Texas 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

WISCONSIN CENTER OF EXCELLENCE IN GENOMICS SCIENCE

GENETICS OF ATHEROSCLEROSIS IN MEXICAN AMERICANS

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

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

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.

MAPPING DRUG RESISTANCE GENES IN PLASMODIUM FALCIPARUM

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.

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

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

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

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

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.

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

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.

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

GENETIC BASIS OF PRAZIQUANTEL RESISTANCE

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

GENETICS OF BONE STRUCTURE AND METABOLISM

CVD IN AMERICAN INDIANS GENETICS CENTER

ESTABLISHMENT OF A SPF RHESUS MACAQUE COLONY

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.

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.

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

ANTIGEN PRESENTATION BY EPITHELIAL STEM CELLS TO PROMOTE LIFE LONG IMMUNITY

RESEARCH TO IMPROVE AND STANDARDIZE MARMOSET NUTRITION AND DIETARY HUSBANDRY

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.

GENETIC DETERMINANTS OF HUMAN TRANSCRIPTIONAL AGING

GENETIC ANALYSIS OF CERCARIAL RELEASE IN SCHISTOSOMES

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.

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.

IDENTIFICATION OF NOVEL MICRORNAS ASSOCIATED WITH BRAIN STRUCTURE AND FUNCTION

LARGE-SCALE METHYLATION PROFILING IN METABOLIC SYNDROME PHENOTYPES

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.

MECHANISM AND EVOLUTION OF FILOVIRAL MONOCLONAL AFFINITY REAGENT SANDWICH ASSAYS

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

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.

QUANTITATIVE TRAIT LOCUS MAPPING IN HUMAN PEDIGREES

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

HIGH PERFORMANCE COMPUTING SYSTEM FOR HUMAN GENOMICS

IDENTIFICATION OF PRE-ECLAMPSIA SUSCEPTIBILITY GENES

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.

SINGLE CELL GENOMICS FOR MALARIA PARASITES

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.

GENETIC ANALYSIS OF HOST SPECIFICITY IN SCHISTOSOMA MANSONI

EXPRESSION-BASED EMPIRICAL CANDIDATE GENES INFLUENCING BODY MASS INDEX

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.

GENETICS OF GALLBLADDER DISEASE IN MEXICAN AMERICANS

COPY NUMBER VARIATION IN MALARIA PARASITES

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

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

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

TB VACCINE DEVELOPMENT IN NONHUMAN PRIMATE MODEL

GENETIC ANALYSIS OF IDIOPATHIC THROMBOSIS

IDENTIFICATION OF REGULATORY VARIANTS IN NOVEL CANDIDATE GENES FOR DIABETES

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

A GENOME SCAN FOR SUSCEPTIBILITY TO HELMINTHIC INFECTION

GENETICS OF CORONARY ARTERY DISEASE IN ALASKA NATIVES (*

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.

CHARACTERIZATION OF A MENDELIAN FORM OF PSYCHOSIS IN A POPULATION ISOLATE

1/5 - GENETICS OF TRANSCRIPTIONAL ENDOPHENOTYPES FOR SCHIZOPHRENIA

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

ENHANCING PULMONARY IMMUNE RECONSTITUTION AND LIMITING VIRAL PERSISTENCE WITH IL-21 THERAPY IN MTB/SIV CO-INFECTION - ABSTRACT TUBERCULOSIS (TB) REMAINS A LEADING CAUSE OF MORTALITY IN PEOPLE LIVING WITH HIV (PLHIV), WITH 161,000 DEATHS IN 2023 DESPITE WIDESPREAD USE OF COMBINATORIAL ANTIRETROVIRAL THERAPY (CART). ALTHOUGH CART EFFECTIVELY SUPPRESSES HIV REPLICATION, IT DOES NOT ELIMINATE THE MARKEDLY ELEVATED RISK OF MYCOBACTERIUM TUBERCULOSIS (MTB) REACTIVATION IN CO-INFECTED INDIVIDUALS. THIS PERSISTENT SUSCEPTIBILITY IS THOUGHT TO STEM FROM INCOMPLETE IMMUNE RECONSTITUTION, INCLUDING IMPAIRED TH1/TH17 BALANCE, REDUCED CD4⁺ EFFECTOR MEMORY T (TEM) CELLS, AND ONGOING IMMUNE ACTIVATION IN THE LUNG. OUR PRELIMINARY STUDIES IN AN ESTABLISHED MTB/SIV RHESUS MACAQUE MODEL DEMONSTRATE THAT CART ALONE FAILS TO RESTORE IL-21 AND STAT1 SIGNALING, BOTH CRITICAL FOR IMMUNE CONTROL OF TB AND HIV. IL-21, A PLEIOTROPIC CYTOKINE PRODUCED BY CD4⁺ T CELLS, REGULATES TH1 AND TH17 RESPONSES AND SUPPORTS MACROPHAGE FUNCTION. PRIOR WORK SHOWS THAT IL-21-IGFC THERAPY IN SIV-INFECTED MACAQUES IS SAFE AND PRESERVES MUCOSAL IMMUNITY. WE PROPOSE THAT IL-21-IGFC, WHEN ADMINISTERED DURING EARLY CART, CAN ENHANCE IMMUNE RECONSTITUTION, REDUCE VIRAL PERSISTENCE, AND IMPROVE TB CONTROL. THE CENTRAL HYPOTHESIS OF THIS STUDY IS THAT ADJUNCTIVE IL-21-IGFC THERAPY WILL RESTORE KEY IMMUNE PATHWAYS IN THE LUNG, LIMIT VIRAL RESERVOIRS, AND IMPROVE MACROPHAGE FUNCTION—THEREBY PREVENTING MTB REACTIVATION IN SIV- INFECTED MACAQUES. TO TEST THIS, WE WILL PURSUE TWO SPECIFIC AIMS: • AIM 1 WILL DETERMINE HOW IL-21-IGFC THERAPY DURING EARLY CART IMPACTS PULMONARY IMMUNE RECONSTITUTION AND MTB-SPECIFIC RESPONSES. WE WILL ASSESS T CELL SUBSETS, STAT1/STAT3 SIGNALING, GRANULOMA INTEGRITY, AND TRANSCRIPTIONAL SIGNATURES USING FLOW CYTOMETRY, SCRNASEQ, AND SPATIAL TRANSCRIPTOMICS. ADDITIONALLY, WE WILL PERFORM CROSS-SPECIES TRANSCRIPTOMIC COMPARISONS BETWEEN NHP AND HUMAN PBMCS FROM CART-TREATED AND UNTREATED COHORTS TO IDENTIFY CONSERVED IMMUNE SIGNATURES AND PATHWAYS ASSOCIATED WITH PROTECTION OR DISEASE PROGRESSION. • AIM 2 WILL EVALUATE THE EFFECT OF IL-21-IGFC THERAPY ON VIRAL PERSISTENCE AND MACROPHAGE FUNCTION. WE WILL MEASURE SIV RESERVOIRS IN LUNG TISSUE, QUANTIFY MACROPHAGE PROLIFERATION, AND ASSESS TISSUE PATHOLOGY TO DETERMINE THE IMPACT OF IL-21 ON HIV-ASSOCIATED INNATE IMMUNE DYSFUNCTION. IMPACT: THESE STUDIES WILL ELUCIDATE MECHANISMS BY WHICH IL-21 ENHANCES LUNG IMMUNITY AND LIMITS PATHOGEN PERSISTENCE IN TB/HIV CO-INFECTION. FINDINGS WILL GUIDE DEVELOPMENT OF IL-21-BASED IMMUNOTHERAPIES, A PROMISING HOST-DIRECTED STRATEGY TO IMPROVE OUTCOMES IN HIGH-BURDEN POPULATIONS WHERE TB AND HIV REMAIN SYNDEMIC.

DENGUE VIRUS DETERMINANTS OF VIRULENCE AND TRANSMISSION

EPITHELIAL CELLS AS MUCOSAL ADJUVANT FOR LIFE LONG IMMUNITY

DONOR-SPECIFIC ANTI-HIV/SIV IMMUNITY MEDIATED BY APOBEC3 ENZYMES - 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 APOBEC3 ENZYMES AND/OR DIFFERENT VARIANTS OF THE SAME APOBEC3 ENZYME 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, WHICH IS MOSTLY EXPRESSED IN EUROPEAN AND ASIAN POPULATIONS, INDUCES LITTLE/NO CHANGES IN THE HIV-1 GENOME. OUR ANALYSIS OF ALL REPORTED HIV-1 SEQUENCES FROM ~37,000 PATIENTS SHOW THAT THE EXTENT AND PATTERN OF VIRAL HYPERMUTATION IS HIGHLY PATIENT-SPECIFIC. ADDITIONALLY, OUR DATA INDICATE THAT HYPERMUTATION PATTERNS ARE DIFFERENT BETWEEN HUMAN AND NONHUMAN PRIMATE MODELS SUCH AS RHESUS MACAQUE. WE HYPOTHESIZE THAT VARIATIONS IN BOTH APOBEC3 AND HIV-1 GENES ARE RESPONSIBLE FOR THE OBSERVED DIFFERENTIAL HYPERMUTATION PATTERNS. OUR PRELIMINARY DATA SUGGEST THESE VARIATIONS CREATE A COMPLEX CASCADE OF PATIENT-SPECIFIC INTERACTIONS BETWEEN HIV- 1 AND APOBEC3 ENZYMES. IN SUPPORT OF THIS HYPOTHESIS, WE HAVE IDENTIFIED A PATIENT-SPECIFIC INTERACTION BETWEEN APOBEC3H AND ONE OF THE OTHER APOBEC3 ENZYMES. THIS INTERACTION IS INDUCED BY THE HIV-1 PROTEASE PROCESSING OF APOBEC3H HAPLOTYPE II SPLICE VARIANT SV200. ADDITIONALLY, WE HAVE GENERATED DATA THAT POINT TO A SIGNIFICANT DEFECT IN APOBEC3G MRNA SPLICING IN NON-HUMAN PRIMATES USED FREQUENTLY AS HIV-1 MODELS. FURTHERMORE, OUR DATA INDICATE THAT VARIATIONS IN MULTIPLE HIV-1 PROTEINS, NOT ONLY VIF, CONTRIBUTE TO PATIENT-SPECIFIC HYPERMUTATION PATTERNS. WE PROPOSE TO COMBINE COMPUTATIONAL AND EXPERIMENTAL TECHNIQUES TO DETERMINE THE LINK BETWEEN THESE VIRAL AND HOST VARIATIONS AND DIFFERENTIAL HYPERMUTATION PROFILES.

SINGLE CELL TRANCRIPTOMICS TO IDENTIFY LTBI REACTIVATION MARKERS IN TB/HIV CO-INFECTION - PROJECT SUMMARY THE GLOBAL CONTROL OF TUBERCULOSIS (TB) IS COMPOUNDED BY CO-INFECTION WITH HUMAN IMMUNODEFICIENCY VIRUS (HIV). THOSE INFECTED WITH HIV ARE AT HIGH RISK OF REACTIVATING LATENT TB INFECTION (LTBI). VIRAL-INDUCED CHRONIC IMMUNE ACTIVATION RATHER THAN MERE DEPLETION OF CD4+ T CELLS CORRELATES WITH LTBI REACTIVATION DUE TO SIV CO-INFECTION. FURTHER, COMBINATORIAL ANTIRETROVIRAL THERAPY (CART) FAILS TO RESTORE T-EFFECTOR FUNCTIONS AND THUS PREVENT LTBI REACTIVATION IN A NONHUMAN PRIMATE MODEL OF TB/SIV CO-INFECTION. INVESTIGATING THE IMPACT OF CART ON CHRONIC IMMUNE ACTIVATION IN A RELEVANT CO-INFECTED PRECLINICAL MODEL AT THE SINGLE CELL LEVEL WILL LEAD TO THE IDENTIFICATION OF KEY BIOMARKERS PREDICTING LTBI REACTIVATION. WE WILL SYNERGIZE THE RESEARCH PROPOSED HERE WITH A FUNDED K01-AWARD AIMS TO COLLECT COHESIVE DATA FROM ARCHIVED CONTROL SAMPLES (UNINFECTED, LTBI, SIV INFECTED, MTB/SIV CO-INFECTED BUT CART NAÏVE; ARCHIVED SAMPLES) AND MTB/SIV CO-INFECTED SHORT-TERM, LONG-TERM CART TREATED (K-AWARD) RHESUS MACAQUES. BASED ON OUR FINDINGS FROM THE CO-INFECTION STUDY, WE HYPOTHESIZE THAT WHILE INITIATING CART AT PEAK VIREMIA IN MTB/SIV CO- INFECTION CAUSES A SIGNIFICANT DECLINE IN IMMUNE ACTIVATION AND MACROPHAGE TURNOVER, IT FAILS TO RESCUE THE SKEWED CD4+T EFFECTOR MEMORY RESPONSES LEADING TO LTBI REACTIVATION. THUS, IN AIM 1, WE WILL IDENTIFY THE SPECIFIC LINEAGE MARKERS IN THE INTERPLAY BETWEEN MACROPHAGES AND CD4+ T CELLS THAT REMAIN IMPAIRED DESPITE CART IN MTB/SIV CO-INFECTED RHESUS MACAQUES. IN AIM 2, WE WILL INVESTIGATE THE EARLIEST EVENTS OF SIV-DRIVEN CHRONIC IMMUNE ACTIVATION AND COMPARE OUR FINDINGS TO CART TREATED COHORTS. FOR THIS, I) WE WILL COMPARE THE ACTIVATION STATUS OF PLASMACYTOID DENDRITIC CELLS (PDCS) AND NK CELLS IN MTB/SIV CO-INFECTION, II) WE WILL STUDY THE IMPACT OF CART ON PLASMACYTOID DENDRITIC CELL (PDC)-DRIVEN TYPE-I INTERFERON (IFN) PRODUCTION AND ITS IMPACT ON DOWNSTREAM EFFECTOR RESPONSES COMPARED TO UNTREATED CONTROLS. OVERALL, WE WILL BE ABLE TO DETERMINE IF I) MACROPHAGE TURNOVER INTERFERES WITH CD4+ T CELL RESTORATION AND FUNCTION, II) IF CART EFFECTIVELY CONTROLS THE EARLIEST EVENTS OF VIRUS-DRIVEN IMMUNE ACTIVATION BY IMPROVING NK CELL FUNCTION AND III) THE IMPACT OF CART ON TYPE-I IFN RESPONSE IN DRIVING THE IMMUNE ACTIVATION IN MTB/SIV CO- INFECTION. STUDYING THE IMPACT OF CART ON IMMUNE ACTIVATION IN MTB/SIV CO-INFECTION IS CRITICAL TO IDENTIFYING BIOMARKERS FOR THERAPEUTICS AND VACCINE DESIGN TO PREVENT LTBI REACTIVATION.

PIGTAIL MACAQUE MODEL OF HUMAN-SIMIAN IMMUNODEFICIENCY VIRUS INFECTION - ABSTRACT/SUMMARY: COMMONLY USED ANIMAL MODELS OF HIV-1 INCLUDE INFECTION OF MACAQUES WITH SIMIAN IMMUNODEFICIENCY VIRUS (SIV) OR SIMIAN-HUMAN IMMUNODEFICIENCY VIRUS (SHIV) CONTAINING HIV ENVELOPE (ENV) OR REVERSE TRANSCRIPTASE. THESE ANIMAL MODELS HAVE BEEN EXTREMELY USEFUL IN UNDERSTANDING HIV PATHOGENESIS AND DISEASE PROGRESSION, AS WELL AS UNDERSTANDING THE EFFICACY OF VACCINES AND DRUGS. HOWEVER, THE GENETIC DIFFERENCE BETWEEN HIV-1 AND SIV, AND THE ABSENCE OF OTHER HIV-1 GENES SUCH AS GAG, POL, VIF, VPR, AND NEF IN SHIV LIMITS THE UTILITY OF THESE MODELS IN VACCINE STUDIES. IDEALLY, GOOD ANIMAL MODEL OF HIV-1 INFECTION/AIDS WOULD BE INFECTION OF MACAQUES WITH HIV-1. HOWEVER, HIV-1 DOES NOT REPLICATE IN MACAQUE CELLS DUE TO THE PRESENCE OF RETROVIRAL RESTRICTION FACTORS. HIV-1 CAN BE MADE TO REPLICATE BY SUBSTITUTING ITS ACCESSORY GENES WITH SIV GENES SUCH AS VIF, VPX, VPR, AND NEF, WHICH CAN COUNTERACT INTERFERON-INDUCED RESTRICTION FACTORS IN MACAQUE CELLS. INDEED, WE HAVE PREVIOUSLY REPORTED THAT HUMAN-SIMIAN IMMUNODEFICIENCY VIRUS GENERATED BY SUBSTITUTION OF HIV-1NL4-3 VIF WITH SIV SUBSTITUTION (NAMED HSIV-VIFNL4-3) CAN REPLICATE PERSISTENTLY IN PIGTAIL MACAQUES (PTMS). HOWEVER, INFECTION DID NOT RESULT IN HIGH PEAK VIREMIA AND SETPOINT VIRAL LOADS AS OBSERVED DURING SIV OR SIMIAN-HUMAN IMMUNODEFICIENCY VIRUS (SHIV) INFECTION OF MACAQUES. TO FURTHER ADAPT HSIV, WE PERFORMED SERIAL IN VIVO PASSAGING TO ENHANCE INFECTIVITY OR REPLICATIVE CAPACITY. WE CONDUCTED ANIMAL-TO-ANIMAL TRANSFER OF INFECTED BLOOD IN 3 IMMUNOCOMPETENT PTMS WITH STARTING INITIAL INOCULUM CONTAINING A MIXTURE OF CXCR4- (HSIV-VIFNL4-3 RECOVERED FROM PREVIOUSLY INFECTED MACAQUE) AND CCR5-TROPIC HSIV (HSIV-VIF DERIVATIVE BASED ON PNL-AD8 AND BRU-YU2). INTERESTINGLY, ALL THE MACAQUES SHOWED PEAK VIREMIA CLOSE TO OR ABOVE 105 COPIES/ML AND VIRUS REPLICATION PERSISTED FOR MORE THAN 20 WEEKS. WE HAVE RECOVERED THREE CXCR4-TROPIC INFECTIOUS MOLECULAR CLONES (IMCS) FROM PASSAGE 3 MACAQUE (HSIV-P3 IMCS) WITH INTERESTING MUTATIONS THROUGHOUT THE GENOME, PERHAPS SUGGESTING ADAPTATION TO PTMS. WE HYPOTHESIZE THAT FURTHER IN VIVO PASSAGING OF HSIV-P3 IMCS WILL GENERATE PATHOGENIC VARIANTS WITH ENHANCED REPLICATION CAPACITY. WE PROPOSE TO CONDUCT SERIAL IN VIVO PASSAGING IN OLDER PTMS, WHICH MAY SUPPORT BETTER VIRUS REPLICATION COMPARED TO JUVENILE/YOUNGER MACAQUES. SINCE WE RECOVERED ONLY CXCR4-TROPIC HSIV, WE ALSO PROPOSE TO USE HSIV-P3 IMCS AS BACKBONES TO DEVELOP CCR5-TROPIC HSIV. THE RESULTS FROM THIS STUDY WILL PROVIDE VALUABLE INSIGHTS INTO DEVELOPMENT OF BIOLOGICALLY RELEVANT ANIMAL MODEL OF HIV-1 INFECTION FOR PRECLINICAL EVALUATION OF VACCINE PREVENTION OF HIV TRANSMISSION.

EFFECT OF IL-21 TREATMENT CONCURRENT TO ART AND 3HP IN MTB/SIV CO-INFECTION - ABSTRACT THE GLOBAL CONTROL OF TB IS COMPOUNDED BY CO-INFECTION WITH HIV. THOSE INFECTED WITH HIV ARE AT HIGH RISK OF REACTIVATING LATENT TB INFECTION (LTBI). DECADES OF RESEARCH HAS NOT YIELDED A SUCCESSFUL VACCINE FOR THE TB/HIV SYNDEMIC. WHILE THERAPEUTICS CONTROL MYCOBACTERIUM TUBERCULOSIS (MTB) AND HIV BURDEN, THEY HAVE A POTENTIAL DISADVANTAGE OF SELECTING FOR RESISTANCE AND NOT RESCUING FROM THE IMMUNE DYSFUNCTION. TARGETING THE HOST IMMUNE RESPONSE VIA A HOST-DIRECTED IMMUNOTHERAPY (HDT) PROVIDES AN OPPORTUNITY TO AUGMENT IMMUNITY DURING THE SHORT-WINDOW OF ACUTE HIV-1 CO-INFECTION OF MTB. TREATMENT OF LTBI IN PEOPLE LIVING WITH HIV (PLHIV) REDUCES THE RISK OF ACTIVE TB BY ONLY 35% AND IS DEPENDENT ON THE CD4 COUNTS AT THE TIME OF INITIATION. THOUGH COMBINATORIAL ANTIRETROVIRAL THERAPY (CART) CONCURRENTLY GIVEN WITH ANTI-TB THERAPY IMPROVES THE CLINICAL AND MICROBIOLOGICAL PARAMETERS, IT LEADS TO INSUFFICIENT RECONSTITUTION OF PROTECTIVE TH1, TH17 AND CD4+ TEM LEVELS IN THE LUNGS AND FAILS TO RESCUE FROM VIRUS-DRIVEN IMMUNE ACTIVATION. ADDITIONALLY, THE LONG-TERM STERILIZATION OF BACTERIA AND IMMUNE RECONSTITUTION IN THE LUNGS HAS NOT BEEN SHOWN IN THE INDIVIDUALS TREATED CONCURRENTLY WITH CART AND 3HP. HIV INFECTION DEPLETES IL-21 PRODUCING CD4+ T CELLS AND CART ONLY PARTIALLY RESTORES IL-21 IN HUMANS. WE HYPOTHESIZE THAT SUPPLEMENTING CART AND ONCE-WEEKLY, THREE-MONTH ISONIAZID AND RIFAPENTINE (3HP) TREATMENT WITH IL-21-IGFC FUSION PROTEIN IN MTB/SIV CO-INFECTION WILL RESULT IN A LONG-TERM RECONSTITUTION OF TH1 RESPONSE AND FUNCTIONAL CD4+T EFFECTOR MEMORY (TEM) RESPONSE IN THE LUNG. FURTHER, IL-21-IGFC FUSION PROTEIN ADJUNCTIVE TO CART AND 3HP, WILL AUGMENT CYTOTOXIC FUNCTION OF NK CELLS, THUS SIGNIFICANTLY REDUCING THE LIKELINESS OF LTBI REACTIVATION IN MTB/SIV CO-INFECTION. HENCE, WE AIM TO EXPLORE A HDT THAT UTILIZES IL-21-IGFC FUSION PROTEIN SUPPLEMENTATION OF CART+3HP TO MITIGATE VIRUS-DRIVEN IMMUNE DYSREGULATION AND THUS REDUCE INCIDENCE OF LTBI REACTIVATION. BY CONCURRENTLY TREATING MTB/SIV CO-INFECTED MACAQUES WITH CART+3HP+IL-21-IGFC FUSION PROTEIN, OUR GOAL IS TO SIGNIFICANTLY REDUCE LTBI REACTIVATION BY PROVIDING LONG-TERM IMMUNE RECONSTITUTION CONCURRENT TO BACTERIAL CONTROL BY I) INDUCTION OF AN ADEQUATE TH1 RESPONSE VIA STAT1, II) GENERATION OF CD4+ TEM CELLS, III) MAINTENANCE OF TH17 RESPONSES VIA STAT3 AND IV) PROMOTION OF CYTOTOXIC FUNCTION OF NK CELLS. THE SIGNIFICANCE OF OUR NOVEL, TRANSLATIONAL APPROACH IS UNDERSCORED BY OUR STRONG PRELIMINARY DATA, AND A REPRODUCIBLE, HUMAN-LIKE MODEL SYSTEM. THE DATA FROM THE PROPOSED STUDY WILL BE CRITICAL TO THE DEVELOPMENT OF AN IMMUNE-BASED INTERVENTION ALONG WITH CART AND ANTI-TB THERAPY TO CONTROL DYSREGULATED IMMUNE RESPONSES GENERATED DURING EARLY EVENTS OF HIV CO-INFECTION OF LTBI AND PROVIDE LONG-TERM IMMUNE RECONSTITUTION. THIS WILL LEAD TO THE FUTURE CLINICAL TESTING OF SUCH HOST-DIRECTED THERAPEUTIC APPROACHES TO CONTROL TB/HIV CO-INFECTION RELATED MORTALITY AND MORBIDITIES.

IMPACT OF PHYSICAL ACTIVITY ON STRIATAL REINNERVATION IN A NONHUMAN PRIMATE MODEL OF PARKINSON'S DISEASE

RHESUS BREEDING COLONY IN NEPAL AND IMPORTATION TO USA

NANOPARTICLE ANALYSIS OF ENVELOPED VIRUS ENTRY PATHWAYS

PATHOGENESIS OF AND HOST RESPONSE TO CHIKUNGUNYA VIRUS INFECTION OF THE CENTRAL NERVOUS SYSTEM

CHARACTERIZING IL-22 DRIVEN CHRONIC IMMUNE ACTIVATION IN HIV/TB CO-PANDEMIC - PROJECT SUMMARY THE HUMAN IMMUNODEFICIENCY VIRUS (HIV) AND TUBERCULOSIS (TB) CO-PANDEMIC POSES A MAJOR HEALTHCARE BURDEN IN RESOURCE-LIMITED COUNTRIES. HIV CO-INFECTION PREDISPOSES THE HOST TO REACTIVATION OF LATENT TUBERCULOSIS INFECTION (LTBI) RESULTING IN WORSENING OF DISEASE CONDITIONS AND MORTALITY. THE MOST WELL CHARACTERIZED IMPACT OF HIV IS THE CD4+ T CELL DEPLETION IN LYMPHOID TISSUES AND PERIPHERAL BLOOD. HOWEVER, STUDIES USING THE NONHUMAN PRIMATE (NHP) MODEL OF M. TUBERCULOSIS (MTB)/SIV CO-INFECTION HAVE REVEALED PROTECTIVE CD4+ T CELL-INDEPENDENT IMMUNE RESPONSES THAT SUPPRESS LTBI REACTIVATION. RECENT WORK SHOWS THAT THE MERE DEPLETION OF CD4+ T CELLS IS INSUFFICIENT TO CAUSE LTBI REACTIVATION IN SIV CO-INFECTED MACAQUES. INSTEAD, CHRONIC IMMUNE ACTIVATION APPEARS TO BE THE KEY CORRELATE FOR REACTIVATION. FURTHER, HIGHLY EFFECTIVE COMBINATORIAL ANTIRETROVIRAL THERAPY (ART), WHILE EFFECTIVE IN REDUCING VIRAL LOADS IN THE PERIPHERY AND LUNGS OF MTB/SIV CO-INFECTED MACAQUES, FAILS TO REDUCE THE RATE OF REACTIVATION OF LTBI. THUS, UNDERSTANDING THE DRIVING FORCES BEHIND CHRONIC IMMUNE ACTIVATION IN A RELEVANT CO-INFECTED PRECLINICAL MODEL UNDERSCORES THE DISCOVERY OF KEY BIOMARKERS AND DEVELOPMENT OF INTERVENTION STRATEGIES. WE THEREFORE AIM TO GAIN INSIGHT INTO THE MECHANISM OF MUCOSAL DAMAGE, A PARAMOUNT FACTOR IN CHRONIC IMMUNE ACTIVATION, DURING SIV/TB CO-INFECTION. TOWARDS THIS END, WE WILL INVESTIGATE THE ROLE OF IL-22, A KEY CYTOKINE IN PROTECTION FROM HIV AND RESPIRATORY DISEASES INCLUDING TB. WE HYPOTHESIZE THAT IL-22 PROTECTS THE LUNG FROM DAMAGE DURING LTBI AND SIV CO-INFECTION ABROGATES THIS PROTECTION BY IL-22 PERTURBATION. WE AIM TO IDENTIFY THE MECHANISM BY WHICH IL-22 DISARMS THE HOST IMMUNE RESPONSE LEADING TO SIV-DRIVEN IMMUNE ACTIVATION AND ULTIMATELY, THE REACTIVATION OF LTBI, IN A MACAQUE MODEL OF SIV/TB CO-INFECTION. IN AIM 1 WE WILL STUDY THE FUNCTIONAL ROLE OF IL-22 IN MTB/HIV CO-INFECTION. WE WILL DETERMINE THE LEVELS OF IL-22 PRODUCTION BY CD4+ AND CD8+ T CELLS AS WELL AS NON-T-CELL POPULATIONS, NKP44+ CELLS, AND CD103+ DENDRITIC CELLS IN THE BLOOD, LYMPH NODE AND COLORECTUM OF CO-INFECTED MACAQUES AND IN RESPONSE TO ART INTERVENTION (1A). WE WILL ALSO IDENTIFY THE FREQUENCY OF POLYFUNCTIONAL TH17 CELLS PRODUCING IFN, IL-17, IL-22, TNFA, PRODUCTION OF ANTIMICROBIAL PEPTIDES; DEFENSIN-SS, REG-3 PROTEINS AND S100A8/A9 PROTEINS IN THE BRONCHOALVEOLAR LAVAGE FLUID, COLORECTUM, LUNG TISSUE LYSATES OF CO-INFECTED MACAQUES AND IN RESPONSE TO ART INTERVENTION (1B). IN AIM 2, WE WILL STUDY THE IMPACT OF SIV-INDUCED IL-22 LOSS ON LTBI REACTIVATION. WE WILL FIRST DETERMINE WHETHER THE PRESENCE OF IL- 22R EXPRESSING MACROPHAGES IN NON-NECROTIC GRANULOMAS OF LTBI MACAQUES CORRELATES WITH PROTECTIVE CONTROL OF MTB (2A). WE WILL THEN PERFORM TRANSCRIPTIONAL PROFILING ON SORTED CELL SUBSETS AND MONONUCLEAR CELLS ISOLATED FROM BLOOD, LYMPH NODES, LUNGS AND COLORECTAL MUCOSA BEFORE AND AFTER SIV INFECTION TO DETERMINE THE IMPACT OF IL-22 LOSS ON IMMUNE ACTIVATION AND SUBSEQUENT LTBI REACTIVATION (2B). UNDERSTANDING THE MECHANISM OF IL-22 PERTURBATION IN HIV/TB WILL LEAD TO IDENTIFICATION AND DEVELOPMENT OF ANTIBODY-BASED THERAPEUTICS OR USE OF RECOMBINANT IL-22 TO PREVENT REACTIVATION OF LTBI IN COINFECTED COHORTS.

MODELING CHIKUNGUNYA VIRUS NEUROINVASION AND NEUROPATHOGENESIS IN MICE - PROJECT SUMMARY THE ALPHAVIRUS CHIKUNGUNYA VIRUS (CHIKV) REPRESENTS A RE-EMERGING PUBLIC HEALTH CONCERN AS MOSQUITO VECTORS SPREAD TO NEW TERRITORIES. WHILE CHIKV TYPICALLY INDUCES AN ARTHRITOGENIC DISEASE, UP TO 16% OF CASES RESULT IN NEUROLOGICAL DISEASE SUCH AS ENCEPHALITIS AND MENINGITIS. SURVIVING PATIENTS ARE OFTEN LEFT WITH LONG-TERM NEUROLOGICAL DEFICITS, INCLUDING IMPAIRED MOTOR CONTROL, EMOTIONAL AND BEHAVIORAL DISINHIBITION, AND COGNITIVE DISORDERS. DESPITE NUMEROUS PUBLISHED CLINICAL CASE AND PUBLIC HEALTH IMPACT REPORTS, VERY LITTLE IS KNOWN ABOUT THE NEUROPATHOGENESIS OF CHIKV INFECTION. THE LIMITED ACCESSIBILITY AND MINIMALLY REGENERATIVE NATURE OF THE CENTRAL NERVOUS SYSTEM (CNS) CREATE AN INSURMOUNTABLE BARRIER TO STUDYING THE COURSE OF NEUROLOGICAL CHIKV INFECTION IN HUMANS AND REQUIRE THE USE OF ANIMAL MODELS. MICE HAVE BEEN USED EXTENSIVELY TO STUDY ARTHRITOGENIC DISEASE INDUCED BY CHIKV AND NEUROLOGICAL DISEASE INDUCED BY OTHER ALPHAVIRUSES, INCLUDING LONG-TERM NEUROLOGICAL SEQUALAE. HOWEVER, ALMOST ALL MOUSE STUDIES EXAMINING NEUROLOGICAL CHIKV INFECTION HAVE UTILIZED NEONATAL OR IMMUNOCOMPROMISED MICE, WHICH DO NOT ACCURATELY REFLECT THE NEURODEVELOPMENTAL OR IMMUNOLOGICAL STATUS OF SUSCEPTIBLE HUMAN POPULATIONS. FURTHERMORE, BOTH ARTHRITOGENIC AND NEUROLOGICAL DISEASE INDUCED BY ALPHAVIRUSES HAVE BEEN SHOWN TO PRIMARILY BE MEDIATED BY THE IMMUNE RESPONSE RATHER THAN BY THE VIRUS ITSELF, FURTHER NECESSITATING THE USE OF IMMUNOCOMPETENT MICE. PRELIMINARY WORK BY OUR GROUP HAS FOUND THAT WHILE YOUNG ADULT C57BL/6J MICE ARE SUSCEPTIBLE TO INFECTION AND REPLICATE THE CNS PATHOLOGY REPORTED IN HUMANS FOLLOWING INTRACRANIAL INFECTION, CNS INFECTION CANNOT BE CONSISTENTLY ACHIEVED FOLLOWING PERIPHERAL INOCULATION. IN CONTRAST, CC041 MICE, A COLLABORATIVE CROSS STRAIN, CONSISTENTLY DEMONSTRATE INFECTION OF THE BRAIN AND SIGNS OF NEUROLOGICAL DISEASE FOLLOWING SUBCUTANEOUS INOCULATION, PRESENTING A POTENTIAL MOUSE MODEL BY WHICH TO EVALUATE NEUROLOGICAL DISEASE INDUCED BY CHIKV FOLLOWING PERIPHERAL INFECTION. THIS PROPOSAL AIMS TO CHARACTERIZE NEUROLOGICAL CHIKV INFECTION IN CC041 MICE FOLLOWING PERIPHERAL INOCULATION AND DETERMINE WHETHER CC041 MICE DEVELOP THE NEUROLOGICAL DISEASE AND CNS PATHOLOGY SEEN IN HUMANS. IN AIM 1, WE WILL DETERMINE THE MECHANISM AND DYNAMICS BY WHICH CHIKV INFECTS THE BRAIN IN CC041 MICE. IN AIM 2, WE WILL DETERMINE WHETHER CC041 MICE DEVELOP CNS PATHOLOGY AND LONG-TERM NEUROLOGICAL SEQUELAE SIMILAR TO HUMAN PATIENTS FOLLOWING NEUROLOGICAL CHIKV INFECTION. OUR STUDIES WILL ESTABLISH A NEW NEURODEVELOPMENTALLY- APPROPRIATE, IMMUNOCOMPETENT MOUSE MODEL OF PERIPHERAL CHIKV INFECTION, ALLOWING FOR FUTURE IN DEPTH EVALUATION OF THE MECHANISMS DRIVING NEUROLOGICAL DISEASE DEVELOPMENT AND BY WHICH TO TEST NEW POTENTIAL VACCINES, ANTIVIRALS, AND HOST-DIRECTED TREATMENTS FOR NEUROLOGICAL INFECTION WITH CHIKV AND OTHER ALPHAVIRUSES.

ROLE OF THE MICROBIOME OF THE SCHISTOSOME SNAIL HOST - PROJECT SUMMARY THE MICROBIOME – THE COMMUNITY OF MICROORGANISMS THAT INHABIT BOTH THE HUMAN BODY AND THAT OF MULTIPLE OTHER ANIMAL PHYLA – IS INCREASINGLY RECOGNIZED TO PLAY AN IMPORTANT ROLE IN MANY ASPECTS OF BIOLOGY AND AS A KEY DETERMINANT OF HEALTH AND DISEASE. IN INVERTEBRATES, SYMBIOTIC BACTERIA ARE INVOLVED IN SYNTHESIZING ESSENTIAL AMINOACIDS OR VITAMINS, IN DIGESTING CELLULOSE, IN PROTECTION AGAINST PARASITES, IN FEMINIZATION AND PARTHENOGENESIS, AND EVEN IN SPECIATION. IN INSECT VECTORS, THERE ARE MULTIPLE EXAMPLES OF INTERACTIONS BETWEEN RESIDENT MICROBIOMES AND PARASITES: THE SANDFLY GUT MICROBIOME MEDIATES LEISHMANIA TRANSMISSION, WOLBACHIA BACTERIA CAN DISRUPT MALARIA TRANSMISSION IN MOSQUITOES, WHILE THE MICROBIOME OF BOTH REDUVIID BUGS AND TSETSE FLIES MEDIATES TRYPANOSOME INFECTION. OUR GOAL IS TO STUDY THE MICROBIOME OF THE FRESHWATER SNAIL BIOMPHALARIA, VECTOR OF SCHISTOSOMIASIS: SIX LINES OF EVIDENCE SUGGEST A POSSIBLE ROLE FOR THE MICROBIOME IN THE BIOMPHALARIA - SCHISTOSOMA MANSONI INTERACTION: (I) THE SNAIL HEMOLYMPH (BLOOD), TO WHICH SPOROCYST STAGE SCHISTOSOMES ARE CONTINUOUSLY EXPOSED, HARBORS A DIVERSE MICROBIOME, (II) THIS MICROBIOME IS DIFFERENTIATED FROM OTHER ORGAN MICROBIOMES AND SHOWS THE GREATEST MICROBIAL DIVERSITY WITHIN SNAILS, (III) DIFFERENT LABORATORY SNAIL POPULATIONS HAVE A DISTINCTIVE MICROBIOME COMPOSITION, (IV) CO-CULTURE OF DIFFERENT SNAIL POPULATIONS RESULTS IN RAPID CHANGE IN MICROBIOME COMPOSITION, (V) SNAIL GENOTYPES FROM A SINGLE POPULATION THAT ARE RESISTANT OR SENSITIVE TO SCHISTOSOME INFECTION HAVE DISTINCTIVE MICROBIOMES AND (VI) SCHISTOSOME INFECTION RESULTS IN ALTERED MICROBIOME COMPOSITION. IN SPECIFIC AIM 1 WE WILL DIRECTLY TEST THE IMPACT OF MICROBIOME ON SNAIL HOST DEVELOPMENT AND SCHISTOSOME PARASITE SUSCEPTIBILITY. THIS WILL BE DONE BY GENERATING GERM-FREE SNAILS, JUST AS GERM-FREE MICE HAVE BEEN USED TO UNDERSTAND THE DIRECT ROLE OF THE MICROBIOME IN OBESITY. MICROBIOMES ARE DYNAMIC AND MAY VARY IN COMPOSITION AND ABUNDANCE DUE TO ENVIRONMENTAL FACTORS. THESE MICROBIOME MODIFICATIONS CAN EVEN LEAD TO HOST HEALTH AND FITNESS ALTERATIONS. SPECIFIC AIM 2 WILL FOCUS ON THE IMPACT OF MICROBIOME MODIFICATION ON SNAIL HOST FITNESS USING LONG TERM CO-CULTURE (OR COMMON GARDEN) OF SNAIL POPULATIONS CARRYING DISTINCTIVE MICROBIOMES. WE WILL CHARACTERIZE SAMPLED MICROBIOMES USING NEXT GENERATION SEQUENCING OF THE 16S RIBOSOMAL DNA. THIS PROJECT WILL (I) PROVIDE FUNDAMENTAL INFORMATION ABOUT MICROBIOME DYNAMICS AND COMPOSITION, (II) DETERMINE THE ROLE OF THE MICROBIOME IN THE SNAIL-SCHISTOSOME INTERACTIONS AND SNAIL FTINESS, AND (III) HAS STRONG POTENTIAL TO REVEAL NEW ASPECTS OF THE SNAIL-MICROBIOME-PARASITE INTERACTION THAT CAN BE TARGETED BY NOVEL INTERVENTIONS.

NOVEL ANTIVIRAL STRATEGY OFFERING FORWARD CAPABILITY AND REDUCED RISK OF ESCAPE - ABSTRACT THE CURRENT CORONAVIRUS (COV) PANDEMIC, SEASONAL INFECTIONS BY OTHER COV AND OTHER “COLD” VIRUSES, PLUS THE NEED FOR ANNUAL INFLUENZA VACCINATIONS EXEMPLIFY THE CHALLENGES POSED BY VIRAL ANTIGENIC DRIFT AND SHIFT. TARGETING LANDSCAPES OF VIRAL STRUCTURAL PROTEINS DISPLAYED ON THE SURFACES OF VIRUS PARTICLES AND OR THE SURFACES OF INFECTED CELLS HAS BEEN THE PRIMARY BASIS FOR DEVELOPING ANTIBODY-BASED THERAPEUTICS. ALTHOUGH GREAT ADVANCES HAVE BEEN MADE IN TRYING TO IDENTIFY REGIONS OF THESE SURFACE DISPLAYED PROTEINS THAT ARE CONSERVED AND LESS PRONE TO “ESCAPE” ANTIBODY BINDING, IT APPEARS TO BE A CONTINUAL BATTLE OF CAT AND MOUSE AS WE ARE SEEING WITH CONTINUAL EMERGENCE OF COV “VARIANTS OF CONCERN”. IN CONTRAST, VIRAL STRUCTURAL PROTEINS THAT REMAIN INSIDE VIRUS PARTICLES AND CELLS, AVOID THE IMPACT OF CYCLICAL ANTIBODY SELECTION, AND TEND TO BE FAR MORE CONSERVED. OLIGOMERIC ASSEMBLIES OF THESE PROTEINS CAN ALSO BLUNT THE IMPACT OF ANTIVIRAL ESCAPE MUTATIONS OWING TO THE MIX OF MUTANT AND WILD-TYPE MONOMERS PRESENT IN THE PARENT CELL. FURTHERMORE, THE STOICHIOMETRY REQUIRED OF AN ANTIVIRAL TO IMPEDE OLIGOMER FUNCTION NEED NOT BE NECESSARILY 1 ANTIVIRAL TO 1 MONOMER SINCE, ESPECIALLY IF THE ANTIVIRAL WERE A CROSSLINKER, ITS IMPACT WOULD BE RELAYED BEYOND THE IMMEDIATE CONTACT TO NEIGHBORING OLIGOMERS. OUR LONG-TERM HYPOTHESIS IS THAT AFFINITY REAGENTS CAPABLE OF BINDING AN INTERNAL OLIGOMERIC STRUCTURAL PROTEIN OF ALL SPECIES OF A VIRAL GENUS UNIFORMLY WILL IMPEDE VIRAL ASSEMBLY WHEN PRESENT AS DIMERIC CROSSLINKERS IN A MANNER THAT IS BOTH FORWARD CAPABLE AND HAS MUCH REDUCED SUSCEPTIBILITY TO VIRAL ESCAPE. WE WILL EXPLORE THE ANTIVIRAL POTENTIAL OF DIMERIC CROSSLINKERS USING VIRUSES OF THE GENUS EBOLAVIRUS AS OUR MODEL AND A NOVEL, RARE NANOBODY MANIFESTING UNIFORM REACTIVITY TO NUCLEOPROTEIN OF ALL 6 SPECIES. OUR TWO SPECIFIC AIMS ARE: (1) WE WILL ENGINEER MAMMALIAN CELL EXPRESSION VECTORS ENCODING NANOBODY HOMODIMERS AND ASSESS ANTIVIRAL ACTIVITY USING VIRUS LIKE PARTICLE SURROGATES AT BSL-2 FOLLOWING PLASMID TRANSFECTION TO DRIVE INTRABODY EXPRESSION, (2) WE WILL ENGINEER E. COLI EXPRESSION VECTORS ENCODING NANOBODY HOMODIMERS FUSED TO CELL PENETRATING PEPTIDES AND GLYCOSAMINOGLYCAN BINDING MOTIFS AND ASSESS ANTIVIRAL ACTIVITY FOLLOWING PROTEIN TRANSDUCTION OF VIRUS INFECTED CELLS AT BSL-4. SUCCESS WILL DEMONSTRATE A NOVEL ANTIVIRAL STRATEGY THAT CAN THEN BE THOROUGHLY EXPLORED FOR THE PROPENSITY TO SELECT ESCAPE MUTANTS RELATIVE TO AN EXISTING NEUTRALIZING ANTIBODY REGIME TO TEST WHETHER THE STRATEGY IS MORE “ESCAPE-PROOF”. THE OVERALL APPROACH SHOULD BE APPLICABLE TO OTHER HUMAN VIRAL PATHOGENS BY CAREFULLY RETUNING THE AFFINITY REAGENT, WITH ADEQUATE TIME AND RESOURCES, TO MAXIMIZE BROAD LONG-TERM IMPACT IN HELPING TO SAFE GUARD HUMAN HEALTH.

TUNNELING NANOTUBES AS AN ALTERNATE ROUTE OF EBOLA VIRUS DISSEMINATION - EBOLA VIRUS (EBOV) IS AN EMERGING, DANGEROUS VIRUS THAT CAUSES INCREASINGLY MORE FREQUENT OUTBREAKS OF A SYSTEMIC, HEMORRHAGIC DISEASE IN HUMAN POPULATIONS. APPROVED COUNTERMEASURES TO PREVENT OR TREAT EBOV DISEASE ARE CURRENTLY LIMITED. MACROPHAGES ARE THE INITIAL CELLS TARGETED BY EBOV, AND DUE TO THEIR MIGRATORY PROPERTIES ARE BELIEVED TO RAPIDLY DISSEMINATE THE VIRUS TO DISTANT TISSUES AND ORGANS DESPITE THE LACK OF EXPERIMENTAL EVIDENCE. IN CURRENT MODELS, EBOV PROPAGATES INFECTION THROUGH THE CELL-FREE FORM, WHERE VIRUS PARTICLES ENTER THE CELL, REPLICATE THE GENOME, AND THEN ASSEMBLE/EGRESS TO CHALLENGE NEIGHBORING CELLS. WE HAVE PRELIMINARY DATA SUGGESTING THAT EBOV MAY EXPLOIT AN ALTERNATIVE MODE TO SPREAD INFECTION (IN PARALLEL WITH THE ESTABLISHED MODEL): VIRAL NUCLEOCAPSIDS VIA TUNNELING NANOTUBES (TNTS), AN ACTIN-BASED INTERCELLULAR COMMUNICATION SYSTEM THAT ALLOWS DIRECT EXCHANGE OF CYTOPLASMIC MATERIAL BETWEEN CONNECTING CELLS. EBOV INFECTION INDUCES FORMATION OF INTERCELLULAR CONNECTIONS CONTAINING VIRUS NUCLEOCAPSID PROTEIN IN PRIMARY HUMAN ENDOTHELIAL CELL AND MACROPHAGE POPULATIONS. THESE CONNECTIONS SUPPORT CELL-TO-CELL TRANSFER OF THE NUCLEOCAPSID PROTEIN IN THE ABSENCE OF THE VIRUS. THE DATA ALSO SHOW THAT EBOV CAN EFFICIENTLY REPLICATE IN ENDOTHELIAL CELLS DEVOID OF FACTORS CRITICAL FOR VIRUS ENTRY, AFTER INITIAL RETARDATION, AND THAT THE REPLICATION IS COMPROMISED IN CELLS DEPLETED OF HOST M-SEC, A CENTRAL FACTOR FOR TNT FORMATION. THIS PROPOSAL AIMS TO INTERROGATE THE INTERACTIONS BETWEEN EBOV AND TNTS THROUGH TWO SPECIFIC AIMS. IN AIM 1, WE WILL DETERMINE IF TNTS ARE THE INTERCELLULAR CONNECTIONS INDUCED BY EBOV TO SPREAD INFECTION IN HUMAN ENDOTHELIAL CELLS AND MACROPHAGES. IN AIM 2, WE WILL DETERMINE IF EBOV SPREADS INFECTION THROUGH INTERCELLULAR TRANSFER OF NUCLEOCAPSIDS. OUR DISCOVERIES WILL ESTABLISH AN ALTERNATE MODEL OF EBOV DISSEMINATION WITHIN THE HOST, LAYING THE GROUNDWORK FOR FURTHER INVESTIGATIONS INTO PATHOGENESIS OF FILOVIRUSES. IMPORTANTLY, THESE FINDINGS MAY LEAD TO DEVELOPMENT OF NOVEL STRATEGIES TO TARGET EBOV AND RELATED VIRUSES.

THE ROLE OF MACROPHAGE PODOSOMES IN EBOLA VIRUS PATHOGENESIS - PROJECT SUMMARY EBOLA VIRUS (EBOV) IS AN EMERGING, HIGHLY PATHOGENIC VIRUS ASSOCIATED WITH INCREASINGLY MORE FREQUENT OUTBREAKS OF HEMORRHAGIC DISEASE IN HUMAN POPULATIONS. APPROVED COUNTERMEASURES TO PREVENT OR TREAT EBOV DISEASE ARE CURRENTLY LIMITED. MACROPHAGES ARE THE INITIAL CELLS TARGETED BY EBOV, AND YET, LITTLE IS KNOWN ABOUT THE EXACT NATURE OF EBOV-MACROPHAGE SURFACE INTERACTIONS AND SUBSEQUENT UPTAKE INTO THE CELL. DUE TO THEIR MIGRATORY PROPERTIES, MACROPHAGES ARE ALSO BELIEVED TO RAPIDLY DISSEMINATE THE VIRUS TO DISTANT TISSUES AND ORGANS DESPITE THE LACK OF EXPERIMENTAL EVIDENCE. WE HAVE PRELIMINARY DATA SHOWING THAT EBOV DEPENDS ON PODOSOMES, MECHANOSENSITIVE ADHESIVE STRUCTURES USED BY MACROPHAGES TO MIGRATE THROUGH TISSUES AND SAMPLE ANTIGENS, TO ENTER MACROPHAGES. THE DATA ALSO SHOWS THAT EBOV REPLICATION INCREASES MACROPHAGE LOCOMOTION THROUGH A FIBRILLAR 3D MATRIX AND REDUCES PODOSOME NUMBER, SUGGESTING THAT THE VIRUS ACTIVELY TRANSFORMS INFILTRATION OF TISSUES BY THESE CELLS. THIS PROPOSAL AIMS TO EXAMINE THE INTERACTIONS BETWEEN EBOV AND PODOSOMES. IN AIM 1, WE WILL DETERMINE WHETHER PODOSOMES SERVE AS PORTS FOR EBOV ENTRY INTO HUMAN MACROPHAGES. IN AIM 2, WE WILL CHARACTERIZE MIGRATORY AND INVASIVE PROPERTIES OF MACROPHAGES CHALLENGED WITH EBOV. IN AIM 3, WE WILL ASSESS HOST RESISTANCE TO SYSTEMIC INFECTION WITH EBOV IN A MOUSE MODEL OF EBOV DISEASE DEVOID OF FUNCTIONAL MACROPHAGES. OUR FINDINGS WILL ESTABLISH A NEW MODEL OF INTERACTIONS BETWEEN EBOV AND MACROPHAGES, LAYING THE GROUNDWORK FOR FURTHER INVESTIGATIONS INTO PATHOGENESIS OF FILOVIRUSES. IMPORTANTLY, THESE DISCOVERIES MAY LEAD TO NEW AREAS OF DEVELOPMENT OF NOVEL COUNTERMEASURES TARGETING EBOV AND RELATED VIRUSES.

ENGINEERING PATHOGEN TRIGGERED BIOMINERALIZATION TO ENABLE A NEW GENERATION OF POINT-OF-CARE TESTS

NANOBODY TOOLKIT FOR HUMAN CORONAVIRUS CLASSIFICATION - ABSTRACT THE ENDEMIC HUMAN CORONAVIRUSES (HCOV) NL63, OC43, 229E AND HKU1 HAVE TYPICALLY BEEN ASSOCIATED WITH RELATIVELY MILD AND SELF-LIMITING RESPIRATORY DISEASE RESEMBLING THE COMMON COLD IN HEALTHY INDIVIDUALS. HOWEVER, THE NEWLY EMERGING ZOONOTIC CORONAVIRUSES (SARS-COV, MERS-COV AND SARS-COV-2) CAN CAUSE MORE SEVERE RESPIRATORY DISTRESS, ESPECIALLY IN THOSE WITH UNDERLYING HEALTH CONDITIONS AND CAN BE FATAL. CORONAVIRUSES ARE DIVERSE AND SPILLOVER FROM ZOONOTIC RESERVOIRS EITHER DIRECTLY TO HUMANS OR VIA AN INTERMEDIATE HOST. SPILLOVER IS LIKELY TO BE MORE FREQUENT DUE TO THE EVER-EXPANDING HUMAN POPULATION AND ECOSYSTEM EROSION, THEREBY ENSURING MORE COLLISIONS BETWEEN NATURAL AND UNNATURAL HOST. OUR EXPLORATORY RESEARCH PROPOSAL SEEKS TO DEVELOP A SMALL TOOLKIT OF IMMUNOREAGENTS THAT WILL FORM THE BASIS OF A LOGIC GATE TO IDENTIFY AND CLASSIFY CORONAVIRUSES. A SINGLE IMMUNOREAGENT, WITH BROAD RECOGNITION OF OVERALL SHARED PROTEIN ARCHITECTURE, WILL ALLOW THE IDENTIFICATION OF THE CORONAVIRUS GENERA. SEPARATE IMMUNOREAGENTS WITH ABSOLUTE SPECIFICITY FOR UNIQUE AMINO-ACID VARIATION BETWEEN THE HUMAN AND ZOONOTIC CORONAVIRUSES WILL ALLOW THE FURTHER CLASSIFICATION TO A KNOWN PATHOGEN. SUCH A TOOLKIT WILL QUICKLY ESTABLISH IF A NEWLY EMERGING VIRUS IS A CORONAVIRUS WE ARE FAMILIAR WITH OR IS A NOVEL GENUS OR LINEAGE THAT HAS NEVER BEFORE SPILLED OVER INTO THE HUMAN POPULATION AND MAY DEMAND URGENT FOLLOW UP FOR EPIDEMIC OR PANDEMIC POTENTIAL. OUR IMMUNOREAGENTS ARE BASED ON LLAMA SINGLE DOMAIN ANTIBODIES OR NANOBODIES THAT ARE INEXPENSIVE TO PRODUCE AT HIGH YIELDS IN E. COLI AND HIGHLY MODULAR, ALLOWING FUSION TO ENZYMATIC REPORTER ENZYMES FOR ONE-STEP PROBING OF VIRUS INFECTED CELLS AND FACILE ESTABLISHMENT OF SANDWICH ASSAYS. IMMORTALIZATION OF NANOBODY SEQUENCES IN SILICO MEANS THESE ARE IMMEDIATELY AVAILABLE TO THE WIDER RESEARCH COMMUNITY FOLLOWING DEPOSITION IN GENBANK UNLIKE POLYCLONAL SERA OR HYBRIDOMAS SECRETING IGG. SINGLE-POT LIBRARY TECHNOLOGY ENABLES A RAPID RESPONSE TO BE MOUNTED AGAINST ANY NEWLY EMERGED CORONAVIRUS IN THE FUTURE, ENSURING WE ARE MORE PRO-ACTIVE THAN RE-ACTIVE TO HELP SAFEGUARD HUMAN HEALTH.

EFFECT OF HIV INFECTION ON M.TUBERCULOSIS GRANULOMA FORMATION AND EVOLUTION

ESTABLISHING A MIRNA BIOMARKER SIGNATURE FOR BRAIN STRUCTURAL VARIATION IN A NON-HUMAN PRIMATE MODEL

ATTENUATION OF LASSA VIRUS VIA CODON DEOPTIMIZATION?

FUNCTIONAL IMPACT OF LONG NON-CODING RNA EXPRESSION ON HIV CONTROL

RECOMBINANT PAPILLOMAVIRUS-BASED HIV VACCINE TARGETING GENITAL MUCOSA

NOVEL BROAD SPECTRUM INHIBITORS OF FILOVIRUS INFECTION

IMPACT OF TAT-BINDING CELLULAR LNCRNAS ON HIV REPLICATION

RAPID LIGAND PAIRING STRATEGY TO SIMPLIFY DIAGNOSTIC IMMUNOASSAY ASSEMBLY

DEFINING THE NORMAL RANGE OF POSTPRANDIAL METABOLIC RISK: MULTI-OMIC AND MULTI-TISSUE ANALYSIS AFTER A MIXED MEAL

RENOVATION OF FACILITIES FOR MACAQUE HOUSING AT THE SNPRC

HIV- INDUCED LONG NON-CODING RNAS IN VIRAL REPLICATION AND IMMUNE RESPONSE

SIGNIFICANT UPGRADE AND EXPANSION OF THE SOUTHWEST NATIONAL PRIMATE RESEARCH CENTER PATHOLOGY UNIT

NONHUMAN PRIMATE CAGING FOR THE SNPRC

NIH-OWNED CHIMPANZEE RESEARCH RESOURCE AT THE SNPRC

HEAT STABLE FILOVIRAL DIAGNOSTICS

IL-10 AS A HOST MODIFIER OF PZA ACTIVITY

MOLECULAR DETERMINANTS OF HIV HYPERMUTATION

IMPROVEMENT OF NONHUMAN PRIMATE GROUP HOUSING

EFFICIENT LINKAGE MAPPING METHODS FOR SCHISTOSOMA MANSONI

RENOVATION OF BUILDING 4 TO PROVIDE BSL-2 HOUSING FOR INFECTIOUS DISEASE RESEARCH

IMPROVED TUMOR TARGETING OF SALMONELLA VNP20009 VIA ICE-LLAMA ANTIBODY GUIDANCE

DETERMINING THE EFFICACY OF A NOVEL TB DIAGNOSTIC TEST TO MONITOR TREATMENT SUCCESS IN DRUG RESISTANT TB PATIENTS

IMPROVEMENT OF FACILITIES FOR NONHUMAN PRIMATE RESEARCH

BD FACSYMPHONY

MAPPING THE MARMOSET: CHARACTERIZING THE IMPACTS OF CHIMERISM ON MARMOSET DEVELOPMENT AND AGING - PROJECT SUMMARY THE MARMOSET MONKEY HAS EMERGED RECENTLY AS A HIGHLY TRACTABLE NONHUMAN PRIMATE MODEL OF AGING. MARMOSETS HAVE RELATIVELY SHORT LIFESPANS, AND EXHIBIT SIMILAR AGING-RELATED DECLINES TO HUMANS, INCLUDING CHANGES IN BLOOD PRESSURE, IMMUNE FUNCTION, AND COGNITION. HOWEVER, THERE IS A BIOLOGICAL “BLACK BOX” IN MARMOSET BIOLOGY, THE IMPLICATIONS OF WHICH FEW RESEARCHERS MENTION. MARMOSETS ARE NATURAL CHIMERAS, WITH SIBLINGS EXCHANGING GENETIC MATERIAL IN UTERO. SOME REPORTS LIMIT CHIMERISM TO ONLY HEMATOPOIETIC TISSUES (E.G., BLOOD, SPLEEN, BONE MARROW) WHEREAS OTHERS SUGGEST THAT OTHER TYPES OF TISSUE CAN BE CHIMERIC, INCLUDING GERMLINE TISSUES, WITH IMPLICATIONS FOR HERITABILITY. IT REMAINS UNCLEAR HOW THE PRESENCE OF MULTIPLE GENOMES IN AN INDIVIDUAL IMPACTS MARMOSET PHYSIOLOGY AND THE RISK OF AGE-RELATED MORBIDITIES. THE RATIONALE FOR THIS STUDY IS THAT TO UNDERSTAND THE BENEFITS AND LIMITATIONS OF THE MARMOSET MODEL WE MUST CHARACTERIZE THE EMERGENCE AND IMPACTS OF CHIMERISM ON AN INDIVIDUAL. THESE EFFORTS ARE IMPERATIVE TO THE OVERALL OBJECTIVE OF BEING ABLE TO PREDICT AND DETERMINE OUTCOMES OF CHIMERISM. WITH THE INCREASING APPRECIATION THAT CHIMERISM IN HUMANS IS BOTH WIDESPREAD AND A MAJOR DRIVER OF HUMAN HEALTH, MARMOSET MODELS OF CHIMERISM MAY BE A PIVOTAL TOOL IN AGING RESEARCH. THEREFORE, THE SPECIFIC AIMS FOR THIS PROPOSAL TO 1) DEFINE AND MAP CHIMERISM IN MARMOSETS, 2) CHARACTERIZE THE DEVELOPMENTAL DYNAMICS OF MARMOSET CHIMERISM, AND 3) EVALUATE THE COSTS OF CHIMERISM FOR AGING MARMOSETS.

DEVELOPMENT OF NONHUMAN PRIMATE MODELS FOR PSA BIOLOGY STUDIES

MOLECULAR PATHOLOGY OF ORAL IMMUNE DYSREGULATION IN HIV/SIV INFECTION

DEVELOPMENT OF A VACCINE TO PROTECT MONKEYS FROM HERPES B VIRUS INFECTION

LINKING RECEPTOR-MEDIATED PHAGOCYTOSIS AND CAMP PATHWAYS IN MACROPHAGE RESPONSES TO TUBERCULOSIS

RESOLUTION OF ACUTE HEPATITIS B VIRUS INFECTIONS IN CHIMPANZEES

DEVELOPMENT OF A VACCINE TO PROTECT MONKEYS FROM HERPES B VIRUS INFECTION

DETERMINING THE FUNCTIONAL SIGNIFICANCE OF MUTATIONS OBSERVED IN ENVELOPE PROTEIN FOLLOWING SERIAL IN VIVO PASSAGING OF HUMAN-SIMIAN IMMUNODEFICIENCY VIRUS - ABSTRACT/SUMMARY: COMMONLY USED ANIMAL MODELS OF HIV-1 INCLUDE INFECTION OF MACAQUES WITH SIMIAN IMMUNODEFICIENCY VIRUS (SIV) OR SIMIAN-HUMAN IMMUNODEFICIENCY VIRUS (SHIV) CONTAINING HIV ENVELOPE (ENV) OR REVERSE TRANSCRIPTASE. THESE ANIMAL MODELS HAVE BEEN EXTREMELY USEFUL IN UNDERSTANDING HIV PATHOGENESIS AND DISEASE PROGRESSION, AS WELL AS UNDERSTANDING THE EFFICACY OF VACCINES AND DRUGS. HOWEVER, THE GENETIC DIFFERENCE BETWEEN HIV-1 AND SIV, AND THE ABSENCE OF OTHER HIV-1 GENES SUCH AS GAG, POL, VIF, VPR, AND NEF IN SHIV LIMITS THE UTILITY OF THESE MODELS IN VACCINE STUDIES. IDEALLY, GOOD ANIMAL MODEL OF HIV-1 INFECTION/AIDS WOULD BE INFECTION OF MACAQUES WITH HIV-1. HOWEVER, HIV-1 DOES NOT REPLICATE IN MACAQUE CELLS DUE TO THE PRESENCE OF RETROVIRAL RESTRICTION FACTORS. HIV-1 CAN BE MADE TO REPLICATE BY SUBSTITUTING ITS ACCESSORY GENES WITH SIV GENES SUCH AS VIF, VPX, VPR, AND NEF, WHICH CAN COUNTERACT INTERFERON-INDUCED RESTRICTION FACTORS IN MACAQUE CELLS. HUMAN-SIMIAN IMMUNODEFICIENCY VIRUS (HSIV) IS AN HIV-1NL4-3 DERIVATIVE WITH SIV VIF GENE SUBSTITUTION (NAMED HSIV-VIFNL4-3) THAT CAN REPLICATE PERSISTENTLY IN PIGTAIL MACAQUES (PTMS). HOWEVER, INFECTION DID NOT RESULT IN HIGH PEAK VIREMIA AND SETPOINT VIRAL LOADS AS OBSERVED DURING SIV INFECTION OF MACAQUES. SERIAL IN VIVO PASSAGING IN PTMS WAS PERFORMED TO ENHANCE INFECTIVITY OR REPLICATIVE CAPACITY OF HSIV. THREE ROUNDS OF ANIMAL-TO-ANIMAL TRANSFER OF INFECTED BLOOD IN 3 IMMUNOCOMPETENT PTMS WITH STARTING INITIAL INOCULUM CONTAINING A MIXTURE OF CXCR4- (HSIV- VIFNL4-3 RECOVERED FROM PREVIOUSLY INFECTED MACAQUE) AND CCR5-TROPIC HSIV (HSIV-VIF DERIVATIVE BASED ON PNL- AD8 AND BRU-YU2) WAS CONDUCTED TO GENERATE PATHOGENIC VARIANTS. INTERESTINGLY, ALL THE MACAQUES SHOWED PEAK VIREMIA CLOSE TO OR ABOVE 105 COPIES/ML AND VIRUS REPLICATION PERSISTED FOR MORE THAN 20 WEEKS. FOLLOWING IN VIVO PASSAGING, THREE INFECTIOUS MOLECULAR CLONES (IMCS) WERE RECOVERED FROM PASSAGE 3 MACAQUE (HSIV-P3 IMCS). SEQUENCING OF HSIV-P3 IMCS SHOWED SEVERAL INTERESTING MUTATIONS THROUGHOUT THE GENOME, PERHAPS SUGGESTING ADAPTATION TO PTMS. THESE MUTATIONS COULD HELP THE VIRUS IN OVERCOMING RESTRICTION FACTORS, OR BETTER UTILIZATION OF HOST DEPENDENCY FACTORS, OR THEY COULD HELP THE VIRUS ESCAPE HOST IMMUNE RESPONSES. FOCUS OF THIS GRANT APPLICATION IS TO DETERMINE THE FUNCTIONAL SIGNIFICANCE OF MUTATIONS OBSERVED IN ENVELOPE GENE. THE RESULTS FROM THIS STUDY WILL PROVIDE VALUABLE INSIGHTS INTO THE ROLE OF ENVELOPE GENE IN CROSS-SPECIES TRANSMISSION OF HIV-1 TO PIGTAILED MACAQUES.

RATIONALIZING THE MOLECULAR MECHANISMS OF HIGH SENSITIVITY AND SPECIFICITY OF ANTI-FILOVIRAL SINGLE DOMAIN ANTIBODIES

HUMAN ORIGINS: GENETIC AND MOLECULAR BASIS OF CRANIOFACIAL EVOLUTION

DEVELOPMENT OF NONHUMAN PRIMATE MODELS FOR PSA BIOLOGY STUDIES

DUAL BECKMAN COULTER DXH 690T AND BECKMAN COULTER DXH 560 AL - ABSTRACT TEXAS BIOMEDICAL RESEARCH INSTITUTE (TEXAS BIOMED) IS A NON-PROFIT, INDEPENDENT RESEARCH INSTITUTE, FOCUSED ON INFECTIOUS DISEASE RESEARCH. IT IS ALSO THE HOST INSTITUTION FOR THE SOUTHWEST NATIONAL RESEARCH PRIMATE CENTER (SNPRC), ONE OF THE SEVEN NIH-FUNDED NPRCS IN THE NATION. SNPRC HOUSES ~ 3000 NONHUMAN PRIMATES (NHPS). THE SNPRC PATHOLOGY CORE SUPPORTS DIAGNOSTIC, ANATOMIC, MORPHOLOGIC, MOLECULAR AND CLINICAL PATHOLOGY NEEDS OF > 100 INTERNAL AND EXTERNAL SCIENTISTS. MANY INTERNAL FACULTY MEMBERS ALSO EXTENSIVELY UTILIZE RODENTS FOR RESEARCH THAT IS SUPPORTED BY THE PATHOLOGY CORE STAFF. WE REQUEST S10 SUPPORT TO PURCHASE A DUAL BECKMAN COULTER DXH 690T AND BECKMAN COULTER DXH 560 AL FOR HEMATOLOGY ANALYSES OF EXPERIMENTAL AND COLONY HEALTH DATA OF NHPS AND RODENTS. THE REQUESTED REPLACEMENT UNIT WILL REPLACE AND UPGRADE THE CLINICAL DIAGNOSTIC SERVICES THAT ARE CURRENTLY PROVIDED BY END-OF-LIFE EQUIPMENT AT TEXAS BIOMED. THE DXH 690T IS A SUITABLE, COST-EFFECTIVE REPLACEMENT FOR, AND IS CAPABLE OF REPRODUCING ALL ANALYSIS PARAMETERS OF AN EXISTING DXH 800. THE DXH 560L PERFORMS HEMATOLOGY ANALYSIS USING MINISCULE VOLUMES OF BLOOD, WHICH MAKES HEMATOLOGY ANALYSIS POSSIBLE FOR STUDIES INVOLVING RODENTS AND SMALL NHPS.

ELUCIDATING VIRAL AND HOST FACTORS THAT DRIVE CHIKUNGUNYA VIRUS NEUROVIRULENCE AND NEUROPATHOGENESIS USING MICE MODELS - PROJECT SUMMARY THE ALPHAVIRUS CHIKUNGUNYA VIRUS (CHIKV) PRESENTS A RE-EMERGING PUBLIC HEALTH THREAT AS THE GEOGRAPHIC RANGES OF MOSQUITO VECTORS EXPAND. LARGE-SCALE OUTBREAKS HAVE OCCURRED ACROSS INDIA, CHINA, AND THE AMERICAS, INCLUDING LOCAL TRANSMISSION IN TEXAS AND FLORIDA. CHIKV TYPICALLY INDUCES ARTHRITOGENIC DISEASE, BUT UP TO 25% OF CASES RESULT IN NEUROLOGICAL DISEASE, SUCH AS ENCEPHALITIS. WHILE THE FDA RECENTLY APPROVED A CHIKV VACCINE, TREATMENT FOLLOWING INFECTION IS STILL LIMITED TO SYMPTOMATIC CARE. A SIGNIFICANT CHALLENGE FOR DEVELOPING TREATMENTS PATIENTS DEVELOP NEUROLOGICAL SYMPTOMS AFTER VIRUS IS CLEARED. THE IMMUNE RESPONSE, PARTICULARLY T CELLS, HAS BEEN SHOWN TO MEDIATE DEVELOPMENT OF JOINT PATHOLOGY IN CHIKV PATIENTS. THEREFORE, HOST-DIRECTED THERAPIES THAT TARGET THE IMMUNE RESPONSE, WHETHER ALONE OR IN COMBINATION WITH ANTIVIRALS, REPRESENT PROMISING APPROACHES FOR COMBATTING CHIKV-INDUCED DISEASE. THEREFORE, AN UNDERSTANDING OF HOW HOST AND VIRAL FACTORS CONTRIBUTE TO CHIKV-INDUCED NEUROLOGICAL DISEASE IS CRITICAL FOR DEVELOPING EFFECTIVE COUNTERMEASURES. OUTBREAKS CAUSED BY CERTAIN CHIKV STRAINS ARE ASSOCIATED WITH HIGHER RATES OF NEUROLOGICAL DISEASE THAN OTHERS, SUGGESTING THAT VIRAL FACTORS CONTRIBUTE TO CHIKV NEUROPATHOGENESIS. HOWEVER, CLINICAL FEATURES OF NEUROLOGICAL CHIKV INFECTION VARY BY OUTBREAK AND GEOGRAPHICAL REGION, SUGGESTING HOST FACTORS PLAY A ROLE IN CHIKV ENCEPHALITIS. THE OBJECTIVE OF THIS PROPOSAL IS TO DETERMINE THE VIRAL AND HOST FACTORS THAT MEDIATE CHIKV- INDUCED NEUROLOGICAL DISEASE. THE LIMITED ACCESSIBILITY AND MINIMALLY REGENERATIVE NATURE OF THE CENTRAL NERVOUS SYSTEM (CNS) CREATE AN INSURMOUNTABLE BARRIER TO STUDYING THE COURSE OF NEUROLOGICAL CHIKV INFECTION IN HUMANS AND REQUIRE THE USE OF ANIMAL MODELS. MICE HAVE BEEN USED EXTENSIVELY TO STUDY ARTHRITOGENIC DISEASE INDUCED BY CHIKV AND NEUROLOGICAL DISEASE CAUSED BY OTHER ALPHAVIRUSES, INCLUDING LONG-TERM NEUROLOGICAL SEQUELAE. HOWEVER, ALMOST ALL MOUSE STUDIES EXAMINING NEUROLOGICAL CHIKV INFECTION HAVE UTILIZED NEONATAL OR IMMUNOCOMPROMISED MICE, WHICH DO NOT ACCURATELY REFLECT THE NEURODEVELOPMENTAL OR IMMUNOLOGICAL STATUS OF SUSCEPTIBLE HUMAN POPULATIONS. OUR LAB’S PRELIMINARY STUDIES HAVE SHOWN THAT YOUNG ADULT C57BL/6J MICE ARE SUSCEPTIBLE TO INFECTION, BUT DEVELOP MILD, NON-LETHAL CHIKV-INDUCED DISEASE WHEN INFECTED INTRACRANIALLY (IC) WITH SL15649. IN CONTRAST, IC SL15649-INFECTED CC041 MICE, A COLLABORATIVE CROSS STRAIN, DEVELOP BRAIN MODERATE TO SEVERE, LETHAL DISEASE. IN AIM 1, I WILL USE RECOMBINANT CHIKV VIRUSES TO INVESTIGATE WHAT VIRAL PROTEIN(S) ARE DRIVING NEUROINVASION AND NEUROVIRULENCE. IN AIM 2, I WILL USE B6 AND CC041 MICE TO DISSECT IMMUNE COMPONENTS THAT CONTRIBUTE TO CHIKV-INDUCED PATHOLOGY. TOGETHER, THESE DATA WILL HELP UNCOVER THE MECHANISMS THAT DRIVE CHIKV NEUROPATHOGENESIS, AND THE SKILLS AND TECHNIQUES GAINED THROUGH THIS PROJECT WILL PROVIDE ME WITH THE FOUNDATION NEEDED TO ESTABLISH MY INDEPENDENT RESEARCH PROGRAM FOCUSED ON HOST-PATHOGEN INTERACTIONS AND DEVELOPING THERAPEUTICS.

30TH ANNUAL SYMPOSIUM ON NONHUMAN PRIMATE MODELS FOR AIDS

37TH ANNUAL SYMPOSIUM ON NONHUMAN PRIMATE MODELS FOR AIDS

ELUCIDATE THE ADVERSE IMPACT OF MITOCHONDRIA-INDUCED OXIDATIVE STRESS IN MOLECULAR AND CELLULAR DETERMINANTS IN THE AGING LUNG, DRIVING SUSCEPTIBILITY TO MYCOBACTERIUM TUBERCULOSIS INFECTION - PROJECT ABSTRACT THE AGING POPULATION WILL DOUBLE TO 2 BILLION BY 2050. NATURAL LUNG AGING IS ASSOCIATED WITH PROGRESSIVE CHANGES AT MOLECULAR AND PHYSIOLOGICAL LEVELS, CAUSING A DECLINE IN LUNG FUNCTION AND IMPAIRED IMMUNOLOGICAL RESPONSES. TO AVOID CUMULATIVE DAMAGE, LUNG-RESIDENT CELLS RELY ON A ROBUST HOMEOSTATIC BALANCE OF STRESS RESPONSE PATHWAYS; HOWEVER, AT A CERTAIN TIPPING POINT(S) (POINT OF NO RETURN), AGING FINALLY OVERWHELMS THESE CONTROL MECHANISMS LEADING TO AN INCREASED OXIDATIVE ENVIRONMENT AND IRREVERSIBLE DAMAGES. OUR DATA INDICATE THAT LUNG TISSUE IN THE ELDERLY (IN HUMANS AND MICE) HAS HIGH INFLAMMATION AND OXIDATIVE STRESS BASELINES, LEADING TO DYSFUNCTION OF CRITICAL INNATE SOLUBLE AND CELLULAR COMPONENTS DRIVING HOST SUSCEPTIBILITY TO RESPIRATORY INFECTIONS [E.G., TUBERCULOSIS (TB) AND CORONAVIRUS DISEASE 2019 (COVID-19)]. DEFINING WHEN AND HOW THESE CHANGES OCCUR IN THE LUNG AT THE CELLULAR AND MOLECULAR LEVELS IS CRITICAL TO UNDERSTANDING AGE-ASSOCIATED LUNG-SPECIFIC PATHOLOGIES AND AGING IN GENERAL. OUR DATA LINK MITOCHONDRIAL DYSFUNCTION TO CUMULATIVE OXIDATIVE STRESS IN THE LUNG OF THE ELDERLY, WHERE INTERVENTIONS THAT REDUCE LUNG OXIDATIVE STRESS CAN REVERSE SUSCEPTIBILITY TO RESPIRATORY DISEASES. MITOPHAGY (MITOCHONDRIAL AUTOPHAGY) IS ALSO IMPAIRED AT THIS STAGE, RESULTING IN INCREASED ACCUMULATION OF OXIDATIVE STRESSORS IN CELLS. WE NOW HYPOTHESIZE THAT AGING-ASSOCIATED MITOCHONDRIAL DYSFUNCTION AND IMPAIRED MITOPHAGY IS CENTRAL TO THE COLLAPSE IN PULMONARY CONTROL OF MYCOBACTERIA. USING THE WELL-ACCEPTED MOUSE MODEL OF AGING, THIS APPLICATION AIMS TO DETERMINE WHETHER AGING-ASSOCIATED MITOCHONDRIAL DYSFUNCTION DRIVES INCREASED OXIDATIVE STRESS IN LUNG CELLS, GENERATING A PERMISSIVE LUNG ENVIRONMENT FOR RESPIRATORY INFECTIONS SUCH AS MYCOBACTERIUM TUBERCULOSIS, THE CAUSATIVE AGENT OF TB. COMPLETING THE F99 PHASE WILL FACILITATE MY TRANSITION TO THE POSTDOCTORAL PHASE (K00 PHASE) BY PROVIDING ROBUST INTELLECTUAL AND TECHNICAL TRAINING AND, CONSEQUENTLY, CONTRIBUTING TO MY GOAL OF BECOMING AN INDEPENDENT RESEARCHER IN THE BIOLOGY OF AGING FIELD.