BRAIN STIMULATION SUCH AS TRANSCRANIAL ALTERNATING CURRENT STIMULATION (TACS), WHICH INVOLVES ADMINISTERING SMALL ELECTRICAL CURRENTS THROUGH SCALP ELECTRODES. TACS DELIVERS SINUSOIDAL CURRENTS SIMILAR TO THOSE PRODUCED BY INTRINSIC NEURONAL OSCILLATIONS – THAT IS, BRAIN RHYTHMS. BECAUSE OF THIS, TACS CAN BE USED TO ENHANCE A GIVEN BRAIN RHYTHM. THIS IS IMPORTANT BECAUSE BRAIN RHYTHMS MEDIATE A BROAD RANGE OF BRAIN FUNCTIONS, INCLUDING PROCESSING OF SENSORY STIMULI, ATTENTION, MEMORY AND THE GENERATION OF MOTOR OUTPUT, AS WELL AS TO CONTRIBUTE TO ACTIVITY-INDUCED SYNAPTIC PLASTICITY. THEY ALSO PLAY A CAUSAL ROLE IN LINKING DISTRIBUTED BRAIN REGIONS INTO FUNCTIONAL NETWORKS. TESTING THE CAUSAL EFFECTS OF NEURONAL OSCILLATIONS IN SUD USING TACS COULD MARK A SIGNIFICANT STEP FORWARD FOR SUD RESEARCH AND TREATMENT. OUR PILOT DATA SUGGEST THAT WITHIN MOTIVATIONAL INTERVIEWING (MI) TREATMENT SESSIONS, BRAIN RHYTHMS IN THE BETA FREQUENCY BAND (15-25 HZ) ARE STRONGER DURING CLIENT STATEMENTS SUPPORTING BEHAVIOR CHANGE (“CHANGE TALK”) THAN DURING CLIENT STATEMENTS SUPPORTING THE STATUS QUO (“SUSTAIN TALK”). A LARGE BODY OF RESEARCH IN MI INDICATES THAT THE QUANTITY OF THE CLIENT’S CHANGE AND SUSTAIN TALK DURING THEIR INTERVENTION SESSION IS RELATED TO THEIR SUD OUTCOMES. OUR DATA SUGGEST THAT USING TACS TO ENHANCE BETA BAND BRAIN RHYTHMS MAY BOOST THE EFFECTS OF CLIENT CHANGE TALK AND REDUCE THOSE OF CLIENT SUSTAIN TALK. THEREFORE, IN THE UG3 PHASE WE PROPOSE TO TEST THE EFFECTS OF AN MI AND TACS INTERVENTION IN A RANDOMIZED TRIAL TO MODULATE CHANGE TALK AND OSCILLATORY BRAIN ACTIVITY. IN THE UH3 PHASE, WE WILL TEST THE EFFECTS OF THE MI AND TACS INTERVENTION ON BRAIN FUNCTIONAL NETWORKS AND SUBSTANCE USE OUTCOMES. IF SUCCESSFUL, THIS UG3-UH3 PHASED INNOVATION STUDY WILL PROVIDE A NEW AND ACCESSIBLE MEANS OF BOOSTING THE EFFICACY OF MOTIVATIONAL INTERVIEWING, IMPROVING OUTCOMES FOR PEOPLE WITH A SUBSTANCE USE DISORDER.