Lundquist Institute For Biomedical Innovation At Harbor-Ucla Medical

GENERAL CLINICAL RESEARCH CENTER

SYSTEMS IMMUNOLOBIOLOGY OF ANTIBIOTIC-PERSISTENT MRSA INFECTION

CONSTRUCTION - CHRONIC DISEASE CLINICAL RESEARCH CENTER

SYSTEMS EPIGENOMICS OF PERSISTENT BLOODSTREAM INFECTION - PROJECT ABSTRACT PERSISTENT BLOODSTREAM INFECTIONS ARE LIFE-THREATENING INFECTIOUS DISEASE EMERGENCIES POSING SIGNIFICANT CHALLENGES TO EFFECTIVE TREATMENT. SUCH INFECTIONS OCCUR WHEN A PATHOGEN IS SUSCEPTIBLE TO AN ANTI-INFECTIVE AGENT IN VITRO BUT IS NOT CLEARED FROM THE BLOODSTREAM IN VIVO WHEN THAT ANTI-INFECTIVE AGENT IS USED APPROPRIATELY. AS A RESULT, ANTI-INFECTIVE USAGE INCREASES, ACCELERATING ALARMING INCREASES IN ANTI-INFECTIVE RESISTANCE. THIS VICIOUS CYCLE OF PERSISTENCE DRIVING ANTI- INFECTIVE ESCALATION DRIVING RESISTANCE IS AN NIH HIGH–PRIORITY CONCERN. BLOODSTREAM INFECTIONS CAUSED BY STAPHYLOCOCCUS AUREUS (SA) OR CANDIDA ALBICANS (CA) ARE INCREASINGLY COMMON. OF URGENT CONCERN, UP TO 35% OF PATIENTS WITH METHICILLIN-RESISTANT SA (MRSA) PERSISTENT BACTEREMIA SUCCUMB EVEN ON GOLD-STANDARD THERAPY. LIKEWISE, IN PATIENTS WITH HEMATOGENOUSLY DISSEMINATED CANDIDIASIS (HDC), MORTALITY IS 39% OVERALL AND 47% IN THOSE IN THE INTENSIVE CARE UNIT, DESPITE APPROPRIATE TREATMENT. A DISEASE MYSTERY IS CENTRAL TO SUCH INFECTIONS: THE CAUSATIVE PATHOGEN IS SUSCEPTIBLE TO ANTIMICROBIALS IN LABORATORY TESTING—BUT NOT IN THE HUMAN BEING. IMPORTANTLY, PERSISTENCE REFLECTS A UNIQUE TYPE OF TREATMENT-REFRACTORY INFECTIONS DISTINCT FROM CLASSICAL ANTIBIOTIC RESISTANCE. RATHER, PERSISTENT MRSA OR CA ARE ELUSIVE: THEY ADAPT TO HOST IMMUNE RESPONSES AND ANTIBIOTIC STRESSES UNIQUELY IN VIVO AND THEN REVERT QUICKLY IN VITRO. PRESENTLY, THERE ARE FEW THERAPEUTIC OPTIONS FOR PERSISTENT MRSA OR CA BLOODSTREAM INFECTIONS. HENCE, THERE IS A CRITICAL, UNMET NEED TO UNDERSTAND THE UNIQUE INTERACTIONS OF THE HUMAN, PATHOGEN AND THERAPEUTIC FACTORS DRIVING PERSISTENCE OUTCOMES. BASED ON OUR EXTENSIVE PRELIMINARY DATA, WE BELIEVE THAT PERSISTENT INFECTIONS CAUSED BY MRSA AND CA RESULT FROM A THREE-WAY INTERACTION OF THE PATHOGEN, HOST IMMUNE RESPONSE AND ANTIMICROBIAL AGENT IN VIVO. WE HYPOTHESIZE THAT PERSISTENT ISOLATES: 1) HAVE SPECIFIC EPIGENOMES TO ENABLE PERSISTENCE; 2) SUBVERT INNATE IMMUNE PROGRAMMING AND MEMORY FOR IMMUNE EVASION; 3) EVOKE NON-PROTECTIVE OR MALADAPTIVE IMMUNE RESPONSES; AND 4) EXPLOIT CONTEXTUAL IMMUNITY AS PERSISTENCE RESERVOIRS. WE FURTHER POSIT THAT CONVENTIONAL APPROACHES TO STUDY THIS CLINICALLY URGENT PHENOMENON ARE INSUFFICIENT TO UNDERSTAND IT. WE HAVE DEVELOPED THREE INDEPENDENT BUT SYNERGISTIC RESEARCH PROJECTS TO OVERCOME THESE LIMITATIONS. EACH PROJECT BRINGS PROVEN STRENGTHS AND INNOVATIVE APPROACHES TO BEAR ON SPECIFIC AIMS THAT SYNERGIZE VIA A SYSTEMS-BASED APPROACH SUPPORTED BY OUTSTANDING TECHNOLOGY, BIOINFORMATICS AND COMPUTATIONAL CORES. HERE, WE WILL USE STATE-OF-THE-ART TECHNOLOGIES TO COMPREHENSIVELY ANALYZE THE GENETICS AND EPIGENETICS OF PATHOGENS AND THE HOST IMMUNE SYSTEM IN CONTEXT OF ANTIMICROBIAL THERAPY IN LABORATORY STUDIES AND EXPERIMENTAL MODELS OF INFECTION. IN TURN, THESE DATA WILL BE ANALYZED USING POWERFUL BIOINFORMATICS AND COMPUTATIONAL METHODS TO DETECT HIDDEN PATTERNS WITHIN LARGE COMPLEX DATASETS. BY UNDERSTANDING THESE FACTORS AND THEIR INTERACTIONS, NEW APPROACHES TO IDENTIFY AND TREAT HIGH RISK PATIENTS CAN BE DEVELOPED AND APPLIED TO IMPROVE AND SAVE LIVES. THESE GOALS ARE IDEALLY ALIGNED WITH PRIORITIES OF THE NATIONAL INSTITUTES OF HEALTH AND CENTERS FOR DISEASE CONTROL & PREVENTION.

CENTER FOR MALE REPRODUCTIVE EPIGENOMICS

IRON UPTAKE AND MUCORMYCOSIS PATHOGENESIS

CROSS-KINGDOM VACCINE TARGETING HEALTHCARE-ASSOCIATED PRIORITY PATHOGENS

SHORT AND LONG TERM OUTCOMES OF DOXYCYCLINE VERSUS TMP-SMX FOR SSTI TREATMENT

DECIPHERING IMMUNOPATHOGENESIS OF MUCORMYCOSIS TO ADVANCE RISK STRATIFICATION, DIAGNOSIS AND MANAGEMENT OF THE DISEASE - PROJECT SUMMARY/ABSTRACT MUCORALES FUNGI CAUSE MUCORMYCOSIS (MCM), A LETHAL INFECTION IN SUSCEPTIBLE HOSTS SUFFERING FROM DIABETIC KETOACIDOSIS (DKA), NEUTROPENIA, UNDERGOING HEMATOPOIETIC STEM CELL OR SOLID ORGAN TRANSPLANT, OR RECEIVING CORTICOSTEROIDS. MCM HAS POOR PROGNOSIS, WITH OVERALL MORTALITY RATE OF 50% THAT APPROACHES 100% IN CERTAIN PATIENT POPULATIONS. THIS POOR SURVIVAL IS DUE TO LIMITED KNOWLEDGE ABOUT MCM PATHOGENESIS AND THE IMMUNE RESPONSE TO THE INFECTION. THERE IS ALSO LACK OF UNDERSTANDING OF THE MOLECULAR FACTORS THAT LEAD TO MCM AND/OR PREDICT RESPONSE TO THERAPY AS WELL AS A PAUCITY OF EFFECTIVE ASSAYS THAT CAN INFORM EARLY DIAGNOSIS AND RESPONSE TO ANTIFUNGAL THERAPY. IMPORTANTLY, RESEARCH ADVANCES MADE IN THE FIELD HAVE LARGELY BEEN SLOW, AND IN SILO. THIS PROGRAM PROJECT, MUCORMYCOSIS TO ADVANCE RISK STRATIFICATION, DIAGNOSIS, AND MANAGEMENT OF THE DISEASE (MUCOR-ADVANCE), CONSISTS OF THREE INTEGRATED, COMPLEMENTARY PROJECTS SUPPORTED BY THREE CORES. THE GOAL OF MUCOR-ADVANCE IS TO DECIPHER THE IMMUNOPATHOGENESIS OF MCM AND USE THIS KNOWLEDGE TO DEVELOP NEW DIAGNOSTIC ASSAYS AND APPROACH TO THERAPY. MUCOR-ADVANCE IS LED BY WORLD LEADERS IN MCM RESEARCH, WHO HAVE COLLABORATED FOR >20 YEARS IN THE FIELD. THIS MULTIDISCIPLINARY PROPOSAL WILL UNCOVER KEY FUNGAL AND HOST FACTORS THAT MEDIATE THE PATHOGENESIS OF MCM AND THAT WILL BE USED TO AS A BASIS FOR INNOVATIVE APPROACHES FOR EARLY DIAGNOSIS OF MCM AND HOST- AND PATHOGEN-DIRECTED ADJUNCTIVE IMMUNOTHERAPEUTIC STRATEGIES TO ATTENUATE MUCORALES VIRULENCE, REVERSE IMMUNOMETABOLIC DEFECTS, AND HARNESS PHYSIOLOGICAL IMMUNE RESPONSES AGAINST MCM. PROJECT 1 WILL DELINEATE THE ROLE OF UNIQUE MUCORALES TOXINS AND INVASINS IN THE IMMUNOPATHOGENESIS OF MCM AND ASSESS THEIR USE AS TARGETS FOR ADJUNCTIVE IMMUNOTHERAPY AND DIAGNOSIS. PROJECT 2 WILL EXAMINE THE ROLE OF METABOLIC ABNORMALITIES IN IMPAIRING SPECIALIZED HOST DEFENSE MECHANISMS AGAINST MUCORALES WITH EMPHASIS ON IMMUNOMETABOLIC STRATEGIES FOCUSED ON FREE FATTY ACIDS AND BIOACTIVE LIPIDS THAT MEDIATE THE ANTI- MUCORALES ACTIVITY OF PHAGOCYTES. PROJECT 3 WILL USE PATIENT SAMPLES TO IDENTIFY SURROGATES OF DYSFUNCTIONAL INNATE EFFECTOR RESPONSES AND THEIR PROGNOSTIC SIGNIFICANCE IN MCM PATIENTS AND WILL ASSESS MAINLY FDA-APPROVED NON- CELLULAR IMMUNOMODULATORS (IMMUNE CHECKPOINT INHIBITORS, HEMATOPOIETIC GROWTH FACTORS, AND CYTOKINES) AS NOVEL ADJUNCTIVE THERAPIES IN CLINICALLY RELEVANT MURINE MODELS OF MCM. ALL AIMS WILL VALIDATE THE TRANSLATABILITY OF THE IN VITRO AND IN VIVO MODELS BY USING SPECIMEN PROSPECTIVELY COLLECTED BY THE CLINICAL CORE FROM MCM PATIENTS. THE MOLECULAR IMMUNOPATHOGENESIS STUDIES OF MCM WILL BE FACILITATED BY ADVANCED SINGLE CELL RNA-SEQ, DUAL RNASEQ AND BIOINFORMATIC ANALYSES PROVIDED BY THE GENOMIC/TRANSCRIPTOMIC CORE. FINALLY, AN ADMINISTRATIVE CORE WILL ENABLE THE EFFICIENT, COST-EFFECTIVE IMPLEMENTATION OF THE GOALS OF THE PROGRAM. THE OUTCOME OF THE MUCOR-ADVANCE PROGRAM PROJECT WILL IDENTIFY EARLY DIAGNOSTIC AND PROGNOSTIC BIOMARKERS FOR IMPROVED DISEASE MANAGEMENT AND INTRODUCE NOVEL IMMUNE-BASED ADJUNCTIVE THERAPIES. THESE THERAPIES INCLUDE FDA- APPROVED DRUGS WITH THE POTENTIAL TO RAPIDLY ENHANCE TREATMENT OF THIS LETHAL DISEASE.

IND-ENABLEMENT OF KINOCIDIN GAMMA-RP-1 FOR MDR GRAM-NEGATIVE INFECTIONS

TRANSCRIPTIONAL NETWORKS GOVERNING A. FUMIGATUS VIRULENCE - ABSTRACT INVASIVE INFECTIONS DUE TO ASPERGILLUS FUMIGATUS ARE INCREASING AND ARE STILL ASSOCIATED WITH UNACCEPTABLY HIGH MORTALITY, EVEN WITH NEW THERAPIES. OUR UNDERSTANDING OF A. FUMIGATUS INFECTION BIOLOGY IS LIMITED. AMONG 10,180 PREDICTED GENES IN THE A. FUMIGATUS GENOME, OVER 95% ARE UNCHARACTERIZED, AND FEWER THAN 100 GENES HAVE DEMONSTRATED ROLES IN VIRULENCE. IT IS CRITICAL TO IDENTIFY GENES THAT GOVERN VIRULENCE AND THE PATHWAYS IN WHICH THEY ACT BECAUSE THEY CAN POINT TO HIGH PRIORITY TARGETS FOR THERAPEUTIC AND DIAGNOSTIC DEVELOPMENT. WE HAVE IDENTIFIED A TRANSCRIPTIONAL REGULATOR IN A. FUMIGATUS, WRPA, THAT SHARES LIMITED HOMOLOGY WITH CANDIDA ALBICANS WOR1 AND HISTOPLASMA CAPSULATUM RYP1. OUR PRELIMINARY DATA INDICATE THAT WRPA GOVERNS THE CAPACITY OF A. FUMIGATUS TO WITHSTAND MACROPHAGE KILLING, GROW UNDER HYPOXIC CONDITIONS, AND INVADE AND DAMAGE PULMONARY CELLS IN VITRO. WRPA DELETION MUTANTS HAVE HIGHLY ATTENUATED VIRULENCE IN THE MOUSE MODEL OF INVASIVE ASPERGILLOSIS. USING RNA-SEQ, WE FOUND THAT WRPA GOVERNS THE EXPRESSION OF ~15% OF GENES IN THE A. FUMIGATUS GENOME, INCLUDING MULTIPLE TRANSCRIPTION FACTOR GENES. OUR PREMISE IS THAT THE WRPA IS A MASTER REGULATOR THAT GOVERNS HOST CELL INTERACTIONS AND VIRULENCE. IN SUPPORT OF THIS PREMISE, OUR INITIAL INVESTIGATIONS OF THE WRPA REGULON HAVE ALREADY REVEALED NOVEL PATHOGENICITY-RELATED FUNCTIONS OF THREE WRPA-DEPENDENT TRANSCRIPTION FACTORS, SRBB, FCR1, AND NDT80. OUR GOAL IS TO CHARACTERIZE THE WRPA REGULON IN A. FUMIGATUS AND TO IDENTIFY DOWNSTREAM EFFECTOR GENES WHOSE PRODUCTS MEDIATE PATHOGENICITY BY: 1) IDENTIFYING THE TRANSCRIPTION FACTORS THAT ARE DIRECTLY REGULATED BY WRPA AND DETERMINING THEIR ROLES IN PATHOGENICITY; 2) ANALYZING SELECTED WRPA- DEPENDENT TRANSCRIPTION FACTORS AND IDENTIFYING THEIR DOWNSTREAM TARGET GENES; AND 3) DETERMINING THE FUNCTION OF EFFECTOR GENES CONTROLLED BY THE WRPA REGULON AND INVESTIGATING THEIR ROLES IN VIRULENCE. THE RESULTS OF THE EXPERIMENTS DESCRIBED IN THIS PROPOSAL WILL ENABLE US TO CHARACTERIZE A KEY TRANSCRIPTIONAL REGULATOR THAT GOVERNS A. FUMIGATUS PATHOGENICITY AND THEN USE THIS INFORMATION TO IDENTIFY DOWNSTREAM EFFECTOR GENES, THE PRODUCTS OF WHICH MEDIATE HOST CELL INTERACTIONS AND VIRULENCE. THE RESULTS OF THIS WORK WILL NOT ONLY PROVIDE FOUNDATIONAL UNDERSTANDING OF A. FUMIGATUS VIRULENCE MECHANISMS, BUT ALSO HOLD PROMISE TO IDENTIFY NEW DIAGNOSTIC, THERAPEUTIC, AND VACCINE TARGETS.

TYPE I IFN SIGNALING DURING LUNG DEVELOPMENT IN DOWN SYNDROME - PROJECT SUMMARY/ABSTRACT DOWN SYNDROME (DS), ALSO REFERRED TO AS TRISOMY 21, IS THE MOST COMMON HUMAN CHROMOSOMAL ANOMALY, AFFECTING 1 IN 700 LIVE BIRTHS. ALTHOUGH DS CAN AFFECT MANY ORGAN SYSTEMS, LUNG AND HEART DISEASE ARE THE LEADING CAUSES OF MORBIDITY AND MORTALITY. SEVERAL CONGENITAL LUNG ANOMALIES ARE REPORTED IN INDIVIDUALS WITH DS INCLUDING AIRWAY BRANCHING DEFECTS, WITH A 25% DECREASE IN THE NUMBER OF BRANCHES AND REDUCED UPPER AIRWAY MUSCLE TONE WITH DYSPHAGIA AND/OR BRONCHOMALACIA. THESE COMPLICATIONS REMAIN CONSTANT INTO ADULTHOOD, AS OPPOSED TO BECOMING EXACERBATED, AND ARE HENCE LIKELY DUE TO DEVELOPMENTAL INSUFFICIENCY. WHILE ABNORMAL PULMONARY STRUCTURE AND FUNCTION IN PEDIATRIC AND ADULT DS SUBJECTS HAS BEEN DESCRIBED, THERE IS LIMITED DATA DEFINING THE ONTOGENY OF THESE ABNORMALITIES. WE POSTULATED THAT SOME DEVELOPMENTAL DIFFERENCES COULD INITIATE PRENATALLY. PRELIMINARY DATA DEVELOPED FOR THIS APPLICATION SHOWS THAT DS FETAL LUNGS, STARTING AS EARLY AS 16 WEEKS GESTATION, PRESENT WITH PRONOUNCED DILATATION OF TERMINAL AIRWAYS/ACINAR TUBULES, DILATED LYMPHATICS AND MUSCULARIZED ARTERIES. IN ADDITION, WE FIND INCREASED LUNG EXPRESSION OF TYPE I IFN SIGNALING TARGET GENES SUCH AS MX1 AND IFI27 IN DS COMPARED TO NON-DS FETAL LUNGS. IFN SIGNALING PLAYS A CRITICAL ROLE IN CELL DIFFERENTIATION, PROLIFERATION, APOPTOSIS, AND ECM PRODUCTION, IMPORTANT EVENTS FOR LUNG DEVELOPMENT. FINALLY, OUR PRELIMINARY DATA SHOW THAT DS LUNGS EXHIBIT ALTERED EXPRESSION OF ECM AFFILIATED PROTEINS, REGULATORS, AND SECRETED FACTORS (FN1, COL6, ETC.). THEREFORE, WE HYPOTHESIZE THAT LUNG DEFECTS IN DS CAN INITIATE DURING FETAL DEVELOPMENT, AND THAT IFN-DEPENDENT CHANGES IN CELL PROLIFERATION, DIFFERENTIATION AND ECM PRODUCTION CONTRIBUTE TO THESE DEFECTS. TO TEST THIS HYPOTHESIS, WE WILL 1) TEST THE HYPOTHESIS THAT MORPHOLOGICAL, CELLULAR AND MOLECULAR ABNORMALITIES ARE INITIATED DURING THE LATE PSEUDOGLANDULAR/EARLY CANALICULAR STAGES OF FETAL DEVELOPMENT IN DS LUNGS USING HISTOPATHOLOGICAL ANALYSES AND SINGLE CELL SEQUENCING, 2) TEST THE HYPOTHESIS THAT EXCESSIVE TYPE I IFN SIGNALING DISRUPTS CELL DIFFERENTIATION AND AIRWAY BRANCHING DURING FETAL LUNG DEVELOPMENT IN DS, AND 3) TEST THE HYPOTHESIS THAT EXCESSIVE IFN SIGNALING DISRUPTS ECM PRODUCTION DURING FETAL LUNG DEVELOPMENT IN DS. FOR AIMS 2 AND 3, WE WILL USE OUR PUBLISHED FETAL LUNG EXPLANTS CULTURE MODEL AS WELL AS EPITHELIAL ORGANOID CULTURES ALONE OR CO-CULTURED WITH MESENCHYMAL CELLS TO TEST THE EFFECT OF GAIN AND LOSS OF FUNCTION OF TYPE I IFN SIGNALING ON CELL DIFFERENTIATION AND ECM PRODUCTION. THESE STUDIES HAVE THE PROMISE TO IMPROVE OUR UNDERSTANDING OF THE KEY MOLECULAR AND CELLULAR DIFFERENCES, AND THE MECHANISMS CONTROLLING BRANCHING DEFECTS IN DS DEVELOPING LUNGS. THE INNOVATIVE ASPECTS OF THIS WORK ARE LIKELY TO HAVE GREAT OVERALL IMPACT AND FACILITATE THE TRANSLATIONAL POTENTIAL FOR THERAPIES FOR DS INDIVIDUALS, AS WELL AS OTHER LUNG CONGENITAL DEFECTS DEMONSTRATING HYPOPLASTIC LUNGS.

STAPHYLOCOCCAL ADAPTATIONS TO PLATELET MICROBICIDAL PROTEIN

CORONARY ARTERY CALCIUM, AORTIC CALCIFICATION AND DENSITY IN MESA

ULTRA-EARLY SURGERY FOR ACUTE SPINAL CORD INJURY: A PROSPECTIVE TRIAL OF TIMING OF SURGICAL DECOMPRESSION

C. ALBICANS INVASION AND PROLIFERATION DURING ORAL INFECTION

ENDOTHELIAL INVASION BY YEAST-PHASE CANDIDA

2/2-TREATMENT OF SUICIDAL AND SELF-INJURIOUS ADOLESCENTS WITH EMOTIONAL DYSREGULA

PROJECT TITLE WEARABLE MULTIPLEX BIOSENSORS TO MONITOR EXACERBATION RISK IN CHRONIC OBSTRUCTIVE PULMONARY DISEASE

ORAL COMMENSAL FUNGI AND STRUCTURAL IMMUNITY - PROJECT SUMMARY WHILE THE FUNGUS CANDIDA ALBICANS HAS LARGELY BEEN STUDIED AS A PATHOGEN, ITS PRIMARY LIFESTYLE IS AS A COMMENSAL ON MUCOSAL SURFACES, INCLUDING THE ORAL CAVITY. MICROORGANISMS THAT OCCUPY MUCOSAL SURFACES ARE CRITICAL FOR APPROPRIATELY TUNING IMMUNE RESPONSES TO ENSURE EFFICIENT RESPONSES TO INVADING PATHOGENS WHILE LIMITING RESPONSES DIRECTED TOWARDS HOST TISSUES. IN THIS CONTEXT, COMMENSAL FUNGI PLAY IMPORTANT ROLES IN HOST IMMUNITY AND INFLAMMATION. FOR INSTANCE, C. ALBICANS COLONIZATION INDUCES CROSS-REACTIVE T CELLS AND INNATE MEMORY IN IMMUNE CELLS, A MECHANISM TERMED TRAINED IMMUNITY. ACCUMULATING EVIDENCE FROM OUR GROUP AND OTHERS HAVE CONVINCINGLY DEMONSTRATED THAT ORAL COMMENSAL COLONIZATION WITH C. ALBICANS INDUCES SPECIFIC IMMUNE RESPONSES, THEREFORE CONTRIBUTING TO MUCOSAL IMMUNE SYSTEM PLASTICITY. HOWEVER, THE FUNDAMENTAL IMMUNOLOGICAL CONSEQUENCES OF ORAL C. ALBICANS COLONIZATION ARE STILL NOT WELL UNDERSTOOD. HOST TISSUES MAINTAIN TRACES OR MEMORY AFTER MICROORGANISM ENCOUNTER THAT EXTEND BEYOND ADAPTIVE RESPONSES OF T AND B CELLS. THESE CHANGES INCLUDE PROFOUND STRUCTURAL REMODELING AND EPIGENETIC ALTERATIONS. ‘STRUCTURAL IMMUNITY’, THE IMMUNE RESPONSE WITHIN A TISSUE FRAMEWORK CREATED BY STRUCTURAL CELLS, SUCH AS ENDOTHELIAL AND EPITHELIAL CELLS, IS ESSENTIAL FOR MAINTAINING TOLERANCE AND THE DEVELOPMENT OF APPROPRIATE PROTECTIVE AND CONTROLLED IMMUNE RESPONSES. IN PROBING FOR STRUCTURAL PROCESSES THAT MAINTAIN EFFECTIVE MUCOSAL IMMUNITY IN THE ORAL CAVITY, WE IDENTIFIED TWO MECHANISMS WHICH ARE TUNED DURING ORAL COMMENSAL C. ALBICANS COLONIZATION, ORAL EPITHELIAL PROLIFERATION AND ANGIOGENESIS. OUR CENTRAL HYPOTHESIS IS THAT COMMENSAL C. ALBICANS EXPOSURE IN THE ORAL CAVITY ESTABLISHES MUCOSAL ‘STRUCTURAL IMMUNITY’ BY INDUCING ORAL EPITHELIAL TRAINED IMMUNITY, EPITHELIAL REMODELING, AND ANGIOGENESIS. THESE TISSUE REMODELING AND INNATE PRIMING EVENTS SERVE TO MAINTAIN EFFECTIVE MUCOSAL IMMUNITY, THUS PREVENTING ORAL FUNGAL OUTGROWTH OF THIS IMPORTANT COMMENSAL ORGANISM. THIS PROPOSAL WILL EVALUATE THE MOLECULAR MECHANISMS BY WHICH ORAL COMMENSAL C. ALBICANS INDUCE EPITHELIAL PROLIFERATION (AIM 1), AND ANGIOGENESIS (AIM1 AND 2). IN AIM1 WE WILL DELINEATE THE ROLE OF IL-22 IN MEDIATING ORAL STRUCTURAL IMMUNITY IN RESPONSE TO COMMENSAL C. ALBICANS COLONIZATION. IN AIM 2 WE WILL DETERMINE THE ROLE OF VEGF AS A MEDIATOR OF TRAINED AND STRUCTURAL IMMUNITY IN THE ORAL CAVITY.

MECHANISMS CONTROLLING EARLY HUMAN LUNG DEVELOPMENT

TRANSCRIPTIONAL REGULATION OF A. FUMIGATUS VIRULENCE

MECHANISMS AND CIRCUMVENTION OF DAPTOMYCIN RESISTANCE IN STREPTOCOCCUS MITIS

C. ALBICANS-EPITHELIAL INTERACTIONS & OROPHARYNGEAL DISEASE

VACCINE STRATEGIES FOR DISSEMINATED CANDIDIASIS

NOVEL CARDIOPULMONARY EXERCISE TESTING VARIABLES TO DIFFERENTIATE NEUROMUSCULAR DECONDITIONING FROM DISEASE - PROJECT SUMMARY CARDIOPULMONARY EXERCISE TESTING (CPET) IS AN OBJECTIVE, NON-INVASIVE, MEASURE OF THE INTEGRATED FUNCTION OF THE PULMONARY, CARDIOVASCULAR AND NEUROMUSCULAR SYSTEMS. CPET EVALUATES SUBMAXIMAL AND PEAK EXERCISE RESPONSES, INFORMING ON CAUSES OF DYSPNEA AND/OR FATIGUE, DISEASE PROGNOSIS AND/OR PROGRESSION, PRE- OR POST- SURGICAL RISK STRATIFICATION, OR EXERCISE TRAINING PRESCRIPTION. CLINICAL USE OF CPET IS ESTIMATED TO HAVE INCREASED BY 81% BETWEEN 2005 AND 2015. THE MOST COMMON PROBLEM FOR CPET INTERPRETATION IS DISCRIMINATING BETWEEN CARDIOVASCULAR DISEASE AND NEUROMUSCULAR DECONDITIONING E.G., LOW MUSCLE MASS, WEAKNESS AND/OR FATIGABILITY. WE HAVE DEVELOPED A SOLUTION TO THIS PROBLEM, USING A MODIFIED CPET WITH INTEGRATED ISOKINETIC (IK) MEASUREMENTS OF MUSCLE POWER, WHICH INDEPENDENTLY ASSESSES NEUROMUSCULAR PERFORMANCE WITHOUT AFFECTING MEASUREMENT OF STANDARD CPET VARIABLES. THE MODIFIED CPET ADDS <10MIN TO THE STANDARD CPET AND USES COMMERCIALLY AVAILABLE EQUIPMENT. THE MODIFIED CPET PRODUCES 4 NEW IK VARIABLES: 1) BASELINE PEAK ISOKINETIC POWER: DECONDITIONED MUSCLES ARE WEAK; 2) TOLERANCE INDEX: THE FRACTION OF PEAK IK POWER AT VO2PEAK. AEROBIC DECONDITIONING REDUCES THE AVAILABLE PEAK IK POWER THAT CAN BE SUPPORTED BY AEROBIC METABOLISM; 3) FATIGUE INDEX: THE LOSS OF PEAK IK POWER FOR A GIVEN WORK RATE. AEROBIC DECONDITIONING INCREASES FATIGABILITY; 4) POWER RESERVE: THE CAPACITY FOR ACUTE POWER INCREASE AT VO2PEAK. A POWER RESERVE INDICATES THAT NEUROMUSCULAR PERFORMANCE IS NOT LIMITING TO EXERCISE TOLERANCE. THE MODIFIED CPET IS REPRODUCIBLE AND WELL TOLERATED BY ELDERLY, NORMAL SUBJECTS AND ATHLETES, AND THOSE WITH HEART FAILURE OR COPD. THE MODIFIED CPET CAN DIFFERENTIATE NEUROMUSCULAR DECONDITIONING AS A CAUSE OF EXERCISE LIMITATION FROM CARDIOVASCULAR OR PULMONARY LIMITATIONS (ASSESSED BY STANDARD CPET). TO INFORM CLINICAL INTERPRETATION, THIS PROJECT WILL DEVELOP PREDICTIVE MODELS DESCRIBING NORMAL REFERENCE VALUES FOR IK VARIABLES IN MEN AND WOMEN AGED 18-80YR. THRESHOLD VALUES OF NEUROMUSCULAR PERFORMANCE (WITH CORRESPONDING SENSITIVITY AND SPECIFICITY) WILL BE DEVELOPED AND OPTIMIZED TO DISCRIMINATE A DECONDITIONED POPULATION. VARIABLES THAT MODERATE NEUROMUSCULAR PERFORMANCE MEASURES ASSOCIATED WITH DECONDITIONING WILL BE IDENTIFIED (E.G. VO2PEAK, DIASTOLIC DYSFUNCTION, VASCULAR REACTIVITY, LEG LEAN MASS, LEG STRENGTH AND POWER, MUSCLE OXIDATIVE CAPACITY AND MUSCLE BIOPSY MORPHOLOGY). FINALLY, THE DISCRIMINATIVE ABILITY OF IK VARIABLES FOR DECONDITIONING WILL BE TESTED BY MEASURING SENSITIVITY OF IK VARIABLES IN RESPONSE TO EXERCISE TRAINING IN DECONDITIONED INDIVIDUALS AND THE SPECIFICITY OF IK VARIABLES TO DISCRIMINATE PERIPHERAL VS. CENTRAL HEMODYNAMIC EXERCISE LIMITATIONS USING INVASIVE CPET IN PATIENTS WITH HEART FAILURE AND PRESERVED EJECTION FRACTION. THIS STUDY WILL TRANSFORM THE UTILITY AND DIAGNOSTIC CAPABILITIES OF CPET AND IMPROVE CLINICAL DECISION-MAKING AND SEVERITY STRATIFICATION ACROSS A WIDE RANGE OF CHRONIC DISEASE STATES WHERE DECONDITIONING IS A COMMON FEATURE.

DECIPHERING FUNCTIONS OF THE ATR-METTL3-BRCA1 AXIS IN GENOME INSTABILITY AND TUMORIGENESIS - PROJECT SUMMARY /ABSTRACT A DEFECT IN DNA DAMAGE REPAIR TRIGGERS GENOME INSTABILITY AS A HALLMARK OF CANCER THAT IS EXPLOITED BY TREATMENTS SUCH AS IONIZING RADIATION, PLATINUM CHEMOTHERAPY AND POLY (ADP-RIBOSE) POLYMERASE INHIBITORS (PARPI). PATHOGENIC GERMLINE AND SOMATIC VARIANTS IN THE DNA REPAIR GENE BRCA1 ARE FREQUENTLY DETECTED IN TRIPLE NEGATIVE BREAST CANCER (TNBC) AND BRCA1-MUTATED TNBC CAN BE TARGETED WITH PARPI TO ACHIEVE A CLINICAL RESPONSE. ELUCIDATING THE MECHANISM(S) BY WHICH BRCA1 AND ITS ASSOCIATED PARTNERS RESOLVE DNA LESIONS WILL PROVIDE NEW INSIGHTS INTO HOW GENOME INSTABILITY PROMOTES CELLULAR TRANSFORMATION AND IDENTIFY NEW THERAPEUTIC TARGETS. IN THIS PROPOSAL, WE WILL INVESTIGATE FUNCTIONS OF THE ATR-METTL3-BRCA1 AXIS IN R-LOOP ASSOCIATED DNA DAMAGE TO SUSTAIN STABLE GENOME. PERSISTENT AND UNSCHEDULED R-LOOPS CREATE STRUCTURAL BARRIERS WHEN THEY COLLIDE WITH COLLAPSED REPLICATION FORKS OR DOUBLE STRAND BREAKS (DSB) TO ELICIT GENOME INSTABILITY. OUR PRELIMINARY DATA FOUND THAT METTL3, A PROTEIN PROMOTING RNA M6A MODIFICATION WHEN R-LOOPS FORM DURING TRANSCRIPTION, IS RECRUITED TO REPAIR DNA LESIONS AT ACTIVE TRANSCRIPTION SITES. METTL3 IS PHOSPHORYLATED BY ATR, A KEY KINASE IN R-LOOP ASSOCIATED DNA DAMAGE SIGNALING, AND THIS PHOSPHORYLATION PROMOTES ASSOCIATION WITH BRCA1. WE FOUND THAT BRCA1 IS BOTH NECESSARY AND SUFFICIENT FOR METTL3 RECRUITMENT TO DNA DAMAGE SITES. WE IDENTIFIED SEVERAL BREAST CANCER ASSOCIATED METTL3 MUTATIONS AND FOUND THAT TWO CATALYTICALLY INACTIVE MUTATIONS IMPEDE HR REPAIR BY ABOLISHING THE ASSOCIATION WITH BRCA1. OUR PROTEOMIC ANALYSIS HAS IDENTIFIED A DNA/RNA HELICASE THAT RESOLVES R-LOOPS AS THE EFFECTOR OF THE ATR-METTL3-BRCA1 AXIS. FURTHERMORE, INHIBITION OF METTL3 CATALYTIC ACTIVITY WITH A SMALL MOLECULE INDUCES A DEFECT IN HOMOLOGOUS RECOMBINATION AND ENHANCES PARPI SENSITIVITY IN TNBC CELL LINES. WE HYPOTHESIZE THAT THE ATR-METTL3-BRCA1 AXIS IS CRITICAL FOR THE R-LOOP- ASSOCIATED DNA DAMAGE REPAIR AND BREAST CANCER DEVELOPMENT AND OFFERS A NEW THERAPEUTIC TARGET FOR TNBC. WE WILL TEST THIS HYPOTHESIS IN THE FOLLOWING SPECIFIC AIMS. IN AIM 1, WE WILL ELUCIDATE MECHANISMS BY WHICH METTL3 SERINE 43 PHOSPHORYLATION PROMOTES DNA/RNA HELICASE RECRUITMENT TO RESOLVE R-LOOP ASSOCIATED DNA DAMAGE. IN AIM 2, WE WILL DETERMINE THE ROLES OF THE ATR-METTL3-BRCA1 AXIS IN BREAST CANCER DEVELOPMENT WITH MOUSE MODELS. IN AIM 3, WE WILL TARGET THE ATR-METTL3-BRCA1 AXIS WITH A METTL3 INHIBITOR IN TNBC. OUR STUDIES WILL UNRAVEL A NOVEL FUNCTION FOR METTL3 IN DNA REPAIR AND IDENTIFY METTL3 AS A PROMISING THERAPEUTIC TARGET IN TNBC.

THE ROLE OF PURINE BIOSYNTHESIS AND STRINGENT RESPONSE IN PERSISTENT MRSA ENDOVASCULAR INFECTIONS

HORMONAL MECHANISMS OF SLEEP RESTRICTION

E-CIGARETTE VAPING DURING PREGNANCY AND LACTATION, GERM CELL EPIGENETIC MEMORY, AND TRANSGENERATIONAL ASTHMA

ADD FUNDING IN THE AMOUNT OF 657257.81

MATERNAL OBESITY PROGRAMS OFFSPRING HYPOTHALAMIC NEUROGENESIS AND APPETITE: MECHANISMS AND PREVENTION OF HYPERPHAGIA-MEDIATED CHILDHOOD OBESITY

TRYPTOPHAN METABOLISM AND ITS ROLE IN FIBROID PATHOGENESIS - ABSTRACT IN THE COURSE OF OUR PROFILING FOR NON-CODING RNAS IN FIBROIDS WE DISCOVERED HIGHLY ABERRANT OVEREXPRESSION OF TRYPTOPHAN 2,3 DIOXYGENASE (TDO2) AND INDOLEAMINE 2,3-DIOXYGENASE (IDO1) IN FIBROIDS. WE CONFIRMED THIS FINDING BY BOTH QRT-PCR AND WESTERN BLOT ANALYSIS, AND FOUND A CONSISTENTLY ELEVATED EXPRESSION OF TDO2 AND MORE VARIABLE OVEREXPRESSION OF IDO1 IN OUR TISSUE SPECIMENS. THE INCREMENT IN TDO2 EXPRESSION IN FIBROIDS WAS HIGHER AS COMPARED TO IDO1, AND THE EXPRESSION OF IDO2 WAS BARELY DETECTED. RELEVANT TO THE PATHOGENESIS OF FIBROIDS WAS THAT THE INCREMENT IN TDO2 BUT NOT IDO1 WAS RACE DEPENDENT, WITH THE INCREMENT BEING SIGNIFICANTLY HIGHER IN AFRICAN AMERICANS AS COMPARED WITH CAUCASIANS. FURTHERMORE, THE EXPRESSION OF TDO2 WAS SIGNIFICANTLY INCREASED IN MED12 MUTATION BEARING LEIOMYOMAS, AND ITS INHIBITION BY PHARMACOLOGIC BLOCKADE IN VITRO LED TO DECREASED PROLIFERATION AND EXPRESSION OF GENES RELATED TO EXTRACELLULAR MATRIX, INFLAMMATION AND CELL GROWTH IN LEIOMYOMA SMOOTH MUSCLE CELLS (LSMC) SPHEROIDS. ABERRANT EXPRESSION OF THESE ENZYMES IN FIBROIDS RESULTED IN INCREASED LEVELS OF KYNURENINE (KYN), A METABOLIC BYPRODUCT OF TRYPTOPHAN DEGRADATION AND A KNOWN ENDOGENOUS LIGAND FOR ARYL HYDROCARBON RECEPTOR (AHR). BASED ON THIS PRELIMINARY DATA WE HYPOTHESIZED THAT DYSREGULATION OF TRP METABOLISM AS CHARACTERIZED BY MARKED OVEREXPRESSION OF TDO2 IS FUNDAMENTAL TO THE PATHOGENESIS OF FIBROIDS AND CORRECTION OF TRP METABOLIC DYSREGULATION BY INHIBITION OF TDO2 AND NORMALIZATION OF KYNURENINE LEVELS WILL INHIBIT FIBROID GROWTH AND PROGRESSION AND POTENTIALLY TUMOR ESTABLISHMENT. WE PROPOSE TO THIS HYPOTHESIS IN 3 AIMS. IN AIM1 WE WILL CHARACTERIZE TRP METABOLISM IN MYOMETRIUM AND FIBROID TUMORS AND EXPLANTS, AND DETERMINE THE MECHANISM(S) UNDERLYING THE MARKED OVEREXPRESSION OF TDO2 IN FIBROIDS USING AN IN VITRO APPROACH. IN AIM 2 WE WILL EXAMINE THE IMPACT OF KYNURENINE AND ITS ACTIVATION OF AHR ON DOWNSTREAM GENES REGULATING EXTRACELLULAR MATRIX (ECM), INFLAMMATION AND CELL PROLIFERATION. AIM 3 IS DESIGNED TO DETERMINE THE UTILITY OF A PHARMACOLOGICAL INHIBITOR OF TDO2 AND WITH LENTIVIRUS BEARING SHRNA TO KNOCK DOWN TDO2 ON FIBROID ESTABLISHMENT AND PROGRESSION IN IN VIVO MOUSE MODELS OF FIBROIDS. THIS NOVEL METABOLIC MECHANISM FOR FIBROID PATHOGENESIS HAS GREAT TRANSLATIONAL SIGNIFICANCE AS IT OPENS THE WAY FOR NOVEL THERAPIES AIMED AT CORRECTION OF TRYPTOPHAN METABOLISM IN FIBROIDS.

DEFINING CEREBELLAR PATHOPHYSIOLOGY IN ATAXIA TELANGIECTASIA - PROJECT SUMMARY ATAXIA TELANGIECTASIA (A-T) IS A RARE (~1 IN EVERY 100,000 LIVE BIRTHS) BUT CATASTROPHIC DISEASE THAT CAUSES PREMATURE DEATH BETWEEN THE AGES OF 10 AND 30 YEARS. THERE IS NO CURE OR DISEASE ALTERING THERAPY AVAILABLE. IT IS CHARACTERIZED BY A PROGRESSIVE AND SEVERE LOSS OF MOTOR COORDINATION (ATAXIA) AND CEREBELLAR NEURODEGENERATION. IMMUNE DEFICIENCY AND CANCER ARE ALSO PREVALENT SYMPTOMS. IN 1995, DEFICIENCY IN THE A-T MUTATED (ATM) PROTEIN WAS IDENTIFIED AS THE UNDERLYING CAUSE OF A-T. HOWEVER, RESEARCHERS HAVE YET TO DETERMINE HOW A DNA REPAIR PATHWAY PROTEIN LIKE ATM PREFERENTIALLY INDUCES CEREBELLAR PATHOLOGY AND ATAXIA. THIS KNOWLEDGE GAP ARISES FROM THE LACK OF A SUITABLE ANIMAL RESEARCH MODEL THAT RECAPITULATES THE CLINICAL ATAXIA AND CEREBELLAR PATHOLOGY—MORE THAN SIX DIFFERENT ATM DEFICIENT MOUSE VARIANTS HAVE BEEN GENERATED BUT ALL FAIL TO DEVELOP A CLINICAL PHENOTYPE. WE RECENTLY OVERCAME THIS ROADBLOCK BY ENHANCING THE RATE OF GENOTOXIC STRESS IN THE MOUSE USING A GENETIC DOUBLE-HIT STRATEGY. OUR NOVEL DOUBLE KNOCKOUT (DKO) MOUSE LACKS EXPRESSION OF BOTH ATM AND APRATAXIN (APTX), A RELATED DNA REPAIR GENE. WE FIND THE DKO MICE DEVELOP A CLINICAL PHENOTYPE, INCLUDING PROGRESSIVE AND SEVERE ATAXIA ALONG WITH CEREBELLAR NEURODEGENERATION THAT IS NOT PRESENT IN EITHER THE ATM OR APTX SINGLE KNOCKOUTS. IN ATAXIC DKO MICE, WE FIND THAT A SUBSET OF GENES CONTROLLING PURKINJE NEURON FUNCTION ARE PREFERENTIALLY DOWNREGULATED (E.G., ITPR1 AND CAR8) DUE A DISTINCT CHROMATIN ARCHITECTURE THAT LEAVES THEM PREFERENTIALLY OPEN TO DNA DAMAGE. CONSISTENT WITH THIS, OUR INITIAL EXAMINATION SHOWS THAT PURKINJE NEURONS IN DKO MICE DISPLAY SIGNIFICANT SIGNS OF PHYSIOLOGICAL AND MORPHOLOGICAL ABNORMALITIES. WE THEREFORE HYPOTHESIZE THAT PROGRESSIVE ATAXIA IN A-T RESULTS FROM DNA DAMAGE INDUCED DOWNREGULATION OF GENES THAT CONTROL PN PHYSIOLOGY LEADING TO INCREASED PN DYSFUNCTION AND ABNORMAL CEREBELLAR CIRCUIT ACTIVITY AND OUTPUT TO MOTOR CENTERS. IN THREE AIMS, WE PROPOSE EXPERIMENTS TO ESTABLISH A TEMPORAL LINK BETWEEN ATAXIA SEVERITY AND THE MAGNITUDE OF PROTEIN DOWNREGULATION, PN PATHOPHYSIOLOGY, AND ABNORMAL CEREBELLAR CIRCUIT ACTIVITY AT THE MOLECULAR, CELLULAR, CIRCUIT, AND SYSTEM LEVELS. IN AIM 1, WE WILL CORRELATE THE TEMPORAL PROGRESSION (EARLY TO LATE) OF INCREASING ATAXIA TO A SPECIFIC SET OF PURKINJE NEURON PATHOLOGIES USING IN VITRO ELECTROPHYSIOLOGICAL, CA2+ IMAGING, HISTOLOGY, AND MEASUREMENTS OF GENE EXPRESSION. IN AIM 2, WE WILL ESTABLISH THE PATHOGENIC NEURAL CIRCUIT ACTIVITY CHANGES THAT ARISE IN A-T BOTH WITHIN THE CEREBELLUM AND ITS DOWNSTREAM PROJECTIONS (E.G., MOTOR THALAMUS) USING A CUTTING-EDGE, IN VIVO MULTIELECTRODE ARRAY RECORDING TECHNIQUE IN AWAKE BEHAVING MICE. FINALLY, IN AIM 3, WE WILL ASSESS THE PIVOTAL ROLE PURKINJE NEURONS PLAY IN A-T ETIOLOGY BY TESTING WHETHER LOSS OF ATM EXPRESSION ALONE IN THESE CELLS IS SUFFICIENT TO GENERATE ATAXIA AND CEREBELLAR DEFECTS. MOREOVER, IN AIM 3, WE WILL TEST IF SELECTIVE CHEMOGENETIC AND PHARMACOLOGICAL MODULATION OF PATHOLOGICAL PURKINJE NEURONS CAN ALLEVIATE ATAXIA. RESULTS WILL PROVIDE NEW THERAPEUTIC TARGETS AND A BROAD UNDERSTANDING OF HOW ATM DEFICIENCY CAN ALTER CEREBELLAR STRUCTURE/FUNCTION AND CIRCUIT TO CAUSE A RELENTLESS ATAXIA.

EPITRANSCRIPTOMIC REGULATION OF SPERMATOGENESIS AND MALE FERTILITY

BICARBONATE-MEDIATED ENHANCEMENT OF BETA-LACTAM-MRSA KILLING: MECHANISMS AND CLINICAL TRANSLATABILITY

CELLULAR SENESCENCE IN TRISOMY 21 LUNG DISEASE - SUMMARY TRISOMY 21 (T21) OCCURS IN ~1 IN 700 LIVE BIRTHS RESULTING IN DOWN SYNDROME (DS), WHERE PULMONARY COMPLICATIONS ARE THE MAIN CAUSE OF HOSPITALIZATIONS IN CHILDREN AND ADULTS. THESE INCLUDE TRACHEOBRONCHIAL DISEASES PRESENTING AS DECREASED UPPER AIRWAY MUSCLE TONE, TRACHEOBRONCHIAL MALACIA AND AIRWAY OBSTRUCTION WITH WHEEZING, PARTICULARLY IN BABIES AND CHILDREN. THERE IS CURRENTLY LIMITED TO NO INFORMATION REGARDING THE PATHOGENESIS OF PRIMARY LUNG DISEASE IN DS. THUS, UNDERSTANDING EARLY, PERINATAL FACTORS LEADING TO LUNG DISEASE IN NEONATES AND CHILDREN WITH DS BECOMES CLINICALLY RELEVANT AND INNOVATIVE WITH POTENTIAL FOR SUBSTANTIAL LIFELONG IMPACT. SENESCENT (SEN) CELLS IN G1 CELL CYCLE ARREST SECRETE FACTORS (SEN-ASSOCIATED SECRETORY PHENOTYPE; SASP) THAT MEDIATE PARACRINE EFFECTS ON NAÏVE CELLS. SEN CAN BE INDUCED BY DNA DAMAGE, OXIDATIVE STRESS AND MITOCHONDRIAL DYSFUNCTION, ALL OF WHICH ARE INCREASED IN T21. WHILE SEN CAN BE BENEFICIAL IN GROWTH AND REPAIR, IT BECOMES DETRIMENTAL WHEN SEN CELL NUMBERS EXCEED IMMUNE CLEARANCE (DEFECTIVE IN T21), AND SECRETE A PRO- INFLAMMATORY, PRO-FIBROTIC SASP THAT MAINTAINS AND EXPANDS DETRIMENTAL SEN. MAINTENANCE OF SASP, KNOWN AS LATE SEN, IS MODULATED BY IFN-I SIGNALING, A PATHWAY CONSISTENTLY ACTIVATED IN T21. THUS, IF DS IS SEEN AS A FORM OF PROGERIA (EARLY AGING) INVOLVING SEN, THERE IS STRONG PREMISE TO LINK SEN, IFN-I AND T21. THESE RELATIONSHIPS ARE ENTIRELY UNKNOWN IN THE DS LUNG, ESPECIALLY IN PRENATAL/EARLY POSTNATAL LIFE WHEN THE POTENTIAL FOR IMPACTFUL INTERVENTION IS HIGH. HERE, DRUGS THAT KILL SEN CELLS (SENOLYTICS) SUCH AS DASATINIB + QUERCETIN (D+Q), OR FISETIN (FLAVONOID IN STRAWBERRIES), OR EVEN SASP MODULATORS (SENOMORPHICS) REPRESENT NOVEL THERAPIES THAT ARE ONLY RECENTLY BEING TESTED IN LUNG DISEASES. OUR KEY PRELIMINARY DATA SHOW INCREASED BASELINE SEN IN T21 FETAL LUNG TISSUES AND MESENCHYMAL CELLS (FLMC). WE HYPOTHESIZE THAT DETRIMENTAL SEN/SASP PRESENT IN T21 INITIATES IN UTERO AND PROMOTES LUNG DISEASE IN DS; EFFECTS ALLEVIATED BY SENOLYTICS AND SENOMORPHICS. OUR APPROACH WILL BE TO USE DEVELOPING HUMAN LUNG TISSUES, MESENCHYMAL CELLS AND EX VIVO EXPLANT MODELS FROM T21 AND NON-T21 FETAL, NEONATAL AND PEDIATRIC SAMPLES TO DETERMINE THE IMPACT OF SEN IN T21, THE INTERPLAY BETWEEN T21 AND IFN- I AND THE POSSIBLE EFFECTS OF SENOLYTICS AND SENOMORPHICS TOWARDS THERAPIES FOR LUNG DISEASE IN DS. OUR PROPOSAL REPRESENTS A NOVEL APPROACH TO UNDERSTANDING AND TREATING LUNG DISEASE IN A HIGHLY VULNERABLE POPULATION WHERE PULMONARY DISORDERS REPRESENT A MAJOR HEALTHCARE ISSUE, YET WITH LITTLE TO NO INFORMATION REGARDING DISEASE PATHOGENESIS.

DEVELOPMENTAL NICOTINE EXPOSURE & TRANSGENERATIONALLY ALTERED LUNG PHENOTYPE

SPATIOTEMPORAL REGULATION OF HUMAN ISLET ORGANOGENESIS - SUMMARY HUMAN PANCREATIC ISLETS ARE THE KEY REGULATOR OF WHOLE-BODY LIPID AND GLUCOSE HOMEOSTASIS. THE DYSFUNCTION OF HORMONE SECRETION FROM ISLETS, AND/OR LOSS OF Β CELL MASS IS AN IMPORTANT PART OF THE PATHOGENESIS OF DIABETES, OFTEN REQUIRING INSULIN OR ISLET REPLACEMENT. HUMAN INDUCED PLURIPOTENT STEM CELLS (HIPSCS) PROVIDE A POTENTIAL ALTERNATIVE SOURCE TO CADAVERIC HUMAN PANCREATIC ISLETS, BUT THE GENERATED ISLETS FROM HIPSCS IN VITRO COMMONLY SHOWS FUNCTIONAL HETEROGENEITY (BATCH-TO-BATCH DIFFERENCE) AND LIMITED SCALABILITY. IN ADDITION, EVEN WITH AUTOLOGOUS DELIVERY STRATEGIES, HIPSC DERIVED ISLETS (HIPSC-ISLETS) STILL REQUIRE LIFE-LONG IMMUNE SUPPRESSION BECAUSE OF A HYPER-ACTIVE IMMUNE REACTION ESPECIALLY IN TYPE 1 DIABETES (T1D) PATIENTS. THEREFORE, UNIVERSAL HUMAN PSCS THAT EVADE IMMUNE DETECTION CONSTITUTE A MAJOR GOAL OF TRANSLATIONAL RESEARCH IN T1D RESEARCH. OUR RESEARCH PROPOSAL FOCUSES ON INVESTIGATING METHODS FOR THE FUNCTIONAL SELECTION, MASS PRODUCTION, AND IMMUNE PROTECTION OF HIPSC-ISLETS. THE RATIONALE IS THAT IDENTIFYING SUCH A PATHWAY IS NECESSARY FOR LARGE- SCALE ISLET CELL THERAPY FOR DIABETES. OUR PRELIMINARY RESULTS REVEALED THAT 1) THE MINERAL ABSORPTION FXYD2 PATHWAY IS A NOVEL HALLMARK OF FUNCTIONAL HETEROGENEITY OF HIPSC-ISLETS. 2) NEWLY DEVELOPED GIANT ISLET (GISLETS) TECHNOLOGY PROVIDES BETTER SCALABILITY THAN TRADITIONAL HIPSC-ISLETS. 3) PD-L1 PROVIDES IMMUNE EVASIVE FUNCTION IN HIPSC-ISLETS. WE HYPOTHESIZE THAT FXYD2 ENRICHED GISLETS WITH PD-L1 EXPRESSION PROVIDES LONG-TERM VIABILITY AND FUNCTIONALITY IN IMMUNE COMPETENT DIABETIC CONDITION. THIS HYPOTHESIS WILL BE TESTED WITH THE FOLLOWING SPECIFIC AIMS. AIM 1: IDENTIFICATION OF FXYD2 AS FUNCTIONAL STEM CELL DERIVED Β CELL MARKER. USING A NEWLY DEVELOPED THREE-DIMENSIONAL PROTOCOL TO MIMIC HUMAN ISLET ORGANOGENESIS, WE WILL TEST THE ROLE OF FXYD2 IN ISLET FUNCTIONALITY, A CORE MINERAL ABSORPTION PATHWAY USING GENETIC AND PHYSIOLOGICAL APPROACHES. AIM 2: EXPLORE THE IN VIVO EFFICACY OF FXYD2 ENRICHED GISLETS. WE HYPOTHESIZED THAT FXYD2 CAN BE SELECTION MARKER OF FUNCTIONAL GISLETS. WE WILL ISOLATE FXYD2 ENRICHED GISLETS (FGISLETS) BY FXYD2 PROMOTER DRIVEN REPORTER SYSTEM AND WILL EXPLORE THE EFFICACY FOR GLYCEMIC CONTROL OF FGISLETS IN T1D MODEL MICE. AIM 3: EXPLORE THE LONG-TERM EFFICACY OF IMMUNE EVASIVE GISLETS IN ALLOGENIC AND AUTOIMMUNE REACTION. TO INVESTIGATE WHETHER OVEREXPRESSION OF PD-L1 WITH HLA-A&B DELETION WILL LIMIT THE IMMUNE RESPONSE OF FGISLETS UPON TRANSPLANTATION AND PROLONG SURVIVAL IN HUMAN PATIENTS, WE WILL USE HUMANIZED CD34+ MICE (CD34+-NGS-SGM3, JACKSON LABORATORY) AS AN IN VIVO GRAFT REJECTION MODEL. THE SIGNIFICANCE OF THE PROPOSED STUDY IS IN ITS FOCUS ON THE MECHANISMS INVOLVED IN HUMAN ISLET MATURATION, TISSUE ENGINEERING OF PRACTICAL SCALING OF FUNCTIONAL INSULIN PRODUCING CELLS BY GISLETS. THE SUCCESS OF THIS STUDY WILL ENABLE US TO CREATE MORE ROBUST, REPRODUCIBLE, AND FUNCTIONAL MULTI-CELLULAR ISLET-ORGANOIDS FROM HIPSCS. THIS RELIES ON OUR INNOVATIVE CONCEPTS OF FUNCTIONAL SELECTION, MASS PRODUCTION AND IMMUNE PROTECTION FOR CELL THERAPY IN DIABETES.

ROLE OF CTCF IN EGF-INDUCED CORNEAL EPITHELIAL GROWTH

ACTIVATED TARGETED KILLER (ATAK) CELLS FOR INVASIVE FUNGAL INFECTIONS

CANDIDA INVASION OF ENDOTHELIUM AND VIRULENCE

MECHANISM OF LONG NON-CODING RNAS ACTION IN LEIOMYOMA

IMMUNOMODULATION IN MELANOMA THERAPY

RACIAL AND ETHNIC APPROACHES TO COMMUNITY HEALTH US

FIBROBLAST CELL SIGNALING IN UTERO NICOTINE-INDUCED LUNG INJURY

DETERMINANTS OF 5 YEAR PROGRESSION OF MUSCLE DYSFUNCTION AND INACTIVITY IN COPDGENE PARTICIPANTS.

DIFFERENTIAL EFFECTS OF CORNEAL HYPOXIA ON LIMBAL STEM AND EPITHELIAL CELL FATES

NEUROIMAGING AND NEUROPATHOLOGY OF MUCOPOLYSACCHARIDOSIS I

TARGETING THE REGULATORY MECHANISM OF HYPHAE TO LATERAL YEAST GROWTH AS A NOVEL THERAPEUTIC APPROACH AGAINST CANDIDIASIS

MECHANISMS OF PROGRAMMED GESTATIONAL HYPERPHAGIA

MECHANISMS OF ALS VACCINE PROTECTION AGAINST S. AUREUS

PULMONARY NEUROIMMUNE DETERMINANTS OF IMMUNOGLOBULIN PRODUCTION AGAINST PNEUMONIA - PROJECT SUMMARY. COMMUNITY ACQUIRED PNEUMONIA CASES ARE PREDOMINANTLY CAUSED BY INFECTIONS FROM STREPTOCOCCUS PNEUMONIAE. EVIDENCE SHOWS THAT EFFECTIVE ADAPTIVE IMMUNITY IN THE FORM OF ADEQUATE ANTIBODY PRODUCTION IS NECESSARY TO PROTECT AGAINST PULMONARY S. PNEUMONIAE INFECTION. EMERGING EVIDENCE DEMONSTRATES THAT THE SENSORY NERVOUS SYSTEM REGULATES INNATE AND CELLULAR IMMUNITY. OUR DATA AND OTHERS SHOW THAT SENSORY NEURONS ARE NECESSARY FOR ANTIBODY PRODUCTION. YET, THE NEURONAL INVOLVEMENT IN ADAPTIVE AND HUMORAL IMMUNITY IS UNDER-INVESTIGATED. THEREFORE, THERE IS AN URGENT NEED TO UNDERSTAND THE NEURAL REGULATION OF HUMORAL IMMUNITY. THIS APPLICATION AIMS TO INVESTIGATE THE ROLE OF SENSORY NEURONS IN B CELL MATURATION, IMMUNOGLOBULIN PRODUCTION INCLUDING CLASS SWITCH RECOMBINATION, AND INTERACTION WITH NEUTROPHILS IN RESPONSE TO PULMONARY S. PNEUMONIAE INFECTION. OUR PRELIMINARY DATA HAVE LED US TO HYPOTHESIZE THAT IN RESPONSE TO PULMONARY S. PNEUMONIAE INFECTION, SENSORY NEURONS RELEASE SELECT NEUROPEPTIDES TO ACTIVATE B CELL IMMUNOGLOBULIN RELEASE, TO INTERACT WITH NEUTROPHILS TO CLEAR INFECTION. SECONDLY, WE HYPOTHESIZE THAT THIS DOWNSTREAM EFFECT RESULTS FROM SENSORY NEURONS BEING ABLE TO DETECT SPECIFIC ANTIGENS. WE SHOW THAT 1) SENSORY NEURON NEUROPEPTIDE RELEASE IS DIRECTLY INVOLVED IN MATURING AND STIMULATING B CELLS IN THE LUNGS; 2) SENSORY NEUROPEPTIDES STIMULATE IMMUNOGLOBULIN PRODUCTION IN VITRO AND IN VIVO; 3) SENSORY NEURONS APPEAR TO ASSIST WITH B CELL-NEUTROPHIL-INTERACTION TO ASSIST WITH BACTERIAL CLEARANCE. WITH REGARDS TO NEURONAL ANTIGEN SENSING, WE SHOW THAT 1) SENSORY NEURONS HARBOR RNA REQUIRED FOR ANTIGEN SENSING; 2) ANTIGEN RECEPTOR PROTEINS ARE UPREGULATED IN SENSORY NEURONS WITH REPEAT S. PNEUMONIAE EXPOSURE; 3) SENSORY NEURON STIMULATION WITH ANTIGEN INCREASES NEURONAL EXCITATION AND SELECT NEUROPEPTIDE RELEASE WHICH HAS IMMUNOGENIC PROPERTIES AND; 4) NEURONS SEQUESTER IMMUNOGLOBULINS FROM B CELLS. WE WILL INVESTIGATE THIS NOVEL NEURONAL CIRCUIT OF B CELL IMMUNOGLOBULIN PRODUCTION AND NEUTROPHIL INTERACTION WITH TWO AIMS. STUDIES IN AIM 1 WILL INVESTIGATE B CELL DEVELOPMENT AND MATURATION USING FLOW CYTOMETRY, CLASS-SWITCH RECOMBINATION UTILIZING FLOW CYTOMETRY AND TANDEM SINGLE-CELL RNASEQ AND V(D)J REPERTOIRE ANALYSIS, AND NEUTROPHIL INTERACTION USING IN VIVO AND IN VITRO ASSAYS, INCLUDING SINGLE-CELL RNASEQ AND FLOW CYTOMETRY. STUDIES IN AIM 2 WILL USE ELECTROPHYSIOLOGICAL RECORDING, CALCIUM IMAGING, AND FLOW CYTOMETRY TO ASSESS A NOVEL SENSORY NEURON ANTIGEN RECOGNITION SIGNALING PATHWAY AND WHAT THIS STIMULATION MEANS REGARDING NEUROPEPTIDE RELEASE. SUBSEQUENT MOLECULAR ASSAYS WILL DELINEATE THE EXACT DOWNSTREAM EFFECTOR MOLECULE TO IDENTIFY POTENTIAL DRUG TARGETS FOR FUTURE STUDY. FINALLY, TARGETED ANTIGEN RECEPTOR DELETION IN SENSORY NEURONS WITH GENETIC MODELS WILL ESTABLISH THE NECESSITY OF NEURONAL ANTIGEN SENSING TO CONTRIBUTE TO HUMORAL IMMUNITY. DISEASE SEVERITY IN ALL AIMS WILL BE ASSESSED BY ENUMERATION OF BACTERIAL BURDEN AND IMMUNOGLOBULIN ANALYSIS. THIS CONTRIBUTION IS SIGNIFICANT BECAUSE IT IS EXPECTED TO ELUCIDATE THE ROLE OF SENSORY NEURONS IN PULMONARY ADAPTIVE IMMUNITY.

ARF6 INHIBITION AS NOVEL TREATMENT FOR MULTIDRUG RESISTANCE GRAM NEGATIVE INFECTIONS

PROGRAMMED ADIPOGENESIS AND LIPID DYSREGULATION

MECHANISMS OF ENVIRONMENTAL STRESS AFFECTING CORNEAL EPITHELIAL WOUND HEALING

SYNTHETIC LUNG SURFACTANT OPTIMIZED FOR BIOMEDICAL APPLICATION

SYNTHETIC SURFACTANT AND TOXIC CHEMICAL LUNG INJURY

MITIGATING RESISTANCE & VIRULENCE IN MRSA

ENDOCRINOLOGY, METABOLISM AND NUTRITION TRAINING GRANT

FACULTY DEVELOPMENT IN PRIMARY CARE

VENTRICULAR SIZE AND VALVE CALCIFICATION MEASURES BY CT

INTRATHECAL ENZYME THERAPY FOR MUCOPOLYSACCHARIDOSIS I

ALCOHOLIC HEPATITIS PATHOGENESIS AS DETERMINED FROM HUMAN LIVER TISSUE ANALYSIS

MICROBICIDAL PROTEINS FROM PLATELETS

USING HTS TO IDENTIFY INHIBITORS OF R155H-P97/VCP MUTANT TO TREAT IBMPFD/ALS

SCREENING FOR JERVELL AND LANGE-NIELSEN SYNDROME

LIPID PEROXIDATION IN ALCOHOLIC LIVER DISEASE

TRAJECTORIES OF ADHERENCE TO CARDIOVASCULAR MEDICATIONS IN PATIENTS ON DIALYSIS

EXAMINING RACIAL AND CARDIOVASCULAR PARADOXES IN CHRONIC KIDNEY DISEASE

NON-INVASIVE DETECTION OF CORONARY INFLAMMATION USING COMPUTED TOMOGRAPHY IN THE MULTI-ETHNIC STUDY OF ATHEROSCLEROSIS - IN THE CURRENT HEALTHCARE ENVIRONMENT, IMPROVING YIELD FROM PERFORMED TESTS TO OBTAIN INCREMENTAL INFORMATION WILL RESULT IN REDUCING COSTS BY BETTER RISK STRATIFYING PATIENTS FOR ATHEROSCLEROTIC CARDIOVASCULAR DISEASE (ASCVD) RISK IS A LAUDABLE GOAL. THE INTERPLAY BETWEEN EPICARDIAL FAT DENSITY AND PERICORONARY FAT ATTENUATION AND ASCVD EVENTS HAS NOT BEEN PREVIOUSLY STUDIED LONGITUDINALLY IN A PRIMARY PREVENTION OR POPULATION-BASED COHORT. UNDERSTANDING RELATIONSHIPS BETWEEN THESE FAT BEDS AND EARLY CVD MAY PERMIT DEVELOPMENT OF STRATEGIES TARGETING INFLAMMATION TO REDUCE CARDIOMETABOLIC AND CVD DEVELOPMENT. THIS STUDY POSES TO LEVERAGE THE MULTI-ETHNIC STUDY OF ATHEROSCLEROSIS (MESA) AS A RIGOROUSLY COMPILED AND HIGHLY VALIDATED DATASET USING A COMPREHENSIVELY PHENOTYPED, MULTI-ETHNIC COHORT. WE WILL UTILIZE THE CONSIDERABLE RESOURCES ALREADY INVESTED IN THE COLLECTION OF ALMOST 20,000 WELL PERFORMED CT SCANS TO DETERMINE PREVALENCE, RISK FACTOR ASSOCIATIONS, AND PROGNOSTIC VALUE USING OVER 20 YEARS OF CAREFUL ADJUDICATED ASCVD FOLLOW UP AND REPEAT IMAGING. SPECIFICALLY, WE WILL MEASURE EPICARDIAL FAT DENSITY AND PERI-CORONARY FAT ATTENUATION IN 17,138 NON-CONTRAST CARDIAC CT SCANS: 6,814 PARTICIPANTS IN THE BASELINE STUDY, ALONG WITH CAC SCANS FROM EXAM 2 (N=2908), EXAM 3 (N=2764), EXAM 4 (N=1388) AND EXAM 5 (N=3264). EACH EXAM WAS APPROXIMATELY 2 YEARS AFTER THE PRIOR EXAM. THE SCIENTIFIC PREMISE IS THAT THESE FAT BEDS (BOTH AROUND THE HEART – EPICARDIAL, AND AROUND THE CORONARIES – PERI-CORONARY) MAY PORTEND RISK FOR CVD INDEPENDENT OF CAC, AND THAT THESE NON-CONTRAST CT SCANS CAN SERVE AS A VIABLE SOURCE OF BOTH CAC AND MEASURES OF INFLAMMATION/VISCERAL FAT MEASUREMENT. THE HOPE IS THAT THIS MAY SERVE AS A WAY TO FURTHER PERSONALIZE MEDICINE TO DELIVER APPROPRIATE PREVENTATIVE TREATMENT, EMPHASIZING THERAPEUTIC INTERVENTION FOR THOSE WHO ARE AT HIGHER PREDICTED RISK. THESE ASPECTS ARE HIGHLY INNOVATIVE, AS IS THE EMPHASIS ON EVALUATING FAT VOLUME AND DENSITY AS A RISK ENHANCING, EFFECT MODIFIER FOR ASYMPTOMATIC INDIVIDUALS IS UNIQUE TO OUR KNOWLEDGE. WITH ACHIEVEMENT OF THE PROPOSED STUDY AIMS, A VALUABLE EPIDEMIOLOGICAL DATABASE WILL BE DEVELOPED, AND THE OPPORTUNITY TO UTILIZE EXISTING CAC SCANS FOR ROUTINE VISCERAL FAT VOLUME AND DENSITY AND PERICORONARY FAT CVD RISK STRATIFICATION WILL MOVE CLOSER TO REALITY. THERE ARE ALREADY MOVES TO AUTOMATE THIS PROCESS, SO IF THIS STUDY IS SUCCESSFUL, CAN LEAD TO MORE SCALABLE AND AUTOMATED DETECTION ON THE MILLIONS OF HEART SCANS OBTAINED EACH YEAR, AS WELL AS THE TENS OF MILLIONS OF ANNUAL THORACIC CT SCANS. WE FEEL THE PROPOSAL IS HIGHLY SIGNIFICANT WITH CONSIDERABLE POTENTIAL TO EXERT A DIRECT, POSITIVE IMPACT ON CVD PREVENTION AND PROVIDE MECHANISTIC INSIGHTS AND TARGETS TO THE ROLE OF INFLAMMATION AND CVD EVENTS.

PLANT-FOCUSED NUTRITION IN PATIENTS WITH DIABETES AND CHRONIC KIDNEY DISEASE (PLAFOND STUDY): A PILOT/FEASIBILITY STUDY - SUMMARY CHRONIC KIDNEY DISEASE (CKD) AFFECTS 10-15% OF US ADULTS INCLUDING 30-40% OF PERSONS WITH DIABETES MELLITUS (DM), IS ASSOCIATED WITH POOR OUTCOMES, AND OFTEN PROGRESSES TO REQUIRING DIALYSIS OR TRANSPLANTATION. HALF OF ALL AMERICANS WITH CKD ALSO HAVE DM. WHILE TRADITIONAL AND EMERGING PHARMACOTHERAPIES ARE OFTEN USED IN CKD WITH DIABETES (CKD/DM), THE SYNERGISTIC ROLE OF DIETARY INTERVENTIONS HAS NOT BEEN WELL EXAMINED. LOW-CARBOHYDRATE LOW-FAT DIETS ARE OFTEN RECOMMENDED IN DM, WHEREAS LOW-PROTEIN DIETS (LPDS) ARE RECOMMENDED FOR NON-DIABETIC CKD WITH INCREASING EMPHASIS ON PLANT-BASED PROTEIN SOURCES. EVIDENCE SUGGESTS THAT HIGH-PROTEIN DIETS WITH GREATER ANIMAL PROTEIN CONTENT MAY LEAD TO GLOMERULAR HYPERFILTRATION AND FASTER DECLINE IN RENAL FUNCTION IN PATIENTS WITH CKD/DM. THERE REMAINS MAJOR CONTROVERSY REGARDING THE POTENTIAL RISKS VS. BENEFITS OF PLANT-BASED DIETS IN CKD/DM, FOR WHICH GUIDELINES REMAIN BASED ON EXPERT OPINION. GIVEN CONVENTIONAL DIETARY RESTRICTIONS FOR THE MANAGEMENT OF DM, THERE IS CONCERN THAT PLANT-BASED LPDS MAY LEAD TO PROTEIN-ENERGY WASTING AND HYPERKALEMIA, WHEREAS THESE DIETS MAY INDEED BE MOST BENEFICIAL IN PATIENTS WITH CKD/DM GIVEN THEIR FASTER RATES OF CKD PROGRESSION AS COMPARED TO NON-DIABETICS. AT PRESENT, CLINICAL PRACTICE GUIDELINES PROVIDE CONFLICTING RECOMMENDATIONS REGARDING THE AMOUNT (LOW VS. HIGH) AND SOURCE (PLANT VS. ANIMAL) OF DIETARY PROTEIN INTAKE (DPI) IN CKD/DM. GIVEN THAT PRIOR DIETARY TRIALS IN CKD SUCH AS THE 1994 MODIFICATION OF DIET IN RENAL DISEASE (MDRD) STUDY EXCLUDED CKD/DM AND DID NOT EXAMINE THE OPTIMAL PROPORTION OF PLANT VS. ANIMAL-BASED PROTEINS, THERE IS URGENT UNMET NEED FOR A RIGOROUS DIETARY INTERVENTION STUDY TO EXAMINE THE EFFICACY AND SAFETY OF PATIENT-CENTERED PLANT-BASED DIETS IN CKD/DM. IN THE SPIRIT OF PAS 20-160 (SMALL R01S FOR CLINICAL TRIALS TARGETING NIDDK DISEASES) WE PROPOSE A PILOT FEASIBILITY RANDOMIZED CONTROLLED TRIAL, CONDUCTED IN PARALLEL WITH PATIENTS' ROUTINE FOLLOW-UP VISITS AT TWO CKD CLINICS WITHIN THE UNIVERSITY OF CALIFORNIA IRVINE, TO TEST THE FEASIBILITY AND SAFETY OF IMPLEMENTING A PLANT- FOCUSED NUTRITION IN CKD/DM (PLAFOND) DIET WITH A DPI OF 0.6-0.8 G/KG/D COMPRISED OF >2/3 PLANT-BASED SOURCES, VS. STANDARD-OF-CARE RENAL DIET WITH <1/3 PLANT-SOURCES AND LOW-POTASSIUM CONTENT, ADMINISTRATED BY DIETITIANS, OVER A 6-MONTH PERIOD IN 120 PATIENTS WITH CKD/DM STAGE 3-5. WE WILL DETERMINE WHETHER THE PLAFOND DIET VS. THE STANDARD-OF-CARE RENAL DIET CAN BE ADHERED TO WITH CONSISTENT SEPARATION IN DIETARY PROTEIN AND PLANT-BASED PROPORTIONS AT 3- AND 6-MONTHS. WE WILL ALSO EXAMINE NUTRITIONAL STATUS, PHYSICAL PERFORMANCE, AND BODY COMPOSITION, AS WELL AS GLYCEMIC MEASURES USING TRADITIONAL METRICS AND CONTINUOUS GLUCOSE MONITORING, WHILE OTHER BIOCHEMICAL PARAMETERS AND PATIENT-REPORTED OUTCOMES INCLUDING CKD-RELATED SYMPTOMS WILL ALSO BE STUDIED. IN ADDITION TO PROVIDING THE REQUISITE FEASIBILITY AND SAFETY DATA OF PATIENT- CENTERED DIETARY REGIMENS NEEDED FOR THE DESIGN AND CONDUCT OF FUTURE MULTI-CENTER TRIALS, OUR PROPOSED STUDY WILL HAVE MAJOR IMMEDIATE IMPACT BY REINVIGORATING THE CRITICAL ROLE OF DIETARY MANAGEMENT OF CKD/DM.

EQUIPMENT ACQUISITION

IND-ENABLING STUDIES TO DEVELOP TRIPTONIDE INTO THE FIRST NONHORMONAL MALE PILL - PROJECT SUMMARY OUR TWELVE YEARS OF RESEARCH HAS LED TO THE DISCOVERY OF TRIPTONIDE (TN) AS A PROMISING NON- HORMONAL MALE CONTRACEPTIVE AGENT (NATURE COMMUNICATIONS, 2021, 12:1253). IN ADDITION TO ITS EXCELLENT POTENCY AND PROVEN ORAL BIOAVAILABILITY, TN DOES NOT CAUSE DISCERNABLE TOXIC EFFECTS IN EITHER MICE OR MONKEYS AT ITS MINIMUM EFFECTIVE DOSE. ALL AVAILABLE DATA STRONGLY SUGGEST THAT TN IS WORTH FURTHER INVESTMENT TO DEVELOP IT AS A VIABLE NON-HORMONAL MALE CONTRACEPTIVE DRUG CANDIDATE. TAKING ADVANTAGE OF THE R61/R33 FUNDING MECHANISM, WE PROPOSE TO CONDUCT SEVERAL INVESTIGATIONAL NEW DRUG (IND)-ENABLING STUDIES TO COLLECT DATA REQUIRED BY THE FDA FOR OFFICIAL IND APPLICATION. SPECIFICALLY, WE WILL CONDUCT NON-GLP TOXICITY AND SAFETY PHARMACOLOGY STUDIES TO IDENTIFY THE MAXIMUM TOLERATED DOSE (MTD) FOLLOWED BY A SIX-MONTH REPEAT DOSE TOXICITY STUDY USING MTD AND MINIMUM EFFECTIVE DOSES (MED) IN MALE SPRAGUE-DAWLEY RATS IN THE R61 PHASE. IN ADDITION, DEEP LEARNING-BASED PATHOLOGICAL AND RNA-SEQ-BASED TRANSCRIPTOMIC ANALYSES WILL ALSO BE PERFORMED. THE DATA WILL NOT ONLY GUIDE AND CORROBORATE THE GLP-COMPLIANT TOXICITY AND SAFETY PHARMACOLOGY STUDIES PROPOSED IN THE R33 PHASE, BUT ALSO PROVIDE MOLECULAR INSIGHTS FOR ANY POTENTIAL PHENOTYPES OBSERVED. IN THE R33 PHASE, WE WILL WORK WITH PHARMARON INC. TO PERFORM NON-GLP TOXICITY STUDIES IN DOGS, IN VITRO SECONDARY PHARMACOLOGY, AND GLP-COMPLIANT REPEAT-DOSE TOXICITY AND SAFETY PHARMACOLOGY STUDIES ALL AS PART OF A PACKAGE OF STUDIES INTENDED TO MOVE TRIPTONIDE INTO A PHASE I CLINICAL TRIAL.

CONFOCAL MICROSCOPE - LEICA STELLARIS - SUMMARY: THE LUNDQUIST INSTITUTE (TLI) IS REQUESTING FUNDS TO PURCHASE A LEICA STELLARIS CONFOCAL MICROSCOPE TO BE HOUSED IN ITS ESTABLISHED, CENTRALLY MANAGED CORE FACILITY. THIS NEW SYSTEM IS INTENDED TO REPLACE AN AGING 12- YEAR-OLD LEICA SP8 MICROSCOPE THAT NO LONGER MEETS THE EVOLVING NEEDS OF OUR RESEARCH COMMUNITY. A BROAD USER GROUP OF 12 INVESTIGATORS (10 OF WHOM ARE NIH-FUNDED), WHO ARE ALL MAKING SIGNIFICANT AND PIONEERING CONTRIBUTIONS TO CROSS- DISCIPLINARY RESEARCH AT THE INTERFACE BETWEEN DEVELOPMENTAL BIOLOGY, CELL BIOLOGY, MOLECULAR BIOLOGY, CANCER, ENDOCRINOLOGY, NEUROBIOLOGY, IMMUNOLOGY, AND HOST-PATHOGEN INTERACTIONS, WILL IMMEDIATELY BENEFIT FROM THE TRANSFORMATIVE IMAGING CAPABILITIES OF THE INSTRUMENT. THE STELLARIS SYSTEM OFFERS MAJOR ADVANCEMENTS IN CONFOCAL IMAGING TECHNOLOGY, INCLUDING A TUNABLE WHITE LIGHT PULSED LASER FOR FLUORESCENCE LIFETIME IMAGING MICROSCOPY (FLIM), INTEGRATED WITH THE HIGH-SPEED FALCON FLIM PLATFORM AND CAPABLE OF MULTIPLEXING UP TO 11 SPECTRAL CHANNELS. THESE FEATURES PROVIDE USERS WITH QUANTITATIVE IMAGING MODALITIES TO MONITOR COMPLEX DYNAMIC PROCESSES IN LIVE AND FIXED SAMPLES. THE INSTRUMENT ALSO INCLUDES LIGHTNING SUPER-RESOLUTION CAPABILITIES BASED ON ADAPTIVE DECONVOLUTION, EXPANDED SPATIAL COVERAGE, AND LEICA'S PROPRIETARY HYD DETECTORS WITH TUNABLE SPECTRAL SENSITIVITY (1-NM PRECISION, 400–850 NM), ENABLING HIGH- RESOLUTION, LOW-PHOTOTOXICITY IMAGING ACROSS A WIDE RANGE OF FLUOROPHORES. ACQUISITION OF THIS SYSTEM WILL ENSURE CONTINUED ACCESS TO STATE-OF-THE-ART IMAGING TECHNOLOGY, ENABLING INVESTIGATORS TO GENERATE HIGH-QUALITY, MULTIDIMENSIONAL DATASETS AND ADDRESS INCREASINGLY COMPLEX BIOLOGICAL QUESTIONS. THIS INSTRUMENT WILL DIRECTLY ENHANCE THE RIGOR, REPRODUCIBILITY, AND COMPETITIVENESS OF NIH-SUPPORTED RESEARCH AT TLI BY FACILITATING TRANSFORMATIVE INSIGHTS INTO MOLECULAR AND CELLULAR MECHANISMS OF HEALTH AND DISEASE.

SLEEP LOSS AND CIRCADIAN MISALIGNMENT - MECHANISMS OF INSULIN RESISTANCE - SLEEP RESTRICTION, THE REDUCTION IN SLEEP RESULTING FROM INSUFFICIENT OR INTERRUPTED SLEEP, AFFECTS A THIRD OF ADULTS IN THE US. IT ARISES FROM SHORTENED SLEEP PERIODS, ENVIRONMENTAL DISTURBANCES, AND DISRUPTED SLEEP FROM OBSTRUCTIVE SLEEP APNEA, MOVEMENT DISORDERS, OR OTHER PATHOLOGY. NIGHT SHIFT WORK, IN ADDITION TO MISALIGNING THE TIMING OF BEHAVIORAL RHYTHMS WITH THE ENDOGENOUS CIRCADIAN RHYTHM, INTERFERES WITH THE HOMEOSTATIC REGULATION OF SLEEP, LEADING TO SLEEP LOSS. NIGHT SHIFT WORKERS COMPRISE 10-15% OF THE WORKFORCE, EXPERIENCE BOTH SLEEP LOSS AND CIRCADIAN MISALIGNMENT, AND HIGHER RATES OF METABOLIC DISEASES. WHEREAS SLEEP LOSS ALONE TYPICALLY WORSENS INSULIN RESISTANCE BY 20%, EXPERIMENTAL STUDIES SHOW THAT ADDING CIRCADIAN MISALIGNMENT TO SLEEP LOSS DOUBLES THIS EFFECT. INSULIN RESISTANCE TRIGGERS THE DEVELOPMENT OF TYPE 2 DIABETES MELLITUS (T2DM) – A DISORDER THAT AFFECTS 38 MILLION AMERICANS AT AN ESTIMATED COST OF $413 BILLION/YEAR. THE MECHANISMS UNDERLYING INDUCTION OF INSULIN RESISTANCE (IR) FROM SLEEP RESTRICTION REMAIN UNKNOWN, ALTHOUGH THE MAJOR STRESS HORMONE, CORTISOL, APPEARS TO PLAY A MAJOR ROLE. UNTIL THE PRECISE MECHANISMS UNDERLYING RISING IR ARE UNVEILED, TARGETED THERAPIES CANNOT BE DEVELOPED. THIS PROPOSAL AIMS TO ESTABLISH THAT THE SHAPE AND THE TIMING OF CORTISOL RHYTHMS UNDERLIE IR DEVELOPMENT IN MEN AND WOMEN. WE WILL ALSO CHARACTERIZE THE DOWNSTREAM 24- HOUR METABOLOMIC AND ENDOCRINE SIGNATURES THAT IDENTIFY METABOLIC, INFLAMMATORY AND OTHER PATHWAYS. INTERVENTIONS TO MODIFY RELEASE, TIMING OR ACTION OF CORTISOL ARE EMINENTLY FEASIBLE, AND WILL PROVIDE A MEANS TO ULTIMATELY ALTER THE FORMIDABLE INCREASING INCIDENCE OF T2DM IN THIS COUNTRY, BUT THE ROLE OF CORTISOL AND ITS DOWNSTREAM PATHWAYS MUST INITIALLY BE OUTLINED. WE WILL CONDUCT 2 RANDOMIZED PLACEBO CONTROLLED TRIALS (RCT) IN 48 ADULTS (24 WOMEN) AGED 22-45 YEARS ACROSS BOTH STUDIES. STUDIES WILL BE OF 7 DAYS DURATION AND SLEEP WILL BE RESTRICTED TO 4 HOURS/NIGHT FOR 3 NIGHTS FOLLOWED BY A 24-HOUR CONSTANT ROUTINE DURING WHICH NO SLEEP OCCURS. THE FIRST RCT SIMULATES DAY SHIFT WORK AND WILL COMPARE THE EFFECT OF SLEEP RESTRICTION UNDER CONDITIONS WHERE THE SHAPE OF THE CORTISOL RHYTHM BECOMES ALTERED VERSUS CONDITIONS WHERE THE SHAPE IS FIXED. THE SECOND RCT SIMULATES NIGHT SHIFT WORK (TO INDUCE CIRCADIAN MISALIGNMENT) AND WILL COMPARE THE EFFECT OF SLEEP RESTRICTION WITH CIRCADIAN MISALIGNMENT UNDER CONDITIONS WHERE THE TIMING OF THE CORTISOL RHYTHM IS MISALIGNED OR ALIGNED. SINCE THERE ARE SEX DIFFERENCES IN THE REGULATION OF CORTISOL AND SLEEP ARCHITECTURE, WE WILL MAXIMIZE POWER TO UNVEIL UNDERLYING IR MECHANISMS THAT DO OR DO NOT DIFFER BY SEX BY POOLING DATA FROM BOTH DAY AND NIGHT SHIFT WORK SCHEDULES. THE GOAL IS TO UNDERSTAND HOW SHORTENED AND MISALIGNED SLEEP LEAD TO METABOLIC ILL-HEALTH, AND TO UNVEIL THE UNDERPINNING ENDOCRINE/METABOLOMIC MECHANISMS. THE STUDIES HAVE THE POTENTIAL TO EXERT A MAJOR IMPACT ON UNDERSTANDING THE MECHANISMS CONTRIBUTING TO T2DM, WHICH IS CLOSELY ASSOCIATED WITH SLEEP-DISORDERED BREATHING, NIGHT SHIFT WORK AND OTHER CONDITIONS WHICH ALTER SLEEP AT SUBSTANTIAL NATIONAL HEALTH COST.

NOVEL CANDIDA ALBICANS HOST RECEPTORS DURING INFECTION AND IMMUNE RESPONSE

INNATE EFFECTORS OF RALS3P-N ANTI-CANDIDA VACCINE

NEWBORN SCREENING FOR CRITICAL CONGENITAL HEART DISEASE IN U.S.: ASSESSMENT OF IMPLEMENTATION OBSTACLES AND APPLICATION OF COST-EFFECTIVENESS TO GAUGE IF ACTION NEEDED FOR IMPROVED IMPLEMENTATION

HORMONAL REGULATION OF LUNG DEVELOPMENT; PTHRP BIOLOGY

A RANDOMIZED CLINICAL TRIAL TO PREVENT RECURRENT CA-MRASA INFECTION

PPAR GAMMA SIGNALING AND LUNG FIBROBLAST TRANSDIFFERENTIATION

CANDIDA ADHERENCE & PENETRATION OF VASCULAR ENDOTHELIUM

A TYPE 2 HYBRID EFFECTIVENESS-IMPLEMENTATION STUDY OF TIME LIMITED TRIALS TO REDUCE POTENTIALLY NON-BENEFICIAL TREATMENTS FOR CRITICALLY ILL PATIENTS WITH ADVANCED MEDICAL ILLNESSES - PROJECT SUMMARY OVERUTILIZATION OF INVASIVE INTENSIVE CARE UNIT (ICU) TREATMENTS FOR PATIENTS WITH ADVANCED MEDICAL ILLNESSES MAY LEAD TO MEDICAL CARE THAT PROVIDES MINIMAL BENEFIT AND PROLONGS SUFFERING. ALTHOUGH THE FACTORS THAT LEAD TO OVERUSE OF POTENTIALLY NON-BENEFICIAL TREATMENTS ARE COMPLEX, AN IMPORTANT CONTRIBUTOR IS POOR COMMUNICATION BETWEEN CLINICIANS, PATIENTS AND SURROGATE DECISION MAKERS. TIME LIMITED TRIALS (TLT) INVOLVE DETAILED DISCUSSIONS OF PATIENTS’ PREFERENCES FOR CARE AND PROGNOSIS FOLLOWED BY AGREEMENTS BETWEEN CLINICIANS AND PATIENTS/SURROGATE DECISION-MAKERS TO USE CERTAIN MEDICAL THERAPIES FOR DEFINED PERIODS OF TIME. TLTS PROMOTE REGULAR STRUCTURED DIALOGUE BETWEEN PROVIDERS, PATIENTS AND FAMILIES, REASSURE THEM THAT ALL INDICATED INTERVENTIONS HAVE BEEN PURSUED, AND SET RATIONAL BOUNDARIES TO TREATMENTS BASED ON PATIENTS’ GOALS OF CARE. IN A PILOT STUDY OUR TEAM SHOWED THAT UTILIZING TLTS AS THE DEFAULT COMMUNICATION APPROACH FOR CRITICALLY-ILL PATIENTS WITH ADVANCED MEDICAL ILLNESSES IMPROVED SHARED DECISION-MAKING AND REDUCED UTILIZATION OF POTENTIALLY NON-BENEFICIAL ICU TREATMENTS. HOWEVER, KEY GAPS IN OUR KNOWLEDGE INCLUDE WHETHER THE TLT INTERVENTION CAN BE SCALED-UP PRAGMATICALLY IN REAL-LIFE HEALTHCARE SYSTEMS AND UNDERSTANDING THE FACTORS THAT AFFECT IMPLEMENTATION AND EFFECTIVENESS. WE WILL CONDUCT A PRAGMATIC TYPE 2 HYBRID EFFECTIVENESS- IMPLEMENTATION STEPPED WEDGE CLINICAL TRIAL OF THE TLT INTERVENTION IN THE ICUS OF THE LOS ANGELES COUNTY DEPARTMENT OF HEALTH SERVICES (LAC DHS) AND SOUTHERN CALIFORNIA KAISER PERMANENTE (KPSC) HOSPITALS. LAC DHS IS THE SECOND LARGEST PUBLIC HEALTHCARE SYSTEM IN THE UNITED STATES, AND KPSC IS THE LARGEST INTEGRATED HEALTHCARE SYSTEM IN SOUTHERN CALIFORNIA. THE PATIENT POPULATION IN THESE HEALTHCARE SYSTEMS ARE RACIALLY AND CULTURALLY DIVERSE, AND MANY HAVE HIGH RISK FOR BREAKDOWNS IN SHARED DECISION-MAKING DUE TO SOCIOECONOMIC DISADVANTAGES, LANGUAGE DISCORDANCE, AND LOW HEALTH LITERACY. THE TLT INTERVENTION STRATEGY WAS CO-DESIGNED WITH STAKEHOLDERS IN LAC DHS AND KPSC. THE SPECIFIC AIMS ARE TO: (1) EXAMINE WHETHER THE TLT INTERVENTION REDUCES ICU LENGTH OF STAY AND UTILIZATION OF INVASIVE TREATMENTS, (2) IDENTIFY CHARACTERISTICS OF THE INTERVENTION, PARTICIPANTS, AND HEALTHCARE ENVIRONMENT THAT AFFECT IMPLEMENTATION USING THE CONSOLIDATED FRAMEWORK FOR IMPLEMENTATION RESEARCH (CFIR) AND (3) PERFORM A MIXED METHODS EVALUATION OF IMPLEMENTATION OUTCOMES USING THE RE-AIM FRAMEWORK. THE PROJECT TEAM HAS THE TRANSDISCIPLINARY EXPERTISE, ESTABLISHED PARTNERSHIPS WITH STAKEHOLDERS IN LAC DHS AND KPSC, AND INSTITUTIONAL SUPPORT TO COMPLETE THE AIMS OF THIS PROJECT. BY EXAMINING THE EFFECTIVENESS AND IMPLEMENTATION, THIS STUDY HAS THE POTENTIAL TO IMPROVE THE QUALITY OF ICU CARE IN TWO ESSENTIAL HEALTHCARE SYSTEMS IN SOUTHERN CALIFORNIA AND GUIDE IMPLEMENTATION OF APPROACHES TO IMPROVE SHARED DECISION-MAKING IN OTHER HEALTHCARE ENVIRONMENTS.

EFFECT OF PRE AND POSTNATAL NUTRITIONAL PROGRAMMING ON SCD1 ACTIVITY AND OBESITY

COMMUNITY-BASED PREVENTION OF YOUTH VIOLENCE IN LATINOS

MENTORSHIP IN THE ENDOCRINOLOGY OF SLEEP

RAP1 - KRIT1 REGULATION OF ENDOTHELIAL PERMEABILITY

MODIFIED NEURAL STEM CELLS FOR SLYCOSYLATION-INDEPENDENT TREATMENT OF MPS IIIB

SNP HAPLOTYPING TO DETECT DIABETIC NEPHROPATHY RISK

MODIFIED NEURAL STEM CELLS FOR GLYCOSYLATION-INDEPENDENT TREATMENT OF MPS IIIB

RESIDENCY TRAINING IN PRIMARY CARE

ATAXIA TELANGIECTASIA: ELUCIDATING DISEASE PATHOGENESIS AND TESTING NEW TREATMENTS

OPTIMIZING SMALL MOLECULE READ-THROUGH COMPOUNDS FOR TREATING ATAXIATELANGIECTASIA - PROJECT SUMMARY ATAXIA-TELANGIECTASIA (A-T) IS A RARE (~ 1 IN EVERY 100,000) BUT CATASTROPHIC AND DEADLY DISEASE THAT CAUSES PROGRESSIVE LOSS OF MOTOR FUNCTION AND DEATH BETWEEN THE AGES OF 10 AND 30 YEARS. IN ABOUT ONE-THIRD OF A-T CASES, THE CAUSE IS A NONSENSE MUTATION IN THE ATM (ATAXIA-TELANGIECTASIA MUTATED) GENE THAT ENCODES A PREMATURE TERMINATION CODON (PTC). NO EFFECTIVE TREATMENTS ARE AVAILABLE. HOWEVER, OUR GROUP HAS BEEN DEVELOPING AND TESTING A SERIES OF COMPOUNDS THAT EFFECTIVELY READTHROUGH PTCS IN THE TRANSCRIBED MRNA. PUBLISHED AND UNPUBLISHED STUDIES DEMONSTRATE THE CAPABILITY OF THESE “SMRT” COMPOUNDS (FOR SMALL MOLECULE READTHROUGH) TO READTHROUGH PTCS AND RESTORE TRANSLATION OF FUNCTIONAL ATM PROTEIN IN IN VIVO AND EX VIVO EXPERIMENTS. THE RATIONALE TO FURTHER DEVELOP SMRT COMPOUNDS IS STRENGTHENED BY PROMISING THERAPEUTIC RESULTS IN MOUSE MODELS FOR DUCHENNE MUSCULAR DYSTROPHY AND HEREDITARY PULMONARY ARTERIAL HYPERTENSION BY OUR COLLABORATORS. HOWEVER, SUCCESS IN THESE MODELS DOES NOT ENSURE SUCCESS IN A MULTISYSTEM, NEUROLOGICAL DISORDER LIKE A-T, IN PART BECAUSE A-T UNIQUELY REQUIRES THE COMPOUND TO CROSS THE BLOOD-BRAIN BARRIER IN ADEQUATE AMOUNTS. WE HAVE MADE CONSIDERABLE PROGRESS IN OUR PRECLINICAL STUDIES. WE PRESENT DATA INDICATING THAT SMRT COMPOUNDS ELICIT SYNTHESIS OF FULL-LENGTH FUNCTIONAL PROTEIN THAT PENETRATES THE BRAIN. WE ALSO RECENTLY SOLVED A MAJOR HURDLE IN THE FIELD OF A-T RESEARCH: LACK OF AN ANIMAL MODEL THAT FAITHFULLY REFLECTS CLINICAL DISEASE. IN NINDS-SUPPORTED STUDIES, WE USED A DOUBLE HIT STRATEGY TO GENERATE A MOUSE HARBORING BOTH A CLINICALLY RELEVANT PTC IN THE ATM GENE AND A KNOCKOUT OF A RELATED DNA REPAIR GENE CALLED APRATAXIN (APTX). CHARACTERIZATION OF THIS MOUSE MODEL, INCLUDING THE PROFOUND, PROGRESSIVE ATAXIA THAT IS A HALLMARK OF A-T IS COMPLETE. NOW, WE MOVE TO NEXT LOGICAL PHASE: TO USE A RATIONALIZED AND COMPREHENSIVE APPROACH TO OPTIMIZE OUR TOP CANDIDATE SMRT COMPOUND VIA MEDICINAL CHEMISTRY. IN AIM 1 WE WILL DEVELOP AND VALIDATE A NOVEL POTENCY ASSAY SPECIFICALLY DESIGNED TO EVALUATE POTENCY ACROSS THE 10 MOST COMMON A-T CAUSING NONSENSE MUTATIONS. IN AIM 2 WE WILL DEVELOP AND VALIDATE AN ASSAY DESIGNED TO CONFIRM ATM FUNCTION IN HUMAN CELLS TAKING ADVANTAGE OF OUR UNIQUE REPOSITORY OF A-T PATIENT DERIVED CELL LINES. FINALLY, IN AIM 3, THESE ASSAYS (ALONG WITH THOSE ALREADY STANDARD IN OUR LABS TO ASSESS SOLUBILITY, PROTEIN BINDING, BLOOD BRAIN BARRIER PERMEABILITY, AND TOXICITY) WILL BE UTILIZED TO CONDUCT A MEDICINAL CHEMISTRY OPTIMIZATION CAMPAIGN TO GENERATE A SMALL SET OF CANDIDATE COMPOUNDS WITH PROPERTIES THAT MAXIMIZE THEIR CHANCES OF SUCCESS IN FOLLOW-ON EFFICACY STUDIES IN OUR NEW A-T MOUSE MODEL. OUR WORK REPRESENTS THE FIRST REAL HOPE FOR KIDS SUFFERING FROM A-T'S DEVASTATING EFFECTS.

TYPE I INTERFERON REGULATES ANGIOGENESIS IN DOWN SYNDROME - PROJECT SUMMARY TRISOMY 21 (T21) IS THE MOST PREVALENT CHROMOSOMAL ABNORMALITY WORLDWIDE, RESULTING IN DOWN SYNDROME (DS) [1, 2], A MULTISYSTEM SYNDROME ASSOCIATED WITH CARDIOPULMONARY DISORDERS. PULMONARY DISORDERS INCLUDE PULMONARY HYPERTENSION, PULMONARY HYPOPLASIA, PULMONARY HEMOSIDEROSIS AND PULMONARY CAPILLARITIS, ALL OF WHICH CAN RESULT FROM A DYSREGULATED LUNG ENDOTHELIUM. IN ADDITION, T21 LUNGS ARE CHARACTERIZED BY THE PERSISTENCE OF AN IMMATURE DOUBLE CAPILLARY NETWORK SYSTEM SURROUNDING THE ALVEOLI AND REPRESENTATIVE OF THE SACCULAR STAGE OF DEVELOPMENT [3]. THIS SUGGESTS THAT ENDOTHELIAL DEFECTS IN INDIVIDUALS WITH DS ARE LIKELY INITIATED IN UTERO. SINGLE CELL TRANSCRIPTOMICS DATA DEMONSTRATED THE PRESENCE OF DIVERSE ENDOTHELIAL CELL POPULATIONS IN THE HUMAN ADULT LUNG [4, 5]. WHILE OTHERS AND WE HAVE REPORTED IMPAIRED ENDOTHELIAL NETWORKS IN THE DEVELOPING T21 LUNG CHARACTERIZED BY CONGESTED CAPILLARIES, LYMPHATIC DILATATION, AND MUSCULARIZED ARTERIES [6], THE NATURE OF THESE DEFECTS AND THE DIFFERENT ENDOTHELIAL CELL POPULATIONS CONTRIBUTING TO SUCH DEFECTS ARE YET TO BE UNDERSTOOD. MOREOVER, DEVELOPING T21 HUMAN LUNGS DISPLAY INCREASED EXPRESSION OF ANTI-ANGIOGENIC FACTORS [3, 6-8]. A DECREASE IN ENDOTHELIAL PROGENITOR CELLS IS FOUND IN T21 PERIPHERAL BLOOD [9]. FURTHERMORE, T21 IPSC- DERIVED ENDOTHELIAL CELLS EXHIBIT DECREASED CELL PROLIFERATION, IMPAIRED SPROUTING AND TUBE FORMATION COMPARED TO EUPLOID CELLS [10]. ONE OF THE MAJOR PATHWAYS CONSISTENTLY ACTIVATED IN DS IS THE TYPE I INTERFERON (IFN) PATHWAY [6]. ELEVATED TYPE I IFN CAUSES A DECREASE OF ENDOTHELIAL PROGENITOR CELLS [9] AND INHIBITS VEGF INDUCED DEVELOPMENT OF CAPILLARY LIKE STRUCTURES [11, 12], THUS PLAYING AN ANTI-ANGIOGENIC ROLE. ADDITIONALLY, TYPE I INTERFERONOPATHIES ARE LINKED TO PULMONARY HYPERTENSION AND PULMONARY VASCULOPATHY [13]. SC-RNASEQ DATA OF T21 AND EUPLOID LUNG CELLS DEMONSTRATED THE PRESENCE OF SEVERAL ENDOTHELIAL CELL POPULATIONS DISPLAYING INCREASED EXPRESSION OF TYPE I IFN SIGNALING PATHWAY GENES IN T21. THEREFORE, WE HYPOTHESIZE THAT DEFECTIVE LUNG ANGIOGENESIS OCCURS IN T21 DURING DEVELOPMENT AND IS A RESULT OF TYPE I IFN-DEPENDENT CHANGES IN ENDOTHELIAL CELL PROLIFERATION, DIFFERENTIATION AND MIGRATION. TO TEST THIS HYPOTHESIS, WE WILL 1) DETERMINE ENDOTHELIAL CELL DEFECTS IN T21 VS EUPLOID HUMAN DEVELOPING LUNGS BY DEFINING THE DIFFERENT ENDOTHELIAL CELL POPULATIONS AND THEIR SPATIOTEMPORAL DISTRIBUTION USING COMBINATORIAL IN SITU HYRIDIZATION WITH IF STAININGS AND ASSESSING DEFECTS IN ENDOTHELIAL CELL PROLIFERATION AND SPROUTING OF EACH GROUP VIA IF AND 3D IMAGE QUANTIFICATIONS AND 2) DEFINE THE ROLE OF TYPE I IFN SIGNALING IN THE ENDOTHELIAL DEFECTS IN T21 BY DETERMINING THE EFFECT OF GAIN AND LOSS OF TYPE I IFN SIGNALING ON PROLIFERATION, MIGRATION, PERMEABILITY AND TUBE FORMATION IN PRIMARY AND IPSC-DERIVED ENDOTHELIAL CELL CULTURES OF T21 AND EUPLOID CELLS. IMPROVED STRATEGIES TO PREVENT, AMELIORATE OR REVERSE ENDOTHELIAL RELATED LUNG DISEASE IN DS WILL BE CONTINGENT UPON A DETAILED UNDERSTANDING OF THE DEFECTS AND PATHWAYS DRIVING THESE DEFECTS THAT MAY IDENTIFY NOVEL THERAPEUTIC TARGETS FOR INTERVENTION.

GENETICS OF SUSCEPTIBILITY TO CANDIDIASIS

PURPOSE:THE LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER (TLI) WILL USE THE FUNDING TO ACHIEVE THREE OBJECTIVES: INCREASING THE NUMBER OF UNDERGRADUATE INTERNS FROM LOCAL UNIVERSITIES LIKE CALIFORNIA STATE UNIVERSITY DOMINGUEZ HILLS (CSUDH) AND CHARLES DREW UNIVERSITY (CDU) IN TLI RESEARCH LABS, SUPPORTING GRADUATE STUDENTS IN TLI'S PHD PROGRAM IN TRANSLATIONAL RESEARCH, AND ENHANCING RECRUITMENT OF UNDERSERVED STUDENTS. THESE EFFORTS FOCUS ON ADVANCING STUDENTS OF COLOR IN SOUTH LOS ANGELES, ALIGNING WITH THE COMMUNITY SERVED BY HARBOR-UCLA MEDICAL CENTER.ACTIVITIES TO BE PERFORMED:TLI WILL RECRUIT UNDERGRADUATE STUDENTS FROM CSUDH, CDU, AND OTHER LOCAL UNIVERSITIES AS RESEARCH INTERNS. GRADUATE STUDENTS IN TLI'S PHD PROGRAM IN TRANSLATIONAL RESEARCH WILL RECEIVE TRAINING THAT COMBINES SCIENTIFIC RESEARCH WITH ENTREPRENEURIAL SKILLS. ADDITIONALLY, OUTREACH INITIATIVES WILL TARGET UNDERSERVED STUDENTS IN THE COMMUNITY. THROUGHOUT THESE PROGRAMS, TLI RESEARCHERS AND HARBOR-UCLA PHYSICIANS, WHO ARE EXPERIENCED IN THEIR RESPECTIVE FIELDS, WILL MENTOR THE STUDENTS, PROVIDING THEM WITH VALUABLE GUIDANCE AND SUPPORT FOR THEIR RESEARCH AND CAREER DEVELOPMENT.EXPECTED OUTCOMES:THESE INITIATIVES ARE EXPECTED TO INCREASE THE PARTICIPATION OF UNDERREPRESENTED STUDENTS IN SCIENTIFIC RESEARCH, CONTRIBUTING TO A MORE DIVERSE TALENT PIPELINE. THE PROGRAMS AIM TO ENHANCE DIVERSITY IN BIOMEDICAL RESEARCH BY PROVIDING EQUAL OPPORTUNITIES FOR STUDENTS OF COLOR IN SOUTH LOS ANGELES, ENSURING THAT THEIR UNIQUE PERSPECTIVES AND EXPERIENCES ARE REPRESENTED IN THE FIELD. PHD STUDENTS' RESEARCH WILL RESULT IN PUBLISHED FINDINGS ON HEALTH ISSUES LIKE HOSPITAL-ACQUIRED INFECTIONS, STEM CELL THERAPIES, AND TYPE I DIABETES, FURTHER CONTRIBUTING TO THE DIVERSITY OF RESEARCH IN THESE AREAS.INTENDED BENEFICIARIES:THE PRIMARY BENEFICIARIES INCLUDE UNDERGRADUATE STUDENTS FROM LOCAL UNIVERSITIES AND GRADUATE STUDENTS IN TLI'S PHD PROGRAM. THE BROADER SOUTH LOS ANGELES COMMUNITY, PARTICULARLY STUDENTS OF COLOR AND UNDERSERVED POPULATIONS, WILL BENEFIT FROM EXPANDED BIOMEDICAL RESEARCH AND ENTREPRENEURSHIP CAREER OPPORTUNITIES. THESE INITIATIVES AIM TO BUILD A ROBUST, DIVERSE TALENT PIPELINE SUPPORTING LONG-TERM COMMUNITY GROWTH.SUBRECIPIENT ACTIVITIES: NONE

LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY

IMMUNOMODULATION IN MELANOMA: TOLL LIKE RECEPTORS AND THERAPEUTIC VACCINES

IMMUNOMETABOLIC AND EPIGENETIC DYSREGULATION OF ORAL MUCOSAL IMMUNITY IN DIABETES-ASSOCIATED CANDIDIASIS - PROJECT SUMMARY/ABSTRACT DIABETES INCREASES SUSCEPTIBILITY TO INFECTIOUS DISEASES, INCLUDING OROPHARYNGEAL CANDIDIASIS (OPC, OR THRUSH), AN OPPORTUNISTIC INFECTION CAUSED BY THE COMMENSAL FUNGUS CANDIDA ALBICANS. BOTH THE INCIDENCE AND SEVERITY OF OPC ARE SUBSTANTIALLY HIGHER IN INDIVIDUALS WITH POORLY CONTROLLED DIABETES. DESPITE ITS CLINICAL SIGNIFICANCE, THERE ARE NO TARGETED THERAPIES FOR DIABETIC OPC, LARGELY BECAUSE THE MECHANISMS BY WHICH HYPERGLYCEMIA INCREASE SUSCEPTIBILITY TO OPC REMAIN UNKNOWN. THE ORAL MUCOSAL TISSUE ACTS AS BOTH A PHYSICAL BARRIER AND AN ACTIVE IMMUNE SENTINEL AGAINST C. ALBICANS. HOWEVER, HOW CHRONIC METABOLIC DISEASE, ESPECIALLY DIABETES, ALTERS ORAL MUCOSAL IMMUNITY REMAINS INCOMPLETELY UNDERSTOOD. THIS CRITICAL KNOWLEDGE GAP LIMITS OUR UNDERSTANDING OF HOST–FUNGAL INTERACTIONS IN THE ORAL MUCOSA AND HINDERS THE DEVELOPMENT OF EFFECTIVE THERAPEUTIC INTERVENTIONS. OUR LONG-TERM GOAL IS TO DEFINE THE MOLECULAR MECHANISMS UNDERLYING OPC SUSCEPTIBILITY IN DIABETES AND TO IDENTIFY THERAPEUTIC TARGETS THAT RESTORE MUCOSAL IMMUNITY AND MITIGATE INFECTION RISK. THE OBJECTIVE OF THIS RESEARCH IS TO DETERMINE HOW HYPERGLYCEMIA ALTERS EPITHELIAL IMMUNOMETABOLISM AND EPIGENETIC REGULATION TO DRIVE BARRIER DYSFUNCTION AND SUSCEPTIBILITY TO ORAL C. ALBICANS INFECTION. OUR CENTRAL HYPOTHESIS, SUPPORTED BY STRONG PRELIMINARY DATA, IS THAT HYPERGLYCEMIA ENHANCES OPC VULNERABILITY BY SUPPRESSING HYPOXIA-INDUCIBLE FACTOR 1-ALPHA (HIF-1Α) SIGNALING AND IMPAIRING PROLINE CATABOLISM, LEADING TO EPITHELIAL BARRIER DYSFUNCTION, ALTERED MUCOSAL IMMUNITY, AND INCREASED PYROPTOSIS DURING C. ALBICANS INFECTION (AIM 1 AND 2). IN AIM 1, WE WILL DELINEATE THE MECHANISMS BY WHICH HYPERGLYCEMIA- INDUCED SUPPRESSION OF HIF-1Α DRIVES EPITHELIAL BARRIER DYSFUNCTION AND PYROPTOSIS. OUR WORKING HYPOTHESIS IS THAT HYPERGLYCEMIA-INDUCED ACCUMULATION OF TCA CYCLE INTERMEDIATES, PARTICULARLY Α-KETOGLUTARATE, SUPPRESSES HIF-1Α SIGNALING, RESULTING IN BARRIER DISRUPTION AND ACTIVATION OF PYROPTOSIS DURING C. ALBICANS INFECTION. IN AIM 2, WE WILL DETERMINE HOW HYPERGLYCEMIA DYSREGULATES PROLINE METABOLISM AND IMPAIRS EPITHELIAL MEMORY, A NOVEL NON-HEMATOPOIETIC MECHANISM OF TRAINED IMMUNITY, DURING MUCOSAL CANDIDIASIS. OUR WORKING HYPOTHESIS IS THAT HYPERGLYCEMIA IMPAIRS PROLINE CATABOLISM AND EPITHELIAL MEMORY, AS EVIDENCED BY ELEVATED INTRACELLULAR PROLINE LEVELS AND REDUCED PRODH EXPRESSION DURING INFECTION. THE PROPOSED RESEARCH IS SIGNIFICANT BECAUSE IT WILL REVEAL HOW HYPERGLYCEMIA INCREASES OPC SUSCEPTIBILITY AND IDENTIFY THERAPEUTIC TARGETS TO RESTORE ORAL MUCOSAL IMMUNITY. THESE FINDINGS WILL FILL A CRITICAL KNOWLEDGE GAP IN OUR UNDERSTANDING OF HOST–FUNGAL INTERACTIONS WITHIN THE ORAL MUCOSA IN DIABETIC PATIENTS AND PROVIDE A FOUNDATION FOR DEVELOPING TARGETED INTERVENTIONS TO PREVENT OPPORTUNISTIC INFECTIONS IN THE GROWING DIABETIC POPULATION.

KILLED BUT METABOLICALLY ACTIVE LEISHMANIA: A NOVEL PROTOZOAN VACCINE TECHNOLOGY

PILOT/PHASE I STUDY OF INTRATHECAL LARONIDASE FOR SPINAL CORD COMPRESSION IN MUCO

SUICIDAL BEHAVIOR AMONG ADOLESCENTS EXPERIENCING HOMELESSNESS IN THE US DURING THE COVID-19 PANDEMIC - PROJECT SUMMARY SUICIDE IS A MAJOR PUBLIC HEALTH PROBLEM IN THE U.S., RANKING AS THE SECOND LEADING CAUSE OF DEATH AMONG TEENS, WITH A RATE OF 7.1 PER 100,000 POPULATION IN THIS AGE GROUP. PREVIOUS STUDIES HAVE REVEALED SEASONAL TRENDS AND DAY-OF-THE-WEEK EFFECTS IN ADOLESCENT SUICIDAL BEHAVIOR, SUCH AS HIGHER INCIDENCE IN SPRING, ON SUNDAYS, AND ON MONDAYS, AND LOWER OCCURRENCES IN SUMMER, ALIGNING WITH THE SCHOOL CALENDAR. A SIGNIFICANT GAP IN UNDERSTANDING SUICIDAL BEHAVIOR EXISTS AMONG ADOLESCENTS EXPERIENCING HOMELESSNESS (AEH), WHO SHOW HIGHER RATES AND DISTINCT SEASONAL PATTERNS AND DAY-OF-THE-WEEK EFFECTS COMPARED TO THEIR DOMICILED COUNTERPARTS. OUR PREVIOUS RESEARCH SHOWED THAT AEH FACE INCREASED RISK WHEN AWAY FROM SCHOOL, EVIDENCED BY AN ATTENUATED SUMMER DECLINE IN SUICIDE ATTEMPTS AND PEAK SUICIDE DEATHS IN SUMMER AND THE BEGINNING OF THE WEEKEND (I.E., FRIDAYS AND SATURDAYS). THE PANDEMIC BROUGHT NEW CHALLENGES, SUCH AS ECONOMIC INSTABILITY, LIMITED ACCESS TO ESSENTIAL SERVICES, AND INCREASED SOCIAL ISOLATION, WHICH MAY HAVE DISPROPORTIONATELY AFFECTED THE VULNERABLE POPULATION OF AEH. WE HYPOTHESIZED THAT THE PANDEMIC HAS EXACERBATED EXISTING DISPARITIES BETWEEN AEH AND THEIR DOMICILED COUNTERPARTS. ADDITIONALLY, RECENT STUDIES REPORTED DECREASED RATES IN SUICIDAL BEHAVIOR AMONG ADOLESCENTS IN SPRING 2020, COINCIDING WITH NATIONWIDE IMPLEMENTATION OF SCHOOL CLOSURES AS A PANDEMIC CONTROL MEASURE. HOWEVER, NO RESEARCH HAS FOCUSED ON AEH DURING THIS TIME. EXPANDING ON OUR PRIOR STUDY THAT SHOWED A HIGHER RISK FOR AEH WHEN THEY ARE AWAY FROM SCHOOL, WE HYPOTHESIZE THAT SUICIDAL BEHAVIOR INCREASED AMONG AEH IN SPRING 2020. TO TEST THESE INNOVATIVE HYPOTHESES, WE WILL ANALYZE THREE LINKED DATASETS: 1] HOMELESS MANAGEMENT INFORMATION SYSTEM (HMIS) DATA; 2] HOSPITAL DISCHARGE RECORDS; AND 3] DEATH RECORDS FROM FIVE JURISDICTIONS, WHICH COLLECTIVELY REPRESENT 32.5% OF AEH IN THE U.S. AND 10.8% OF THE U.S. ADOLESCENT POPULATION. ADOLESCENTS AGED 13 TO 17 WILL BE ANALYZED DUE TO DISTINCT SUICIDAL BEHAVIOR MECHANISMS WITHIN THIS AGE GROUP COMPARED TO YOUNGER CHILDREN AND OLDER ADOLESCENTS. FOR EXAMPLE, ADOLESCENTS YOUNGER THAN 18 ARE SUBJECT TO DISTINCT STRESSORS RELATED TO SCHOOL ENROLLMENT, WHEREAS SOME INDIVIDUALS ABOVE 18 ARE NOT IN SCHOOL. THE CIRCUMSTANCES SURROUNDING HOMELESSNESS MAY DIFFER AS WELL, WITH THOSE BELOW 18 OFTEN ACCOMPANIED BY PARENTS AND THOSE ABOVE 18 OFTEN LIVING INDEPENDENTLY. AIM 1 WILL INVESTIGATE DIFFERENCES IN ANNUAL RATES OF SUICIDAL BEHAVIOR AMONG AEH AND DOMICILED ADOLESCENTS BETWEEN 2016 AND 2022 TO DETERMINE WHETHER THE PANDEMIC HAS EXACERBATED EXISTING DISPARITIES BETWEEN THE TWO GROUPS. AIM 2 WILL ASSESS MONTHLY TRENDS IN SUICIDAL BEHAVIOR BEFORE, DURING, AND AFTER THE PANDEMIC TO DETERMINE WHETHER SUICIDAL BEHAVIOR INCREASED AMONG AEH IN SPRING 2020. INVESTIGATING TRENDS IN SUICIDAL BEHAVIOR AMONG AEH DURING THE PANDEMIC WILL INFORM WHETHER SUICIDE PREVENTION MEASURES TAILORED TO AEH ARE NEEDED, ESPECIALLY BEFORE AND DURING SCHOOL BREAKS. THE FINDINGS WILL ALSO ENHANCE PREPAREDNESS FOR POTENTIAL SCHOOL CLOSURES DUE TO FUTURE PANDEMICS.

BEHAVIORAL AND BRAIN NETWORK EFFECTS OF DYSFUNCTION IN THE COGNITIVE CEREBELLUM - PROJECT SUMMARY CEREBELLAR DYSFUNCTION HAS BEEN IMPLICATED IN VARIOUS COGNITIVE DISORDERS (E.G., AUTISM SPECTRUM DISORDER, SCHIZOPHRENIA, AND ATTENTION DEFICIT AND HYPERACTIVITY DISORDER) ASSOCIATED WITH THE INABILITY TO ADAPTIVELY ALTER PREVIOUSLY LEARNED BEHAVIORS. SEVERAL INDEPENDENT STUDIES POINT TO DISEASE RELATED CEREBELLAR DYSFUNCTION AS A CAUSAL OR AT LEAST CONTRIBUTING FACTOR IN THIS BEHAVIORAL DEFICIT AS EXPERIMENTAL DISRUPTION OF THE CEREBELLUM DECREASES THE ABILITY OF MICE TO ADAPTIVELY CHANGE PREVIOUSLY LEARNED BEHAVIORS IN THE FACE OF A CHANGING ENVIRONMENT. MOREOVER, CERTAIN NEURONS IN THE COGNITIVE CEREBELLUM (I.E., PURKINJE NEURONS) ARE CONSISTENTLY FOUND TO BE DAMAGED IN COGNITIVE DISORDERS WHERE BEHAVIORAL INFLEXIBILITY IS A PROMINENT FEATURE. THE FIELDS WORKING HYPOTHESIS IS THAT DYSFUNCTION OF THE “COGNITIVE CEREBELLUM” (E.G., CRUS I AND LOBULE VI) CAUSES ABNORMAL STATES OF COMMUNICATION BETWEEN THE CEREBELLUM AND FOREBRAIN AREAS INVOLVED IN FLEXIBLE BEHAVIOR (E.G. PREFRONTAL CORTEX). THERE REMAINS HOWEVER MAJOR GAPS IN OUR UNDERSTANDING OF THE CEREBELLUM'S ROLE IN FLEXIBLE AND INFLEXIBLE BEHAVIOR, THIS INCLUDES: 1) WHAT TYPES OF ABNORMAL CEREBELLAR ACTIVITY CAN CAUSE INFLEXIBLE BEHAVIOR; 2) WHICH SPECIFIC ANATOMICAL/FUNCTIONAL SUB-REGIONS OF THE CEREBELLAR CORTEX ARE INVOLVED; 3) WHAT INFORMATION DOES THE CEREBELLUM ENCODE PERTINENT TO BEHAVIORAL FLEXIBILITY; 4) WHAT DOWNSTREAM FOREBRAIN REGIONS COMMUNICATE WITH THE CEREBELLUM DURING FLEXIBLE BEHAVIOR, AND ARE THESE THE SAME REGIONS IMPACTED BY CEREBELLAR DYSFUNCTION; AND 5) WHAT IS THE EFFECT OF ABNORMAL COMMUNICATION ON DOWNSTREAM FOREBRAIN REGIONS AND NETWORK ACTIVITY AND DOES IT MATCH ABNORMAL BRAIN STATES ASSOCIATED WITH MENTAL DISORDER. IN AIM 1 WE WILL ADDRESS QUESTIONS 1 & 2 BY DISRUPTING DEFINED SUBREGIONS OF THE CEREBELLUM (CRUS I, CRUS II, AND LOBULE VI) USING DREADD TECHNOLOGY AND THEN MEASURING FLEXIBLE BEHAVIOR IN A 2-CUE REWARD-ASSOCIATION PARADIGM IN MICE. WE WILL ALSO ADDRESS QUESTION 3 BY RECORDING FROM THE CEREBELLUM USING DENSE-ELECTRODE ARRAYS DURING FLEXIBLE BEHAVIOR TO ESTABLISH WHAT INFORMATION THE CEREBELLUM ENCODES TO SUPPORT ADAPTIVE REVERSAL OF PREVIOUSLY LEARNED STIMULUS-REWARD ASSOCIATIONS. IN AIM 2, WE WILL ADDRESS QUESTIONS 4 & 5 BY COMBINING CHEMO-GENETIC DISRUPTION OF THOSE SAME DEFINED SUBREGIONS OF THE CEREBELLUM WITH WHOLE-BRAIN NEUROIMAGING, SPECIFICALLY RESTING-STATE FUNCTIONAL MAGNETIC RESONANCE IMAGING (RS-FMRI) IN MICE. HERE, WE PROPOSE TWO DISTINCT APPROACHES THAT WILL ALLOW US TO ESTABLISH MECHANISTIC HYPOTHESES RELATED TO QUESTIONS 1 - 5 THAT WILL SET THE STAGE FOR MULTIPLE FOLLOW-ON STUDIES. OUR OVERALL GOAL IS TO DETERMINE HOW DISPARATE BRAIN REGIONS COLLABORATE TO INFLUENCE NORMAL AND ABNORMAL COGNITIVE BEHAVIORS, PROVIDE CLUES AS TO HOW NEUROCOGNITIVE DYSFUNCTION ARISES, AND EXPLORE HOW DISEASE DEVELOPMENT IMPACTS—OR IS IMPACTED BY— ABNORMAL BRAIN NEUROCIRCUITRY.

DELINEATING HOST RESPONSE TO CENTRAL VENOUS CATHETER ASSOCIATED CANDIDA ALBICANS BIOFILM INFECTIONS, AND DEVELOPMENT OF NOVEL THERAPEUTICS TO COMBAT DRUG RESISTANT BIOFILMS - ABSTRACT CANDIDA ALBICANS CAUSES HALF OF ALL INVASIVE FUNGAL INFECTIONS IN HUMANS. IT ADHERES TO INDWELLING MEDICAL DEVICES AND DEVELOPS INTO DRUG RESISTANT MULTILAYERED COMMUNITY OF CELLS CALLED BIOFILMS, WHICH SERVE AS A RESERVOIR OF CONTINUING INFECTIONS. WHILE HOST RESPONSE TO MUCOSAL AND DISSEMINATED CANDIDIASIS IS WELL STUDIED, LITTLE IS KNOWN ON THE IMMUNE RESPONSE TO C. ALBICANS BIOFILMS ON CATHETERS. WE HYPOTHESIZE THAT DISCOVERING VIRULENCE FACTORS OF BIOFILM CELLS, AND DELINEATION OF THE HOST IMMUNE RESPONSE TO THE BIOFILM, CAN INFORM THERAPEUTIC STRATEGIES AIMED AT ATTENUATION OF BIOFILM-ASSOCIATED DISSEMINATED CANDIDIASIS IN THE HOST. FOR THIS WE WILL USE A RAT CENTRAL VENOUS CATHETER MODEL OF BIOFILM INFECTION. TO UNDERSTAND THE BIOFILM ENVIRONMENT OF CATHETER- INFECTED CARDIOVASCULAR TISSUE, WE WILL UTILIZE A NEW TECHNOLOGY CALLED VISIUM 10X SPATIAL GENOMICS, THAT PROVIDES A SPATIAL “ATLAS” OF ALL GENES (HOST AND PATHOGEN) EXPRESSED DURING AN ACTIVE INFECTION THAT CAN BE VISUALIZED AND ANALYZED SIMULTANEOUSLY, WITHOUT COMPROMISING TISSUE ARCHITECTURE. FOLLOWING THIS, ROLE OF CERTAIN GENES AND PROTEINS WILL BE VALIDATED IN IN VITRO, AND EX VIVO ASSAYS USING ENDOTHELIAL CELL LINES, BY IMMUNOHISTOCHEMISTRY, AND EVALUATED IN VIVO FOR THEIR ROLE IN BIOFILM CLEARANCE. FURTHERMORE, WE WILL ASSESS THE EFFICACY OF TWO REPURPOSED FDA-APPROVED SMALL MOLECULES (THAT HAVE PREVIOUSLY DEMONSTRATED ACTIVITY IN MUCOSAL MODELS), FOR THEIR PROMISE AS ANTI-BIOFILM DRUGS. INDEED, BIOMARKERS OF HOST RESPONSE IDENTIFIED FROM AIM 1 WILL BE MEASURED POST DRUG TREATMENT, TO CORRELATE ERADICATION OF INFECTION WITH THE HOST RESPONSE. OVERALL, OUR STUDIES WILL UNEARTH THE HOST RESPONSE TO CENTRAL VENOUS CATHETER ASSOCIATED BIOFILMS, AND INVESTIGATE THE POTENCY OF TWO PROMISING COMPOUNDS IN ERADICATING BIOFILMS.

REGULATION OF CANDIDA ALBICANS CENTRAL VENOUS CATHETER BIOFILMS BY TRANSCRIPTION FACTOR NETWORKS - PROJECT SUMMARY/ABSTRACT BIOFILMS CONTRIBUTE TO OVER 80% OF MICROBIAL INFECTIONS. CANDIDA ALBICANS BIOFILMS, PARTICULARLY THOSE FORMING ON INDWELLING MEDICAL DEVICES SUCH AS CENTRAL VENOUS CATHETERS (CVCS), ARE A MAJOR SOURCE OF DISSEMINATED BLOODSTREAM INFECTIONS. WHILE IN VITRO STUDIES HAVE DEFINED CORE BIOFILM REGULATORS, THEY FAIL TO RECAPITULATE THE COMPLEXITY OF CLINICAL ENVIRONMENTS WHERE CVCS ARE EXPOSED TO CONTINUOUS BLOOD FLOW AND VASCULAR HOST SURFACES. CONSEQUENTLY, KEY REGULATORS OF BIOFILM FORMATION UNDER THESE CONDITIONS REMAIN ELUSIVE. TO ADDRESS THIS GAP, WE UTILIZE A CLINICALLY RELEVANT JUGULAR VEIN CATHETER MOUSE MODEL, IN WHICH C. ALBICANS BIOFILMS FORM UNDER PHYSIOLOGICAL BLOODSTREAM CONDITIONS. COMPLEMENTING THIS, WE HAVE DEVELOPED A HUMAN-RELEVANT EX VIVO VASCULARIZED CATHETER SYSTEM USING SILICONE CATHETER MATERIAL PRECONDITIONED WITH PLASMA AND SEEDED WITH HUMAN ENDOTHELIAL CELLS TO SIMULATE THE HOST-DEVICE INTERFACE. PRELIMINARY SCREENING OF C. ALBICANS TRANSCRIPTION FACTOR (TF) MUTANTS REVEALED THAT SEVERAL CANONICAL REGULATORS OF IN VITRO BIOFILM FORMATION (BCR1, BRG1, ROB1) ARE DISPENSABLE IN VIVO. NOTABLY, WE IDENTIFIED NOVEL REGULATORS, INCLUDING PHO4 AND CAS5, THAT ARE ESSENTIAL FOR VASCULAR BIOFILM GROWTH AND WERE NOT PREVIOUSLY LINKED TO THIS PROCESS. WE HYPOTHESIZE THAT DISTINCT, CONTEXT- SPECIFIC TRANSCRIPTIONAL CIRCUITS GOVERN CVC BIOFILM FORMATION IN VIVO AND CAN ONLY BE UNCOVERED THROUGH DIRECT INVESTIGATION IN HOST-RELEVANT SYSTEMS. IN AIM 1, WE WILL CONDUCT A COMPREHENSIVE SCREEN OF ~155 BARCODED TF MUTANTS IN VIVO TO IDENTIFY REGULATORS CRITICAL FOR BIOFILM GROWTH ON CVCS, FOLLOWED BY SECONDARY VALIDATION IN THE VASCULARIZED EX VIVO MODEL. IN AIM 2, WE WILL DEFINE THE DOWNSTREAM EFFECTOR GENES AND REGULATORY CIRCUITS CONTROLLED BY THESE TFS USING TRANSCRIPTOMIC PROFILING AND TARGETED MUTANT ANALYSIS. IMPORTANTLY, THIS TIERED STRATEGY PRIORITIZES VALIDATION IN HUMAN-RELEVANT MODELS AND LIMITS IN VIVO EXPERIMENTS, IN FULL ALIGNMENT WITH THE NIH’S POLICY PROMOTING NEW APPROACH METHODOLOGIES (NAMS) TO REDUCE ANIMAL USE. THIS RESEARCH WILL YIELD HIGH-RESOLUTION INSIGHTS INTO VASCULAR BIOFILM REGULATION AND UNCOVER NOVEL, CLINICALLY ACTIONABLE TARGETS TO PREVENT OR TREAT CATHETER-ASSOCIATED BLOODSTREAM INFECTIONS.

ROLE OF IMMUNOMODULATORS IL-33 AND SPRR IN OROPHARYNGEAL CANDIDIASIS - PROJECT SUMMARY/ABSTRACT FUNGAL INFECTIONS OF THE ORAL MUCOSA ARE A MAJOR COMPLICATION FOR THE IMMUNOCOMPROMISED HOST, INCLUDING IN NEONATES, CANCER PATIENTS, AND PATIENTS WITH HIV/AIDS. CANDIDA ALBICANS, AN OTHERWISE BENIGN COMMENSAL OF HUMAN ORAL CAVITY IS THE FOREMOST CAUSATIVE AGENT OF OROPHARYNGEAL CANDIDIASIS (OPC). DESPITE ANTIFUNGAL THER- APY, OPC CAUSES CONSIDERABLE MORBIDITY, NUTRITIONAL DEFICITS, AND INCREASED PREVALENCE OF ESOPHAGEAL CANCER. STUDIES ON THE IMMUNOLOGICAL MECHANISMS OF PROTECTION FROM OPC HAVE LARGELY FOCUSED ON THE PRO-INFLAMMATORY INNATE IMMUNE RESPONSE. SIGNALING IN ORAL EPITHELIAL CELLS THROUGH TH-17 CYTOKINES IL-17A, IL-17F, IL-22 AND STAT3 DRIVE CRITICAL MUCOSAL RESPONSES AGAINST CANDIDA, RECRUITING NEUTROPHILS, TRIGGERING MORE CYTOKINES, CHEM- OKINES, AND ANTIMICROBIAL PEPTIDES (AMP) RESULTING IN ACUTE TISSUE INFLAMMATION. LITTLE IS KNOWN ABOUT THE ANTI- INFLAMMATORY OR IMMUNOMODULATORY SIGNALS THAT REGULATE INFLAMMATION, AND AID IN TISSUE REPAIR. THUS, OUR KNOWLEDGE ON THE GLOBAL IMMUNITY WITHIN THE FUNGI-INFECTED MUCOSA REMAINS INCOMPLETE. TO OBTAIN A COMPREHENSIVE UNDERSTANDING OF THE HOST-PATHOGEN RESPONSES DURING OPC, WE USED A NEXT-GENER- ATION SEQUENCING TECHNOLOGY CALLED VISIUM 10X SPATIAL GENOMICS WHICH PROVIDED A “VISUAL” LANDSCAPE OF GLOBAL GENE EXPRESSION CHANGES IN INFECTED TONGUE SECTIONS. WHILE WE DID FIND THE TH-1 AND TH-17 ASSOCIATED PRO- INFLAMMATORY GENES HIGHLY UPREGULATED, AS EXPECTED; WE ALSO DISCOVERED THE PRESENCE OF TYPE-2 IMMUNE RE- SPONSES MARKED BY THE PRESENCE OF ANTI-INFLAMMATORY M2 MACROPHAGES, AND THE IMMUNOMODULATORY IL-33/ST2 AXIS. FURTHERMORE, A NOVEL FAMILY OF EPIDERMALLY PRODUCED SMALL PROLINE-RICH PROTEINS (SPRRS) WERE REMARKABLY UPREGULATED DURING OPC. EXOGENOUS TREATMENT WITH RECOMBINANT (REC) RECIL-33 OR RECSPRR COULD ERADICATE OPC IN MICE. IN FACT, RECSPRR PROTEINS DIRECTLY KILLED CANDIDA IN VITRO BY PERMEABILIZING THEIR CELL MEMBRANES. THE ROLE OF M2 MACROPHAGES, IL-33 OR SPRRS HAVE NEVER BEEN EXAMINED IN THE CONTEXT OF MUCOSAL CANDIDIASIS. IN AIM 1 WE WILL INVESTIGATE THE MECHANISTIC CONTRIBUTIONS OF IL-33 BOTH AS A PLEOTROPIC ALARMIN WITH ABILITIES TO TRIGGER INFLAMMATORY NEUTROPHILS, AS WELL AS A TYPE-2 CYTOKINE THAT CAN POLARIZE MACROPHAGES TO THE ANTI-INFLAM- MATORY M2 TYPE. IL-33’S EFFECT ON KEY DRIVERS OF PROTECTION INCLUDING: TH17, TH1, IMMUNOSUPPRESSIVE CYTOKINES AND TH2 WILL BE TESTED IN RECIL-33 TREATED VERSUS WT AND IL-33 -/- INFECTED MICE. IL-33 INCREASED UPON RECSPRR2A TREATMENT, INDICATING THAT SPRR LIKELY REGULATES IL-33 SECRETION FROM ORAL EPITHELIAL CELLS. TO INVESTI- GATE THE ROLE OF SPRRS IN THE HOST RESPONSE TO OPC (AIM 2), WE WILL DETERMINE THE ORAL FUNGAL BURDEN, CYTOKINE- CHEMOKINE RESPONSE (INCLUDING IL-33), AND THE EXTENT OF INFILTRATING NEUTROPHILS AND MONOCYTES IN WT AND SPRR- /- MICE. TO UNRAVEL THEIR POTENTIAL EPISTASIS WITH SPRR IN ORAL INFECTIONS, WE WILL EVALUATE IL-17/IL-23, IL-6/STAT3, AND IL-4, IL-13 IN SPRR-/- AND IN RECSPRR2A TREATED WT MICE AND USE KNOCKOUT ORAL KERATINOCYTE CELL LINES OF CYTOKINES PRIORITIZED FROM THE IN VIVO STUDIES. PROPOSED STUDIES WILL ADVANCE OUR KNOWLEDGE ON PROTECTIVE IMMUN- ITY DURING OPC THAT COULD BE MANEUVERED FOR FUTURE DEVELOPMENT OF NOVEL IMMUNOTHERAPEUTIC INTERVENTIONS.

MOUSE MODEL OF NEONATAL FUNGAL COLONIZATION TO STUDY HOMEOSTASIS AND DISEASE - SUMMARY WHILE THE FUNGUS CANDIDA ALBICANS HAS LARGELY BEEN STUDIED AS A PATHOGEN, ITS PRIMARY LIFESTYLE IS AS A COMMENSAL ON MUCOSAL SURFACES, INCLUDING THE ORAL CAVITY. MICROORGANISMS THAT OCCUPY MUCOSAL SURFACES ARE CRITICAL FOR APPROPRIATELY TUNING IMMUNE RESPONSES TO ENSURE EFFICIENT RESPONSES TO INVADING PATHOGENS WHILE LIMITING RESPONSES DIRECTED TOWARDS HOST TISSUES. IN THIS CONTEXT, COMMENSAL FUNGI PLAY IMPORTANT ROLES IN HOST IMMUNITY AND INFLAMMATION. FOR INSTANCE, C. ALBICANS COLONIZATION INDUCES CROSS-REACTIVE T CELLS AND INNATE MEMORY IN IMMUNE CELLS, A MECHANISM TERMED TRAINED IMMUNITY. ACCUMULATING EVIDENCE FROM OUR GROUP AND OTHERS HAVE CONVINCINGLY DEMONSTRATED THAT ORAL COMMENSAL COLONIZATION WITH C. ALBICANS INDUCES SPECIFIC IMMUNE RESPONSES, THEREFORE CONTRIBUTING TO MUCOSAL IMMUNE SYSTEM PLASTICITY. HOWEVER, OUR KNOWLEDGE OF ANTIFUNGAL IMMUNITY IS BASED ON DATA OF ADULT ANIMALS. IMPORTANTLY, HUMANS ARE COLONIZED WITH C. ALBICANS DURING BIRTH. FURTHERMORE, THE NEONATAL IMMUNE SYSTEM HAS LONG BEEN REGARDED IMMATURE WHEN COMPARED TO THE ADULT IMMUNE SYSTEM. HOWEVER, IS BECOMES CLEAR THAT NEONATAL IMMUNITY IS SPECIFICALLY ADAPTED TO PREVENT PATHOGEN INVASION, WHILE SUPPORTED ITS DEVELOPMENT. HOWEVER, THE FUNDAMENTAL IMMUNOLOGICAL CONSEQUENCES OF C. ALBICANS COLONIZATION IN NEONATES ARE STILL UNKNOWN. WE DEVELOPED A NOVEL MOUSE MODEL OF COMMENSAL CANDIDA COLONIZATION IN NEONATES. THIS PROPOSAL WILL CHARACTERIZE ORAL MUCOSAL IMMUNE CELL POPULATIONS IN NEONATES IN RESPONSE TO COMMENSAL CANDIDA COLONIZATION (AIM 1) AND ELUCIDATE AGE RELATED DIFFERENCES IN IL- 17 AND IL-22 IMMUNITY DURING ORAL FUNGAL COLONIZATION (AIM 2).

EFFECTIVENESS OF HOSPITAL-BASED PROGRAM FOR VACCINATION OF BIRTH MOTHERS AND HOUS

DEVELOPMENT OF IMMUNOTHERAPEUTIC STRATEGIES TO OVERCOME NEONATAL CANDIDIASIS

TARGETING EVOLUTIONARILY ACQUIRED INSERTION SEQUENCES IN CANDIDA SPECIES, FOR DEVELOPMENT OF ANTIFUNGAL DRUGS - ABSTRACT CANDIDA ALBICANS CAUSES HALF OF ALL INVASIVE FUNGAL INFECTIONS IN HUMANS. ITS SUCCESS AS A PATHOGEN IS DUE TO ITS ABILITY TO ADAPT AND THRIVE IN VARIOUS MICROENVIRONMENTS IN THE HOST. USING A FORWARD GENETICS SCREEN, WE IDENTIFIED AN UNCHARACTERIZED GENE NDU1 WHOSE LOSS CAUSED C. ALBICANS TO LOSE VIABILITY ON ALTERNATIVE CARBON SOURCES AND MADE THE ORGANISM COMPLETELY AVIRULENT IN VIVO. WE FOUND THAT CANDIDA-NDU1 PROTEIN POSSESSES THREE EVOLUTIONARILY ACQUIRED STRETCHES OF AMINO ACID INSERTS (~64 AMINO ACIDS), ABSENT FROM ALL OTHER EUKARYOTES INCLUDING HUMANS. WE REPORTED FOR THE FIRST TIME THAT ~17% OF THE CANDIDA PROTEOME POSSESS “ADDITIONAL” SETS OF EVOLUTIONARILY ACQUIRED AMINO-ACID INSERTS, THAT ARE ABSENT FROM ALL NON-CTG CLADE EUKARYOTES, INCLUDING HUMANS. THE ROLE OF INSERT REGIONS IN C. ALBICANS PROTEOME REMAINS TO BE UNRAVELED. WE HAVE SHOWN THAT THE NDU1 INSERTION SEQUENCES ARE VITAL AND REQUIRED FOR NDU1’S ACTIVITY. IN AIM 1, THE MECHANISTIC CONTRIBUTION OF NDU1 INSERTS WILL BE INVESTIGATED BY PROTEIN BIOCHEMISTRY STUDIES. THE ROLE OF INSERT REGIONS IN OTHER CANDIDA PROTEINS WILL ALSO BE EVALUATED , TO UNDERSTAND IF INSERTS ARE UNIVERSALLY IMPORTANT FOR FUNCTION. CONSIDERING NDU1 IS IMPORTANT FOR GROWTH AND VIRULENCE IN VIVO, WE TARGETED NDU1 AND ITS INSERT REGIONS FOR DEVELOPMENT OF ANTIFUNGAL MOLECULES. WE IDENTIFIED A REPURPOSED FDA APPROVED DRUG NICLOSAMIDE (NCL), AND DEVELOPED EUDRAGIT EPO NANOPARTICLE FORMULATIONS OF THIS MOLECULE TO ENHANCE ITS SOLUBILITY AND BIOAVAILABILITY. NCL NANOPARTICLES (NCL-EPO-NP) COULD PREVENT GROWTH OF CANDIDA IN ALTERNATIVE CARBON SOURCES, PENETRATE AND DISMANTLE MATURE BIOFILMS IN VITRO, AND ERADICATE IT IN VIVO IN BIOFILM MODELS OF MUCOSAL CANDIDIASIS. THE ACTIVITY OF NCL-NP WAS 6-10 FOLD HIGHER THAN THE GENERIC DRUG ALONE. IN AIM 2, WE FURTHER CHARACTERIZE AND EVALUATE NCL-EPO-NP FOR THEIR PHARMACOKINETICS AND ORAL BIODISTRIBUTION, TOXICITY PROFILES AND IN VIVO EFFICACY IN AN ORAL REGIMEN FOR TREATMENT OF ORAL CANDIDIASIS, AS STANDALONE OR ADJUNCT THERAPY WITH OTHER ANTIFUNGAL DRUGS.

UNRAVELING TRANSCRIPTOMIC AND FUNCTIONAL CHANGES TO IMMUNE SENSING NEURONAL GANGLIA IN RESPONSE TO ALLERGY AND BACTERIA - PROJECT SUMMARY ALLERGIES AFFECT MILLIONS OF AMERICANS AND ARE EXACERBATED BY BACTERIAL INFECTION. DESPITE MAINSTAY THERAPIES THAT SUPPRESS IMMUNE RESPONSES TO ALLERGY OR BACTERIA, THESE HEALTH ISSUES CONTINUE TO PERSIST. AN UNEXPLORED TARGET FOR NEW THERAPIES LIES WITH NEURAL SURVEILLANCE AND REGULATION OF IMMUNITY. HOWEVER, THIS NEURAL INFLUENCE IS POORLY UNDERSTOOD AND SEEMINGLY PARADOXICAL: IMMUNE RESPONSES ARE EXACERBATED IN ALLERGY, BUT IN RESPONSE TO BACTERIAL INFECTION, THE NEURAL RESPONSE REDUCES INFLAMMATION. HOW MIGHT THIS HAPPEN? ONE POSSIBILITY IS THAT THE PERIPHERAL NEURAL-IMMUNE SENSING APPARATUS MIGHT BE ALTERED BY SUCH FACTORS AS IMMUNOGLOBULIN PROFILES, AND GENDER, IMPLYING THE PARTICIPATION OF SEX HORMONES. WE HYPOTHESIZE THAT SUCH VARIABLES AFFECT THE TRANSCRIPTOME AND THEREBY THE EXCITABILITY OF THE VAGUS AND CAROTID BODIES. STUDIES IN AIM 1 WILL USE SINGLE-CELL RNASEQ ANALYSIS OF VAGAL AND CAROTID BODY GANGLIA FROM MALE AND FEMALE MICE WITH ASTHMA, PNEUMONIA TO IDENTIFY HOW THESE NERVES ARE CHANGED IN RESPONSE TO EACH CONDITION. THEN, WE WILL ASSESS NERVE ACTIVITY IN RESPONSE TO IMMUNOGLOBULIN STIMULI OF EACH GANGLIA USING STATE-OF-THE-ART ELECTROPHYSIOLOGICAL RECORDING TECHNIQUES. THIS PROJECT WILL REVEAL WHICH NEURAL CLUSTER (VAGUS VS. CAROTID BODY) IS THE PREDOMINANT SENSORY GANGLIA INVOLVED IN REGULATING ALLERGIC VS. BACTERIAL IMMUNE RESPONSES AND HOW SEX MAY AFFECT NEURO- IMMUNE SIGNALLING. STUDIES IN AIM 2 WILL UTILIZE DTR (DIPHTHERIA TOXIN RECEPTOR) TECHNOLOGY IN MICE TO SELECTIVELY ABLATE CELLS WHERE DTR EXPRESSION IS DRIVEN BY TRPV1 (TRANSIENT RECEPTOR POTENTIAL VANILLOID 1) OR TH (TYROSINE HYDROXYLASE). USE OF THESE TRANSGENIC MICE WILL ALLOW ABLATION OF EITHER VAGAL (TRPV1-DTR) OR CAROTID BODY (TH- DTR) CENTERS BY DIRECT NEURAL MICROINJECTION OF DIPHTHERIA TOXIN (DTX) INTO VAGAL OR CAROTID BODY GANGLIA USING SALINE INJECTIONS AS CONTROL. WE WILL THEN EXPOSE MICE TO EITHER ALLERGEN OR BACTERIAL INFECTIONS. DTX INJECTIONS WILL SELECTIVELY ABOLISH NEURONAL FUNCTION AND REVEAL WHICH GANGLION IS THE PREDOMINANT NEURO-IMMUNE SENSOR IN THE SPECIFIED CONDITION IN MALE AND FEMALE MICE. THESE DATA WILL: 1) IDENTIFY HOW THE RESPONSE TO ALLERGIC OR BACTERIAL STIMULI ALTERS RECEPTOR GENE EXPRESSION IN ORDER TO IDENTIFY THE PHENOTYPIC SWITCH FROM ANTI- TO PRO-INFLAMMATORY NEURAL-IMMUNE SIGNALLING; 2) ESTABLISH A MODEL AND SEX DIFFERENCES THAT WILL SET THE STAGE FOR FURTHER INVESTIGATION INVOLVING TARGETED KNOCKDOWN WITHIN NEURO-IMMUNE SENSING GANGLIA TO DISCERN FUNCTION; 3) VALIDATE OUR MODELS AND METHODS FOR FUTURE INVESTIGATIONS INTO THE ROLE OF NEURAL-IMMUNE SENSING IN DISEASE STATES.

PSYSTART RAPID TRIAGE OF PTSD RISK IN PEDIATRIC INJURY PATIENTS

MONOCLONAL ANTIBODY PASSIVE VACCINATION TO TREAT XDR ACINETOBACTER INFECTIONS

BISPHENOL A EFFECTS ON THE PERIPHERAL MECHANISMS OF PENILE ERECTION

PREVENTION OF OBESITY?IN INFANTS OF OVERWEIGHT AND OBESE WOMEN - THROUGHOUT THE WORLD, OBESITY AND METABOLIC SYNDROME ARE AN ONGOING EPIDEMIC CRISIS, PRESENTING MAJOR PUBLIC HEALTH CHALLENGES AND SIGNIFICANT ECONOMIC BURDENS. THE DEVELOPMENTAL PROGRAMMING EFFECTS OF THE IN-UTERO ENVIRONMENT IS DEMONSTRATED BY THE INCREASED RISK OF LARGE FOR GESTATIONAL AGE (LGA) NEWBORNS AND CHILDHOOD AND ADULT OBESITY IN OFFSPRING OF OVERWEIGHT/OBESE (OW/OB) MOTHERS. ANIMAL STUDIES HAVE CONFIRMED THAT MATERNAL OBESITY AND HIGH FAT DIET PROGRAMS THE OFFSPRING APPETITE CENTER, THE ARCUATE NUCLEUS, RESULTING IN OFFSPRING HYPERPHAGIA AND RAPID WEIGHT GAIN, CONTRIBUTING TO THE GENERATIONAL CYCLE OF OBESITY. COMPOUNDING THE EFFECT OF PROGRAMMED HYPERPHAGIA IS THE IMPACT OF BREAST MILK CALORIC CONTENT. DESPITE EVIDENCE THAT BREASTFEEDING REDUCES THE INCIDENCE OF CHILDHOOD OBESITY, BREASTFEED LGA INFANTS’ CHILDHOOD OBESITY RATES ARE NEARLY 50% GREATER THAN FORMULA FED APPROPRIATE FOR GESTATIONAL AGE INFANTS. MATERNAL BMI AND DIETARY INTAKE ARE HIGHLY CORRELATED WITH MILK TOTAL CALORIE AND FAT CONTENT. OUR LABORATORY STUDIES SHOW THAT OFFSPRING BORN TO AND NURSED BY OBESE RAT DAMS EXHIBIT HYPERPHAGIA AND DEVELOP EARLY LIFE AND ADULT OBESITY. IN CONTRAST, IF THESE OFFSPRING ARE CROSS FOSTERED AND NURSED BY NON-OBESE DAMS (MATCHED FOR GENDER AND LITTER-SIZE), THE OFFSPRING GROW TO NORMAL WEIGHT AND BMI AS ADULTS. THESE STUDIES SUGGEST THAT DESPITE PROGRAMMED OFFSPRING HYPERPHAGIA AT BIRTH, THE RESTRICTION TO CONTROL DAM MILK PREVENTS EXCESSIVE NEWBORN WEIGHT GAIN AND OBESITY. WE PROPOSE THAT NORMALIZED CALORIC INTAKE AND NEWBORN GROWTH WILL PREVENT THE ONSET OF INFANT OBESITY, HYPERLEPTINEMIA (AND LEPTIN RESISTANCE), AND THUS NORMALIZE APPETITE THROUGHOUT CHILDHOOD. THE PROPOSED STUDIES WILL 1) CONFIRM THAT OW/OB MOTHERS’ MILK HAS INCREASED TOTAL CALORIE CONTENT AND THAT INFANTS OF OW/OB MOTHERS HAVE INCREASED MILK INTAKE, AND AS AN INTERVENTION 2) DETERMINE IF TITRATED HUMAN MILK OR FORMULA CALORIE INTAKE PREVENTS EXCESSIVE INFANT WEIGHT GAIN IN THE FIRST SIX MONTHS OF LIFE. THIS HIGHLY INNOVATIVE STUDY WILL EXAMINE THE POTENTIAL EFFICACY OF A NOVEL PREVENTATIVE STRATEGY TO PREVENT THE DEVELOPMENT OF INFANT OBESITY AND INTERRUPT THE GENERATIONAL CYCLE OF OBESITY.

DISPARITIES IN THE EFFECT OF STATE POLICY FOR THE NEWBORN SCREENING FOR CRITICAL CONGENITAL HEART DISEASES - INEQUITIES IN HEALTH OUTCOMES FOR INFANTS FROM LOWER SOCIAL ECONOMIC GROUPS; UNINSURED, PUBLICLY INSURED, OR RURAL FAMILIES, AND MARGINALIZED RACIAL AND ETHNIC GROUPS RESULT IN DISTURBING MORTALITY STATISTICS, ESPECIALLY AMONG BLACK INFANTS. THEREFORE OUR PROPOSED STUDY LOOKS AT CRITICAL CONGENITAL HEART DISEASE (CCHD), THE MOST SEVERE FORMS OF CONGENITAL HEART DISEASE, REQUIRING SURGERY OR CATHETER-BASED INTERVENTIONS WITHIN THE FIRST YEAR. OCCURRING IN 7,200 NEWBORNS AND CAUSING ABOUT 1,260 INFANT DEATHS IN THE U.S EACH YEAR, MORTALITY FOR CONGENITAL HEART DISEASE IS BETWEEN 35-40% HIGHER AMONG BLACK INFANTS IN THEIR FIRST YEAR THAN THEIR WHITE COUNTERPARTS, DEPENDING ON SUBTYPE AND STATE HEALTHCARE MODEL. CURRENTLY, ADVANCES IN ULTRASOUND AND ECHOCARDIOGRAPHY, DIAGNOSTIC TOOLS THAT ARE EXPENSIVE AND REQUIRE ADVANCED SKILLS, ALLOW CERTAIN TYPES OF CCHD TO BE DIAGNOSED PRENATALLY AMONG WOMEN. YET, INEQUITIES ARE SUBSTANTIAL IN PRENATAL DETECTION RATES, WHICH ARE 71-100% IN TEACHING HOSPITALS, AND 0-39% IN NON-TEACHING HOSPITALS. ALL 50 STATES AND D.C. NOW MANDATE NEWBORN SCREENING USING PULSE OXIMETRY (POX), A SCREENING ADDED TO THE RECOMMENDED UNIFORM SCREENING PANEL IN 2011 WHICH CAN COST-EFFECTIVELY SCREEN FOR CCHD WITH A SIMPLE AND NON-INVASIVE PROCEDURE. FULL ACCESS AND CORRECT USE OF POX PROVIDES THE LAST BEST OPPORTUNITY FOR PROMPT DIAGNOSIS TO PREVENT MORTALITY AND SECONDARY MORBIDITIES IN UNDER-RESOURCED AND MARGINALIZED COMMUNITIES. FOR OUR PROPOSED STUDY, WE HYPOTHESIZE THAT HEALTH OUTCOMES WILL IMPROVE AND HEALTHCARE UTILIZATION DECREASE MORE FOR HISTORICALLY UNDERSERVED AND MARGINALIZED POPULATIONS THAN THEIR MORE PRIVILEGED COUNTERPARTS AFTER IMPLEMENTATION OF STATE POX SCREENING MANDATES, AND ESPECIALLY AMONG BLACK INFANTS, GIVEN THE HIGH MORTALITY INEQUITIES. WE WILL EXAMINE HOSPITAL, BIRTH, AND DEATH RECORDS FROM TEN STATES (FL, GA, MA, MI, MO, NE, PA, SC, TN, WA) AND ONE CITY (NYC), COVERING OVER ONE THIRD OF U.S. LIVE BIRTHS, TO ASSESS DIFFERENCES IN THE EFFECT OF STATE-MANDATED POX SCREENING POLICIES ON TRADITIONALLY UNDERSERVED AND MARGINALIZED POPULATIONS AND THEIR MORE PRIVILEGED COUNTERPARTS. OUTCOMES WILL INCLUDE [1] INFANT DEATHS DUE TO CCHD, [2] INFANT DEATHS DUE TO MISSED OR LATE DIAGNOSES OF CCHD, AND [3] USE OF HEALTHCARE RESOURCES DURING INFANCY AMONG CCHD PATIENTS, SUCH AS HOSPITALIZATION COST AND LENGTH OF STAY. A DIFFERENCE-IN-DIFFERENCES MODEL, WHICH IS COMMONLY USED TO EVALUATE THE CHANGES IN OUTCOMES ASSOCIATED WITH HEALTH CARE POLICY IMPLEMENTATIONS, WILL BE USED. IF MANDATORY SCREENING NARROWS GAP TO IMPROVED OUTCOMES AMONG UNDER- RESOURCED AND MARGINALIZED POPULATIONS, AND ESPECIALLY BLACK INFANTS, THE NEXT STEP WILL BE A MIXED METHODS R01 TO LINK MOTHER AND CHILD HEALTH DATA, INCLUDING REGISTRIES, TO DETERMINE CHARACTERISTICS THAT INCREASE LIKELIHOOD OF DEVELOPING CCHDS. USING IN-DEPTH INTERVIEWS AND A QUESTIONNAIRE SURVEY, WE WILL DEVELOP INTERVENTIONS TARGETED TO MARGINALIZED FAMILIES TO PROMOTE PRENATAL CARE AND PRENATAL CHD DETECTION, AND THE EXPANSION OF POX SCREENING.

UNDERSTANDING SOURCES OF BIAS AND OUTCOMES IN PERITONEAL DIALYSIS PATIENT RESEARC

ROLE OF MULTI-DRUG RESISTANT CANDIDA AURIS HYR1 / IFF-LIKE PROTEINS IN VIRULENCE AND THEIR POTENTIAL AS VACCINE TARGETS - PROJECT SUMMARY: CANDIDATE’S CAREER DEVELOPMENT: DR. SHAKTI SINGH, AN ACCOMPLISHED IMMUNOLOGIST, PRESENTLY HOLDS THE POSITIONS OF INVESTIGATOR AND ASSISTANT PROFESSOR AT THE LUNDQUIST INSTITUTE (TLI) AND THE UNIVERSITY OF CALIFORNIA, LOS ANGELES (UCLA), RESPECTIVELY. OVER THE PAST SEVEN YEARS AT TLI, DR. SINGH HAS EXHIBITED REMARKABLE PRODUCTIVITY, PUBLISHED OVER 20 RESEARCH ARTICLES, AND HAS ASCENDED FROM A POSTDOCTORAL TRAINEE TO A FACULTY RANK. THIS APPLICATION OUTLINES A PROJECT LEVERAGING HIS EXISTING EXPERTISE TO FILL GAPS IN HIS TRAINING IN THE FUNGAL FIELD, WITH THE ULTIMATE GOAL OF ACHIEVING FULL INDEPENDENCE. MENTORS/ENVIRONMENT: DR. SINGH WILL BE GUIDED BY A DIVERSE TEAM OF ACCOMPLISHED SCIENTISTS WITH EXPERTISE IN FUNGAL PATHOGENESIS, IMMUNOLOGY, VACCINE DEVELOPMENT, ANIMAL MODELING, AND MRNA VACCINE TECHNOLOGY. THIS TEAM IS WELL-POSITIONED TO PROVIDE HANDS-ON EXPERIENCE AND SUPPORT, DR. SINGH, IN ACHIEVING THE OUTLINED TRAINING GOALS: 1) FILLING GAPS IN RESEARCH EXPERTISE, 2) DEVELOPING PROFESSIONAL AND LEADERSHIP SKILLS, AND 3) CONDUCTING RESEARCH RESPONSIBLY, ETHICALLY, AND EFFICIENTLY. THE COLLABORATIVE AND ENRICHING ENVIRONMENT, COUPLED WITH THE ABUNDANT RESOURCES AT TLI, HARBOR-UCLA MEDICAL CENTER, AND UCLA, OFFERS AN OPTIMAL SETTING FOR COMPREHENSIVE PROFESSIONAL TRAINING. RESEARCH: C. AURIS IS A RECENTLY DISCOVERED CANDIDA SPECIES THAT CAUSES LIFE-THREATENING BLOODSTREAM INFECTIONS (BSI) IN IMMUNOSUPPRESSED PATIENTS AND HAS A VERY HIGH (~60%) MORTALITY RATE. C. AURIS EASILY COLONIZES AND SPREADS IN HEALTHCARE SETTINGS AND ADHERES AND FORMS BIOFILMS ON MEDICAL DEVICES. THE MAJORITY (~88%) OF C. AURIS BSI CASES CAN BE ATTRIBUTED TO THE INVOLVEMENT OF CATHETERS AND OTHER INVASIVE MEDICAL DEVICES. C. AURIS INFECTIONS ARE HARD TO TREAT BECAUSE C. AURIS CAN RESIST ALL CLASSES OF CLINICALLY AVAILABLE ANTIFUNGAL DRUGS. DUE TO THESE REASONS, THE WORLD HEALTH ORGANIZATION AND CENTER FOR DISEASE PREVENTION AND CONTROL (CDC) HAVE CATEGORIZED C. AURIS AS A “CRITICAL THREAT” AND “URGENT THREAT” PATHOGEN, RESPECTIVELY. TO ADDRESS THIS URGENT PUBLIC HEALTH CHALLENGE, WE HAVE IDENTIFIED THREE HYR1/IFF FAMILY (CAU-HIL) PROTEINS THAT DEMONSTRATE UNIVERSAL PRESENCE ACROSS ALL CLADES OF C. AURIS. USING A MONOCLONAL ANTIBODY TARGETED AGAINST AN EPITOPE IN THE C. ALBICANS HYR1 PROTEIN, CONSERVED IN CAU-HIL, WE HAVE ASCERTAINED THESE PROTEINS AS POTENTIAL THERAPEUTIC TARGETS. HOWEVER, CAU-HIL PROTEINS EXHIBIT ONLY ~35% SEQUENCE IDENTITY WITH CHARACTERIZED PROTEINS IN OTHER CANDIDA SPECIES, AND THEIR SPECIFIC ROLE IN C. AURIS VIRULENCE REMAINS UNCHARACTERIZED. WE HYPOTHESIZE THAT THESE CAU-HIL PROTEINS WOULD BE SUPERIOR VACCINE CANDIDATES AGAINST C. AURIS INFECTION COMPARED TO C. ALBICANS ANTIGENS, SIMPLY BECAUSE OF THEIR C. AURIS ORIGIN. THEREFORE, WE PROPOSE TO: 1. STUDY THE ROLE OF HIL PROTEINS IN C. AURIS VIRULENCE, 2. STUDY THEIR POTENTIAL AS A VACCINE AGAINST C. AURIS USING AN INNOVATIVE MRNA PLATFORM AND CLINICALLY RELEVANT ANIMAL MODELS.

ROLE OF MIR29C AND MIR200C IN LEIOMYOMA PATHOGENESIS

EMSC TARGETED ISSUE GRANTS

EARLY AGR ACTIVATION IS A KEY PATHOGENIC SIGNATURE IN PERSISTENT MRSA BACTEREMIA

AN FTR1 VACCINE TO ABROGATE MUCORMYCOSIS

TWO-DIMENSIONAL 3 TESLA MRS OF THE BRAIN IN HIV- INFECTED YOUTH

MITIGATING RESISTANCE & VIRULENCE IN MRSA

MOLECULAR BASIS OF CHILDHOOD ASTHMA FOLLOWING PERINATAL VITAMIN D DEFICIENCY

EFFECTS OF ACUPUNCTURE-INDUCED NNOS-NO IN DORSAL MEDULLA ON SENSORY NEUROPATHY

IN UTERO NICOTINE EXPOSURE & TRANSGENERATIONAL TRANSMISSION OF ASTHMA

EFFECTS OF ADVANCED EA ON HYPERTENSION: ROLE OF NNOS-NO IN THE DORSAL MEDULLA

ARF6 INHIBITION AS NOVEL TREATMENT FOR MULTIDRUG RESISTANCE GRAM NEGATIVE INFECTIONS

DEVELOPMENTALLY PROGRAMMED HYPERPHAGIA AND OBESITY VIA BPA ENHANCED NEUROGENESIS

GLYCOSYLATION-INDEPENDENT ENZYME THERAPY OF THE BRAIN IN SANFILIPPO B SYNDROME

IGF::OT::IGF BAA-11-01-HPW SUBDISSOCIATIVE DOSE KETAMINE FOR TREATMENT OF ACUTE PAIN IN SUBJECTS WITH CHRONIC PAIN: A RANDOMIZED CONTROLLED TRIAL

ASTHMATIC INFLAMMATION REQUIRES NEURONAL UPREGULATION OF B-CELLS - PROJECT SUMMARY ASTHMA AND ASTHMATIC SYMPTOMS CONTINUE TO PERSIST AND ARE A GROWING HEALTH BURDEN DEMONSTRATING THE NEED TO UNDERSTAND HOW THE MAIN ASTHMATIC EFFECTOR MOLECULE, IMMUNOGLOBULIN (IG) E (IGE), IS PRODUCED. ASTHMATIC PATHOLOGY OCCURS BY IGE BEING PRODUCED IN RESPONSE TO WHAT ARE TYPICALLY INNOCUOUS SUBSTANCES. WHILE IGE PRODUCTION CAN LEAD TO PROTECTIVE IMMUNITY IN RESPONSE TO VENOMS AND MULTICELLULAR PATHOGENS, ITS OVERPRODUCTION IN RESPONSE TO BENIGN SUBSTANCES UNDERLIES ITS ROLE IN ASTHMATIC AND ALLERGIC INFLAMMATION. IGE IS SECRETED FROM B LINEAGE CELLS IN THE PRESENCE OF COGNATE ANTIGEN AND ACTIVATES MAST CELLS AND BASOPHILS TO RELEASE POTENT INFLAMMATORY MOLECULES WHICH ACT DIRECTLY ON THE AIRWAY WALL AND ATTRACT INFLAMMATORY CELLS TO THE LUNGS. EMERGING EVIDENCE DEMONSTRATES THAT ALLERGIC INFLAMMATION SENSITIZES ANOTHER CELL TYPE- NEURONS. ONCE SENSITIZED, NEURONS RELEASE A HOST OF NEUROTRANSMITTERS WHICH ENHANCE ALLERGIC INFLAMMATION, AND RECENT DATA INDICATE THE NEURONAL ABILITY TO ENHANCE B-CELL MEDIATED IGE OVERPRODUCTION. HOWEVER, WHAT SENSITIZES THESE NEURONS AND WHAT EFFECT SENSITIZED NEURONS HAVE ON ASTHMATIC PATHOLOGY, IN PARTICULAR THE OVERPRODUCTION OF IGE, REQUIRES FURTHER INVESTIGATION. THE OVERALL OBJECTIVE OF THIS APPLICATION IS TO INVESTIGATE THE ROLE OF THE SENSORY AND AUTONOMIC NEURONS IN IGE OVERPRODUCTION IN ALLERGIC ASTHMA. OUR PRELIMINARY DATA HAS LED US TO HYPOTHESIZE THAT SENSORY NEURONS STIMULATE SYMPATHETIC NEURONS TO STIMULATE B-CELL OVERPRODUCTION TO INCREASE IGE RELEASE. WE SHOW THAT SENSORY NEURONS CAN SENSE ANTIGEN ALONGSIDE ANTIGEN-SENSING IMMUNE CELLS. THIS STIMULATION OF SENSORY NEURONS BY ANTIGEN THEN INCREASES B-CELL PRODUCTION BY ELICITING SYMPATHETIC REFLEX ACTIVATION. ONCE THE SURPLUS B-CELL RESERVOIR REACHES THE ALLERGIC LUNG, STIMULATED SENSORY NEURONS THEN RELEASE THEIR NEUROTRANSMITTERS DURING ANTIGEN RECOGNITION WHICH STIMULATES THE OVERPRODUCTION OF IGE. WE WILL INVESTIGATE THIS NOVEL NEURONAL CIRCUIT OF IGE OVERPRODUCTION IN TWO AIMS. STUDIES IN AIM 1 WILL USE ROBUST ELECTROPHYSIOLOGICAL RECORDING AND CALCIUM IMAGING TECHNIQUES TO ASSESS A NOVEL SIGNALING PATHWAY OF SENSORY NEURON ANTIGEN RECOGNITION, EXCITATION, AND HYPERSENSITIZATION. SUBSEQUENT MOLECULAR ASSAYS WILL DELINEATE THE EXACT DOWNSTREAM MOLECULE EFFECTOR WITH THE HOPES TO IDENTIFY POTENTIAL DRUG TARGETS FOR FUTURE STUDY. STUDIES IN AIM 2 WILL USE NEURAL TRACER TECHNOLOGY TO LEARN WHICH BRAIN REGIONS ARE INVOLVED IN THIS NOVEL NEURONAL CIRCUIT. FURTHER STUDIES IN THIS AIM WILL TEST THE NECESSITY AND SUFFICIENCY OF SENSORY NEURONS AND AUTONOMIC NEURONS IN REGULATING B-CELL PRODUCTION USING STATE- OF-THE-ART CHEMOGENETIC TOOLS. ISOLATED CELL EXPERIMENTS WILL DELINEATE WHICH NEURONAL MOLECULES AFFECT B-CELL IGE PRODUCTION. THESE DATA WILL DRIVE FURTHER EXPERIMENTS TO IDENTIFY HOW THE ENTIRE PATHWAY WORKS IN A MOUSE MODEL OF FUNGAL ALLERGIC ASTHMA. THIS CONTRIBUTION IS SIGNIFICANT BECAUSE IT IS EXPECTED TO ELUCIDATE A COMPLETE PICTURE OF HOW IGE IS PRODUCED AND DYSREGULATED IN ASTHMA. SUCH KNOWLEDGE HAS THE POTENTIAL TO INFORM THE DEVELOPMENT OF NEW STRATEGIES THAT WILL HELP TO REDUCE THE GROWING PROBLEM OF ALLERGIC ASTHMA.

EXPLORING THE ROLE OF HEDGEHOG PATHWAY-PRIMARY CILIA AXIS IN HUMAN LUNG ALVEOLOGENESIS AND DISEASE - PROJECT SUMMARY/ ABSTRACT BRONCHOPULMONARY DYSPLASIA (BPD) IS THE MOST COMMON CHRONIC LUNG DISEASE OF PREMATURITY, CHARACTERIZED BY DEFECTIVE GAS EXCHANGE AREA, DUE TO REDUCED ALVEOLAR SURFACE. THE ALVEOLAR EPITHELIUM IS COMPOSED OF SQUAMOUS ALVEOLAR EPITHELIAL TYPE 1 CELLS (AT1), WHICH COVER MORE THAN 90% OF THE ALVEOLAR SURFACE, AND CUBOIDAL ALVEOLAR EPITHELIAL TYPE 2 (AT2) CELLS THAT POSSESS STEM AND PROGENITOR CELL CAPABILITIES AS DEMONSTRATED BY THEIR ABILITY TO SELF-RENEW, MAINTAIN THE AT2 POOL, AND DIFFERENTIATE INTO AT1 DURING ALVEOLARIZATION AND FOLLOWING LUNG INJURY. THE ALVEOLAR EPITHELIUM SERVES AS A PHYSICAL BARRIER AGAINST ENVIRONMENTAL STIMULI. ALVEOLAR FORMATION IN HUMANS OCCUR IN UTERO AND IS REGULATED BY MULTIPLE CELLULAR AND MOLECULAR PROCESSES, INVOLVING PROLIFERATION, DIFFERENTIATION, AND EPITHELIAL-MESENCHYMAL CROSSTALK. THE HEDGEHOG (HH) PATHWAY PLAYS AN IMPORTANT ROLE IN ORCHESTRATING THESE EVENTS AND ITS DEREGULATION CAN RESULT IN DEFECTIVE LUNG DEVELOPMENT, THUS LEADING TO DISEASES SUCH AS BPD. ABERRANT HH SIGNALING IS IMPLICATED IN BPD AS WELL AS VARIOUS ADULT LUNG DISEASES, SUCH AS ASTHMA AND COPD. WE RECENTLY DESCRIBED THAT THE HH SIGNALING PLAYS AN IMPORTANT ROLE IN EARLY HUMAN LUNG DEVELOPMENT. THE HH PATHWAY LIGAND, SONIC HEDGEHOG (SHH) REGULATES PROGENITOR CELL INTERACTIONS IN THE AIRWAY AND ALVEOLAR COMPARTMENTS DURING LUNG DEVELOPMENT. OUR PREVIOUSLY GENERATED SCRNASEQ DATA DEMONSTRATE A CONTINUOUS INCREASE OF SHH EXPRESSION DURING HUMAN LUNG DEVELOPMENT. ADDITIONALLY, THE HH PATHWAY ACTIVATOR (PTCH1) IS HIGHLY EXPRESSED DURING EARLY LUNG DEVELOPMENT, COINCIDING WITH THE EXPANSION OF THE PROGENITOR CELL POOL, WHILE THE INHIBITOR (HHIP) IS HIGHLY EXPRESSED DURING PROGENITOR DIFFERENTIATION. THESE OBSERVATIONS SUGGEST THAT EARLY ACTIVATION OF THE HH PATHWAY IS ESSENTIAL IN EARLY LUNG DEVELOPMENT, WHEREAS ITS INHIBITION PROMOTES ALVEOLOGENESIS. THE HH PATHWAY IS REGULATED BY SENSORY ORGANELLES, CALLED PRIMARY CILIA (PC), WHICH PLAY AN IMPORTANT ROLE IN THE TRANSDUCTION OF THE HH SIGNALING. THE PRESENCE AND PROPER FUNCTION OF PC ARE KNOWN TO BE ESSENTIAL FOR THE ACTIVATION AND MODULATION OF THE HH PATHWAY DURING VARIOUS STAGES OF LUNG DEVELOPMENT. WE RECENTLY DEMONSTRATED A DYSREGULATION OF THE HH PATHWAY AND PC IN COPD, AND OUR PRELIMINARY DATA SUGGEST DISRUPTION OF THESE ELEMENTS IN BPD. THEREFORE, WE HYPOTHESIZE THAT A HH/PRIMARY CILIA SIGNALING AXIS IS CRUCIAL DURING ALVEOLOGENESIS, FACILITATING THE MATURATION OF AT2 CELLS. TO TEST THIS HYPOTHESIS, WE WILL 1) INVESTIGATE THE ROLE OF HH SIGNALING IN THE MATURATION OF ALVEOLAR EPITHELIAL CELLS (K99), 2) INVESTIGATE THE INTERACTION BETWEEN PC AND HH SIGNALING IN ALVEOLOGENESIS (K99/R00), AND 3) ASSESS THE INTERPLAY BETWEEN THE HH PATHWAY AND PC IN BPD (R00). THE PROPOSED CAREER DEVELOPMENT PLAN IS DESIGNED TO EQUIP THE PI WITH UNIQUE SKILL SETS, EXPAND KNOWLEDGE BASE AND RESEARCH EXPERIENCE TO BECOMING A PRODUCTIVE AND INDEPENDENT INVESTIGATOR RESEARCHER IN THE FIELD OF LUNG BIOLOGY, ALVEOLOGENESIS AND BPD.

CELLULAR-MOLECULAR SIGNATURE AND MECHANISM OF BPA EFFECTS ON PENIL ERECTION

DREW_UCLA CONNECT_COMMUNITY PARTNERED RESOURCES TO IMPROVE DEPRESSION OUTCOMES

ISCHEMIC ARREST, CYTOKINES, AND ORGAN DYSFUNCTION

NOVEL CANDIDA ALBICANS HOST RECEPTORS DURING INFECTION AND IMMUNE RESPONSE

DDT-BMQ-000161 ANALYTIC VALIDATION OFNON-CALCIFIED PLAQUE VOLUME (NCPV) FOR REGULATORY QUALIFICATION AS AN ENRICHMENT BIOMARKERIN CORONARY ARTERY DISEASE TRIALS - CORONARY CT ANGIOGRAPHY (CCTA) IMAGING IS A WELL-DEVELOPED, NON-INVASIVE TOOL WHICH VISUALIZES ATHEROSCLEROSIS, SATISFYING CONDITIONS BE BASED ON EARLY ASSESSMENT OF THE ACTUAL DISEASE PROCESS OF ATHEROSCLEROSIS, OR PLAQUE AND HAS BEEN SHOWN TO ACCURATELY RISK STRATIFY INDIVIDUALS FOR EARLY PROGRESSION OF ATHEROSCLEROSIS AS WELL AS LATE ADVERSE EVENTS. HERE WE PROPOSE SEVERAL STUDIES TO STREAMLINE DRUG DEVELOPMENT AND ENCOURAGE INNOVATION BY DETECTING EARLY, BENEFICIAL DRUG RESPONSES ON ATHEROSCLEROSIS; AND IDENTIFY ATHEROSCLEROSIS BIOMARKER(S) TO SERVE AS SURROGATE ENDPOINTS FOR CLINICAL TRIALS. THE OVERARCHING SIGNIFICANCE OF THE CURRENT GRANT IS TO ADDRESS THE UNMET NEED FOR NON-INVASIVE MEASUREMENT OF CORONARY ATHEROSCLEROSIS, BY SUPPORTING INVESTIGATIONS THAT WILL ADVANCE THE QUALIFICATION PLAN APPLICATION FOR QUALIFICATION OF CCTA-DERIVED NCPV AS AN FDA PROGNOSTIC BIOMARKER. THE INTENDED USE OF NCPV IS AS AN ACCURATE, VALIDATED AND WIDELY AVAILABLE ENRICHMENT BIOMARKER THAT CAN OPTIMIZE RISK ASSESSMENT OF CLINICAL TRIAL PARTICIPANTS PRIOR TO ENROLLMENT IN ASCVD STUDIES. AS PART OF THE LARGER EFFORT TO DEVELOP CCTA NCPV ASSESSMENT AS AN ENRICHMENT BIOMARKER, WE WILL PERFORM A SERIES OF STUDIES TO COMPREHENSIVELY ASSESS ANALYTIC VALIDITY. SPECIFICALLY, WE ARE PROPOSING THREE AIMS COMPRISING 5 FIVE MEASUREMENT QUALITY STUDIES TO PROVIDE ANALYTIC VALIDITY FOR NCPV BY STANDARDIZING THE MINIMAL ACCEPTABLE PRECISION, SHORT AND LONG INTERVAL REPEATABILITY, AND ACCURACY OF CCTA NCPV MEASUREMENT. FURTHER WE WILL COMPARE THE RESULTS OBTAINED WITH THE TWO MOST WELL-ESTABLISHED AND WIDELY USED SOFTWARE PACKAGES, TO DEMONSTRATE THE IMPACT OF DIFFERENT TOOLS ON MEASUREMENTS. THESE STUDIES ARE ESSENTIAL FOR SUBMISSION OF AN FDA QUALIFICATION PLAN FOR NCPV AS A PROGNOSTIC MARKER. FINALLY, WE WILL WORK TO ESTABLISH CONSENSUS STANDARDS FOR MEASUREMENT QUALITY THAT IS ACCEPTABLE TO ACADEMIC, CORPORATE AND REGULATORY STAKEHOLDERS. THE ULTIMATE GOAL OF THE PROPOSED WORK IS TO ESTABLISH MEASUREMENT VALIDITY FOR NCPV BIOMARKERS BASED UPON WELL-DEFINED VALIDATION TECHNIQUES USING A DATA DRIVEN CONSENSUS REGARDING MINIMAL ACCEPTABLE QUALITY THRESHOLDS. APPLICATION OF THE NCPV AS A BIOMARKER HAS THE POTENTIAL TO DE-RISK CLINICAL ENDPOINT-BASED TRIALS BY ENRICHING TRIAL POPULATIONS WITH PARTICIPANTS AT HIGHER LIKELIHOOD OF MACE, INCLUDING IN PRIMARY PREVENTION INDIVIDUALS WHO HAVE NOT YET HAD A MYOCARDIAL INFARCTION. IMPORTANTLY, THIS WORK IS AN INTEGRAL PART OF AN INTERNATIONAL CONSORTIUM OF ACADEMICS, PHARMA, DEVICE MANUFACTURERS, SOFTWARE VENDORS, AND REGULATORS, (THE A2D2 CONSORTIUM) TO ADVANCE THE USE OF CCTA AS A DRUG DEVELOPMENT TOOL (DDT). THIS CONSORTIUM PROVIDES THIS WORK WITH ROBUST, CRITICAL SUPPORT INCLUDING ACCESS TO APPROPRIATE IMAGES (STEP A), DEVELOPMENT OF CONSENSUS STANDARDS (STEP E) AND DISSEMINATION AND IMPLEMENTATION (STEP F) TO ENSURE THE ACCEPTABILITY OF THESE STUDIES BY ALL STAKEHOLDERS. WITH COMPLETION OF THE PROPOSED INVESTIGATIONS, WE WILL HAVE PERFORMED A COMPREHENSIVE SET OF IMAGE ASSESSMENTS AND ANALYSES USING WELL ESTABLISHED TECHNIQUES WHICH WILL FIRMLY ESTABLISH THE MEASUREMENT QUALITY AND VALIDITY OF CCTA QUANTITATION OF NCPV. IMPORTANTLY, THIS IS A COMPONENT OF THE IN-PROGRESS QUALIFICATION PROCESS OF NCPV AS A PROGNOSTIC BIOMARKER TO ENRICH STUDY POPULATIONS FOR TRIALS OF INVESTIGATION ATHEROSCLEROSIS THERAPIES (CONTEXT OF USE)

ASTHMATIC LGE RELEASE IS DUE TO SUBSTANCE P RELEASED BY SENSORY NEURONS - PROJECT SUMMARY ASTHMA AND ASTHMATIC SYMPTOMS CONTINUE TO POSE A SIGNIFICANT HEALTH BURDEN TO THE USA, DEMONSTRATING THE NEED TO UNDERSTAND HOW THE PRIMARY ASTHMATIC EFFECTOR MOLECULE, IMMUNOGLOBULIN (IG) E (IGE), IS PRODUCED. WHILE IGE PRODUCTION CAN LEAD TO PROTECTIVE IMMUNITY IN RESPONSE TO VENOMS AND MULTICELLULAR PATHOGENS, ITS OVERPRODUCTION IN RESPONSE TO BENIGN SUBSTANCES UNDERLIES ITS ROLE IN ASTHMATIC INFLAMMATION. IGE IS SECRETED FROM B LINEAGE CELLS IN THE PRESENCE OF COGNATE ANTIGEN AND ACTIVATES MAST CELLS AND BASOPHILS TO RELEASE POTENT INFLAMMATORY MOLECULES THAT ACT DIRECTLY ON THE AIRWAY WALL AND ATTRACT INFLAMMATORY CELLS TO THE LUNGS. EMERGING EVIDENCE DEMONSTRATES THAT ASTHMATIC INFLAMMATION SENSITIZES ANOTHER CELL TYPE, NEURONS. ONCE SENSITIZED, NEURONS RELEASE A HOST OF NEUROPEPTIDES THAT ENHANCE ASTHMATIC INFLAMMATION. THIS APPLICATION AIMS TO INVESTIGATE HOW THE RELEASE OF NEUROPEPTIDES FROM SENSORY NEURONS DRIVES B CELL FATE, MATURATION, AND CLASS SWITCH RECOMBINATION TO IGE. THE OVERALL OBJECTIVE OF THIS APPLICATION IS TO INVESTIGATE HOW SENSORY NEURON RELEASE OF THE NEUROPEPTIDE SUBSTANCE P STIMULATES B CELL IGE RELEASE IN ASTHMA. OUR PRELIMINARY WORK SHOWS THAT SUBSTANCE P IS PREFERENTIALLY RELEASED IN RESPONSE TO ALTERNARIA ALTERNATA INDUCTION OF ASTHMA IN MICE AND THAT SENSORY NEURON DEPLETION REDUCES SUBSTANCE P RELEASE AND IGE PRODUCTION. SUPPLEMENTATION OF SUBSTANCE P BACK INTO THE LUNGS OF SENSORY NEURON-ABLATED MICE RESTORES THE ASTHMATIC PHENOTYPE. ALSO, SUBSTANCE P KNOCKOUT MICE (THOSE THAT DO NOT PRODUCE SUBSTANCE P) PRODUCE LESS IGE IN RESPONSE TO A ALTERNATA COMPARED TO WT LITTERMATES. THIS OCCURS VIA SUBSTANCE P BINDING TO MRGPRA1 RECEPTORS ON B CELLS AND ULTIMATELY CONTROLS TRANSCRIPTION FACTORS TO STEER CELL FATE, MATURATION, AND CLASS SWITCH RECOMBINATION. WE WILL INVESTIGATE THIS NOVEL MECHANISM OF IGE OVERPRODUCTION IN TWO AIMS. AIM 1 WILL TEST HOW SUBSTANCE P REGULATES B CELL CLASS SWITCH RECOMBINATION VIA SPECIFIC TRANSCRIPTIONAL REGULATION IN VITRO WITH PROTEIN ANALYSIS, FLOW CYTOMETRY, AND IGE GERMLINE AND CIRCULAR TRANSCRIPTS. IN ADDITION, THE ASTHMATIC PHENOTYPE IN VITRO USING MURINE AND HUMAN B CELLS AND IN VIVO USING ADOPTIVE TRANSFER TECHNIQUES. AIM 2 WILL INVESTIGATE THE MATURATION OF B CELLS INDUCED BY SUBSTANCE P USING IN VITRO AND IN VIVO PROTEIN ANALYSIS AND FLOW CYTOMETRY. THIS CONTRIBUTION IS SIGNIFICANT BECAUSE IT IS EXPECTED TO ELUCIDATE HOW INTEGRATED NEUROIMMUNE REGULATION AUGMENTS IGE PRODUCTION AND DYSREGULATION IN ASTHMA. SUCH KNOWLEDGE CAN INFORM THE DEVELOPMENT OF NEW STRATEGIES THAT WILL HELP REDUCE THE GROWING PROBLEM OF CHRONIC ASTHMATIC DISEASE, DIRECTLY IN LINE WITH THE MISSION STATEMENT OF THE NIH, TO REDUCE THE BURDEN OF HUMAN DISEASE.

INVESTIGATING NOVEL TARGETABLE VULNERABILITIES OF LAM DISEASE

A NEW TREATMENT FOR HERITABLE PULMONARY ARTERY HYPERTENSION CAUSED BY NONSENSE MUTATIONS

GENETICS OF BIPOLAR DISORDER IN LATINO POPULATIONS

CYCLIC ANTIFUNGAL PEPTIDES FOR DRUG-RESISTANT INVASIVE FUNGAL INFECTIONS - PROJECT SUMMARY ASPERGILLOSIS AND CANDIDIASIS, CAUSED PRIMARILY BY ASPERGILLUS FUMIGATUS AND CANDIDA ALBICANS, ARE THE MOST PREVALENT INVASIVE FUNGAL INFECTIONS (IFIS), PARTICULARLY IMPACTING IMMUNOCOMPROMISED PATIENTS, SUCH AS THOSE WITH HIV/AIDS, CANCER, OR ORGAN TRANSPLANT RECIPIENTS. A. FUMIGATUS AND C. ALBICANS ARE CLASSIFIED AS CRITICAL PRIORITY FUNGAL PATHOGENS BY THE WHO DUE TO THE RISE IN ANTIFUNGAL DRUG RESISTANCE, ESPECIALLY AGAINST THE CURRENT FIRST-LINE THERAPIES, SUCH AS TRIAZOLES FOR INVASIVE ASPERGILLOSIS AND ECHINOCANDINS FOR CANDIDIASIS. RESISTANCE IN THESE PATHOGENS TYPICALLY INVOLVES MUTATIONS IN THE CYP51A GENE IN ASPERGILLUS AND THE ERG11 GENE IN CANDIDA. THIS INCREASING RESISTANCE POSES A SIGNIFICANT THREAT TO GLOBAL HEALTH, EMPHASIZING THE URGENT NEED FOR NOVEL ANTIFUNGAL AGENTS WITH BROAD-SPECTRUM ACTIVITY AND INNOVATIVE MECHANISMS OF ACTION. DESPITE THE GRAVE IMPACT, THE DEVELOPMENT OF NEW ANTIFUNGAL DRUGS HAS LAGGED BEHIND ANTIBACTERIAL DRUG DEVELOPMENT, AND RESISTANCE TO EXISTING TREATMENTS HAS BECOME AN ALARMING ISSUE. OUR RESEARCH FOCUSES ON DEVELOPING NOVEL CYCLIC ANTIFUNGAL PEPTIDES (CAFPS) AS A PROMISING THERAPEUTIC APPROACH TO COMBAT THESE INFECTIONS. PRELIMINARY STUDIES HAVE IDENTIFIED LEAD PEPTIDES, WHICH EFFECTIVELY INHIBIT DRUG-RESISTANT C. ALBICANS AND A. FUMIGATUS ISOLATES. THESE PEPTIDES ACT BY DISRUPTING FUNGAL MEMBRANES, AND THEY ALSO ENHANCE THE EFFICACY OF EXISTING ANTIFUNGAL AGENTS LIKE FLUCONAZOLE. THE MODULAR STRUCTURE OF THESE PEPTIDES OFFERS THE POTENTIAL FOR RAPID OPTIMIZATION AND COMBINATION WITH CURRENT THERAPIES, IMPROVING EFFICACY AND REDUCING RESISTANCE. WE HYPOTHESIZE THAT OPTIMIZING THE CHARGE AND HYDROPHOBICITY OF THESE PEPTIDES WILL IMPROVE THEIR POTENCY, STABILITY, AND SAFETY PROFILES, OFFERING ENHANCED TREATMENT OUTCOMES FOR A. FUMIGATUS AND C. ALBICANS INFECTIONS. IN AIM 1, WE WILL DESIGN A LIBRARY OF NEW CAFPS BASED ON OUR LEAD PEPTIDES TO INVESTIGATE THEIR STRUCTURE-ACTIVITY RELATIONSHIPS, STABILITY, TOXICITY, AND SYNERGY WITH EXISTING ANTIFUNGAL DRUGS. IN AIM 2, WE WILL ASSESS THE IN VIVO TOXICITY AND EFFICACY OF THE TOP PEPTIDES IN MURINE MODELS OF INVASIVE PULMONARY ASPERGILLOSIS AND HEMATOGENOUSLY DISSEMINATED CANDIDIASIS. THESE STUDIES WILL DETERMINE THE SAFETY, PHARMACOKINETICS, AND OPTIMAL DOSING REGIMENS FOR THESE PEPTIDES. IN THE R33 PHASE, WE WILL REFINE THE MOST PROMISING CAFPS FOR PRECLINICAL DEVELOPMENT. THIS WILL INVOLVE EVALUATING THEIR PHARMACOKINETIC AND PHARMACODYNAMIC PROFILES, OPTIMIZING FORMULATIONS, AND TESTING THEIR EFFICACY IN COMBINATION WITH EXISTING ANTIFUNGAL AGENTS IN MURINE MODELS. WE WILL ALSO USE MULTI-OMICS APPROACHES TO ELUCIDATE THE MECHANISM OF ACTION OF THESE PEPTIDES AND THEIR IMPACT ON FUNGAL CELL FUNCTION. OUR GOAL IS TO IDENTIFY NOVEL ANTIFUNGAL AGENTS THAT CAN BE USED ALONE OR IN COMBINATION WITH CURRENT THERAPIES TO COMBAT RESISTANT FUNGAL INFECTIONS, ULTIMATELY REDUCING MORBIDITY AND MORTALITY ASSOCIATED WITH INVASIVE ASPERGILLOSIS AND CANDIDIASIS. THIS RESEARCH AIMS TO CREATE POTENT, BROAD-SPECTRUM ANTIFUNGAL THERAPIES CAPABLE OF OVERCOMING CURRENT TREATMENT LIMITATIONS, MITIGATING RESISTANCE, AND IMPROVING PATIENT OUTCOMES FOR THESE LIFE-THREATENING INFECTIONS.

EXAMINING THE ANDROGENIC AND PROGESTATIONAL EFFECTS OF NOVEL ANDROGENS FOR MALE CONTRACEPTION

NOVEL DETERMINANTS OF PD-L1 PROTEIN TRAFFICKING AND STABILITY IN LUNG ADENOCARCINOMA CELLS

FUNCTION OF LONG NON-CODING RNA MD1 IN LEIOMYOMA PATHOGENESIS - UTERINE LEIOMYOMA (FIBROIDS) DEVELOP DURING REPRODUCTIVE YEARS IN OVER 70% OF WOMEN; HOWEVER, THEIR ETIOLOGY REMAINS UNKNOWN. OUR GROUP HAS IDENTIFIED TWO MIRNAS, MIR-200C AND MIR-29C, WHICH ARE CRITICAL TO REGULATION OF GENES FUNCTIONALLY ASSOCIATED WITH CELL CYCLE, CELLULAR TRANSFORMATION, INFLAMMATION AND EXTRACELLULAR MATRIX ACCUMULATION AND THUS OF GREAT SIGNIFICANCE TO FIBROID PATHOGENESIS. OUR RECENT FINDINGS USING NEXT GENERATION SEQUENCING PROVIDED A COMPREHENSIVE PROFILE OF NON-CODING RNAS INCLUDING LONG NON- CODING RNAS (LNCRNAS) AS WELL AS NOVEL GROUPS OF SMALL NON-CODING RNAS (SNCRNAS). OUR PRELIMINARY FINDINGS HERE HAVE IDENTIFIED ABERRANT EXPRESSION OF LINC-MD1 AND MIR-135 FAMILY IN FIBROIDS AND THUS THE IMPETUS FOR THIS PROPOSAL. USING QRT-PCR WE DETECTED MARKEDLY REDUCED LEVELS OF LINC-MD1 WHILE SIGNIFICANTLY HIGHER EXPRESSION OF MIR-135 FAMILY IN LEIOMYOMAS. THE SEQUENCES OF THE LINC-MD1 AND OUR PRELIMINARY DATA SUGGEST THAT LINC-MD1 CAN ACT AS MIRNA SPONGE FOR MIR-135 FAMILY (MIR-135A/-135B) IN LEIOMYOMA SMOOTH MUSCLE CELLS. THEREFORE, BASED ON THIS PRELIMINARY DATA WE HYPOTHESIZE THAT LEIOMYOMA AS COMPARED TO MYOMETRIUM DISPLAYS REDUCED EXPRESSION OF LINC-MD1 WHICH THROUGH A MIRNA-GUIDED MECHANISM INVOLVING MIR-135 REGULATES THE EXPRESSION OF SPECIFIC TARGET GENES FUNCTIONALLY ASSOCIATED WITH WNT/SS-CATENIN SIGNALING PATHWAY WHICH IS KNOWN TO BE CENTRAL TO LEIOMYOMA PATHOGENESIS. TO TEST OUR CORE HYPOTHESIS WE PROPOSE 2 SPECIFIC AIMS. IN AIM 1 WE WILL DETERMINE THE INTERACTION BETWEEN LINC-MD1 AND MIR-135 FAMILY BY RNA IMMUNOPRECIPITATION AND RNA PULL-DOWN ASSAY. WE WILL OVER OR UNDER EXPRESS LINC-MD1 IN LSMC OR MSMC SPHEROID CELLS AND DETERMINE ITS EFFECT ON MIR-135 FAMILY AND ITS DOWNSTREAM TARGET GENES NAMELY GSK3SS AND APC WHICH ARE KNOWN TO REGULATE SS-CATENIN DEGRADATION. IN THESE EXPERIMENTS SS-CATENIN, ITS PHOSPHORYLATED FORM AND ITS NUCLEAR LOCALIZATION WILL BE DETERMINED IN RESPONSE TO OVEREXPRESSION AND KNOCKDOWN STUDIES IN VITRO. TO ESTABLISH THE CLINICAL RELEVANCE AND THERAPEUTIC POTENTIAL OF LINC-MD1 IN AIM 2 WE WILL DETERMINE THE EFFECT OF LINC-MD1 OVEREXPRESSION IN LEIOMYOMA CELLS ON FIBROID PROGRESSION IN A LEIOMYOMA ANIMAL MODEL. THIS TRANSLATIONAL PROPOSAL ADDRESSES A SIGNIFICANT GAP IN KNOWLEDGE ON THE ROLE OF A NOVEL LNCRNA-MIRNA NETWORK IN THE PATHOGENESIS OF FIBROIDS WHICH IS A PRIORITY AREA OF INVESTIGATION FOR NIH, AND COULD POTENTIALLY BE TARGETED FOR THERAPEUTIC PURPOSES FOR FIBROIDS.

A NEW MOUSE MODEL OF ATAXIA-TELANGIECTASIA FOR TESTING THERAPEUTIC READ-THROUGH COMPOUNDS

GESTATIONAL PROGRAMMING OF NEPHROGENESIS

PROGRAMMED ADIPOGENESIS AND LIPID DYSREGULATION

BCR1P AS A CANDIDA NEUTROPHIL RESISTANCE REGULATOR

ADIPOCYTE RENIN-ANGIOTENSIN SYSTEM AND PROGRAMMING OF HYPERTENSION

A MOLECULAR APPROACH TO PREVENT PULMONARY DYSFUNCTION IN THE INTRAUTERINE GROWTH

MENTORED PATIENT-ORIENTED RESEARCH CAREER DEVELOPMENT AWARD

CURCUMIN TO AUGMENT NEONATAL LUNG INJURY/REPAIR

BISPHENOL A MEDIATED EFFECTS ON OFFSPRING GLUCOCORTICOID HOMEOSTASIS AND OBESITY

BRC MODERNIZATION OF STERILIZING EQUIPMENT - ABSTRACT THIS PROPOSAL IS TO MODERNIZE THE C.W. STEERS BIOLOGICAL RESOURCES CENTER (BRC) TO MEET ITS GROWING STERILIZING NEEDS BY PURCHASING A NEW EFFICIENT STERILIZER WITH A WATER CONSERVATION VACUUM SYSTEM. THIS AUTOCLAVE WILL INCREASE RELIABLE STERILIZATION OF ANIMAL- AND BIOMEDICAL-RELATED RESEARCH TOOLS/MATERIALS. IN NOVEMBER OF 2020, THE PRECLINICAL RESEARCH CENTER (PRC) WAS OPENED FOR THE HOUSING OF RODENTS AT THE BRC. ITS MODERN DESIGN INCREASED THE LUNDQUIST INSTITUTE’S RODENT HOUSING CAPACITY FROM 1,000 MICE CAGES TO 6,000 (WITH SPACE FOR 1,000 CAGES DEDICATED FOR GENETICALLY MODIFIED IMMUNOSUPPRESSED RODENTS). AS THE PANDEMIC SLOWED, RESEARCH RAMPED; IN THE LAST SIX MONTHS, CAGES IN USE GREW FROM 400 TO 1,000. ADDITIONAL PROJECTED GROWTH IS EXPECTED: GRANT AWARDS WITH UPCOMING START DATES WILL MORE THAN DOUBLE THE CURRENT POPULATION CAGE USE TO 2,500 WITHIN THE YEAR. WITH THIS GROWTH, THE DEMAND FOR AUTOCLAVED CAGES TO PROVIDE APPROPRIATE HOUSING FOR SUSCEPTIBLE ANIMALS HAS ALSO INCREASED. AUTOCLAVES ARE INDISPENSABLE IN ANIMAL RESEARCH FOR PREVENTING DISEASE FROM SPREADING WITHIN THE RODENTS. TWO AREAS OF RESEARCH WILL BE SUPPORTED BY A NEW MODERNIZED EFFICIENT AUTOCLAVE: (1) THE INFECTIOUS DISEASE DEPARTMENT, WHOSE STUDIES REQUIRE STERILE CAGING AT ALL TIMES (30,848 RODENTS); AND (2) STUDIES REQUIRING OTHER IMMUNOSUPPRESSED RODENTS (16,694), SUCH AS TRANSGENIC MICE USED FOR STUDIES FOCUSED ON CHRONIC DISEASES (CANCER, DIABETES, MUCOPOLYSACCHARIDOSIS TYPE III, ETC.). IMMUNOSUPPRESSED MICE USED FOR CHRONIC DISEASES STUDIES ARE KEPT IN THE IMMUNOSUPPRESSED HOUSING AREA, WHICH GREW FROM ONE ROOM WITH CAPACITY FOR 100 CAGES TO TWO ROOMS WITH CAPACITY FOR 1,000 CAGES WITH THE MOVE TO PRC. WHILE THE BRC’S CURRENT AUTOCLAVE (2000 STERIS AMSCO EAGLE SV-3043 SCIENTIFIC PREVACUUM STERILIZER IS WELL-MAINTAINED, IT IS BEING USED AT CAPACITY. DUE TO THE INCREASE IN THE NUMBER OF CAGES NEEDING STERILIZATION AT ONE TIME, AN ADDITIONAL AUTOCLAVE IS NEEDED TO TRIPLE THE BRC’S CAPACITY TO STERILIZE CAGES WITH A FASTER CYCLE OF 45-60 MINUTES. THEREFORE, OUR REQUEST IS TO FUND THE PURCHASE OF A NEW EFFICIENT STERILIZER WITH A WATER CONSERVATION VACUUM SYSTEM TO ALLOW FOR RELIABLE STERILIZATION TO MEET THE NEW AND GROWING NEEDS OF INFECTIOUS DISEASE AND IMMUNOSUPPRESSED RODENT STUDIES, ENSURING EFFICIENT WORKFLOWS FOR BOTH IN-HOUSE AND CRO-CONTRACTED INVESTIGATORS. THIS PURCHASE WILL SUPPLEMENT THE CAPACITY OF THE CURRENT OLDER AUTOCLAVE BY TRIPLING THE NUMBER OF DAILY LOADS FROM TWO TO SIX. THE PURCHASE OF THE STATE-OF- THE-ART AUTOCLAVE WILL ALLOW FOR THE FOLLOWING AIMS: AIM 1: SUPPORT HIGH-QUALITY ANIMAL RESEARCH IN THE AREAS OF INFECTIOUS AND CHRONIC DISEASE. AIM 2: MODERNIZE THE FACILITY BY ENABLING ADVANCEMENT IN OPERATIONS AND IMPROVING EFFICIENCIES. AIM 3: IMPLEMENT GREEN TECHNOLOGY.

MULTICENTER BAYESIAN DECISION-THEORETIC CLINICAL TRIALS

THE HUMORAL IMMUNE RESPONSE TO RECOMBINANT ENZYME IN MUCOPOLYSACCHARIDOSIS I

STUDY OF THE TRENDS IN EVALUATION AND MANAGEMENT OF THE YOUNG FEBRILE INFANT AND ITS RELATED HEALTHCARE COSTS AND MORBIDITIES

FETAL MECONIUM PASSAGE MEDIATED VIA CRF PATHWAYS

HYPOPITUITARISM AFTER MODERATE AND SEVERE HEAD INJURY

FUNCTIONAL PROTEOMICS OF LEUKEMIC MARROW STROMAL CELLS

CONFERENCE ON COMPUTERIZED ELECTROCARDIOGRAPHY

US DOHAD SOCIETY: THE 5TH ANNUAL MEETING 2020 - ABSTRACT THE FUNDAMENTAL BASIS OF DOHAD (DEVELOPMENTAL ORIGINS OF HEALTH AND DISEASE) IS THAT “THE BEGINNING OF LIFE PAVES THE WAY FOR FUTURE HEALTH/DISEASE”. THE US DOHAD IS AN ESTABLISHED SOCIETY THAT ENCOMPASSES THIS CONCEPT. ITS ANNUAL MEETING COMBINES ORIGINAL RESEARCH PRESENTATIONS AND GUEST SPEAKERS WHOSE RESEARCH FOCUSES ON THE EVIDENCE AND MECHANISMS BY WHICH ENVIRONMENTAL TOXICANTS, NUTRIENT EXPOSURES, STRESS AND OTHER FACTORS INFLUENCE THE DEVELOPING FETUS AND NEWBORN, AND THEREBY CONTRIBUTE TO THE HEALTH AND DISEASE OF THE OFFSPRING. IN THE US, THIS FIELD HAS GATHERED MOMENTUM AMONGST CLINICIANS, TOXICOLOGISTS, EPIDEMIOLOGISTS, NUTRITIONISTS, PHYSIOLOGISTS AND BASIC SCIENTISTS WHO ARE WORKING IN DIVERSE TOPICS RANGING FROM DEVELOPMENTAL BIOLOGY, NUTRITION, ENVIRONMENTAL TOXICOLOGY, CANCER, STRESS AND HORMONES. THE MANDATE OF US DOHAD, AN AFFILIATE OF THE INTERNATIONAL DOHAD SOCIETY, IS TO BRING TOGETHER THESE MULTIDISCIPLINARY AND/OR INTERDISCIPLINARY INVESTIGATORS IN ORDER TO FOSTER INTERACTIONS AND COLLABORATION ON THESE DIVERSE TOPICS. THIS INDEED REPRESENTS A UNIQUE INTEGRATED MODEL WHICH NO OTHER SOCIETY OFFERS. IN ADDITION TO THE PRESENTATIONS BY ESTABLISHED SCIENTISTS, THE SOCIETY PROVIDES A FORUM FOR GRADUATE STUDENTS, POSTDOCTORAL FELLOWS AND JUNIOR INVESTIGATORS WITH VIEW TO FACILITATE INTERACTION, TRAINING AND CAREER OPPORTUNITIES FOR THEM. THE PAST MEETINGS HAVE BEEN TREMENDOUSLY SUCCESSFUL AND HAVE PROVIDED IMPETUS FOR CONTINUED MEETINGS AND HAVE STRENGTHENED THE US DOHAD SOCIETY WITH GROWING MEMBERS AND PARTICIPANTS. THE FIFTH ANNUAL 2020 MEETING WILL BE HELD AT RIZZO CENTER, CHAPEL HILL. THE THEME IS “WOMB AND CRADLE: KEY DEVELOPMENTAL WINDOWS IN EFFECTING THE TRAJECTORY OF HEALTH AND DISEASE” AND THE PROGRAM INCLUDES A DEDICATED DAY ON CAREER DEVELOPMENT AND GRANT WRITING FOR THE TRAINEES. LASTLY, THE SOCIETY ALSO EMBRACES DIVERSITY AND EQUAL REPRESENTATION OF MALES AND FEMALES AS WELL AS MINORITY GROUPS.

DEVELOPMENTAL ORIGINS OF HEALTH AND DISEASE

CELEBRATING THE INSPIRED AND INSPIRING TESTIS BIOLOGY- THE XXVII NORTH AMERICAN TESTIS WORKSHOP - PROJECT SUMMARY THIS R13 APPLICATION REQUESTS FUNDS TO SUPPORT THE XXVII NORTH AMERICAN TESTIS WORKSHOP WITH A THEMATIC FOCUS ON “CELEBRATING THE INSPIRING AND INSPIRED TESTIS BIOLOGY”, WHICH WILL BE HELD ON MAY 2-5, 2024, AT THE EMBASSY SUITES DENVER, IN DENVER, COLORADO. THE MEETING WILL BE HELD IMMEDIATELY AFTER THE AMERICAN SOCIETY OF ANDROLOGY ANNUAL MEETING, WHICH WILL TAKE PLACE ON APRIL 19 – 2 MAY AT THE SAME LOCATION. SINCE 1972, THE NORTH AMERICAN TESTIS WORKSHOP HAS BEEN THE PREMIER INTERNATIONAL FORUM FOR BASIC AND CLINICIAN SCIENTISTS TO PRESENT AND DISCUSS THEIR RECENT FINDINGS ON TESTIS REGULATION AND FUNCTION. IT ATTRACTS 150-200 ATTENDEES FROM NORTH AMERICA AND AROUND THE WORLD, APPROXIMATELY 30% OF WHOM ARE TRAINEES. THE REQUESTED R13 FUNDS WILL HELP DEFRAY TRAVEL COSTS FOR 10 TRAINEES OR YOUNG INVESTIGATORS SELECTED FOR SHORT TALKS FROM SUBMITTED ABSTRACTS AND PART OF THE HOUSING COSTS FOR THE 19 INVITED SPEAKERS. THE MAIN GOALS FOR THE WORKSHOP ARE AS FOLLOWS: 1) TO PROVIDE A PLATFORM FOR DISSEMINATION AND DISCUSSION OF NEW DISCOVERIES ON TESTIS BIOLOGY, ESPECIALLY NOVEL DATA OBTAINED USING THE LATEST GENOMIC, EPIGENOMIC, AND SINGLE-CELL TECHNOLOGIES. 2) TO CREATE OPPORTUNITIES FOR SCIENTIFIC EXCHANGE AND COLLABORATIONS AMONG PEERS TO FOSTER CAREER GROWTH AND TO ENSURE THE CONTINUED ADVANCE OF THE FIELD OF TESTIS BIOLOGY. 3) TO OFFER A FORUM FOR TRAINEES (FELLOWS, POST-DOCS, AND GRADUATE STUDENTS) AND JUNIOR INVESTIGATORS (WITHIN THREE YEARS OF INDEPENDENCE) TO PRESENT THEIR RESEARCH, RECEIVE FEEDBACK FROM SENIOR INVESTIGATORS, AND BUILD PROFESSIONAL RELATIONSHIPS, ALL OF WHICH ARE KNOWN CRITICAL FOR BOTH INTELLECTUAL AND ACADEMIC GROWTH. 4) TO ENCOURAGE THE PARTICIPATION OF WOMEN AND UNDERREPRESENTED GROUPS, INCLUDING FACULTY AND TRAINEES OF RACIAL AND ETHNIC MINORITIES AND PEOPLE WITH DISABILITIES, IN THIS WORKSHOP TO FOSTER THEIR INTELLECTUAL AND ACADEMIC GROWTH BY PROVIDING THEM WITH OPPORTUNITIES TO PRESENT THEIR WORK, RECEIVE FEEDBACK FROM PEERS, AND BUILD PROFESSIONAL NETWORKS.

THE XXVITH NORTH AMERICAN TESTIS WORKSHOP - PROJECT SUMMARY THIS R13 APPLICATION REQUESTS FUNDS TO SUPPORT THE XXVITH NORTH AMERICAN TESTIS WORKSHOP WITH A THEMATIC FOCUS ON “NEW HORIZONS IN TESTIS BIOLOGY AND MEN'S HEALTH”, WHICH WILL BE HELD ON APRIL 21-24, 2021 AT THE ESTANCIA LA JOLLA HOTEL IN SAN DIEGO, CALIFORNIA. THE MEETING WILL BE HELD RIGHT PRIOR TO THE AMERICAN SOCIETY OF ANDROLOGY ANNUAL MEETING, WHICH WILL TAKE PLACE ON APRIL 24- 28 AT THE SAME LOCATION. SINCE 1972, THE NORTH AMERICAN TESTIS WORKSHOP HAS BEEN THE PREMIERE INTERNATIONAL FORUM FOR BASIC AND CLINICIAN-SCIENTISTS TO PRESENT AND DISCUSS THEIR RECENT FINDINGS ON TESTIS REGULATION AND FUNCTION. IT ATTRACTS 150-200 ATTENDEES FROM NORTH AMERICA AND AROUND THE WORLD, APPROXIMATELY 30% OF WHOM ARE TRAINEES. THE REQUESTED R13 FUNDS WILL BE USED TO HELP DEFRAY TRAVEL COSTS FOR 10 TRAINEES OR YOUNG INVESTIGATORS WHO ARE SELECTED FOR SHORT TALKS FROM SUBMITTED ABSTRACTS, AND PART OF THE HOUSING COSTS FOR THE 19 INVITED SPEAKERS. THE MAIN GOALS FOR THE WORKSHOP ARE AS FOLLOWS: 1) TO PROVIDE A PLATFORM FOR DISSEMINATION AND DISCUSSION OF NEW DISCOVERIES ON TESTIS BIOLOGY, ESPECIALLY NOVEL DATA OBTAINED USING THE LATEST GENOMIC, EPIGENOMIC AND SINGLE CELL TECHNOLOGIES. 2) TO CREATE OPPORTUNITIES FOR SCIENTIFIC EXCHANGE AND COLLABORATIONS AMONG PEERS TO FOSTER CAREER GROWTH AND TO ENSURE CONTINUED ADVANCE OF THE FIELD OF TESTIS BIOLOGY. 3) TO OFFER A FORUM FOR TRAINEES (FELLOWS, POST-DOCS AND GRADUATE STUDENTS) AND JUNIOR INVESTIGATORS (WITHIN THREE YEARS OF INDEPENDENCE) TO PRESENT THEIR RESEARCH, RECEIVE FEEDBACK FROM SENIOR INVESTIGATORS, AND BUILD PROFESSIONAL RELATIONSHIP, ALL OF WHICH ARE KNOWN CRITICAL FOR BOTH INTELLECTUAL AND ACADEMIC GROWTH. 4) TO ENCOURAGE THE PARTICIPATION OF WOMEN AND UNDERREPRESENTED GROUPS, INCLUDING FACULTY AND TRAINEES OF RACIAL AND ETHNIC MINORITIES, AND PEOPLE WITH DISABILITIES, IN THIS WORKSHOP TO FOSTER THEIR INTELLECTUAL AND ACADEMIC GROWTH BY PROVIDING THEM WITH OPPORTUNITIES TO PRESENT THEIR WORK, RECEIVE FEEDBACK FROM PEERS, AND BUILD PROFESSIONAL NETWORKS.

US DOHAD 2019: THE 4TH ANNUAL MEETING

US DOHAD 2018: THE 3RD ANNUAL MEETING