The Methodist Hospital

HEALTH CARE INNOVATION CHALLENGE

DYNAMICS OF COLONIZATION AND INFECTION BY MULTIDRUG-RESISTANT PATHOGENS IN IMMUNOCOMPROMISED AND CRITICALLY ILL PATIENTS (DYNAMITE)

CENTER FOR TRANSPORT ONCOPHYSICS

CENTER FOR SYSTEMATIC MODELING OF CANCER DEVELOPMENT

HEALTH CARE INNOVATION CHALLENGE

CENTER FOR IMMUNOTHERAPEUTIC TRANSPORT ONCOPHYSICS

A GMP/GLP INVESTIGATION OF DEGRADABLE POLYMERIC SHELLS FOR TRAUMATIC OSTEOREGENERATION

TOWARD INDIVIDUALIZED BREAST CANCER THERAPY: LEVERAGING MOLECULAR MEDICINE WITH MULTISTAGE VECTOR TECHNOLOGY

THE STROMAL MICROENVIRONMENT AS A CO-ORGANIZER OF BLADDER CARCINOGENESIS AND PROGRESSION - OVERALL PROJECT SUMMARY BLADDER CANCER (BC) IS THE SECOND MOST COMMON UROLOGIC MALIGNANCY AFFECTING 573,278 PEOPLE WORLDWIDE IN 2020. PATHOLOGICALLY, BC IS DIAGNOSED AS NON-MUSCLE-INVASIVE (NMI) AND MUSCLE-INVASIVE (MI) DISEASE. HERE WE DEFINE EARLY BLADDER LESIONS AS NMIBC. MAJOR CLINICAL GAPS IN NMIBC INCLUDE I) LACK OF MECHANISTIC INSIGHTS DEFINING NMIBC PROGRESSION, AND II) LACK OF PLATFORM FOR RISK STRATIFICATION OF NMIBC THAT RECUR BUT NEVER PROGRESS (“NON-PROGRESSORS”), FROM THOSE THAT PROGRESSES INTO MIBC (“PROGRESSORS”) AND CONSEQUENTLY DEMONSTRATE POOR PROGNOSIS. THE GOAL OF OUR CENTER IS TO TACKLE THIS CLINICAL ISSUE BY DECIPHERING THE UNDERLYING MECHANISMS RESTRAINING OR PROMOTING THE PROGRESSION OF EARLY LESIONS (PROJECT 1 & 2), AND TO LEVERAGE THIS NOVEL BIOLOGY AS CANDIDATE BIOMARKERS TO RISK-STRATIFY AGGRESSIVE NMIBC (PROJECT 3). THIS PROPOSAL SEEKS TO SHIFT THE CURRENT RESEARCH PARADIGM IN THE FIELD OF NMIBC, BY PROPOSING A CONCEPTUALLY INNOVATIVE TUG-OF-WAR BETWEEN A TUMOR- RESTRAINING (PROJECT 1) AND A TUMOR-PROMOTING MECHANISM (PROJECT 2) IN DETERMINING THE OUTCOME OF EARLY BLADDER LESIONS/NMIBC IN BECOMING “PROGRESSORS” OR “NON-PROGRESSORS” (PROJECT 3). CLINICALLY, WHY “NON-PROGRESSORS” OFTEN RECUR BUT SELDOM PROGRESS, AND WHAT ARE THE DRIVING FORCES ADVANCING “PROGRESSORS” INTO MIBC WITH POOR SURVIVAL REMAIN FUNDAMENTAL QUESTIONS IN FIELD. OUR TUG-OF-WAR HYPOTHESIS WITH TWO OPPOSING FORCES IS CONCEPTUALLY DIFFERENT TO MOST OTHER STUDIES, WHICH PRIMARILY FOCUS ON ONE SIDE OF THE COIN. FURTHER, THE INTEGRATION OF KNOWLEDGE FROM PROJECT 1 AND 2 AS A UNIFIED SPATIAL PROTEOMICS AND TRANSCRIPTOMICS MAP BY THE SHARED RESOURCE CORE WILL REVEAL SPATIAL AND TEMPORAL RELATIONSHIPS BETWEEN DISTINCT FIBROBLAST POPULATIONS WITH OPPOSING FUNCTIONS, THEIR PHYSICAL INTERACTIONS WITH TUMOR AND IMMUNE CELL CLUSTERS, AS WELL AS THEIR RELATIONSHIP TO THE BIOMARKERS FROM PROJECT 3. BENCHMARK OF SUCCESS: THE KNOWLEDGE GAINED HERE WILL SHIFT CLINICAL PRACTICE PARADIGM, BY INFORMING FUTURE NIMBC MANAGEMENT THROUGH 1) THE DEVELOPMENT OF NOVEL URINARY PROFILING STRATEGIES THAT COULD RISK STRATIFY AGGRESSIVE NMIBC (PROJECT 1-3); 2) THE IDENTIFICATION OF TARGETS FOR FUTURE PRECISION INTERVENTION, EITHER BY ENHANCING/SUSTAINING THE TUMOR-RESTRAINING MECHANISMS (PROJECT 1) AND/OR INHIBITING THE TUMOR-PROMOTING MECHANISMS (PROJECT 2). THE OVERALL SUCCESS OF OUR PROGRAM IS FURTHER ENSURED BY AN EXTRAORDINARY MULTI-INVESTIGATOR TEAM THAT INTEGRATES THREE “ORGAN-SPECIFIC” BLADDER CANCER INVESTIGATORS WITHIN CEDARS-SINAI MEDICAL CENTER. ALL HAVE ACTIVE R01S AND INDIVIDUAL NCI-FUNDING TRACK RECORD IN PERFORMING BASIC SCIENCE RESEARCH, TRANSLATIONAL BLADDER CANCER RESEARCH, OR LEADING MULTI-CENTER CLINICAL TRIALS ON THE DISCOVERY AND VALIDATION OF BIOMARKERS. FINALLY, THEY PROPOSE TO COLLECT VALUABLE RETROSPECTIVE AND PROSPECTIVE NMIBC COHORTS, WHICH ARE ESSENTIAL TO ADDRESS THE CLINICAL QUESTIONS POSED WITHIN THIS PROPOSAL AND WILL BECOME AVAILABLE TO THE RESEARCH COMMUNITY AS A SHARED RESOURCE TO ADVANCE THE FIELD.

CLINICAL TRIAL READINESS FOR SCA1 AND SCA3

MODULATION OF INNATE IMMUNE CELLS TO CREATE TRANSPLANT TOLERANCE

CANCER-ASSOCIATED FIBROBLAST IN THE REGULATION OF BLADDER CANCER STEM CELLS

A NOVEL EFACE SYSTEM TO PREVENT THE RISKS OF FACIAL DISTORTION AFTER CMF SURGERY

ULTRA-LONG ACTING TRANSCUTANEOUSLY REFILLABLE ISLATRAVIR NANOFLUIDIC IMPLANT FOR HIV PRE-EXPOSURE - ABSTRACT LONG-ACTING (LA) PRE-EXPOSURE PROPHYLACTIC (PREP) STRATEGIES OFFER THE PROMISE OF IMPROVING ADHERENCE TO THERAPEUTIC REGIMEN FOR MAXIMAL HIV PREVENTIVE EFFICACY. KEY ATTRIBUTES OF ZERO-ORDER RELEASE KINETICS, LONG-TERM DRUG DELIVERY, USER-INDEPENDENT DOSING, THERAPEUTIC DISCRETION, SAFETY FOR CHRONIC USE AND RETRIEVABILITY ARE DESIRABLE CRITERIA FOR SUCCESSFUL WIDESPREAD CLINICAL PREP IMPLEMENTATION. TO DATE, LA PREP STRATEGIES DO NOT SUFFICIENTLY ADDRESS THE AFOREMENTIONED CRITERIA. OUR GOAL IS TO ADDRESS THESE LIMITATIONS BY DEVELOPING AN ULTRA- LONG ACTING ISLATRAVIR (ISL) DELIVERY IMPLANT WITH AN UNPRECEDENTED RELEASE DURATION OF AT LEAST 2 YEARS UNINTERRUPTED FOR DURABLE AND SAFE HIV PREVENTION INDEPENDENT OF USER ADHERENCE. TO ACHIEVE THIS GOAL, WE PROPOSE THE NANODDI, A TRANSCUTANEOUSLY REFILLABLE SUBCUTANEOUS NANOFLUIDIC DRUG DELIVERY IMPLANT FOR SUSTAINED AND CONSTANT ISL RELEASE. THE NANODDI COMPRISES A NEWLY PATENTED NANOFLUIDIC MEMBRANE, AND PORTS FOR RAPID, MINIMALLY INVASIVE TRANSCUTANEOUS DRUG REFILLING. REFILLING IS PERFORMED MANUALLY VIA A SYRINGE WITHOUT ANY COMPLEX PUMPS OR EQUIPMENT TO EXTEND IMPLANT USE DURATION BEYOND 2 YEARS. THE NANOFLUIDIC MEMBRANE USE NANOCHANNELS TO CONTROL DRUG RELEASE THROUGH PASSIVE DIFFUSION WITHOUT PUMPING MECHANISMS, PERMITTING DISCRETE, LONG-TERM USER-INDEPENDENT DOSING. UNLIKE INJECTABLES OR OTHER LA POLYMERIC STRATEGIES, NANODDI AVOIDS BURST AND DECAY RELEASE. ZERO-ORDER RELEASE KINETICS IS ACHIEVED INDEPENDENT OF PHYSIOLOGICAL CONDITIONS, REGARDLESS OF INTERINDIVIDUAL HETEROGENEITY. IMPORTANTLY, THE NANODDI ADDRESSES USER PREFERENCES FOR DISCRETION AND LONGER DOSE DURATION. HERE WE WILL TEST THE HYPOTHESIS THAT CONSTANT AND SUSTAINED ISL DELIVERY FROM NANODDI WILL ACHIEVE PREVENTATIVE DRUG LEVELS FOR A 2-YEAR DURATION AND EFFECTIVELY PREVENT SHIV INFECTION IN NON-HUMAN PRIMATES (NHP). THIS PROPOSAL OUTLINES A COMPREHENSIVE PRECLINICAL FRAMEWORK FUNDAMENTAL FOR DEVELOPING NANODDI AS A HIV PREP PLATFORM, LEVERAGING THE TEAM’S EXPERIENCE IN PRE-CLINICAL AND CLINICAL HIV PREP, LONG-ACTING DRUG DELIVERY, DRUG FORMULATION, AND ANTIRETROVIRAL PHARMACOLOGY. WE AIM 1) TO DEVELOP AND OPTIMIZE NANODDI AND ISL FORMULATION FOR SUSTAINED AND CONSTANT RELEASE; 2) TO ASSESS PHARMACOKINETICS (PK), TOLERABILITY, AND SAFETY OF NANODDI-ISL FOR 2 YEARS IN NHP AND EVALUATE EFFECTIVENESS OF TRANSCUTANEOUS DRUG REFILLING; AND 3) TO COMPREHENSIVELY EVALUATE PREP EFFICACY OF NANODDI-ISL IN NHPS USING 4 ROUTES OF SIMIAN HIV TRANSMISSION, NAMELY RECTAL, PENILE, VAGINAL AND INTRAVENOUS. OUR MULTIDISCIPLINARY TEAM HAS A SOLID HISTORY OF COLLABORATIVE HIV PREP STUDIES WITH LONG-ACTING DRUG DELIVERY IMPLANTS AND WILL RECEIVE MERCK’S SCIENTIFIC AND TECHNICAL SUPPORT AND DRUG SUPPLY FOR THIS STUDY. WE WILL USE A MILESTONE-DRIVEN RESEARCH APPROACH TO ADVANCE THE NANODDI-ISL TECHNOLOGY TOWARDS THE ULTIMATE GOAL OF CLINICAL TRANSLATION.

O-FUCOSE MODIFIED NOTCH AS A REGULATOR OF THE HEMATOPOIETIC STEM CELL HOMEOSTASIS

TARGETING ANTIGEN-INDUCED ER STRESS RESPONSE IN B-CELL CHRONIC LYMPHOCYTIC LEUKEM

A NOVEL IMAGING INFORMATICS PLATFORM FOR CRANIOMAXILLOFACIAL SURGERY

VENOUS: A TRANSLATIONAL STUDY OF ENTEROCOCCAL BACTEREMIA

VASCULARIZED NICHE WITH LOCAL IMMUNOSUPPRESSION FOR CELL REPLACEMENT FOR TYPE 1 DIABETES - CELL ENCAPSULATION TECHNOLOGIES ARE POISED TO IMPROVE CONVENTIONAL ISLET TRANSPLANTATION TO MORE EFFECTIVELY MANAGE TYPE I DIABETES. CURRENTLY, LIFELONG WHOLE-BODY IMMUNOSUPPRESSION IS ADMINISTERED TO AVOID IMMUNE REJECTION OF THE TRANSPLANT, DESPITE THE ASSOCIATED LIFE-THREATENING ADVERSE EFFECTS. CLINICAL STUDIES REVEAL THAT TRANSPLANTS EVENTUALLY FAIL DUE TO LACK OF VASCULAR SUPPORT FOR NUTRIENTS AND OXYGEN SUPPLY AND HOST IMMUNE REJECTION. TO ADDRESS ALL THESE CRITICAL NEEDS AND SUPPORTED BY PRELIMINARY STUDIES, WE PROPOSE THE NICHE, AN INNOVATIVE SUBCUTANEOUS VASCULARIZED ENCAPSULATION SYSTEM WITH LOCAL ELUTION OF IMMUNOSUPPRESSANTS TO PROTECT TRANSPLANTED CELLS FROM IMMUNE REJECTION. THE NICHE PRESENTS DUAL TRANSCUTANEOUSLY REFILLABLE RESERVOIRS, FOR DRUG AND CELLS, RESPECTIVELY, SEPARATED BY A NANOPOROUS MEMBRANE. LOCAL IMMUNOSUPPRESSANT DELIVERY CONFINES DRUGS TO THE GRAFT SITE WHERE IMMUNE ATTACK OCCURS, MINIMIZING EXPOSURE TO THE REST OF THE BODY, THUS AVOIDING SYSTEMIC IMMUNOSUPPRESSION AND ASSOCIATED ADVERSE EFFECTS. THE NICHE CELL RESERVOIR IS FULLY VASCULARIZED WITH FUNCTIONAL VESSELS, RECREATING AN IDEAL PHYSIOLOGICAL ENVIRONMENT CONDUCIVE FOR MAINTAINING LONG-TERM VIABILITY AND FUNCTION OF TRANSPLANTED CELLS. WE HYPOTHESIZE THAT THE NICHE WILL PROVIDE A VASCULARIZED ENVIRONMENT WITH LOCAL IMMUNOSUPPRESSANT DELIVERY CONDUCIVE FOR SUCCESSFUL LONG-TERM ISLET ALLOTRANSPLANTATION TO RESTORE EUGLYCEMIA IN DIABETIC NONHUMAN PRIMATES (NHP). IN AIM 1, WE WILL STUDY THE EFFICACY AND SAFETY OF LOCAL IMMUNOSUPPRESSANT COMBINATIONS AS WELL AS CHARACTERIZE THEIR RELEASE FROM THE NICHE VIA IN VITRO STUDIES. THIS WILL BE FOLLOWED IN AIM TO DEFINE THE OPTIMAL LOCAL IMMUNOSUPPRESSION REGIMEN FOR ISLET ALLOTRANSPLANTATION IN NICHE AND ESTABLISH DRUG PHARMACOKINETICS (PK) AND BIODISTRIBUTION IN HEALTHY NHP. IN AIM 3, WE WILL EVALUATE THE CURATIVE EFFICACY OF NICHE WITH ALLOTRANSPLANTED ISLETS TO RESTORE AND MAINTAIN EUGLYCEMIA IN DIABETIC NHPS FOR ONE YEAR. THE PROPOSED STUDIES ARE BASED ON OUR TEAM’S EXTENSIVE EXPERTISE IN IMPLANTABLE DRUG AND CELL DELIVERY SYSTEMS, TISSUE ENGINEERING, RESEARCH AND CLINICAL TRANSPLANTATION, TRANSPLANT IMMUNOLOGY, TYPE 1 DIABETES, AS WELL AS SUPPORTIVE PRELIMINARY DATA AND PREVIOUSLY PUBLISHED WORK. IMPORTANTLY, THE NICHE IS DESIGNED PRIORITIZING CLINICAL CONSIDERATIONS OF EFFICACY, SAFETY AND USER ACCEPTABILITY. TRANSCUTANEOUS CELL AND DRUG REFILLING ALLOW FOR EASE OF DRUG REPLENISHMENT WHEN NEEDED, THUS EXTENDING IMPLANT LIFESPAN POTENTIALLY FOR THE LIFETIME OF PATIENTS. FURTHER, THE THIN AND COMPACT SIZE OF THE NICHE, WHICH IS SMALLER THAN THE ENCAPSULATION IMPLANTS UNDER CLINICAL INVESTIGATION, IS FAVORABLE FOR USER ACCEPTABILITY. SUCCESSFUL COMPLETION OF THE PROPOSED WORK WILL PROVIDE A BROADLY APPLICABLE ENCAPSULATION SYSTEM WITH LOCALIZED IMMUNOSUPPRESSANT DELIVERY FOR LONG- TERM PROTECTION OF TRANSPLANTED ISLETS, AS WELL AS MINIMIZE ADVERSE EFFECTS ASSOCIATED WITH IMMUNOSUPPRESSIVE DRUGS. THIS COULD TRANSLATE TO A CLINICAL BREAKTHROUGH FOR DEPLOYMENT OF CELL THERAPIES BEYOND ISLETS, INCLUDING STEM CELL-DERIVED SS CELLS, TO TREAT DIABETES AS WELL AS OTHER DISEASES.

THE IMPACT OF SLEEP ON NETWORK CODING AND PERCEPTUAL PERFORMANCE

NOVEL STEM CELL IMMUNOTHERAPY FOR MDR-TUBERCULOSIS - ABSTRACT TUBERCULOSIS (TB) KILLS AROUND 2 MILLION PEOPLE EACH YEAR AND IS A MAJOR CAUSE OF MORTALITY FROM A SINGLE INFECTIOUS DISEASE WORLDWIDE. EMERGENCE OF MULTIDRUG-RESISTANT TB (MDR) IS NOW A MAJOR PROBLEM IN MORE THAN 60 COUNTRIES OF THE WORLD. MDR-TB IS THEREFORE A DISEASE, WHERE NOVEL INTERVENTION STRATEGIES INCLUDING ‘THERAPEUTIC VACCINES’ ARE REQUIRED TO SUPPLEMENT AVAILABLE DRUG REGIMEN. A CHARACTERISTIC FEATURE OF LUNG TUBERCULOSIS IS THE GRANULOMA, WHICH IS COLLECTION OF IMMUNE CELLS INCLUDING T CELLS, NEUTROPHILS, DCS, MESENCHYMAL STEM CELLS (MSCS) AND MACROPHAGES (MS) SURROUNDING A CENTRAL CORE OF MYCOBACTERIUM TUBERCULOSIS (MTB) INFECTED MACROPHAGES (MS). GRANULOMAS RESTRICT THE GROWTH OF MTB AND CONTINUOUSLY RECRUIT IMMUNE CELLS. MACROPHAGES ARE IN TURN, THE MAJOR PHAGOCYTIC CELLS WHICH KILL MTB. WE DISCOVERED TWO NOVEL MECHANISMS THROUGH WHICH MTB CAN BE KILLED DURING TUBERCULOSIS. FIRST, WE FOUND THAT HUMAN PRO-INFLAMMATORY M1-MS EXPRESSED NITRIC OXIDE (NO) AND UP-REGULATED AUTOPHAGY TO KILL MTB WHEREAS, ANTI-INFLAMMATORY M2-MS ALLOWED THE GROWTH OF MTB DUE TO A DECREASE IN NO SYNTHESIS AND AUTOPHAGY. SECONDLY, WE DISCOVERED THAT, BCG VACCINE AND MTB INFECTED MESENCHYMAL STEM CELLS (MSCS) REPROGRAMMED NAÏVE HUMAN MACROPHAGES TO BECOME M1 PHENOTYPE AND MORE ACTIVATED TO KILL MTB. INTERESTINGLY, PHASE I TRIALS USING NAÏVE, AUTOLOGOUS MSCS HAVE IMPROVED THE HEALTH OF MDR-TB PATIENTS. IN THIS PROPOSAL, WE WILL LEVERAGE OUR NEW FINDINGS TO INCREASE HUMAN MACROPHAGE ACTIVATION THROUGH STEM CELL-MEDIATED IMMUNOTHERAPEUTIC VACCINE AS FOLLOWS. SPECIFIC AIM- 1: INVESTIGATE THE MOLECULAR MECHANISMS THROUGH WHICH, BCG VACCINE AND MTB INFECTED OR CONDITIONED HUMAN MESENCHYMAL STEM CELLS TO EPIGENETICALLY PROGRAM NAIVE HUMAN MACROPHAGES. SPECIFIC AIM-2: INVESTIGATE THE IN VIVO THERAPEUTIC AND PROPHYLACTIC EFFECTS OF CONDITIONED MSCS DURING EXPERIMENTAL TUBERCULOSIS. WE WILL ANALYZE THE ABILITY OF TRANSFUSED MSCS TO ERADICATE BACTERIA AND RESTORE LUNG FUNCTION. MSCS HAVE BEEN USED IN MORE THAN 300 CLINICAL TRANSFUSION TRIALS TO CONTROL CANCER, AUTOIMMUNE DISEASES AND INFLAMMATION. WE WILL DEVELOP A NEW METHOD OF STEM CELL THERAPEUTIC VACCINATION TO CONTROL MDR-TB.

A NOVEL NANOCHANNEL SYSTEM FOR SUSTAINED DELIVERY OF TENOFOVIR ALAFENAMIDE FUMARATE AND EMTRICITABINE FOR HIV PRE-EXPOSURE PROPHYLAXIS

CONTROL OF TREG EXHAUSTION BY OX40

THE LIAFSR SYSTEM AND ANTIMICROBIAL PEPTIDE RESISTANCE IN ENTEROCOCCI

A NEONATAL MONKEY MODEL FOR TUBERCULOSIS VACCINATION

VEIN OF MARSHALL ETHANOL INFUSION FOR PERSISTENT ATRIAL FIBRILLATION

THERAPEUTIC APPROACHES TO DYSREGULATED HDL METABOLISM

TARGETING LIPID UNSATURATION IN OVARIAN CANCER STEM CELLS

ANH TATRC 2010 PRE CENTER AND SEED GRANTS

INTERVENTION OF IMMUNE TOLERANCE BY SMALL MOLECULES TO ENHANCE IMMUNE THERAPY

MECHANISMS OF JAG1-MEDIATED IMMUNE SUPPRESSION IN THE PANCREATIC CANCER MICROENVIRONMENT - PANCREATIC CANCER (PANCAN) IS NOTORIOUS FOR ITS RESISTANCE TO THERAPY INCLUDING IMMUNOTHERAPY, WHICH IS LARGELY CONTRIBUTED BY A SUPPRESSIVE TUMOR MICROENVIRONMENT. TARGETING MOLECULAR PATHWAYS INTEGRAL TO PANCAN RESISTANCE TO CURRENT THERAPY REMAINS AN UNMET NEED. NOTCH SIGNALING REGULATES PANCAN GROWTH, METASTASIS, AND THE TUMOR MICROENVIRONMENT. WHILE OTHER STUDIES HAVE SHOWN THAT NOTCH SIGNALING IN MACROPHAGE AND MYELOID DERIVED SUPPRESSOR CELLS PLAYS AN IMMUNE-SUPPRESSIVE ROLE IN BREAST CANCER AND LUNG CANCER, FOR EXAMPLE, OUR PROPOSAL WILL STUDY PREVIOUSLY UNRECOGNIZED ROLE OF JAG1 IN THE REGULATION OF THE METABOLIC FITNESS OF CONVENTIONAL DENDRITIC CELLS (CDCS) THAT UNDERLIES THE POOR ANTI-TUMOR RESPONSES TO IMMUNOTHERAPY. THE OVERARCHING OBJECTIVE OF THIS APPLICATION IS TO DEFINE IF TARGETING JAGGED1 (JAG1), A NOTCH LIGAND, WHOSE EXPRESSION CORRELATES WITH A WORSE PROGNOSIS AND INVERSELY CORRELATES WITH BATF3, ESSENTIAL FOR TYPE I CDC (CDC1) DEVELOPMENT, COULD REVERSE THE IMMUNOTHERAPY RESISTANCE IN TUMORS WITH POOR TUMOR-INFILTRATING CDC1 AND T CELLS. CDC1 CELLS PRESENT TUMOR ANTIGENS TO TUMOR-KILLING T CELLS AND THUS PLAY A CRITICAL ROLE IN SUPPORTING T CELL ANTI- TUMOR IMMUNITY IN IMMUNOTHERAPY. HOWEVER, DENDRITIC CELLS ARE PRESENT IN LOW NUMBERS AND OFTEN DISPLAY FUNCTIONAL SUPPRESSION IN PANCREATIC CANCER PATIENTS. OUR RECENT WORK REVEALED THAT JAG1 FUNCTIONS AS AN INHIBITORY NOTCH LIGAND FOR CDC1 DEVELOPMENT AS WELL AS CDC METABOLIC FITNESS. WE ALSO FOUND THAT ABLATING JAG1 INDUCED MARKED TUMOR REGRESSION AND PROLONGED TUMOR-BEARING MICE LONG-TERM SURVIVAL BY INCREASING AND REVIVING TUMOR-INFILTRATING DCS AND NOTCH-ACTIVATED TUMOR-KILLING T CELLS. WE WILL TEST OUR CENTRAL HYPOTHESIS THAT JAG1 PROMOTES PANCAN THERAPY RESISTANCE BY SUPPRESSING CDC1 METABOLIC FITNESS AND CD8 T CELL ANTI-TUMOR RESPONSE THROUGH THREE INTERRELATED AIMS. IN AIM 1, WE WILL TEST IF ANTI-JAG1 BLOCKING ANTIBODY COULD SENSITIZE RESISTANT TUMORS TO IMMUNOTHERAPY. IN AIM 2, WE WILL INVESTIGATE THE MECHANISM BY WHICH JAG1 IMPAIRS CDC METABOLIC FITNESS ESSENTIAL FOR ANTI-TUMOR ACTIVATION. AIM 3 WILL FOCUS ON THE MECHANISM OF ANTI-TUMOR T CELL REINVIGORATION MEDIATED BY JAG1 ABLATION/BLOCKADE. WE WILL USE A COMBINATION OF THE AUTOCHTHONOUS KPC MOUSE MODEL, IMMUNOPHENOTYPICALLY DEFINED KPC ORTHOTOPIC TUMORS, HUMAN PANCAN ORGANOIDS/MYELOID CELL COCULTURE, AND HUMAN PANCAN SPECIMENS. WE WILL EMPLOY CUTTING-EDGE MASS CYTOMETRY TO INVESTIGATE THE INTERPLAY BETWEEN TUMOR CELLS AND THE SURROUNDING IMMUNE CELLS AND THE TUMOR STROMA. WE WILL INTEGRATE THE IMMUNOPHENOTYPING AND THE SPATIAL TRANSCRIPTOMICS OF RARE IMMUNE CELL POPULATIONS WITH CLINICAL AND HISTOPATHOLOGICAL INFORMATION TO CORRELATE MOLECULAR FINDINGS WITH PATIENT OUTCOMES. THIS WORK IS INNOVATIVE AS WE ILLUMINATE A NOVEL FUNCTION OF JAG1 IN THE MAINTENANCE OF PANCAN THERAPY RESISTANCE AND EXPLORE ITS POTENTIAL AS A NOVEL IMMUNOMODULATORY TREATMENT FOR THIS DEADLY DISEASE.

LONG-ACTING MULTI PREVENTION IMPLANT FOR 2-YEAR CONTRACEPTION AND HIV PREP - PROJECT SUMMARY/ABSTRACT MATERNAL MORTALITY AND AIDS ARE AMONG THE LEADING CAUSES OF DEATH IN REPRODUCTIVE-AGE WOMEN GLOBALLY DUE TO UNINTENDED PREGNANCY AND THE DISPROPORTIONATE BURDEN OF HIV INFECTION. WHILE EFFECTIVE PREVENTION METHODS EXIST, ONLY 21% OF WOMEN HAVE ACCESS TO MODERN CONTRACEPTIVES AND ADHERENCE TO HIV PREP REGIMEN IS POOR IN MANY HIV-ENDEMIC REGIONS. MULTIPURPOSE PREVENTION TECHNOLOGIES (MPT) AIM TO SYNCHRONIZE CONTRACEPTIVE DELIVERY WITH HIV PREP, OFFERING A PATIENT-CENTERED APPROACH TO PROVIDE DUAL COVERAGE TO SEXUALLY ACTIVE WOMEN IN A SINGLE PRODUCT. A LONG-ACTING UNIFIED DELIVERY PRODUCT THAT CAN BE ADMINISTERED IN A DISCREET MANNER COULD REDUCE STIGMA AROUND CONTRACEPTIVES AND HIV PREP, THEREBY IMPROVING UPTAKE AND ADHERENCE TO COMBINATION PREVENTION PRODUCTS. OUR GOAL IS TO DEVELOP A LONG-ACTING DELIVERY IMPLANT OF ETONOGESTREL (ENG) AND ISLATRAVIR (ISL) DRUGAMER WITH AN UNPRECEDENTED 2-YEAR RELEASE DURATION FOR PREGNANCY AND HIV PREVENTION THAT ELIMINATES FURTHER NEED OF ADHERENCE. A NEW SUBCUTANEOUS NANOFLUIDIC MULTIPURPOSE IMPLANT (NANOMPI) WILL BE BROUGHT TOGETHER WITH A NEW POLYMERIC PRODRUG TECHNOLOGY TERMED “DRUGAMER”, TO ACHIEVE THIS INNOVATIVE PRODUCT CANDIDATE THAT DELIVERS ULTRA-LONG ACTING ENG+ISL DRUG DOSING DURATIONS. THE NANOMPI LEVERAGES A NANOFLUIDIC MEMBRANE WITH NANOCHANNELS AND A SINGLE DRUGAMER RESERVOIR TO ACHIEVE ZERO-ORDER RELEASE KINETICS THROUGH PASSIVE DIFFUSION WITHOUT PUMPING MECHANISMS, PERMITTING DISCREET, USER-INDEPENDENT DOSING. KEY PRODUCT ATTRIBUTES WILL INCLUDE DRUG STABILIZATION ON THE MULTI-YEAR TIMEFRAME, MINIMIZED EARLY- AND LATE-STAGE BURST RELEASE AND PHARMACOKINETIC (PK) TAILING, ZERO-ORDER DRUG RELEASE KINETICS TAILORED TO THE INDIVIDUAL DRUG PK/PD (PHARMACODYNAMIC) REQUIREMENTS, USER-INDEPENDENT DOSING, AND RETRIEVABILITY. THE PROJECT IS STRUCTURED AROUND 3 SPECIFIC AIMS: 1) TO DEVELOP OPTIMIZED ENG-ISL DRUGAMER FORMULATIONS THAT STABILIZE THE DRUGS AND EXHIBIT THE DESIRED ZERO ORDER RELEASE PROFILES, IN CONCERT WITH THE OPTIMIZATION OF THE NANOMPI DEVICE FOR 2-YEAR SUSTAINED RELEASE; 2) TO ASSESS PHARMACOKINETICS, TOLERABILITY, AND SAFETY OF THE LEAD ENG-ISL NANOMPI DEVICE FOR 2 YEARS IN PIGTAIL MACAQUES; AND 3) TO EVALUATE CONTRACEPTION AND HIV PREP EFFICACY OF ENG-ISL NANOMPI IN PIGTAIL MACAQUES VIA REPEATED LOW-DOSE VAGINAL CHALLENGE AND DEVELOP A PK/PD MODEL. WE WILL USE A MILESTONE-DRIVEN RESEARCH APPROACH TO ADVANCE THE ENG-ISL NANOMPI TECHNOLOGY TOWARDS THE ULTIMATE GOAL OF CLINICAL TRANSLATION, LEVERAGING THE INTERDISCIPLINARY TEAM’S EXPERIENCE IN DRUGAMER TECHNOLOGY, DRUG DELIVERY, PRE-CLINICAL AND CLINICAL HIV PREVENTION STUDIES, AND HORMONAL CONTRACEPTIVE AND ANTIRETROVIRAL PHARMACOLOGY. IMPORTANTLY, THE NANOMPI ADDRESSES USER PREFERENCES FOR A DUAL PREVENTION PRODUCT, DISCRETION AND LONGER DOSING DURATION. SUCCESSFUL COMPLETION WILL GENERATE A VERSATILE MPT PLATFORM THAT COULD BE ADOPTED FOR OTHER DRUG COMBINATIONS AND PREVENTATIVE STRATEGIES.

CARDIAC AUTONOMIC ACTIVATION IN ATRIAL FIBRILLATION TRIGGERS AND SUBSTRATE - PROJECT SUMMARY/ABSTRACT ATRIAL FIBRILLATION (AF) IS THE MOST COMMON SUSTAINED ADULT ARRHYTHMIA, ASSOCIATED WITH AN INCREASED RISK OF STROKE, HEART FAILURE AND DEMENTIA. WITH INCREASED LONGEVITY IN CHRONIC DISEASES, THE PREVALENCE OF AF – CURRENTLY ESTIMATED AT 46 MILLION WORLDWIDE – IS DRAMATICALLY RISING. CATHETER ABLATION – TISSUE DESTRUCTION – IS THE MOST EFFECTIVE THERAPY BUT IS FRAUGHT WITH PROCEDURAL RISKS AND SUBOPTIMAL EFFICACY. THE CARDIAC AUTONOMIC SYSTEM (CANS) IS KNOWN TO BE INVOLVED IN THE PATHOGENESIS OF AF, BUT NO SPECIFIC DIAGNOSTIC OR THERAPEUTIC APPROACHES HAVE EVOLVED FROM THIS. OUR LONG-TERM GOAL IS TO DEVISE NEUROMODULATORY AF TREATMENT AND PREVENTIVE STRATEGIES AND FILL THE KNOWLEDGE GAP ON THE MECHANISMS BY WHICH CANS NEURONAL AND HUMORAL PARACRINE OUTPUT MODULATE ATRIAL FUNCTION IN HUMANS. WE BREAK THROUGH EXISTING TECHNICAL BARRIERS THAT LIMITED OUR UNDERSTANDING OF INTRINSIC CARDIAC GANGLIA, CAPITALIZING ON THE VEIN OF MARSHALL AS A VASCULAR ROUTE TO SAMPLE ITS ELECTROPHYSIOLOGY AND HUMORAL RESPONSES, COLLECTING ATRIAL CORONARY CIRCULATION BLOOD, AND RECORDING NERVE ACTIVITY FROM THE GANGLIONATED PLEXI (GP). OUR EXTENSIVE PRELIMINARY DATA IN PATIENTS SHOWS THAT APNEA INCREASES GP ACTIVITY MEASURED USING NOVEL PERCUTANEOUS TECHNOLOGY IN AF PATIENTS UNDERGOING ABLATION PROCEDURES. REMARKABLY, WE FOUND THAT SUBSTANCE P (SP) COLLECTED FROM THE CORONARY SINUS IS ELEVATED COMPARED TO UNDETECTABLE LEVELS IN PERIPHERAL BLOOD OF AF PATIENTS SUGGESTING THAT GP ACTIVATION VIA SECRETED SP MAY PLAY A ROLE IN AF SUBSTRATES. IN LARGE ANIMAL MODELS, WE HAVE FOUND THAT SPECIFIC ABLATION OF GP SENSORY NEURONS BLUNTS THE PRO-FIBRILLATORY RESPONSE TO APNEA AND THAT A CRESCENDO GP RESPONSE OCCURS AFTER REPEATED CONSECUTIVE APNEAS. OUR DATA IN HUMAN PLURIPOTENT STEM CELL-DERIVED ATRIAL CARDIOMYOCYTES (HIPSC-ACM) SHOWS THAT CHRONIC SP TREATMENT AFFECTS CARDIOMYOCYTE ELECTROPHYSIOLOGY AND MODIFIES GENE EXPRESSION OF MIR-21 TARGETS. THESE EXCITING OBSERVATIONS, INNOVATIVE METHODS, AND UNIQUE CLINICAL AND BASIC SCIENCE EXPERTISE POSITION OUR TEAM TO DEVELOP THIS PROJECT SUCCESSFULLY. WE PROPOSE THE CENTRAL HYPOTHESIS THAT CANS PRODUCES A SUBSTRATE FOR AF THROUGH NEURAL (NERVE FIRING) AND HUMORAL EFFECTS (SECRETOME), IN WHICH SP – RELEASED BY GP SENSORY NEURONS – PLAYS A MAJOR ROLE IN INCREASING SUSCEPTIBILITY TO AF THROUGH DIRECT ELECTROPHYSIOLOGICAL AND GENOMIC EFFECTS IN ATRIAL CARDIOMYOCYTES. THE CENTRAL HYPOTHESIS WILL BE TESTED BY PURSUING STUDIES 1) IN HUMANS WITH PAROXYSMAL AND PERSISTENT AF AIMING TO MEASURE NERVE ACTIVITY AND SECRETOME OF INTRINSIC GANGLIA THROUGH THE VEIN OF MARSHALL DURING ABLATION PROCEDURES, 2) IN CANINE MODELS OF ACUTE AND PERSISTENT AF TO DETERMINE WHETHER ABLATION OF GP OR SP ANTAGONISM AMELIORATES AF, 3) IN HIPSC-ACM AND ENGINEERED ATRIAL TISSUES TO ELUCIDATE SP ACTIONS. THE PROPOSED RESEARCH IS SIGNIFICANT BECAUSE IT IS EXPECTED TO PROVIDE A MECHANISTIC UNDERSTANDING OF THE RELATIONSHIP BETWEEN CANS AND SLEEP APNEA FOR THE CONTINUED DEVELOPMENT OF EFFECTIVE THERAPIES AGAINST AF. ULTIMATELY, SUCH KNOWLEDGE CAN OFFER NEW OPPORTUNITIES TO DEVELOP INNOVATIVE THERAPIES TO TREAT AF.

AGE-RELATED TDP-43 NEUROPATHOLOGY: USING DISEASE-DRIVING MECHANISMS TO GUIDE CLASSIFICATION

SYSTEMATIC MODELING AND PREDICTION OF CELL-TYPE-SPECIFIC AND SPATIOTEMPORAL CROSSTALK PATHWAYS IN ALZHEIMER'S DISEASE - ABSTRACT ALZHEIMER’S DISEASE (AD) AFFECTS MORE THAN 50 MILLION PEOPLE WORLDWIDE BUT THERE IS NO CLEAR THERAPEUTIC OPTION FOR THE PATIENTS. FOR LAST TWO DECADES, AD RESEARCH HAS BEEN FOCUSING ON A NEURON-CENTRIC BIOCHEMICAL PROCESS THAT LEADS TO SYNAPTIC DEFICITS AND NEURONAL DEGENERATION. HOWEVER, RECENT FAILURES IN CLINICAL TRIALS CLEARLY DEMONSTRATE A GAP IN KNOWLEDGE IN OUR CURRENT UNDERSTANDING OF AD PATHOGENESIS AND CALL FOR STUDIES THAT LEAD TO UNBIASED AND HOLISTIC UNDERSTANDING OF DISEASE PATHWAYS IN DIFFERENT TYPES OF BRAIN CELLS. THIS PROJECT AIMS TO TACKLE THIS IMPORTANT AND URGENT ISSUE BY COMBINING A COMPUTATIONAL SYSTEMS BIOLOGY PLATFORM SINGLE-CELL RESOLUTION BRAIN INTERACTOME (SCRBI) EXPLORER, 3D HUMAN ALZHEIMER’S-IN-A-DISH MODELS, AND THE PUBLICLY AVAILABLE MULTIPLE-OMICS AD DATABASES THROUGH NIH-FUNDED AMP-AD PORTAL. WE WILL EXPAND THE KNOWLEDGE BASE OF SCRBI EXPLORER TO HANDLE SINGLE CELL TRANSCRIPTOMIC AND MULTIPLE OMICS PROFILES FROM 3D CELL MODELS AND HUMAN BRAIN TISSUES, WHICH CAN DETECT ON MULTIPLE LAYERS OF NEURON-GLIA AND GLIA-GLIA CROSSTALK PATHWAYS VIA LIGAND-RECEPTOR INTERACTIONS, CYTOKINE/CHEMOKINE SIGNALING, INTRACELLULAR SIGNALING ACTIVITIES, AND TRANSCRIPTIONAL ACTIVATION. THE CENTRAL HYPOTHESIS IS THAT THE COMBINED USE OF MULTI-CELLULAR SYSTEMS BIOLOGY MODELING AND 3D HUMAN AD CELLULAR MODELS WILL IDENTIFY AD-SPECIFIC NEURON-GLIA AND GLIA-GLIA CROSSTALK PATHWAYS, WHICH WOULD PROVIDE NOVEL THERAPEUTIC TARGETS FOR DRUG REPOSITIONING. WE WILL TEST THIS HYPOTHESIS BY PURSUING THREE SPECIFIC AIMS: 1) DEVELOP A MULTI-CELLULAR CROSSTALK MODEL TO UNCOVER ALTERED NEURON-GLIA AND GLIA-GLIA CROSSTALK PATHWAYS IN AD, 2) IDENTIFY AND VALIDATE AD-SPECIFIC NEURON-GLIA AND GLIA-GLIA CROSSTALK PATHWAYS THAT ARE ENRICHED IN 3D HUMAN AD CELLULAR MODELS AND HUMAN AD BRAIN CELLS, AND 3) EVALUATE THE THERAPEUTIC POTENTIAL OF NEURON-GLIA AND GLIA-GLIA CROSSTALK USING 3D HUMAN NEURAL CELL CULTURE MODELS OF AD. THE POTENTIAL IMPACT OF THIS PROPOSAL IS HIGH BECAUSE THE PROPOSED STUDY, IF SUCCESSFUL, WILL PROVIDE A UNIQUE INTEGRATED BIOINFORMATICS TOOL TO UNBIASEDLY IDENTIFY NEURON-GLIA AND GLIA-GLIA CROSSTALK PATHWAYS IN AD AND EVEN OTHER NEURODEGENERATIVE DISEASES. MORE IMPORTANTLY, IT WILL PROVIDE NOVEL THERAPEUTIC TARGETS BASED ON ALTERED NEURON-GLIA INTERACTION PATHWAYS IN AD AND OPEN UP A NEW VISTA FOR DRUG REPOSITIONING TARGETING CELL-CELL INTERACTIONS IN THE BRAIN OF AD PATIENTS.

CHEMOPREVENTION OF LUNG CANCER WITH MITOCHONDRIA-TARGETED HONOKIOL

MULTISTAGE VECTOR-ENABLED BREAST ONCOLOGY

VALIDATION OF A NOVEL 3D CULTURE PLATFORM FOR TNBC TREATMENT SELECTION - PROJECT SUMMARY THE GOAL OF THIS ACADEMIC [HOUSTON METHODIST RESEARCH INSTITUTE (HMRI) AND MD ANDERSON CANCER CENTER (MDACC)] AND INDUSTRIAL (EMPIRI) COLLABORATIVE PROJECT IS TO VALIDATE A NOVEL, FUNCTIONAL CANCER DIAGNOSTIC ASSAY FOR CLINICAL TRANSLATION TO IMPROVE AND PERSONALIZE THE CARE OF TRIPLE-NEGATIVE BREAST CANCER (TNBC) PATIENTS. THE STUDY WILL VALIDATE A NOVEL 3D TUMOR TISSUE CULTURE METHOD (E-SLICES) INVENTED BY MPI KYUSON YUN (HMRI) AND LICENSED TO EMPIRI, INC., A BIOTECH STARTUP CO-FOUNDED BY DR. YUN AND DAVE GALLUP (MPI). TOGETHER WITH DR. NAOTO UENO (MPI, MDACC), A RENOWNED PHYSICIAN-SCIENTIST SPECIALIZING IN TNBC RESEARCH AND PATIENT CARE, THE TEAM WILL VALIDATE THE PREDICTIVE ACCURACY OF E-SLICES FOR INDIVIDUAL TNBC PATIENT RESPONSES TO RECENTLY APPROVED CHEMOIMMUNOTHERAPY FOR EARLY TNBC PATIENT CARE. THE NEED FOR PERSONALIZED MEDICINE IN ONCOLOGY IS WIDELY ACCEPTED BUT TRANSLATING THIS IMPORTANT CONCEPT INTO CLINICAL PRACTICE HAS BEEN CHALLENGING. CURRENTLY, THE DOMINANT PLATFORM FOR PRECISION MEDICINE UTILIZES GENOMICS/SEQUENCING-BASED ASSAYS TO MEASURE THE EXPRESSION AND/OR MUTATIONAL PROFILES AND THEN INFER RESPONSES TO TARGETED THERAPIES; HOWEVER, THIS APPROACH BENEFITS <10% OF PATIENTS WITH PROFILED TUMORS. RECOGNIZING THE INHERENT LIMITATIONS OF THESE INFERENCE-BASED METHODS, FUNCTIONAL ASSAYS (E.G., ORGANOIDS, PDX MODELS) HAVE BEEN DEVELOPED, BUT THESE APPROACHES HAVE NUMEROUS LIMITATIONS INCLUDING HIGH COST AND TIME REQUIRED TO ESTABLISH THE MODELS, LOW “TAKE RATES”, AND DESTRUCTION OF THE NATIVE TUMOR MICROENVIRONMENT (TME). TO OVERCOME THESE CHALLENGES, EMPIRI DEVELOPED A NOVEL 3D EX VIVO TUMOR CULTURE METHOD (E-SLICES) THAT ENABLES RAPID, PERSONALIZED DRUG SENSITIVITY TESTING IN INTACT PATIENT TUMOR TISSUES. E-SLICES RETAIN THE NATIVE TME AND TISSUE ARCHITECTURE AND ARE CULTURED IN SERUM-FREE DEFINED MEDIA, OVERCOMING THE LIMITATIONS OF OTHER APPROACHES. IN ADDITION, E-SLICES CAN BE GENERATED FROM ANY SOLID TUMOR AND USED FOR TESTING RESPONSES TO CHEMOTHERAPY, TARGETED THERAPY, AND IMMUNOTHERAPY. IMPORTANTLY, E-SLICE ACCURACY HAS BEEN VALIDATED IN A CLINICAL SETTING FOR METASTATIC COLORECTAL CANCER TO ACCURATELY PREDICT INDIVIDUAL PATIENT TREATMENT RESPONSES AND DETECT INTER-PATIENT DIFFERENCES TO THE SAME TREATMENTS IN 4-12 DAYS, PAVING THE WAY FOR NEAR EVIDENCE-BASED PERSONALIZED TREATMENT SELECTIONS. THE TEAM WILL: (I) DETERMINE WHETHER E-SLICES PREDICT PATIENT RESPONSES TO SOC NAC: DOXORUBICIN (ADRIAMYCIN) PLUS CYCLOPHOSPHAMIDE) IN A RETROSPECTIVE STUDY, (II) MEASURE CHEMOIMMUNOTHERAPY (PEMBROLIZUMAB PLUS PACLITAXEL + CARBOPLATIN) RESPONSES IN HUMANIZED PDX SLICES FROM KNOWN RESPONDERS AND NON-RESPONDERS; AND (II) EVALUATE THE CLINICAL UTILITY OF E-SLICES IN PREDICTING TNBC PATIENT RESPONSES TO NEWLY APPROVED STANDARD OF CARE CHEMOIMMUNOTHERAPY IN A PROSPECTIVE STUDY. SUCCESSFUL COMPLETION OF THIS PROJECT WILL PROVIDE THE NECESSARY DATA TO APPLY E-SLICES AS THE FIRST FUNCTIONAL CANCER DIAGNOSTIC TEST SPECIFICALLY DESIGNED TO INFORM TNBC PATIENT CARE.

SUBCELLULAR LOCALIZATION OF NANOPARTICLES

HUMAN ATHEROGENESIS WITH UNDERLYING DYSFUNCTIONAL HDL-FREE CHOLESTEROL - ATHEROSCLEROTIC CARDIOVASCULAR DISEASE (ASCVD) AND PLASMA HIGH-DENSITY LIPOPROTEIN CHOLESTEROL (HDL-C) ARE NEGATIVELY CORRELATED. ONE MODEL ASSIGNS THIS CORRELATION TO THE ROLE OF HDL AS AN ACCEPTOR OF FREE CHOLESTEROL (FC) TRANSFER FROM ARTERIAL-WALL MACROPHAGES (FC EFFLUX), AND IN SOME STUDIES, FC EFFLUX BETTER PREDICTED ASCVD THAN HDL-C CONCENTRATION. HOWEVER, INTERVENTIONS THAT INCREASE PLASMA HDL FAILED TO REDUCE ASCVD, AND, PARADOXICALLY, SEVERAL STUDIES REVEALED HIGHER ASCVD MORTALITY AMONG PATIENTS WITH VERY HIGH HDL. THE SOURCE OF THIS PARADOX IS UNKNOWN, APPROPRIATE THERAPIES HAVE NOT BEEN FORMULATED, AND IN THIS CONTEXT, THE ROLE OF THE REVERSE PROCESS, HDL-FC INFLUX, WHICH MAY BE SUPPORTED BY EXCESS HDL-FC, IS UNKNOWN. MICE DEFICIENT IN THE HDL-RECEPTOR, SCARB1-/- MICE, ARE A ROBUST MODEL OF THE HUMAN HIGH-HDL PHENOTYPE. COMPARED TO WILD-TYPE MICE, SCARB1-/- MICE ARE MORE ATHERO-SUSCEPTIBLE AND HAVE HIGHER PLASMA LEVELS OF HDL THAT IS MORE FC-RICH. THIS PRODUCES A STATE OF HIGH HDL-FC BIOAVAILABILITY, WHICH WE EXPRESS AS AN INDEX: HDL- FCBI = HDL PARTICLE NUMBER (HDL-P) X MOL% HDL-FC. THIS CONCEPTUALLY NEW METRIC WAS FIRST REPORTED BY THIS STUDY TEAM, WHICH REPORTED THAT HDL-FCBI INCREASES AS WILD-TYPE MICE < HUMAN << SCARB1-/- MICE AND THAT MORE FC TRANSFERS FROM HDL FROM SCARB1-/- VS. WILD-TYPE MICE TO MACROPHAGES. THUS, WE HYPOTHESIZE THAT SIMILAR MECHANISMS UNDERLIE ASCVD AMONG HUMANS WITH VERY HIGH PLASMA HDL-C. OUR GOAL IS TO COMPARE PATIENTS WITH POSITIVE (CACS>0) AND NEGATIVE (CACS=0) CORONARY ARTERY CALCIUM SCORES RESPECTIVELY ASSIGNED AS ASCVD AND NON-ASCVD IN THREE SUBGROUPS—THOSE WITH HIGH (HH), INTERMEDIATE (IH), OR OPTIMAL (OH) PLASMA HDL-C CONCENTRATIONS—AND TEST WHETHER ASCVD IS ASSOCIATED WITH A HIGH HDL-FCBI. ACCORDING TO OUR HYPOTHESIS, A) HDL-FCBI WILL BE HIGHER AMONG HH VS. OH PATIENTS AND AMONG ASCVD VS. NON-ASCVD PATIENTS, ESPECIALLY THOSE WITH HIGH PLASMA HDL-C, AND B) THE MAGNITUDE OF FC TRANSFER FROM HDL FROM ASCVD PATIENTS TO MACROPHAGES WILL BE GREATER THAN THAT FROM HDL FROM NON-ASCVD PATIENTS, AGAIN ESPECIALLY AMONG ASCVD PATIENTS WITH HIGH PLASMA HDL-C. THIS HYPOTHESIS IS SUPPORTED BY STUDIES OF SCARB1-/- MICE IN WHICH A COMPONENT OF HDL-FCBI IS REDUCED AND WITH IT, ASCVD—REDUCING HDL-P WITH PROBUCOL OR MOL% HDL-FC BY INCREASED FC ESTERIFICATION SUPPRESSED ASCVD. COMPARISON OF CACS VS. HDL- FCBI OF ALL THREE GROUPS WILL REVEAL A NON-LINEAR FUNCTIONAL RELATIONSHIP. TRADITIONALLY, PHYSICIANS HAVE MEASURED HDL AND LDL IN TERMS OF THEIR TOTAL CHOLESTEROL CONTENT. THESE MEASURES HAVE HAD GOOD BUT IMPERFECT PREDICTIVE VALUE, MAINLY BECAUSE THE TWO COMPONENTS OF TC, FC AND CHOLESTERYL ESTERS, HAVE DISTINCT METABOLIC ITINERARIES THAT MAY DIFFERENTIALLY CONTRIBUTE TO ASCVD. VALIDATION OF THE STUDY-HYPOTHESES IN HUMANS WOULD PROVIDE A COMPELLING RATIONALE FOR MEASURING PLASMA LIPOPROTEIN FC AS AN ASCVD DIAGNOSTIC AND FOR THE FORMULATION OF THERAPIES THAT REDUCE PLASMA- AND ESPECIALLY HDL-FC.

SUDDEN DEATH RISK ASSESSMENT AND MECHANISTIC INSIGHTS IN ARRHYTHMIC MITRAL VALVE PROLAPSE USING CARDIAC MRI AND CIRCULATING PROTEOMIC BIOMARKERS - PROJECT SUMMARY: MITRAL VALVE PROLAPSE (MVP) IS A COMMON VALVULOPATHY AFFECTING 2-3% OF THE POPULATION AND NEARLY 200 MILLION INDIVIDUALS GLOBALLY. WHILE IT IS OFTEN BENIGN, A SUBSET OF PATIENTS WITH MVP MAY DEVELOP ARRHYTHMIAS, INCLUDING SUDDEN CARDIAC DEATH (SCD). ALTHOUGH THE ASSOCIATION BETWEEN MVP AND SCD WAS FIRST REPORTED DECADES AGO, THE RISK WAS INITIALLY BELIEVED TO BE SMALL; CONTEMPORARY OBSERVATIONAL STUDIES SUGGEST THAT SCD MAY BE A MORE COMMON SEQUELA IN MVP, WITH AN ESTIMATED YEARLY INCIDENCE BETWEEN 0.4 AND 1.9%. THE NATIONAL HEART, LUNG, AND BLOOD INSTITUTE RECENTLY CONVENED A WORKSHOP COMPOSED OF SUBJECT MATTER EXPERTS AND STAKEHOLDERS TO IDENTIFY RESEARCH NEEDS AND OPPORTUNITIES TO DEVELOP RECOMMENDATIONS FOR THE IDENTIFICATION AND TREATMENT OF INDIVIDUALS WITH MITRAL VALVE PROLAPSE, INCLUDING SUCH INDIVIDUALS WHO MAY BE AT RISK FOR SUDDEN CARDIAC ARREST OR SUDDEN CARDIAC DEATH. OUR CURRENT PROPOSAL IS DESIGNED TO ALIGN CLOSELY WITH THE PRIORITIES IDENTIFIED IN THE RECENT NIHBI WORKSHOP. SPECIFICALLY, OUR PROPOSAL HAS THREE AIMS: 1) PERFORM DEEP PHENOTYPING OF PATIENTS WITH MVP TO UNDERSTAND CHARACTERISTICS AND MECHANISMS ASSOCIATED WITH VENTRICULAR ARRHYTHMIAS; 2) STUDY A BLOOD BIOMARKERS PANEL (INCLUDING FDA-APPROVED AND NOVEL HIGH THROUGHPUT PROTEOMIC MARKERS) THAT COULD BE USED AS A SENSITIVE, COSTEFFECTIVE SCREENING STRATEGY TO IDENTIFY MVP PATIENTS WITH MYOCARDIAL FIBROSIS WHO ARE AT RISK FOR SCD; 3) BUILD A NOVEL SCD RISK PREDICTION MODEL IN MVP INCLUSIVE OF CMR EVIDENCE OF MYOCARDIAL FIBROSIS BY ASSEMBLING A LARGE OBSERVATIONAL MULTICENTER MVP REGISTRY WITH OVER 2,000 CONTRAST-ENHANCED CMRS AND LONGITUDINAL FOLLOW-UP FOR ARRHYTHMIC EVENTS.

OPTIMAL SELECTIVE THYROID HORMONE ANALOGS FOR METABOLIC SYNDROME

INTEGRATIVE MOLECULAR AND IMAGING APPROACHES FOR RISK OF SUBTYPE SPECIFIC BREAST

XBP1 INHIBITION AND STING ACTIVATION FOR THE TREATMENT OF CANCER - PROJECT SUMMARY THIS RESEARCH PROJECT FOCUSES ON TESTING AND FINDING CURES FOR PATIENTS DIAGNOSED WITH CHRONIC LYMPHOCYTIC LEUKEMIA (CLL), THE MOST COMMON ADULT BLOOD CANCER. CLL IS STILL AN INCURABLE TYPE OF CANCER DERIVED FROM B CELLS, WHICH NORMALLY PRODUCE ANTIBODIES TO PROTECT THE HUMAN BODY FROM INFECTIONS. ALTHOUGH IBRUTINIB AND VENETOCLAX HAVE BEEN APPROVED BY THE FDA TO TREAT CLL, A LARGE GROUP OF CLL PATIENTS SUFFER FROM CHRONIC TOXICITIES AND HAVE TO STOP USING THESE TWO DRUGS. IN ADDITION, SOME CLL PATIENTS DEVELOP DRUG RESISTANCE AND THEIR DISEASE BECOMES DIFFUSE LARGE B CELL LYMPHOMA (DLBCL), WHICH IS EVEN HARDER TO TREAT THAN CLL. WE HAVE SHOWN THAT BOTH MOUSE AND HUMAN CLL CELLS REQUIRE ACTIVATION OF A PROTEIN CALLED SPLICED X BOX-BINDING PROTEIN-1 (XBP-1S) TO SUPPORT THEIR GROWTH AND SURVIVAL. WE HAVE DEVELOPED A DRUG NAMED B-I09 THAT CAN EFFECTIVELY SUPPRESS XBP-1 AND KILL CLL CELLS. IN ADDITION, OUR NEW RESULTS HAVE SHOWN THAT CLL CELLS SIGNIFICANTLY REDUCE THEIR EXPRESSION OF A PROTEIN CALLED STIMULATOR OF INTERFERON GENES (STING). BASED ON OUR RESULTS SUGGESTING THAT REDUCED STING IN CLL CELLS MAY CONTRIBUTE TO THEIR SURVIVAL, WE PROPOSE TO USE ADU- S100, A STING ACTIVATOR CURRENTLY IN CLINICAL TRIALS, TO STIMULATE THE REMAINING STING TO KILL CLL CELLS. BASED ON THE LINK BETWEEN XBP-1S AND STING IN REGULATING THE SAME SURVIVAL MECHANISM, NAMELY, B CELL RECEPTOR (BCR) SIGNALING, WE PROPOSE TO TEST WHETHER B-I09 AND ADU-S100 CAN SYNERGIZE TO KILL CULTURED MOUSE AND HUMAN CLL CELLS. TO ENABLE THESE STUDIES, WE HAVE ENGINEERED A NOVEL MOUSE MODEL IN WHICH CLL CELLS PRODUCE NO STING. WE WILL USE THIS MODEL TO TEST WHETHER ADU-S100 CAN ACTIVATE THE IMMUNE SYSTEM TO SYNERGIZE WITH B-I09 TO EFFECTIVELY ERADICATE CLL. FINALLY, BASED ON OUR RECENT PROGRESS IN SUCCESSFULLY DEVELOPING STIMULI-RESPONSIVE INHIBITORS OF THE IRE-1/XBP-1 PATHWAY, WE PROPOSE TO CONJUGATE B-I09 TO A SINGLE-DOMAIN ANTIBODY (OR NANOBODY) AGAINST HUMAN CD20, AN INTEGRAL MEMBRANE PROTEIN EXPRESSED AT HIGH LEVELS IN CLL CELLS IN ORDER TO TARGET THE DRUG TO THESE CELLS. THE RESULTING NANOBODY-DRUG CONJUGATE (NDC) WILL BE EVALUATED IN CELLS AND IN VIVO FOR SUPPRESSION OF XBP-1S, REDUCTION OF TUMOR BURDEN, AND SYNERGY WITH ADU- S100 IN A NOVEL STING-DEFICIENT, HUMAN CD20-EXPRESSSING CLL MOUSE MODEL. THESE STUDIES WILL SERVE AS THE FOUNDATION FOR INITIATING PHASE I CLINICAL TRIALS TO TREAT HUMAN PATIENTS DIAGNOSED WITH CLL, DLBCL, OR POTENTIALLY OTHER SOLID TUMORS WITH AN INHIBITOR OF XBP-1S IN COMBINATION WITH AN ACTIVATOR OF STING. THE DEVELOPMENT OF THE FIRST NDC TO TARGET XBP-1S WILL OPEN A NEW AND IMPORTANT AVENUE TO ENABLE CELL-SPECIFIC INHIBITION OF XBP- 1S IN VARIOUS CELL TYPES AS LONG AS SUCH CELLS EXPRESS A UNIQUE CELL SURFACE MARKER. THESE NOVEL XBP-1S- TARGETING NDCS CAN ALSO BE USED TO INTERROGATE THE BIOLOGY OF THE IRE-1/XBP-1 PATHWAY IN VARIOUS CELL TYPES, AND AS THERAPEUTICS.

MOLECULAR MECHANISM OF VIRULENCE REGULATION IN STREPTOCOCCUS PYOGENES - PATHOGENIC BACTERIA SURVIVE IN COMPLEX AND HOSTILE ENVIRONMENTS IN THE HOST. SEVERAL HOST- AND MICROBIOTA-DERIVED FACTORS CURB PATHOGEN GROWTH DURING INFECTION. SUCCESSFUL PATHOGENS RESPOND BY EXPLOITING THE CUES IN THEIR IMMEDIATE ENVIRONMENT TO COORDINATE SPATIOTEMPORAL PRODUCTION OF VIRULENCE FACTORS. OUR PRELIMINARY DATA INDICATE THAT THE HUMAN PATHOGEN GROUP A STREPTOCOCCUS (GAS) IS ENGAGED IN ARMS RACE WITH A COMMENSAL BACTERIUM DURING OROPHARYNGEAL INFECTION. THE COMMENSAL BACTERIA PRODUCE A PREVIOUSLY UNKNOWN ANTIMICROBIAL METABOLITE WITH A NOVEL CHEMICAL SCAFFOLD THAT MAY CONTRIBUTE TO HOST DEFENSE AGAINST GAS COLONIZATION IN HUMAN OROPHARYNX. AS A COUNTERMEASURE, GAS EMPLOYS SECRETED CYSTEINE PROTEASE SPEB, A MAJOR VIRULENCE FACTOR, TO OVERCOME COMMENSAL DEFENSES BY PROTEOLYTICALLY DEGRADING THE ANTIMICROBIAL METABOLITES. DESPITE OUR EXPERIMENTAL EVIDENCE SUGGESTING ANTAGONISM BETWEEN GAS AND COMMENSAL BACTERIUM, THE FACTORS AND MECHANISMS THAT REGULATE ANTIMICROBIAL METABOLITE PRODUCTION IN THE COMMENSAL AND THEIR INFLUENCE ON SPEB PRODUCTION BY GAS ARE UNKNOWN. RECENTLY, WE DISCOVERED A NOVEL GAS QUORUM SENSING PATHWAY COMPRISED OF A NEW CLASS OF BACTERIAL QUORUM SENSING SIGNAL, A LEADERLESS SECRETED PEPTIDE, AND AN INTRACELLULAR RECEPTOR THAT CONTROLS THE TEMPORAL EXPRESSION OF SPEB DURING INFECTION. INTERESTINGLY, THE COMMENSAL BACTERIUM ALSO EMPLOYS A LEADERLESS PEPTIDE-DEPENDENT QUORUM SENSING PATHWAY TO CONTROL THE ANTIMICROBIAL METABOLITE PRODUCTION. HOWEVER, OUR PRELIMINARY DATA SUGGEST THAT GAS HIJACKS THE COMMENSAL PEPTIDE SIGNAL TO INDUCE ITS ENDOGENOUS QUORUM SENSING PATHWAY AND ACTIVATE SPEB PRODUCTION. THIS FINDING IS HIGHLY RELEVANT TO GAS PATHOGENESIS AS THE INTERSPECIES SIGNALING FACILITATES VIRULENCE FACTOR PRODUCTION IN A SUBOPTIMAL HOST ENVIRONMENT AND PROMOTES GAS VIRULENCE. USING A MULTIDISCIPLINARY APPROACH COMBINING MICROBIOLOGICAL, GENETIC, BIOCHEMICAL AND IMAGING METHODOLOGIES, AND ANIMAL INFECTION STUDIES, WE WILL DISSECT THE MOLECULAR DETAILS OF INTRA- AND INTER- SPECIES SIGNALING, CHARACTERIZE THE MECHANISM OF ANTAGONISM BETWEEN THE TWO BACTERIAL SPECIES, DETERMINE ITS IMPACT ON GAS PATHOGENESIS, AND ELUCIDATE THE MECHANISM OF INTERCELLULAR SIGNALING BY LEADERLESS PEPTIDES IN FOUR SPECIFIC AIMS. THE RESULTS FROM THIS STUDY WILL ELUCIDATE HOW THE PEPTIDE SIGNALING PATHWAYS ARE TAILORED FOR THE PHYSIOLOGICAL NEEDS OF THE PRODUCING BACTERIA AND HOW A PATHOGEN GAIN SURVIVAL ADVANTAGE BY HIJACKING THE NON-COGNATE SIGNAL FROM A COMMENSAL MICROBE TO TRIGGER VIRULENCE FACTOR PRODUCTION AND CAUSE DISEASE. THE PROPOSED RESEARCH IS SIGNIFICANT AS IT INVESTIGATES A CRITICAL PROCESS IN DISEASE PATHOGENESIS OF A MAJOR HUMAN PATHOGEN AND IS LIKELY TO ELUCIDATE NOVEL TRANSLATIONAL STRATEGIES TO COMBAT GAS INFECTIONS.

ALLIANCE FOR NANOHEALTH COMPETITIVE RESEARCH PROGRAM FY2009

SYSTEMATIC ALZHEIMER'S DISEASE DRUG REPOSITIONING (SMART) BASED ON BIOINFORMATICS-GUIDED PHENOTYPE SCREENING AND IMAGE-OMICS

VASCULARIZED ISLET TRANSPLANTATION NICHE WITH LOCAL IMMUNOSUPPRESSION FOR THE TREATMENT OF TYPE 1 DIABETES - CELL ENCAPSULATION TECHNOLOGIES ARE POISED TO IMPROVE CONVENTIONAL ISLET TRANSPLANTATION TO MORE EFFECTIVELY MANAGE TYPE I DIABETES. CURRENTLY, LIFELONG WHOLE-BODY IMMUNOSUPPRESSION IS ADMINISTERED TO AVOID IMMUNE REJECTION OF THE TRANSPLANT, DESPITE THE ASSOCIATED LIFE-THREATENING ADVERSE EFFECTS. CLINICAL STUDIES REVEAL THAT TRANSPLANTS EVENTUALLY FAIL DUE TO LACK OF VASCULAR SUPPORT FOR NUTRIENTS AND OXYGEN SUPPLY AND HOST IMMUNE REJECTION. TO ADDRESS ALL THESE CRITICAL NEEDS AND SUPPORTED BY PRELIMINARY STUDIES, WE PROPOSE THE NICHE, AN INNOVATIVE SUBCUTANEOUS VASCULARIZED ENCAPSULATION SYSTEM WITH LOCAL ELUTION OF IMMUNOSUPPRESSANTS TO PROTECT TRANSPLANTED CELLS FROM IMMUNE REJECTION. THE NICHE PRESENTS DUAL TRANSCUTANEOUSLY REFILLABLE RESERVOIRS, FOR DRUG AND CELLS, RESPECTIVELY, SEPARATED BY A NANOPOROUS MEMBRANE. LOCAL IMMUNOSUPPRESSANT DELIVERY CONFINES DRUGS TO THE GRAFT SITE WHERE IMMUNE ATTACK OCCURS, MINIMIZING EXPOSURE TO THE REST OF THE BODY, THUS AVOIDING SYSTEMIC IMMUNOSUPPRESSION AND ASSOCIATED ADVERSE EFFECTS. THE NICHE CELL RESERVOIR IS FULLY VASCULARIZED WITH FUNCTIONAL VESSELS, RECREATING AN IDEAL PHYSIOLOGICAL ENVIRONMENT CONDUCIVE FOR MAINTAINING LONG-TERM VIABILITY AND FUNCTION OF TRANSPLANTED CELLS. WE HYPOTHESIZE THAT THE NICHE WILL PROVIDE A VASCULARIZED ENVIRONMENT WITH LOCAL IMMUNOSUPPRESSANT DELIVERY FOR SUCCESSFUL LONG-TERM ISLET ENGRAFTMENT TO RESTORE EUGLYCEMIA IN DIABETIC HOSTS. IN AIM 1, WE WILL STUDY THE ABILITY OF MESENCHYMAL STEM CELLS TO INDUCE VASCULARIZATION WITHIN THE NICHE AS WELL AS MODULATE THE NICHE IMMUNE MICROENVIRONMENT TO BE CONDUCIVE FOR ISLET TRANSPLANTATION IN DIABETIC RATS. THIS WILL BE FOLLOWED IN AIM 2 BY THE BIODISTRIBUTION ANALYSIS OF IMMUNOSUPPRESSANTS LOCALLY ELUTED IN THE NICHE AND THE ASSESSMENT OF IMMUNOMODULATION AND PHARMACOKINETICS IN DIABETIC RATS. IN AIM 3, THE NICHE EFFICACY IN PROVIDING A SUITABLE ENVIRONMENT FOR SUCCESSFUL ISLET ENGRAFTMENT TO RESTORE EUGLYCEMIA IN DIABETIC RATS WILL BE ASSESSED OVER 1 YEAR, IN PARALLEL TO A LONGITUDINAL STUDY OF NICHE MICROENVIRONMENT REMODELING. THE PROPOSED STUDIES ARE BASED ON OUR TEAM’S EXTENSIVE EXPERTISE IN IMPLANTABLE DRUG AND CELL DELIVERY SYSTEMS, TISSUE ENGINEERING, RESEARCH AND CLINICAL TRANSPLANTATION, TRANSPLANT IMMUNOLOGY, TYPE 1 DIABETES, AS WELL AS SUPPORTIVE PRELIMINARY DATA AND PREVIOUSLY PUBLISHED WORK. IMPORTANTLY, THE NICHE IS DESIGNED PRIORITIZING CLINICAL CONSIDERATIONS OF EFFICACY, SAFETY AND USER ACCEPTABILITY. TRANSCUTANEOUS CELL AND DRUG REFILLING ALLOW FOR EASE OF DRUG REPLENISHMENT WHEN NEEDED, THUS EXTENDING IMPLANT LIFESPAN POTENTIALLY FOR THE LIFETIME OF PATIENTS. FURTHER, THE THIN AND COMPACT SIZE OF THE NICHE, WHICH IS SMALLER THAN THE ENCAPSULATION IMPLANTS UNDER CLINICAL INVESTIGATION, IS FAVORABLE FOR USER ACCEPTABILITY. SUCCESSFUL COMPLETION OF THE PROPOSED WORK WILL PROVIDE A BROADLY APPLICABLE ENCAPSULATION SYSTEM WITH LOCALIZED IMMUNOSUPPRESSANT DELIVERY FOR LONG- TERM PROTECTION OF TRANSPLANTED ISLETS, AS WELL AS MINIMIZE ADVERSE EFFECTS ASSOCIATED WITH IMMUNOSUPPRESSIVE DRUGS. THIS COULD TRANSLATE TO A CLINICAL BREAKTHROUGH FOR DEPLOYMENT OF CELL THERAPIES BEYOND ISLETS, INCLUDING STEM CELL-DERIVED SS CELLS, TO TREAT DIABETES AS WELL AS OTHER DISEASES.

REGULATION OF INNATE IMMUNITY BY SELF-ADJUVATED MRNA VACCINES - ABSTRACT MYCOBACTERIUM TUBERCULOSIS (MTB), THE CAUSATIVE AGENT OF PULMONARY AND NON-PULMONARY TUBERCULOSIS (TB), KILLS NEARLY 1.2 MILLION PEOPLE EVERY YEAR, WHEREAS ~ ONE-THIRD OF THE GLOBAL POPULATION IS LATENTLY INFECTED WITH MTB. BACILLUS CALMETTE-GUÉRIN (BCG) IS WIDELY USED AS A VACCINE AGAINST TB, AND OVER ONE BILLION DOSES HAVE BEEN USED SO FAR ALTHOUGH IT ONLY PARTIALLY PROTECTS CHILDREN FROM TB BUT NOT ADULTS. WE DISCOVERED THAT A NOVEL-SELF ADJUVANTED MRNA (SAMR) VACCINE DELIVERING MTB-DERIVED ANTIGEN CAN PROTECT MICE AGAINST TB. IN THIS PROPOSAL, WE PROPOSE TO DEVELOP NEW GENERATION MRNA-BASED VACCINES ROUTE TO PROTECT AGAINST TB USING A MULTI-ANTIGEN-BASED PLATFORM WHICH ALSO INTEGRATES AN ADJUVANT. A ‘SYSTEMS IMMUNOLOGY’ APPROACH WILL BE USED TO EVALUATE INNATE IMMUNITY MECHANISMS THAT CONTROL ADAPTIVE IMMUNE RESPONSES IN MICE VACCINATED USING OUR IMPROVED MRNA VACCINES. OUR GOALS ARE: SPECIFIC AIM-1: TO DEVELOP A NEW GENERATION MRNA-BASED VACCINES EXPRESSING PROTECTIVE ANTIGENIC EPITOPES OF MYCOBACTERIUM TUBERCULOSIS (MTB) IN COMBINATION WITH IN-BUILT ADJUVANT PEPTIDES. WE WILL DEVELOP MULTIPLE MTB PROTEIN ANTIGEN EXPRESSING MRNA VACCINES THAT CONTAIN A UNIQUE SELF-ADJUVANT TO BOOST IMMUNE RESPONSES. WE WILL SYNTHESIZE AND EVALUATE MORE EFFICIENT LINEAR MRNA AND/OR CIRCULAR RNA VACCINES FOR TUBERCULOSIS AND FORMULATE THEM IN NANOPARTICLES. SPECIFIC AIM-2: TO CHARACTERIZE THE MOLECULAR MECHANISMS OF NEW GENERATION MRNA VACCINES. WE FOUND THAT OUR NEW GENERATION SAMR-VACCINES INDUCED A ROBUST ACTIVATION OF INNATE IMMUNITY PATHWAYS (RIG-I NOD2 AND TLR) IN MACROPHAGES AND DENDRITIC CELLS. WE WILL INVESTIGATE THE MOLECULAR MECHANISMS OF PROTEIN ANTIGEN-INDUCED IMMUNOGENICITY AND C5- INDUCED ADJUVANTICITY USING APCS FROM WILD TYPE C57BL/6 MICE, AND TRANSGENIC MICE THAT LACK INNATE NODAL GENES. SPECIFIC AIM-3: TO DETERMINE THE EFFICACY OF SAMR-VACCINES AGAINST TUBERCULOSIS IN WILD TYPE AND IMMUNODEFICIENT MICE. WE FOUND THAT OUR NOVEL VACCINE ACTIVATED TRAINED IMMUNITY GENES IN MACROPHAGES. BECAUSE BCG-INDUCED TRAINED AND ADAPTIVE IMMUNITY DECLINES IN HUMAN CHILDREN BY AGE 5 PREDISPOSING THEM TO TB, WE WILL INVESTIGATE THE BOOSTING EFFECT OF OUR TRAINED IMMUNITY INDUCING SAMR-VACCINES FOR PROTECTION IN BCG VACCINATED MICE. WE WILL USE BOTH C57BL/6 AND OUR OPTIMIZED IMMUNE COMPETENT AND IMMUNODEFICIENT HUMANIZED MODEL TO IDENTIFY PARAMETERS OF PROTECTION RELEVANT TO HUMANS.

AIBP MEDIATES A NOVEL INTERPLAY BETWEEN CHOLESTEROL METABOLISM AND LYMPHANGIOGENESIS

REVERSAL OF HEART FAILURE: ROLE OF VASCULAR RECOVERY

CAR T CELLS FOR ADVANCED THYROID CANCER - PROJECT SUMMARY THE OVERALL INCIDENCE OF THYROID CANCER IN 2016 WAS ESTIMATED BY THE NATIONAL CANCER INSTITUTE TO BE 64,300. PATIENTS WHO PRESENT WITH POORLY DIFFERENTIATED THYROID CANCERS FREQUENTLY ARE REFRACTORY TO STANDARD TREATMENT REGIMENS AND HAVE A MUCH WORSE PROGNOSIS. THE MEDIAN OVERALL SURVIVAL IN THIS PATIENT POPULATION IS LESS THAN A YEAR. UNDIFFERENTIATED OR ANAPLASTIC THYROID CANCERS ARE TYPICALLY NOT AMENABLE TO SURGERY AND ARE HIGHLY RESISTANT TO RAI AND VIRTUALLY ALL OTHER THERAPIES. WE THEREFORE DEVELOPED A CHIMERIC ANTIGEN RECEPTOR (CAR) T CELL THERAPY TARGETING INTERCELLULAR ADHESION MOLECULE-1 (ICAM-1) (LABELED AS AIC100) TO TREAT THIS AGGRESSIVE TYPE OF THYROID CANCER. WHILE A VARIETY OF CELLS IN THE BODY NORMALLY EXPRESS LOW, BASAL LEVELS OF ICAM-1, MANY HUMAN CANCERS HAVE UPREGULATED LEVELS OF EXPRESSION. IN PARTICULAR, BOTH REFRACTORY POORLY DIFFERENTIATED AND ANAPLASTIC THYROID CANCERS HAVE GREATLY INCREASED EXPRESSION OF ICAM-1. THE KEY ASPECT OF OUR CAR T CELL TECHNOLOGY IS ITS ABILITY TO SELECTIVELY KILL TUMORS WITH OVER-EXPRESSED ICAM-1 WHILE SPARING NORMAL CELLS WITH BASAL LEVELS OF ICAM-1 EXPRESSION. TO ASSESS IN VIVO DISTRIBUTION OF CAR T CELLS IN BOTH TARGETED TUMORS AS WELL AS NON-TARGET TISSUES, WE HAVE INTRODUCED THE SOMATOSTATIN RECEPTOR 2 (SSTR2) TO FOLLOW CAR T CELLS OVER TIME USING THE CLINICALLY APPROVED RADIOLABELED TRACER 68GALIUM-DOTATATE (NETSPOT). IN THE PLANNED PHASE I STUDY, THE PRIMARY OBJECTIVE IS TO ASSESS THE SAFETY AND DETERMINE THE RECOMMENDED DOSE OF AIC100 FOR PHASE II STUDY IN PATIENTS WITH RELAPSED/REFRACTORY POORLY DIFFERENTIATED THYROID CANCER AND IN PATIENTS WITH ANAPLASTIC THYROID CANCER. THE SECONDARY AIMS ARE TO EVALUATE THE EFFICACY OF AIC100 IN PATIENTS USING PET/CT AND RECIST (RESPONSE EVALUATION CRITERIA IN SOLID TUMORS) CRITERIA, EVALUATE THE FEASIBILITY OF CAR T CELL IMAGING BY DOTATATE, DETERMINE IF OVERALL TUMOR RESPONSE CORRELATES WITH T CELL DISTRIBUTION IN TUMOR SITES AS MEASURED BY DOTATATE AND OTHER EXPLORATORY BIOMARKERS, AND TO EXAMINE PRE-INFUSION CAR T CHARACTERISTICS AS A PREDICTOR OF CLINICAL RESPONSE. OUR INVESTIGATIONAL NEW DRUG (IND) APPLICATION TO OPEN PHASE I STUDY OF AIC100 AGAINST ADVANCED THYROID CANCERS IS NOW APPROVED BY FDA IN OCTOBER 2019, AND PATIENT ENROLLMENT WILL COMMENCE IN EARLY 2020. AS THE COST FOR CAR T MANUFACTURING AND NON-STANDARD OF CARE CLINICAL COSTS WILL BE SPONSORED BY OUR INDUSTRY COLLABORATOR (AFFYIMMUNE THERAPEUTICS, INC.), THE MAJOR GOALS OF THIS GRANT APPLICATION ARE TO ASSESS THE KINETICS OF T CELL DISTRIBUTION BY PET/CT, AND DETERMINE THE EMERGENCE OF HIGH CLONALITY T CELLS AND ASSOCIATED CELLULAR AND GENE EXPRESSION SIGNATURES WITH RESPECT TO CLINICAL RESPONSE, TOXICITY, AND SURVIVAL; TO CORRELATE INTRINSIC CAR T FITNESS FOR SURVIVAL AND CLONAL EXPANSION WITH CLINICAL RESPONSE; TO OPTIMIZE T CELL MANUFACTURING AND NEXT-GENERATION CAR DESIGNS TO IMPROVE THE FITNESS OF CAR T CELLS TOWARD MORE EFFECTIVE CAR T THERAPIES AGAINST SOLID CANCERS.

CHEMOIMMUNOPREVENTION OF EGFR-DRIVEN NON-SMALL CELL LUNG CANCER

INHIBITION OF ORAL TUMORIGENESIS BY ANTITUMOR B

DUAL TARGETING OF PI3K AND NOS PATHWAYS IN METAPLASTIC BREASTCANCER (MBC) - ABSTRACT METAPLASTIC BREAST CANCER (MPBC) IS A RARE SUBSET ACCOUNTING FOR <5% OF ALL BREAST CANCERS. MPBC IS A SIGNIFICANT HEALTH CHALLENGE AS IT EXHIBITS THE MOST DISMAL PROGNOSIS OF ALL BREAST CANCER SUBTYPES, WORSE THAN NON-MPBC TRIPLE-NEGATIVE BREAST CANCER (TNBC), WITH MEDIAN SURVIVAL RATE OF 8 MONTHS OR LESS IN PATIENTS WITH METASTATIC DISEASE. DUE TO A LACK OF DRUGGABLE TARGETS, THE MAIN THERAPEUTIC OPTION FOR METASTATIC MPBC REMAINS SYSTEMIC CHEMOTHERAPY, DESPITE KNOWN RESISTANCE TO MOST CYTOTOXIC DRUGS. ONE COMMON MOLECULAR ALTERATION IN MPBC IS HYPERACTIVATION OF THE PHOSPHOINOSITIDE 3-KINASE AND PROTEIN KINASE B (PI3K/AKT) PATHWAY. ADDITIONALLY, WE PUBLISHED THAT MPBC ALSO DISPLAYS A GAIN-OF-FUNCTION ONCOGENIC MUTATION IN RIBOSOMAL PROTEIN L39 (RPL39), WHICH IS RESPONSIBLE FOR TREATMENT RESISTANCE, STEM CELL SELF-RENEWAL, AND LUNG METASTASIS. THE MECHANISTIC FUNCTION OF RPL39 IS MEDIATED THROUGH INDUCIBLE NITRIC OXIDE SYNTHASE (INOS)-MEDIATED NITRIC OXIDE PRODUCTION. IN A RECENTLY PUBLISHED CLINICAL TRIAL TARGETING THIS NITRIC OXIDE SYNTHASE (NOS) PATHWAY WITH A PAN-NOS INHIBITOR NG-METHYL-L-ARGININE ACETATE (L-NMMA), HIGH EFFICACY IN CHEMOREFRACTORY TNBC PATIENTS WAS DEMONSTRATED. FURTHERMORE, IN VIVO STUDIES PERFORMED SHOWED A SIGNIFICANT REDUCTION IN TUMOR GROWTH, ASSOCIATED WITH A SIGNIFICANT INCREASE IN APOPTOSIS AFTER THE ALPELISIB/L-NMMA COMBINATORIAL REGIMEN. THEREFORE, WE HYPOTHESIZE THAT THE NOS AND PI3K SIGNALING PATHWAYS MAY EXERT THEIR ONCOGENIC RESPONSES SYNERGISTICALLY TO PROMOTE AGGRESSIVE TUMOR GROWTH. TO TEST THIS HYPOTHESIS, SPECIFIC AIM 1 SEEKS TO DEMONSTRATE THE THERAPEUTIC EFFICACY OF SIMULTANEOUS INHIBITION OF NOS AND PI3K PATHWAYS WITH CHEMOTHERAPY IN MPBC PRE- CLINICAL MODELS ON PRIMARY TUMOR GROWTH AND METASTASIS. SPECIFIC AIM 2 WILL INVESTIGATE THE GLOBAL AND RPL39- SPECIFIC RIBOSOME TRANSLATION LANDSCAPE IN RESPONSE TO NOS/PI3K INHIBITION IN MPBC. IN SPECIFIC AIM 3, THE CELL- CELL INTERACTIONS AMONG TUMOR CELLS, MYELOID CELLS, LYMPHOID CELLS, AND STROMAL CELLS WITHIN THE TUMOR MICROENVIRONMENT AND THEIR ROLE IN SUPPORTING CANCER NICHE POPULATIONS WILL BE EVALUATED AT THE SINGLE-CELL LEVEL USING SPATIAL TRANSCRIPTOMICS, IMMUNOFLUORESCENCE, CYTOF IMAGING SYSTEMS, AND A MULTI-MODAL DATA ANALYSIS MODEL. THIS STUDY THUS PROPOSES A MECHANISTIC INVESTIGATION OF A COMBINATORIAL TARGETED APPROACH AGAINST THE TWO KEY PATHWAYS IN MPBC, IDENTIFIES CELL–CELL INTERACTIONS, AND DEVELOPS UNIQUE CROSSTALK MODELS THAT WILL EFFECTIVELY PREDICT OUTCOME AND TREATMENT RESPONSE AND COMPLEMENT OUR RECENTLY FUNDED U01 CLINICAL TRIAL ON MPBC PATIENTS.

HARNESSING NEUROPLASTICITY OF POSTURAL SENSORIMOTOR NETWORKS USING NON-INVASIVE SPINAL NEUROMODULATION TO MAXIMIZE FUNCTIONAL RECOVERY AFTER SPINAL CORD INJURY - ABSTRACT TRANSCUTANEOUS (TSS) AND EPIDURAL SPINAL STIMULATION (ESS) ARE BOTH ELECTROPHYSIOLOGICAL TECHNIQUES THAT HAVE BEEN USED TO INVESTIGATE THE INTERACTIONS BETWEEN EXOGENOUS ELECTRICAL STIMULI AND SPINAL SENSORIMOTOR NETWORKS THAT INTEGRATE DESCENDING MOTOR SIGNALS WITH AFFERENT INPUTS FROM THE PERIPHERY DURING MOTOR TASKS SUCH AS STANDING AND STEPPING. RECENTLY, PILOT PHASE CLINICAL TRIALS USING EACH OF THESE STIMULATION MODALITIES HAVE DEMONSTRATED RESTORATION OF MOTOR FUNCTIONS THAT WERE PREVIOUSLY LOST DUE TO SPINAL CORD INJURY (SCI). HOWEVER, THE SPINAL NETWORK INTERACTIONS THAT OCCUR IN RESPONSE TO TSS OR ESS WITH REHABILITATION TRAINING HAVE YET TO BE CHARACTERIZED AND DIRECTLY COMPARED. THUS, IT IS IMPERATIVE TO (1) UNDERSTAND THE NEUROPHYSIOLOGICAL PROFILES OF INDIVIDUAL PARTICIPANTS COMPREHENSIVELY, THEREBY DELINEATING THE UNDERLYING MECHANISMS AND (2) ESTABLISH THE RELATIVE EFFECTIVENESS OF TSS VERSUS ESS IN REGAINING MOTOR FUNCTION. OUR CENTRAL HYPOTHESIS IS THAT ESS AND TSS WILL PROMOTE SIMILAR LEVELS OF STIMULATION-AUGMENTED STANDING. WE WILL COMPARE THE IMMEDIATE FUNCTIONAL AND NEUROPHYSIOLOGICAL EFFECTS OF TSS VERSUS ESS IN THE SAME PARTICIPANTS (AIM 1). WE HYPOTHESIZE THAT ESS AND TSS WILL GENERATE COMPARABLE MOTOR OUTPUT DURING STANDING DUE TO SIMILAR LEVEL OF MODULATED EXCITABILITY OF SENSORIMOTOR NETWORKS. THEN, WE WILL COMPARE IMMEDIATE AND DELAYED EFFECTS OF STANDING TRAINING COMBINED WITH TSS VERSUS ESS (AIM 2). WE HYPOTHESIZE THAT BOTH TSS- AND ESS-COMBINED TRAININGS WILL RESULT IN SIMILAR FUNCTIONAL GAIN. WE FURTHER PREDICT THAT AFTER ONE MONTH OF REST FOLLOWING STANDING TRAINING WITH TSS OR ESS, THE EXCITABILITY AND RESPONSIVENESS OF SPINAL AND SUPRASPINAL SENSORIMOTOR NETWORKS WILL DIFFER FROM BASELINE LEVELS AND SHAM. COMPLETION OF THIS STUDY WILL ESTABLISH THE EFFICACY OF BOTH NON-INVASIVE AND INVASIVE SPINAL NEUROMODULATORY APPROACHES TO IMPROVE FUNCTIONS AFTER SCI, IMPACTING BOTH RESEARCH AND CLINICAL COMMUNITIES. FURTHER, IT WILL SHED LIGHT ON THE UNDERLYING MECHANISMS OF SPINAL NEUROMODULATION, ALLOWING OURSELVES AND OTHERS TO REFINE AND IMPROVE STIMULATION PROTOCOLS TO MAXIMIZE FUNCTIONAL RECOVERY.

SIRTUIN -DEPENDENT REGULATION OF TUBERCULOSIS AND HIV INTERACTIONS IN MACROPHAGES - ABSTRACT TUBERCULOSIS CONTINUES TO KILL ABOUT 1 MILLION PEOPLE INCLUDING CHILDREN EACH YEAR WITH 8 MILLION NEW CASES. REGRETTABLY, COINFECTION WITH HIV-1 HAS AGGRAVATED THE PROBLEM BECAUSE HIV-1 DEPLETES CD4 T CELLS, WHICH ARE THE MAIN DEFENSE MECHANISM AGAINST TUBERCULOSIS. FOR THE REASON, BCG, WHICH IS A LIVE ATTENUATED VACCINE AGAINST TUBERCULOSIS, CANNOT BE GIVEN TO PEOPLE LIVING WITH HIV. WE RECENTLY DISCOVERED THAT SIRTUIN TYPE OF PROTEIN AND HISTONE DEACETYLATES PLAY A MAJOR ROLE IN REGULATING THE GROWTH OF BOTH M. TUBERCULOSIS AND HIV-1 IN HUMAN MACROPHAGES AND DRUG TARGETING SIRTUINS COULD CONTROL BOTH PATHOGENS IN CELL CULTURE MODELS AND IN HUMANIZED MICE. THEREFORE, WE PROPOSE TO INVESTIGATE SIRTUIN-DEPENDENT INTERVENTION TO UNDERSTAND HOW TB AND HIV-1 INFECTIONS INDUCE THESE ENZYMES AND HOW WE CAN DEVELOP NOVEL IMMUNOCHEMOTHERAPY FOR CONFECTIONS. SPECIFICALLY: AIM-1: WE WILL ANALYZE THE IMPACT OF EARLY MTB AND HIV INFECTION ON SIRTUIN GENE AND PROTEIN INDUCTION IN MACROPHAGES AND IDENTIFY THEIR EPIGENETIC TARGETS. WE WILL EXAMINE THE HYPOTHESIS THAT MTB-INDUCED SIRTUIN-2 CAN AUGMENT HIV-1 REPLICATION, WHEREAS HIV-1 INDUCED SIRTUIN-2, IN TURN ENHANCES MTB GROWTH IN MФS THEREBY AGGRAVATING COINFECTIONS. WE WILL THEN DEVELOP AN IMMUNOCHEMOTHERAPY (ICT) USING A COMBINATION OF SIRTUIN DRUGS, TB AND HIV TARGETING DRUGS TO KILL AND ERADICATE BOTH HIV-1 AND M. TUBERCULOSIS IN MACROPHAGES. AIM-2: WE WILL INVESTIGATE THE EFFECT OF SIRTUIN DRUGS ON EARLY AND LATE STAGES OF TB-HIV-1 COINFECTION USING HUMANIZED MICE AND DETERMINE WHETHER ICT CAN ERADICATE CONFECTIONS. AIM-3: TUBERCULOSIS GRANULOMAS CAN RESTRICT GROWTH OF M. TUBERCULOSIS PATHOGEN, ALTHOUGH PRIOR INFECTION WITH HIV-1 CAN DEPLETE CD4 T CELLS AND AFFECT GRANULOMA FORMATION. WE WILL USE A TB-HIV COINFECTION MODEL OF HUMANIZED MICE TO DETERMINE IF SIRTUIN-DRUG THERAPY CAN ACTIVATE MACROPHAGES OF GRANULOMAS AND INCREASE HOST-DEFENSE AGAINST TUBERCULOSIS. OUR OVERALL GOAL IS TO DEVELOP INNOVATIVE IMMUNE DEFENSE-BASED INTERVENTION METHODS TO ERADICATE TB-HIV COINFECTIONS.

MODELING TUMOR-STROMA CROSSTALK IN LUNG CANCER TO IDENTIFY TARGETS FOR THERAPY

DECIPHERING THE ACTIVATION MECHANISMS AND IMMUNE FUNCTIONS OF GSDMC - PROJECT SUMMARY THE GASDERMIN (GSDM) FAMILY CONSISTS OF SIX PARALOGOUS GENES ENCODING GSDMA, GSDMB, GSDMC, GSDMD, GSDME AND F (GSDMF ALSO KNOWN AS PJVK OR DFNB59). GSDM PROTEINS PLAY A CRUCIAL ROLE IN INNATE IMMUNITY, PARTICULARLY IN INFLAMMATION AND THE INITIATION OF PYROPTOSIS, A FORM OF PROGRAMMED NECROTIC CELL DEATH. UNLIKE MANY OTHER IMMUNE MODULATORS, TRANSCRIPTIONAL REGULATION OF GSDM FAMILY PROTEINS IS NOT SUFFICIENT TO EXECUTE THEIR BIOLOGICAL FUNCTIONS. GSDMS A-E SHARE HIGHLY CONSERVED N-TERMINAL (N-TER) AND C-TERMINAL (C-TER) DOMAINS SEPARATED BY A VARIABLE LINKER. THE C-TERM EXHIBITS SELF-INHIBITION BY COMPLETELY MASKING THE N-TER HYDROPHOBIC POCKET THAT BINDS LIPIDS. THUS, EVEN IF UPREGULATED TRANSCRIPTIONALLY, GSDM CANNOT BE FUNCTIONAL UNTIL IT IS CLEAVED BY A PROTEASE TO RELEASE THE N-TER FROM THE SELF-INHIBITED C-TERM. THUS, THE KEY FOR UNDERSTANDING GSDM BIOLOGY IS THE IDENTIFICATION OF THE PROTEASE. THIS WORK IS BASED ON OUR ROBUST UNPUBLISHED DATA WHERE WE IDENTIFIED A NEW PROTEASE THAT CAN DIRECTLY CLEAVE BOTH HUMAN AND MURINE GSDMC. ROOTING FROM THIS DISCOVERY, WE FURTHER IDENTIFIED THREE UNIQUE FEATURES OF CLEAVED AND ACTIVATED GSDMCN-TER NOT OBSERVED IN OTHER GSDM FAMILY MEMBERS IN TERMS OF ITS MOLECULAR PROPERTIES, CELLULAR FUNCTIONS AND IN VIVO PHENOTYPES. (1) IN CONTRAST TO OTHER PROTEASE-PROCESSED GSDM FAMILY MEMBERS, SUCH AS GSDM-A, -B, -D AND -E, THE N-TERMINUS OF GSDMC (GSDMCN-TER), PROCESSED BY THE NEWLY IDENTIFIED PROTEASE, DID NOT EFFECTIVELY PROMOTE PYROPTOSIS. (2) THIS IS BECAUSE GSDMCN-TER DOES NOT LOCALIZE TO THE PLASMA MEMBRANE BUT TO OTHER SUBCELLULAR ORGANELLES. (3) GSDMCN-TER HAS AN IMMUNE REGULATORY ROLE IN ANIMAL MODELS VIA AMPLIFYING TYPE-2 IMMUNE RESPONSES, UNRELATED TO PORE FORMATION AT THE PLASMA MEMBRANE (E.G. IL-1 FAMILY MEMBER CYTOKINE RELEASE) AND PYROPTOSIS. ACCORDINGLY, WE HAVE PLANNED THREE AIMS TO UNDERSTAND THE MECHANISMS CONTROLLING THESE FEATURES. IN AIM 1, WE WILL ANALYZE THE NON-CONSERVED AMINO ACIDS BETWEEN GSDMCN-TER AND OTHER GSDMS PROTEINS THAT EXPLAIN THE PYROPTOSIS DEFICIENCY. WE WILL FURTHER DETERMINE THE LIPID BINDING PROFILE OF GSDMCN-TER AND TRY TO UNDERSTAND HOW IT DIFFERS FROM THAT OF OTHER GSDMS PROTEINS. IN AIM 2, WE WILL SYSTEMATICALLY INVESTIGATE THE INTRACELLULAR MEMBRANE STRUCTURES THAT CAN BE TARGETED BY GSDMCN- TER AND ASSESS THE BIOLOGICAL CONSEQUENCES AFTER GSDMCN-TER TARGETING. LAST, IN AIM 3 WE WILL USE TWO ANIMAL MODELS TO DETERMINE HOW GSDMCN-TER AMPLIFIES TYPE 2 IMMUNE RESPONSES. IN SUMMARY, AIM 1 AND 2 WILL UNVEIL NOVEL MECHANISMS OF HOW GSDMC FUNCTIONS DIFFERENT FROM OTHER GSDMS, AS WELL AS ITS NEW ROLES IN CELL BIOLOGY (NOT EFFECTIVELY PROMOTING PYROPTOSIS). FOR AIM 3, WITH OUR UNIQUE ANIMAL MODELS, WE WILL REVEAL NEW MECHANISMS OF ACTIONS FOR GSDMC AS WELL AS THERAPEUTIC STRATEGIES TO BOOST TYPE 2 IMMUNITY.

PROTEOMICS BASED MAPPING OF CARDIAC EXTRACELLULAR MATRIX TO DEFINE SEX AND AGE-DEPENDENT CHANGES - PROJECT SUMMARY THE EXTRACELLULAR MATRIX (ECM) PROVIDES A SCAFFOLD FOR CARDIAC STRUCTURAL INTEGRITY AND PLAYS AN ACTIVE ROLE IN MODULATING CELLULAR RESPONSES. ECM CONSISTS OF A NETWORK OF FIBRILLAR PROTEINS AND PROTEOGLYCANS BINDING CYTO- KINES, GROWTH FACTORS, AND ECM-MODIFYING ENZYMES. THE AGING HEART DEVELOPS INTERSTITIAL FIBROSIS, AND PRELIMI- NARY DATA DEMONSTRATE THAT DIFFERENT PROTEINS ARE ENRICHED IN ECM IN THE MALE AND FEMALE HEART. AN ALTERED ECM CAN AFFECT CELLULAR PHENOTYPES. THE ECM-CELL COMMUNICATION OCCURS VIA CELL SURFACE RECEPTORS SUCH AS INTEGRINS, THROUGH OUTSIDE-IN SIGNALING, WHERE INTEGRIN-BINDING TO ECM PROTEIN DOMAINS TRANSLATES INTO PROLIFERA- TION, DIFFERENTIATION, OR GENE EXPRESSION. ON THE OTHER HAND, ACTIVATION OF THE INTRACELLULAR INTEGRIN TAIL INCREASES THEIR ECM-BINDING AFFINITY, PROMOTING ADHESION, MIGRATION, OR ECM ASSEMBLY (BY INSIDE-OUT SIGNALING). THERE- FORE, ECM DICTATES A CELLULAR PHENOTYPE, BUT CELLS CAN MODIFY ECM IN RESPONSE BY CHANGING ITS COMPOSITION OR ASSEMBLY. BECAUSE OF PROFOUND DIFFERENCES IN ECM COMPOSITION IN MALES AND FEMALES, WE HYPOTHESIZE THAT THE QUALITY OF REPAIR ASSEMBLED AFTER INJURY AND SUBSEQUENT ADVERSE REMODELING IN MALES AND FEMALES WILL ALSO BE DIFFER- ENT, AND THE DIFFERENCES WILL BE ACCENTUATED BY AGING. IN SA1, WE WILL MAP THE DIFFERENCES IN ECM DEPENDENT ON AGE (YOUNG AND OLD ANIMALS WILL BE STUDIED), SEX HORMONE LEVEL (ANIMALS WILL BE SUBJECTED TO CHEMICAL OVARY FAILURE OR CASTRATION) OR LEVEL OF INFLAMMATION (CCL2KO ANIMALS WILL BE USED; THESE MICE HAVE REDUCED LEUKO- CYTE INFILTRATION AND THEREFORE REDUCED FIBROSIS). THE CELLULAR PHENOTYPE OF FIBROBLASTS, ENDOTHELIAL CELLS (ECS), AND SMOOTH MUSCLE CELLS (SMCS) ISOLATED FROM ALL THESE ANIMALS WILL BE STUDIED EX VIVO AND IN 3D CULTURE. THE EFFECT OF ECM ON CELL PHENOTYPE WILL BE TESTED IN 3D CULTURES USING THE MATRIX SWAP APPROACH, WHERE I.E. YOUNG CELLS WILL BE CULTURED ON ECM FROM THE OLD HEARTS AND VICE VERSA. AND THE EFFECT OF MATRIX STIFFNESS ON THE PRO- DUCTION OF ECM PROTEIN WILL BE EXAMINED AS WELL. SIMILARLY, THE PHYSIOLOGY OF HUMAN CELLS WILL BE STUDIED. IN SA2, WE WILL EXAMINE ECM-INTEGRIN DEPENDENT PHENOTYPIC CHANGES IN MOUSE AND HUMAN CELLS. WE WILL FIRST EXAMINE THE EFFECT OF SEX HORMONES, AGE, AND INFLAMMATION ON THE LEVELS OF INTEGRIN EXPRESSION IN FIBROBLASTS, ECS, AND SMCS. THEN WE WILL MANIPULATE THE INTEGRIN EXPRESSION LEVEL (VIA DOWNREGULATION OR OVEREXPRESSION), AND EXAMINE THE CELLULAR PHENOTYPE. FINALLY, WE WILL MANIPULATE ESTROGEN AND ANDROGEN RECEPTORS AND DETER- MINE THE EFFECT OF THESE MANIPULATIONS ON INTEGRIN LEVELS AND ECM PROTEIN SYNTHESIS. IN SA3, WE WILL EXAMINE HOW SEX HORMONE LEVELS AND AGE AFFECT SCAR FORMATION AND ADVERSE REMODELING AFTER ISCHEMIA/REPERFUSION INJU- RY. CHANGES IN ECM WILL BE EXAMINED VIA ELISA, MASS SPECTROMETRY, AND WESTERN BLOT. THE APPROACH IS INNO- VATIVE BECAUSE IT EXAMINES THE ROLE OF THREE SUPERIMPOSED VARIABLES (AGE, SEX HORMONE, AND INFLAMMATION) ON ECM AND CELLULAR PHENOTYPES. SUCCESSFUL COMPLETION OF THESE STUDIES WILL PROVIDE A MECHANISTIC LINK FOR FUTURE THERAPEUTICS TARGETING INSUFFICIENT REPARATIVE AND EXCESSIVE ADVERSE FIBROSIS.

SPATIOTEMPORAL MODELING OF CANCER-NICHE INTERACTIONS IN BREAST CANCER BONE METASTASIS

DEEP LEARNING BASED PHARMACOKINETIC MODEL FOR VANCOMYCIN - ABSTRACT: (30 LINES) VANCOMYCIN IS ONE OF THE MOST COMMONLY USED ANTIMICROBIAL DRUGS IN INPATIENT SETTINGS. NATIONAL GUIDELINES RECOMMEND BAYESIAN MODELS TO MONITOR THE THERAPEUTIC DRUG CONCENTRATION OF VANCOMYCIN, ESPECIALLY FOR METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA), TO MINIMIZE DRUG TOXICITY WHILE MAINTAINING ITS EFFICACY. EXISTING BAYESIAN MODELS, DESPITE BEING CLAIMED AS PATIENT-SPECIFIC PHARMACOKINETIC (PK) MODELS, USE SIMPLE PATIENT FEATURES AND ARE STUDIED IN LIMITED PATIENT POPULATIONS FOR THE POPULATION-BASED PK PARAMETERS (THE BAYESIAN PRIOR). INCREASINGLY AVAILABLE REAL-WORLD ELECTRONIC HEALTH RECORDS (EHR) PROVIDE A WIDE RANGE OF PATIENT-SPECIFIC DATA, INCLUDING DATA ON VANCOMYCIN DOSAGE AND SERUM LEVELS. HOWEVER, THE LIMITED FLEXIBILITY OF THE BAYESIAN MODEL STRUCTURE PROHIBITS THE FULL USE OF THESE RICH DATA. DEEP-LEARNING MODELS, SUCH AS RECURRENT NEURAL NETWORK (RNN), ARE PARTICULARLY ATTRACTIVE FOR PK OF VANCOMYCIN IN EHR, COMPARED TO BAYESIAN MODELS AND OTHER TRADITIONAL MACHINE LEARNING MODELS, BECAUSE DEEP-LEARNING MODELS ENABLE MORE FLEXIBLE PATIENT- SPECIFIC INPUTS AND POSSESS A HIGHER LATENT CAPACITY. THUS, THEY DELIVER BETTER PREDICTIONS FOR A DIVERSE POPULATION. OUR DEEP-LEARNING MODEL FOR VANCOMYCIN (PK-RNN-V) OUTPERFORMS PUBLICLY AVAILABLE BAYESIAN MODELS BUT CAN BE IMPROVED ON VARIOUS ASPECTS. IN AIM 1, WE WILL IMPROVE PK-RNN-V MODEL ARCHITECTURES AND ADD MORE PATIENT-SPECIFIC DATA AND A FINER TIMESTEP. WE WILL CONSTRUCT TWO-COMPARTMENT PK-RNN MODELS TO INCREASE PREDICTIVE POWER IN PATIENTS WITH UNSTEADY STATES. WE WILL AUGMENT PK-RNN-V WITH MED-BERT TO IMPROVE THE EMBEDDING OF CATEGORICAL DATA. WE WILL ALSO DEVELOP MULTI-TRACK ORDINARY DIFFERENTIAL EQUATIONS MODELS FOR SIMULTANEOUS PREDICTION OF SERUM CREATININE AND VANCOMYCIN LEVELS. IN AIM 2, WE WILL USE EHR FROM DIFFERENT SOURCES TO VALIDATE OUR PK-RNN-V MODEL AND IMPROVE THE DATA-EXTRACTION FLOW AND PRE-PROCESSING TO HARMONIZE DATA FROM HEALTHCARE SYSTEMS. WE WILL USE EHR FROM HOUSTON METHODIST HOSPITAL AND MEMORIAL HERMANN HOSPITAL SYSTEM/THE UNIVERSITY OF TEXAS HEALTH SCIENCE CENTER IN HOUSTON, TX, THE UNIVERSITY OF ARIZONA IN PHOENIX, AZ, AND THE PUBLICLY AVAILABLE MIMIC-IV DATABASE (BOSTON, MA). THESE DATABASES CONTAIN DATA FROM MORE THAN 121,007 PATIENTS WHO RECEIVED AT LEAST ONE DOSE OF INTRAVENOUS VANCOMYCIN. IN AIM 3, WE WILL ADD DOSING RECOMMENDATIONS BASED ON PK-RNN-V MODEL PREDICTIONS AS A FEATURE AND VALIDATE OUR MODEL IN SPECIFIC SUBGROUPS WITH CHALLENGING VANCOMYCIN PK TO PREDICT PK LEVELS. THIS PROJECT WILL DELIVER SUBSTANTIAL MODEL IMPROVEMENTS, LEADING DIRECTLY TO THE OPTIMIZATION OF VANCOMYCIN USE IN HOSPITALS, INCREASED IN PATIENT SAFETY BY MINIMIZING ADVERSE EVENTS, AND REDUCED HEALTHCARE COSTS, WHICH ALIGN WITH NIH’S RESEARCH MISSION.

REGULATION OF CELL DEATH IN HIV RESERVOIRS - ABSTRACT: THE COMBINATIONAL ANTIRETROVIRAL THERAPY IS EFFECTIVE IN SUPPRESSING HIV-1 REPLICATION. HOWEVER, LATENT HIV RESERVOIRS CANNOT BE CLEARED BY SUCH ANTIRETROVIRAL THERAPY, AS VIRAL REBOUND DEVELOPS RAPIDLY AFTER THE TREATMENT IS INTERRUPTED IN PEOPLE LIVING WITH HIV. THE SHOCK-AND-KILL APPROACH TO ELIMINATE HIV-1 RESERVOIRS IS TO INDUCE VIRAL REACTIVATION USING LATENCY REVERSAL AGENTS AND TRIGGER CELL DEATH THROUGH VIRUS-MEDIATED CYTOPATHIC EFFECTS OR IMMUNE-MEDIATED CLEARANCE. HOWEVER, SHOCK-AND-KILL BY LATENCY REVERSAL AGENTS HAS NOT BEEN SHOWN TO SUCCESSFULLY REDUCE THE VIRAL RESERVOIRS IN HIV-1 PATIENTS. IT HAS BEEN REPORTED THAT LATENCY REVERSAL AGENTS CAN UP-REGULATE PRO-SURVIVAL AUTOPHAGY AND ANTI-APOPTOTIC MOLECULES THAT COUNTERACT CELL DEATH SIGNALING. INHIBITION OF THESE PRO-SURVIVAL MECHANISMS CAN PROMOTE THE KILLING OF HIV-1-INFECTED CELLS, AND FACILITATE THE KILLING OF HIV-1 RESERVOIR CELLS DURING VIAL RE-ACTIVATION. EXPERIMENTS ARE PROPOSED TO TEST HYPOTHESIS THAT CELLULAR MECHANISMS THAT MAINTAIN THE LONG-TERM PERSISTENCE OF HIV-1 RESERVOIRS ARE IMPORTANT FOR PROTECTING THE RESERVOIR CELLS AGAINST CELL DEATH, AND TARGETING THESE CELLULAR MECHANISMS CAN SENSITIZE HIV-1 RESERVOIRS TO CELL DEATH DURING VIRAL REACTIVATION: 1) TO DETERMINE THE MECHANISMS FOR THE SPECIFIC INDUCTION OF CELL DEATH IN HIV-1 RESERVOIRS IN RESPONSE TO VIRAL REACTIVATION. THE MECHANISMS FOR THE INDUCTION OF APOPTOSIS AND ALTERNATIVE CELL DEATH PATHWAYS IN LATENT HIV-INFECTED T CELLS BY LATENCY REVERSAL AGENTS WILL BE TESTED; 2) TO DELINEATE THE MECHANISMS THAT CONFER THE RESISTANCE TO CELL DEATH IN HIV-1 RESERVOIRS UPON VIRAL REACTIVATION; AND 3) TO TEST WHETHER TARGETING PRO-SURVIVAL MECHANISMS SENSITIZES HIV-1 RESERVOIRS CELLS TO CELL DEATH DURING VIRAL RE-ACTIVATION. THE STUDIES WILL REVEAL NOVEL MOLECULAR PATHWAYS THAT COULD BE TARGETED TO SENSITIZE HIV-1 RESERVOIRS TO CELL DEATH, AND FACILITATE THE DEVELOPMENT OF AN EFFECTIVE APPROACH TO CLEAR HIV-1 RESERVOIRS.

HIGH DENSITY LIPOPROTEIN BIOGENESIS AND SPECIATION

LEARNING-BASED APPROACH FOR PERSONALIZED CRANIOMAXILLOFACIAL SURGICAL PLANNING

MECHANISM OF INTRATUMORAL TRANSPORT OF PARTICULATE DRUGS

TARGETING NOTCH, PI3K-AKT AND OTHER NOVEL PATHWAYS IN BREAST CANCER STEM CELLS

MODULATION OF TUMOR INFLAMMATORY FACTOR FOR IMMUNE THERAPY

NECROTIC SURVIVORS AND PLASMA MEMBRANE INTEGRITY SIGNALING - PROJECT SUMMARY/ABSTRACT AFTER DECADES OF CELL DEATH STUDY, IT IS NOW WELL ACCEPTED THAT CELL DEATH CAN BE HIGHLY PROGRAMMED, INCLUDING SOME LYTIC FORMS. FOR EXAMPLE, THE PLASMA MEMBRANE (PM) PORE-FORMING EXECUTOR FOR NECROPTOSIS IS MLKL, WHEN PHOSPHORYLATED BY RIPK3. FOR PYROPTOSIS, GASDERMIN FAMILY MEMBERS ARE THE PM DAMAGING EXECUTORS, ONCE PROCESSED BY CASPASES OR GRANZYMES. INTRIGUINGLY, THE PM PORE-FORMING IS NOT THE “POINT OF NO RETURN” FOR NECROPTOSIS OR PYROPTOSIS. THIS IS BECAUSE ESCRT- III COMPLEX CAN REPAIR THE DAMAGED PM BY MEMBRANE REMODELING. THE BROKEN PM FRACTIONS CAN BE SHED OFF. THEREFORE, FOR BOTH NECROPTOTIC AND PYROPTOTIC CELLS, THEY CAN TOLERATE A LIMITED LEVEL OF PM DAMAGE AND SURVIVE FOR A LONG TIME IF ESCRT-III REPAIRING CAPACITY IS NOT OVERWHELMED. IN THIS GRANT, WE PRESENTED OUR UNPUBLISHED DATA SHOWING IF A NECROTIC CELL CAN MANAGE TO STAY ALIVE, THE SUB-LETHAL PM DAMAGE IS SUFFICIENT TO INITIATE A SOPHISTICATED SIGNAL TRANSDUCTION NETWORK. WE NAMED THIS SIGNALING TRANSDUCTION PATHWAY AS PLASMA MEMBRANE INTEGRITY SIGNALING (PMI SIGNALING). PMI SIGNALS ARE EFFICIENT IN PROMOTING PRO-TUMOR CHEMOKINES/CYTOKINES SECRETIONS. WE HYPOTHESIZE THAT THE NECROTIC “SURVIVORS” CAN STIMULATE ONCOGENESIS VIA THE PMI SIGNALING PATHWAY AND THE CONSEQUENT CHEMOKINES/CYTOKINES PARACRINE. OUR ULTIMATE GOAL IS TO MECHANISTICALLY UNDERSTAND AND FURTHER TARGET THESE “SURVIVORS” TO IMPROVE CANCER TREATMENT. TO ACHIEVE THIS END, WE WILL FIRSTLY HUNT FOR THE SENSORS FOR PM INTEGRITY LOSS. WE PROPOSED TO TEST A PROMISING ION CHANNEL CANDIDATE THAT IS VERY LIKELY TO BE ONE OF THE SENSORS FOR PM DAMAGE. WE HAVE ALSO PLANNED AN ELEGANT CELL CULTURE SYSTEM TO PERFORM BOTH SIRNA AND SGRNA SCREEN TO SEARCH FOR THE PM DAMAGE SENSORS GENOME-WIDE. NEXT, WE WILL USE BOTH ANIMAL MODELS AND HUMAN SPECIMENS TO PROBE THE NECROTIC “SURVIVORS.” WE WILL FURTHER TEST THE ROLES OF THE CHEMOKINES/CYTOKINES SECRETED BY THESE “SURVIVORS” IN TUMORIGENESIS AND METASTASIS. THIRD, WE WILL TRY TO ELIMINATE THE NECROTIC “SURVIVORS” DIRECTLY. OUR PRELIMINARY DATA LEAD US TO A CANDIDATE RING E3 LIGASE. WE WILL TEST WHETHER TARGETING THIS E3 LIGASE CAN HELP US SUPPRESS THE ESCRT- III REPAIR ACTION AND ERADICATE NECROTIC “SURVIVORS.” AS THE NECROTIC “SURVIVORS” WERE ONLY DISCOVERED NOT LONG AGO (BY US AND OTHERS), THE MASSIVE IMPACTS OF CELL DEATH “SURVIVORS” IN CONTRIBUTING TO CANCERS, TRANSPLANTATION, AND OTHER ILLNESSES ARE STILL FRESH AND NEEDED TO BE FULLY DEFINED. INNOVATIVE RESEARCH, LIKE THIS ONE, WILL PROMOTE THE REVELATION.

INDUCTION OF AUTOSIS TO OVERCOME RESISTANCE IN ADOPTIVE CELL THERAPY FOR SOLID TUMORS - PROJECT SUMMARY ADOPTIVE CELL THERAPY IS A PROMISING APPROACH TO TREAT CANCER, BUT DESPITE TREMENDOUS EFFORTS, RESULTS OF CLINICAL TRIALS IN HUMAN SOLID TUMORS USING ACT WITH TUMOR-SPECIFIC T CELLS EXPRESSING T-CELL-RECEPTORS (TCR) OR CHIMERIC ANTIGEN RECEPTORS (CAR) HAVE NOT DEMONSTRATED THE DESIRED THERAPEUTIC RESPONSES. RECENTLY, WE DEVELOPED TUMOR-SPECIFIC T CELLS LOADED WITH THE MYXOMA VIRUS TO OVERCOME RESISTANCE IN SOLID TUMOR ACT. WE HYPOTHESIZE THAT ANTI-SOLID TUMOR ACTIVITY OF THESE T CELLS IS MAINLY ATTRIBUTED TO A SPECIAL TYPE OF TUMOR CELL DEATH, AUTOSIS, THAT HAS NOT BEEN CONSIDERED A T CELL KILLING MECHANISM BEFORE BUT MAY CONTRIBUTE TO THE OBSERVED EXCITING ANTITUMOR POTENCY BY TARGETING BOTH ANTIGEN-POSITIVE AND ANTIGEN-NEGATIVE TUMOR CELLS. AIM 1 WILL DETERMINE THE UNIQUE MOLECULAR MECHANISM UNDERLYING TUMOR CELL AUTOSIS INDUCED BY THE SYNERGY OF MYXOMA VIRUS-DERIVED FACTOR(S) WITH T CELL-DERIVED SOLUBLE FACTOR(S). AIM 2 WILL DETERMINE THE ROLE OF THESE TUMOR-SPECIFIC T CELLS IN PROMOTING AUTOSIS AND ROBUST HOST IMMUNITY TO ERADICATE SOLID TUMORS WHEN THE TARGETED ANTIGEN IS EXPRESSED BY ONLY ~25% OF TUMOR CELLS. OUR PROPOSED STUDIES WILL IDENTIFY A NOVEL ACT STRATEGY ENDOWED WITH THE CAPACITY TO ELIMINATE SOLID TUMORS WITH VERY HIGH ANTIGEN HETEROGENEITY. THIS TRANSLATIONALLY RELEVANT WORK HOLDS PROMISE TO SIGNIFICANTLY ADVANCE THE THERAPEUTIC INDEX OF ACT IN SOLID TUMORS AND COULD THEN LAY THE FOUNDATION FOR FUTURE CLINICAL TRIALS.

SYSTEMATIC IDENTIFICATION OF ASTROCYTE-TUMOR CROSSTALK REGULATING BRAIN METASTATIC TUMORS

TARGETING TUMOR REPOPULATION AND THE IMMUNE MICROENVIRONMENT TO OVERCOME CHEMORESISTANCE - PROJECT SUMMARY THIS APPLICATION IS IN RESPONSE TO PAR-19-183: BIOLOGY OF BLADDER CANCER. MUSCLE INVASIVE BLADDER CANCER (MIBC) CLAIMS APPROXIMATELY 18,000 DEATHS ANNUALLY IN THE UNITED STATES. FUNDING AND RESEARCH DEVOTED TO THIS CANCER-TYPE ARE SIGNIFICANTLY UNDER-PROPORTIONED. AN UNMET CLINICAL NEED FOR MIBC TREATMENT LIES IN THE POOR PATIENT RESPONSE TOWARDS CHEMOTHERAPY, WITH TREATMENTS PROVIDING ONLY A DISMAL 5% IMPROVEMENT IN OVERALL SURVIVAL. THE LONG-TERM GOAL OF THIS APPLICATION IS TO ADDRESS THIS URGENT NEED FOR ADJUVANT THERAPIES TO IMPROVE CHEMOTHERAPEUTIC RESPONSE. THE SUCCESS OF CHEMOTHERAPY IS HISTORICALLY THOUGHT TO SOLELY DEPEND ON ITS DIRECT CYTOTOXIC EFFECTS ON TUMOR CELLS. HOWEVER, THERE IS GROWING EVIDENCE, AS SHOWN BY OUR OWN RESEARCH AND OTHERS, THAT CHEMOTHERAPEUTIC EFFICACY IS ALSO DEPENDENT ON 1) SUCCESSFUL PREVENTION OF CANCER STEM CELLS IN REPOPULATING RESIDUAL TUMORS AND 2) AN EFFECTIVE ANTI-TUMORAL IMMUNE RESPONSE. THESE TWO PHENOMENA ARE OFTEN INVESTIGATED SEPARATELY BUT THEIR POSSIBLE SYNERGY HAS BEEN OVERLOOKED. OUR RESEARCH PROJECT IS CONCEPTUALLY INNOVATIVE TO EXAMINE A COMMON UPSTREAM PATHWAY THAT REGULATES BOTH TUMOR REPOPULATION AND IMMUNE RESPONSE. WE HYPOTHESIZE THAT THE INHIBITION OF THIS COMMON PATHWAY WILL PROVIDE AN EFFECTIVE THERAPEUTIC TARGET FOR CLINICAL TRANSLATION. OUR SPECIFIC AIMS INCLUDE: AIM 1) DECIPHER THIS PATHWAY BY INVESTIGATING THE NON-CANONICAL DOWNSTREAM MECHANISM LEADING TO THE EXTRACELLULAR RELEASE OF PLEIOTROPIC FACTORS. THIS IS SIGNIFICANT, SINCE THESE EXTRACELLULAR FACTORS CAN MODULATE BOTH TUMOR REPOPULATION AND IMMUNE RESPONSE. AIM 2) EVALUATE HOW THESE EXTRACELLULAR FACTORS AND THEIR COGNATE RECEPTORS DRIVE THE REPOPULATION OF QUIESCENT CANCER STEM CELLS. AIM 3) INVESTIGATE HOW INHIBITION OF THIS UPSTREAM PATHWAY CAN COLLECTIVELY ABROGATE TUMOR REPOPULATION AND IMMUNOSUPPRESSION, AND THUS, ENHANCE CHEMOTHERAPEUTIC RESPONSE. SUCCESS OF THIS PROPOSAL WILL POSE DRUG TARGETS CAPABLE OF AUGMENTING PATIENT RESPONSE TO CHEMOTHERAPY. MOREOVER, THESE FINDINGS WILL PROVIDE INSIGHTS TO HOW THESE DRUGS CAN REESTABLISH AN IMMUNOSTIMULATORY TUMOR MICROENVIRONMENT IN MIBCS. IN SUMMARY, THE STUDIES OUTLINED IN THIS PROPOSAL ARE SIGNIFICANT TO ADDRESS AN UNMET NEED, I.E., TO IMPROVE A DISMAL RESPONSE OF MIBC PATIENTS TO STANDARD CHEMOTHERAPY. THE CONCEPTUAL ADVANCE FROM THIS STUDY WILL LIKELY EXTEND BEYOND MIBC TO BENEFIT PATIENTS FROM OTHER EPITHELIAL MALIGNANCIES.

ADVANCING ENDOVASCULAR PAD TREATMENT: OVERCOMING CRITICAL KNOWLEDGE GAPS WITH MRI-HISTOLOGY - ABSTRACT PERIPHERAL ARTERY DISEASE (PAD) CAUSES SEVERE MORBIDITY AS PLAQUES OBSTRUCT BLOOD FLOW, PREVENTING ADEQUATE PERFUSION TO LIMBS, WHICH MAY RESULT IN AMPUTATION OR DEATH. THERE ARE ONE MILLION INTERVENTIONS TO TREAT PAD PER YEAR, EMPHASIZING THE HIGH PREVALENCE OF THIS DISEASE. THE PREFERRED TREATMENT IS PERCUTANEOUS VASCULAR INTERVENTIONS (PVI), WHERE VESSEL PLAQUE IS PENETRATED AND THREADED (CROSSED) BY A GUIDEWIRE, FOLLOWED BY A BALLOON OVER THE WIRE AND/OR OTHER ADJUNCTIVE DEVICES TO OPEN THE BLOOD VESSEL. UNFORTUNATELY, PVI FAILS IMMEDIATELY FOR 20% OF PATIENTS BECAUSE THE PLAQUE PROVES IMPENETRABLE -- MEANING PATIENTS ARE PUT AT RISK FOR NO HEALTH BENEFIT. FURTHERMORE, AFTER BALLOONING OPEN THERE IS DAMAGE TO THE VESSEL WALL AND VESSELS BLOCK AGAIN WITHIN A YEAR FOR 70% OF PATIENTS WITH BELOW-THE-KNEE PLAQUES. THESE FAILURES THEN REQUIRE ADDITIONAL INVASIVE INTERVENTIONS. THE DIFFICULTIES WITH PVI ARE FRUSTRATING FOR THE CLINICIANS PERFORMING THE SURGERY AND NEEDLESSLY RISKY FOR OUR PATIENTS. THERE IS A FUNDAMENTAL GAP IN KNOWLEDGE IN HOW TO SELECT PATIENTS THAT BENEFIT FROM PVI AND HOW DIFFERENT PREPARATION DEVICES ALTER VESSEL WALL INJURY. WE PROPOSE A TWO-PRONGED APPROACH: 1) IMPROVE PATIENT SELECTION FOR PVI USING A NOVEL MRI-HISTOLOGY BASED ANATOMIC SCORING SYSTEM THAT IDENTIFIES PATIENTS WITH IMPENETRABLE PLAQUES; AND 2) IMPROVE DEVICE SELECTION BY IDENTIFYING DEVICES THAT REDUCE VESSEL WALL INJURY DURING PVI USING HISTOPATHOLOGIC ANALYSIS AFTER PVI IN A CADAVERIC MODEL. OUR PRELIMINARY DATA SHOW THAT OUR IN VIVO MRI-HISTOLOGY METHOD CAN VISUALIZE HARD PLAQUE COMPONENTS (BOTH CALCIUM AND DENSE COLLAGEN) TO DECIPHER INDIVIDUAL PATIENTS' PLAQUE MORPHOLOGY AND DETERMINE WHICH PLAQUES ARE PENETRABLE. FURTHERMORE, OUR CADAVERIC MODEL INDICATES DIFFERENCES IN VESSEL WALL INJURY FOLLOWING DIFFERENT PREPARATION DEVICES USING DETAILED EX VIVO PLAQUE ANALYSIS POST-INTERVENTION. OUR SPECIFIC AIMS ARE: 1. ESTABLISH A NOVEL MRI-HISTOLOGY ANATOMIC SCORING SYSTEM THAT PREDICTS PVI IMMEDIATE TECHNICAL FAILURE TO IMPROVE PATIENT SELECTION FOR PVI. WE WILL IMAGE PATIENTS PRIOR TO THEIR PVI WITH OUR CLINICAL MRI-HISTOLOGY PROTOCOL TO PROSPECTIVELY SCORE INDIVIDUAL PATIENTS’ PLAQUES TO PREDICT SUCCESSFUL GUIDEWIRE CROSSING. 2. DESCRIBE THE BENEFITS OF USING ORBITAL ATHERECTOMY BEFORE BALLOON ANGIOPLASTY VERSUS USING BALLOON ANGIOPLASTY ALONE FOR VARIOUS PLAQUE TYPES IN ARTERIES BELOW THE KNEE. WE WILL RANDOMIZE AMPUTATED LEGS FROM PAD PATIENTS WITH CALCIFIED BELOW-THE-KNEE ARTERIES TO UNDERGO PLAIN BALLOON ANGIOPLASTY VERSUS ATHERECTOMY PREPARATION DEVICE PRIOR TO ANGIOPLASTY. PRE-PROCEDURE 3T MRI-HISTOLOGY, INTRAVASCULAR ULTRASOUND, AND EX VIVO PLAQUE ANALYSIS WITH HISTOLOGY, WILL DETERMINE VESSEL WALL INJURY BETWEEN INTERVENTION GROUPS. WE WILL ESTABLISH A PATIENT SELECTION PROCESS FOR PVI AND DEFINE VESSEL WALL RESPONSE WITH BALLOONING ONLY VERSUS VESSEL PREPARATION WITH ATHERECTOMY THROUGH HISTOLOGIC ANALYSIS OF CADAVERIC LESIONS. OUR STUDY WILL EMPOWER CLINICAL DECISION-MAKING TOWARD THE SAFEST, MOST EFFECTIVE STRATEGIES TO ULTIMATELY SAVE PATIENT LIMBS AND LIVES.

LILRB MODULATES TUMOR MICROENVIRONMENT AND PROMOTES TUMOR PROGRESSION

T CELL FATE DECISIONS AND TRANSPLANT OUTCOMES

ROLE OF TUMOR MICROENVIRONMENT-DERIVED CHOLESTEROL IN CD8+ T-CELL EXHAUSTION

(PQA4) POINT-OF-CARE TEST OF EXPOSURE TO HEPATOCELLULAR CARCINOMA RISK BY V-CHIP

HES1-LOSS PROMOTES DYSREGULATION OF EPITHELIAL HOMEOSTASIS AND INFLAMMATION IN A SERRATED ADENOCARCINOMA MODEL

BIOPHYSICAL ROLES OF PRE-METASTATIC NICHE EVOLUTION ON TRANSPORT OF CIRCULATING TUMOR CELLS

A NOVEL ROLE FOR MAGI1 IN REGULATING NON-CANONICAL LATS SIGNALING AND ATHEROSCLEROTIC PLAQUE FORMATION

DEFINING THE ALTERED FUS-PARP-1-DNA LIGASE III AXIS AND ITS IMPLICATIONS TO NUCLEAR AND MITOCHONDRIAL GENOME DAMAGE RESPONSE IN AMYOTROPHIC LATERAL SCLEROSIS (ALS) AND FRONTOTEMPORAL DEMENTIA (FTD)

MECHANISMS DRIVING CARDIAC DYSFUNCTION IN AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE - CARDIOVASCULAR DISEASE IS A MAJOR CAUSE OF MORBIDITY AND MORTALITY IN PATIENTS WITH AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE (ADPKD). CHARACTERIZED BY PROGRESSIVE RENAL DYSFUNCTION, ADPKD IMPOSES VERY SIGNIFICANT HEALTHCARE AND ECONOMIC BURDENS. IT HAS COMMONLY BEEN ASSUMED THAT PROGRESSIVE RENAL IMPAIRMENT PROMOTES CARDIAC DISEASE; HOWEVER, OUR PRELIMINARY DATA SUGGEST THAT CARDIAC DYSFUNCTION ORIGINATES IN CARDIOMYOCYTES AND MANIFESTS PRIOR TO RENAL FAILURE IN ADPKD. RECENT CLINICAL EVIDENCE SUPPORTS OUR FINDINGS BY SHOWING THAT ADPKD PATIENTS EXHIBIT VENTRICULAR DYSFUNCTION BEFORE THE ONSET OF RENAL FAILURE, EVEN IN NON-HYPERTENSIVE INDIVIDUALS. MUTATIONS IN THE GENE ENCODING POLYCYSTIN-1 (PC1) OCCUR IN 85% OF PATIENTS AND ARE RESPONSIBLE FOR THE MOST SEVERE CASES. IMPORTANTLY, PC1 IS EXPRESSED IN CARDIOMYOCYTES, YET ITS ROLE(S) THERE IS(ARE) POORLY UNDERSTOOD. WE PROPOSE THAT THE MUTANT PC1 – IN A CARDIOMYOCYTE-AUTONOMOUS FASHION – INITIATES AND DRIVES HEART DISEASE IN ADPKD, INDEPENDENT OF RENAL FAILURE. OUR DATA SHOW THAT PC1 CARDIOMYOCYTE-SPECIFIC DELETION PROMOTES SYSTOLIC AND DIASTOLIC DYSFUNCTION IN MICE. FURTHERMORE, USING A MOUSE MODEL HARBORING A CLINICALLY ESTABLISHED ADPKD-CAUSING PC1 MUTATION (RC ALLELE), WE PROVIDE EVIDENCE OF IMPAIRED CALCIUM-CYCLING AND CONTRACTILITY AT THE CARDIOMYOCYTE LEVEL, WHICH OCCUR BEFORE THE ONSET OF RENAL FAILURE. HETEROZYGOUS RC/+ YOUNG MICE MANIFEST ALTERATIONS IN CALCIUM HANDLING/CONTRACTILITY IN ISOLATED CARDIOMYOCYTES, WHICH CORRELATE WITH REDUCED LEFT VENTRICULAR GLOBAL LONGITUDINAL STRAIN AND DIASTOLIC DYSFUNCTION. WE DISCOVERED THAT PC1 REGULATES ACTION POTENTIAL DURATION VIA KV CHANNEL CURRENT REGULATION. PC1 ABLATION SHORTENS ACTION POTENTIAL DURATION AND IMPAIRS BOTH CALCIUM TRANSIENTS AND CONTRACTILITY IN CARDIOMYOCYTES. ADDITIONALLY, PC1 DELETION IMPAIRS SARCOPLASMIC RETICULUM (SR) CALCIUM LOADING THROUGH REDUCED SR CALCIUM-ATPASE (SERCA) ACTIVITY. THESE DATA HAVE LED US TO HYPOTHESIZE THAT ADPKD-CAUSING PC1 MUTATIONS DISRUPT PC1 ACTIONS IN CARDIOMYOCYTES, IMPAIR CARDIAC FUNCTION AND PREDISPOSE THE HEART TO HYPERTENSION-INDUCED HEART FAILURE, INDEPENDENT OF RENAL DYSFUNCTION. TO TEST THIS HYPOTHESIS, WE PROPOSE THREE AIMS: 1) DETERMINE HOW PC1 MUTATIONS AFFECT ACTION POTENTIALS AND KV CHANNEL ACTIVITY AND IMPINGE ON CALCIUM HANDLING AND CONTRACTILITY. 2) ELUCIDATE MECHANISMS WHEREBY PC1 REGULATES SR CALCIUM LOADING AND SERCA TO MAINTAIN CARDIOMYOCYTE FUNCTION AND TEST THE IMPACT OF ADPKD MUTATIONS IN PC1 ON THESE EVENTS. 3) DETERMINE IN VIVO WHETHER ALTERATIONS IN PC1 SIGNALING IN CARDIOMYOCYTES DRIVE CARDIAC DYSFUNCTION AND PREDISPOSE THE HEART TO HYPERTENSION-INDUCED HEART FAILURE. COMPLETION OF OUR STUDIES WILL PROVIDE PARADIGM-SHIFTING INFORMATION REGARDING THE ROLE OF CARDIOMYOCYTE-AUTONOMOUS EVENTS DRIVING HEART DISEASE IN ADPKD, THE LEADING CAUSE OF DEATH IN THESE PATIENTS.

GUT MICROBIOME COMMUNICATION WITH THE BONE MARROW REGULATES INTESTINAL INFLAMMATION

CONVERGENT AI FOR PRECISE BREAST CANCER RISK ASSESSMENT

B-LACTAM MEDIATED SOS RESPONSE AND EXPRESSION OF RESISTANCE IN CLINICAL MRSA

MOLECULAR MECHANISM OF VIRULENCE REGULATION IN STREPTOCOCCUS PYOGENES

MOLECULAR REGULATION OF IMMUNITY TO VIRAL INFECTIONS

A PHASE II MULTI-CENTER TRIAL EVALUATING DUAL TARGETING OF THE PI3K/AKT AND NOS PATHWAYS FOR TREATING METAPLASTIC BREAST CANCER (MPBC) - ABSTRACT METAPLASTIC BREAST CANCER (MPBC) IS A RARE SUBSET ACCOUNTING FOR <1% OF ALL BREAST CANCERS. HOWEVER, MPBC IS A SIGNIFICANT HEALTH CHALLENGE AS IT EXHIBITS THE MOST DISMAL PROGNOSIS OF ALL BREAST CANCERS, EVEN WORSE THAN TRIPLE-NEGATIVE BREAST CANCER (TNBC), WITH A SURVIVAL RATE OF 8 MONTHS OR LESS IN PATIENTS WITH METASTATIC DISEASE. DUE TO A LACK OF DRUGGABLE TARGETS, THE MAIN THERAPEUTIC OPTION FOR METASTATIC MPBC REMAINS SYSTEMIC CHEMOTHERAPY, DESPITE KNOWN RESISTANCE TO MOST CYTOTOXIC DRUGS. ONE COMMON MOLECULAR ALTERATION IN MPBC IS HYPERACTIVATION OF THE PHOSPHOINOSITIDE 3-KINASE AND PROTEIN KINASE B (PI3K/AKT) PATHWAY. ADDITIONALLY, WE RECENTLY PUBLISHED THAT MPBC DISPLAYS A GAIN-OF-FUNCTION ONCOGENIC MUTATION IN RIBOSOMAL PROTEIN L39 (RPL39), WHICH IS RESPONSIBLE FOR TREATMENT RESISTANCE, STEM CELL SELF-RENEWAL, AND LUNG METASTASIS. THE MECHANISTIC FUNCTION OF RPL39 IS MEDIATED THROUGH INDUCIBLE NITRIC OXIDE SYNTHASE (INOS)-MEDIATED NITRIC OXIDE PRODUCTION. IN ADDITION, WE DEMONSTRATED IN A COMPLETED CLINICAL TRIAL THAT INHIBITING THIS NITRIC OXIDE SYNTHASE (NOS) PATHWAY USING PAN-NOS INHIBITOR NG-METHYL-L-ARGININE ACETATE (L-NMMA) MAY REPRESENT A HIGHLY EFFECTIVE THERAPEUTIC OPTION FOR TNBC PATIENTS. THEREFORE, WE HYPOTHESIZE THAT A COMBINATORIAL TARGETED APPROACH OF INHIBITING THE TWO MAJOR ONCOGENIC PATHWAYS IMPLICATED IN MPBC, PI3K/AKT AND NOS, WOULD LEAD TO SIGNIFICANT TUMOR REGRESSION. TO TEST THIS HYPOTHESIS, THIS U01 APPLICATION BRINGS TOGETHER RESEARCH TEAMS FROM HOUSTON METHODIST CANCER CENTER (HMCC), THE UNIVERSITY OF TEXAS MD ANDERSON CANCER CENTER, AND THE NATIONAL CANCER INSTITUTE (NCI). SPECIFIC AIM 1 SEEKS TO DEFINE WHETHER DUAL INHIBITION OF PI3K/AKT USING ALPELISIB AND NOS INHIBITION USING L-NMMA COMBINED WITH NAB-PACLITAXEL WILL INCREASE THE OBJECTIVE RESPONSE RATE AND SURVIVAL IN METASTATIC MPBC PATIENTS. IN SPECIFIC AIM 2, USING BLOOD AND CORE BIOPSY TISSUES COLLECTED IN THE TRIAL, WE WILL IDENTIFY MECHANISMS OF RESPONSE TO THERAPY TO DETERMINE THE EFFICACY OF THE TARGETED PI3K/AKT AND NOS PATHWAY INHIBITORY APPROACH. FURTHERMORE, THE CELL-CELL INTERACTIONS AMONG TUMOR CELLS, MYELOID CELLS, LYMPHOID CELLS, AND STROMAL CELLS WITHIN THE TUMOR MICROENVIRONMENT AND THEIR ROLE IN SUPPORTING CANCER STEM CELL POPULATIONS AND DRUG-RESISTANT CELL DEVELOPMENT DURING TREATMENT WILL BE EVALUATED. THE IMPACT OF DISTINCT CELLULAR LOCALIZATION PATTERNS WITHIN THE TUMOR ECOSYSTEM ON THE PROCESS OF CANCER STEM CELL MAINTENANCE AND MODULATION, AS WELL AS THE DEVELOPMENT OF DRUG RESISTANCE, WILL BE ANALYZED AT THE SINGLE-CELL LEVEL USING SPATIAL TRANSCRIPTOMICS, IMMUNOFLUORESCENCE, CYTOF IMAGING SYSTEMS, AND A MULTI-MODAL DATA ANALYSIS MODEL. THIS STUDY THUS PROPOSES A MECHANISTIC INVESTIGATION OF A COMBINATORIAL TARGETED APPROACH AGAINST THE TWO KEY PATHWAYS IN MPBC, DEVELOPS UNIQUE CROSSTALK MODELS, AND IDENTIFIES BIOMARKERS OF RESISTANCE AND CELL–CELL INTERACTIONS USING SPECIMENS DERIVED FROM MPBC PATIENTS.

MOLECULAR REGULATION OF LONG-TERM IMMUNE MEMORY

CLINICAL IMPACT OF THE CEFAZOLIN INOCULUM EFFECT - ABSTRACT STAPHYLOCOCCUS AUREUS IS A MAJOR HUMAN PATHOGEN RESPONSIBLE FOR A WIDE RANGE OF LIFE-THREATENING INFECTIONS. MANY OF THESE INFECTIONS ARE CAUSED BY METHICILLIN-SUSCEPTIBLE S. AUREUS (MSSA). MSSA REPRESENT A MAJOR BURDEN AMONG S. AUREUS INFECTIONS AND ARE IMPORTANT CONTRIBUTORS TO MORTALITY. FOR DECADES, THE FIRST LINE OF THERAPY FOR SEVERE MSSA INFECTIONS HAVE BEEN THE ISOXAZOLYL-PENICILLINS (ISP, E.G., NAFCILLIN). HOWEVER, RECENT DATA SUGGEST THAT CLINICAL OUTCOMES IN MSSA BACTEREMIA ARE SIMILAR IN PATIENTS TREATED WITH CEFAZOLIN (VS NAFCILLIN), A CEPHALOSPORIN WITH ACTIVITY AGAINST MSSA THAT APPEARS TO BE LESS TOXIC. INDEED, TREATMENT WITH NAFCILLIN SEEMS TO BE ASSOCIATED WITH INCREASED COSTS, MORE DRUG REACTIONS (INCLUDING HEPATOTOXICITY, INTERSTITIAL NEPHRITIS AND NEUTROPENIA) AND, POSSIBLY, HIGHER MORTALITY. DUE TO THESE CONCERNS, AN IMPORTANT SHIFT IN THE TREATMENT OF MSSA IS OCCURRING WHEREBY CLINICIANS ARE NOW USING CEFAZOLIN AS FIRST LINE OF THERAPY FOR SEVERE MSSA INFECTIONS. AN IMPORTANT CONCERN OF USING CEFAZOLIN AND OTHER CEPHALOSPORINS AS PRIMARY THERAPY FOR THESE SERIOUS INFECTIONS IS THE CEFAZOLIN INOCULUM EFFECT (CZIE), DEFINED AS A CEFAZOLIN MINIMAL INHIBITORY CONCENTRATION OF > 16 ΜG/ML WHEN A HIGH INOCULUM (107 CFU/ML) IS USED. THE CZIE HAS BEEN ASSOCIATED WITH FAILURES IN THE TREATMENT OF DEEP-SEATED MSSA INFECTIONS AND WITH THE PRODUCTION OF CERTAIN ISOTYPES OF THE STAPHYLOCOCCAL SS-LACTAMASE. HOWEVER, THE CHARACTERIZATION OF THE CLINICAL IMPACT OF THIS PHENOMENON IN DEEP-SEATED MSSA INFECTIONS IS LIMITED. IN ADDITION, IT IS CURRENTLY NOT POSSIBLE TO DETECT THE CZIE IN A STANDARD CLINICAL MICROBIOLOGY LABORATORY GIVEN THE CUMBERSOME AND EXPENSIVE NATURE OF THE GOLD STANDARD TEST FOR ITS DETECTION. OUR PUBLISHED AND PRELIMINARY CLINICAL DATA SUGGEST THAT THE CZIE IS AN IMPORTANT CONTRIBUTOR TO WORSE CLINICAL OUTCOMES OF SEVERE MSSA INFECTIONS. FURTHERMORE, WE HAVE DEVELOPED AND PUBLISHED A NOVEL COLORIMETRIC NITROCEFIN-BASED RAPID TEST (~3 H) THAT DETECTS THE CZIE WITH HIGH SENSITIVITY AND SPECIFICITY THAT CAN BE INCORPORATED IN THE ROUTINE CLINICAL MICROBIOLOGY LABORATORY. WE POSTULATE THAT, I) THE CZIE NEGATIVELY IMPACTS CLINICAL OUTCOMES IN MSSA BACTEREMIA TREATED WITH CEFAZOLIN AND, II) A RAPID TEST CAN BE READILY IMPLEMENTED FOR THE IDENTIFICATION OF THE CZIE IN S. AUREUS BACTEREMIA AND CAN DETECT PATIENTS AT HIGHER RISK OF POOR OUTCOMES. IN ORDER TO ADDRESS THESE HYPOTHESES, WE WILL TAKE ADVANTAGE OF THE STAPHYLOCOCCUS AUREUS NETWORK ADAPTIVE PLATFORM (SNAP) TRIAL, A MULTICENTER, PRAGMATIC, MULTI-ARM, OPEN-LABEL ADAPTIVE PLATFORM TRIAL ADDRESSING MULTIPLE THERAPEUTIC QUESTIONS IN PATIENTS WITH S. AUREUS BACTEREMIA. WE WILL FOCUS IN THE MSSA “DOMAIN” THAT EVALUATES THE EFFECTIVENESS AND SAFETY OF CEFAZOLIN VS ISP IN A RANDOMIZED FASHION, CURRENTLY ENROLLING IN AUSTRALIA, SINGAPORE, CANADA, ISRAEL, NEW ZEALAND, UNITED KINGDOM, UNITED STATES, COLOMBIA AND CHILE. THE SPECIFIC AIMS OF OUR PROPOSAL ARE: I) TO DEFINE THE CLINICAL IMPACT OF THE CZIE IN MSSA BACTEREMIA AND, II) TO DETERMINE THE CLINICAL VALUE AND FEASIBILITY OF A RAPID TEST TO DETECT THE CZIE. OUR FINDINGS ARE LIKELY TO TRANSFORM THE TREATMENT APPROACH FOR MSSA INFECTIONS AND WILL PROVIDE THE BASIS TO DEVELOP NOVEL DIAGNOSTIC TOOLS TO THE MANAGEMENT OF MSSA INFECTIONS.

CENTRAL AND PERIPHERAL IMMUNE CROSS-TALK IN ALZHEIMER'S DISEASE AND THEIR MODULATION BY A NOVEL IMMUNOTHERAPY - PROJECT SUMMARY/ABSTRACT ALZHEIMER’S DISEASE (AD) GENE-RISK DATA, EPIDEMIOLOGICAL FINDINGS AND ANIMAL MODELS CONVERGE TO INDICATE THE CRITICAL ROLE OF INFLAMMATION FOR THE ONSET AND PROGRESSION OF AD. INFLAMMATION COULD PROVIDE A NEW FOCUS FOR THERAPEUTIC INTERVENTION. HOWEVER, INFLAMMATION BIOMARKERS ARE POORLY CHARACTERIZED IN AD. IN THIS PROJECT, WE WILL MEASURE BLOOD AND CEREBROSPINAL FLUID (CSF) INFLAMMATION BIOMARKERS AND COMPARE THEM TO MEASUREMENTS OF BRAIN GLIAL ACTIVATION OBTAINED BY POSITRON EMISSION TOMOGRAPHY (PET). IN ADDITION, WE WILL DETERMINE THE EFFECT OF LOW-DOSE INTERLEUKIN-2 (IL-2) IMMUNOTHERAPY, GIVEN OVER 22 WEEKS, ON THESE INFLAMMATION BIOMARKERS. FOR THIS PURPOSE, WE WILL MEASURE THESE BIOMARKERS IN AD INDIVIDUALS ENROLLED IN A PHASE 2A SAFETY AND EFFICACY TRIAL OF LOW-DOSE IL-2 THERAPY. REGULATORY T CELLS (TREGS) PLAY A NEUROPROTECTIVE ROLE BY SUPPRESSING INFLAMMATION IN THE BLOOD AND THE BRAIN. WE HAVE SHOWN THAT TREG IMMUNOMODULATORY MECHANISMS ARE COMPROMISED IN AD PATIENTS, RESULTING IN ACTIVATION OF PRO-INFLAMMATORY MONOCYTES AND UPREGULATION OF INFLAMMATORY MEDIATORS. IN A PHASE 1 CLINICAL STUDY, WE SHOWED THAT IL-2 ADMINISTRATION INDUCED TREG EXPANSION AND RESTORATION; THE THERAPY WAS SAFE, AND IT WAS ASSOCIATED WITH IMPROVED COGNITION. SUPPORTED BY A “PART-THE-CLOUD” GRANT FROM THE ALZHEIMER’S ASSOCIATION, WE WILL CONDUCT A FOLLOW UP PHASE 2A STUDY WHICH INCLUDES THE MEASUREMENT OF COGNITION, AS WELL AS CSF T-TAU, P-TAU, ASS42, AND NEUROFILAMENT LIGHT CHAIN (NFL) BEFORE AND AFTER 22 WEEKS OF IL-2 TREATMENT IN THE SUBJECTS WITH MILD TO MODERATE AD. TAKING ADVANTAGE OF THIS NOVEL TRIAL, AND BY EVALUATING THE 40 AD PATIENTS PARTICIPATING IN IT, THE CURRENT STUDY WILL INVESTIGATE SYSTEMIC AND CENTRAL NERVOUS SYSTEM (CNS) BIOMARKERS OF INFLAMMATION AND THEIR MODULATION BY IL-2 ADMINISTRATION. WE WILL ANALYZE THE IMPACT OF IL-2 IMMUNOTHERAPY ON PERIPHERAL IMMUNE BIOMARKERS (AIM 1) AS WELL AS CNS INFLAMMATION, MEASURED THROUGH CSF INFLAMMATION BIOMARKERS (AIM 2) AND BRAIN INFLAMMATION POSITRON EMISSION TOMOGRAPHY (AIM 3). TO DETERMINE WHICH BLOOD BIOMARKERS CORRELATE BEST WITH CENTRAL NERVOUS SYSTEM BIOMARKERS (AIM 4), THE ASSOCIATIONS AMONG BLOOD, CSF, AND BRAIN IMAGING MEASURES OF NEUROINFLAMMATION BEFORE AND AFTER IL-2 THERAPY WILL BE DETERMINED. OUR NOVEL APPROACH WILL EXPLORE THE POTENTIAL LINK BETWEEN SYSTEMIC AND CNS INFLAMMATION IN AD CLINICAL SETTING AND ADVANCE THE USE OF INFLAMMATORY BIOMARKERS IN AD ANTI- INFLAMMATORY CLINICAL TRIALS.

MECHANISMS OF STING IN MALIGNANT PROGRESSION AND THERAPY OF CLL. - PROJECT SUMMARY/ABSTRACT CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) REPRESENTS 30% OF ADULT LEUKEMIA AND IS STILL INCURABLE. ACCORDING TO THE NATIONAL CANCER INSTITUTE, APPROXIMATELY 21,250 NEW CASES OF CLL AND 4,320 DEATHS FROM CLL ARE PROJECTED IN THE UNITED STATES ALONE IN 2021. ALTHOUGH THE BRUTON'S TYROSINE KINASE INHIBITOR (BTKI) IS AN EFFECTIVE TARGETED THERAPY FOR CLL, APPROXIMATELY 50% OF BTKI-TREATED CLL PATIENTS HAVE DROPPED OUT OF THE THERAPY DUE TO CHRONIC ADVERSE EFFECTS. ADDITIONALLY, SIGNIFICANTLY INCREASED NUMBERS OF CLL PATIENTS TREATED WITH BTKI OR OTHER KINASE INHIBITORS HAVE DEVELOPED THE MORE MEDICALLY CHALLENGING DIFFUSE LARGE B CELL LYMPHOMA. NOVEL THERAPY THAT CAN DIRECTLY TARGET CLL OR BE RATIONALLY COMBINED WITH BTKI IS HIGHLY DESIRABLE. THE STIMULATOR OF INTERFERON GENES (STING) IS AN ENDOPLASMIC RETICULUM (ER)-RESIDENT PROTEIN CRITICAL FOR SENSING CYTOPLASMIC DNA AND PROMOTING PRODUCTION OF TYPE I INTERFERONS, THEREBY BOOSTING IMMUNE RESPONSES. STING AGONISTS, SUCH AS ADU-S100, HAVE BEEN USED AS COMBINATION IMMUNOTHERAPY WITH PEMBROLIZUMAB IN CLINICAL TRIALS TO TREAT ADVANCED SOLID TUMORS AND LYMPHOMAS, AND THESE THERAPEUTIC APPLICATIONS OF STING AGONISTS ARE BASED ON THE MAIN KNOWN FUNCTION OF STING IN ELICITING ANTI-TUMOR IMMUNITY. WE WERE THE FIRST TO SHOW THAT STING AGONISTS DIRECTLY INDUCE POTENT MITOCHONDRIA-MEDIATED APOPTOSIS IN B CELL-DERIVED MALIGNANCIES INCLUDING CLL, WHILE THESE AGONISTS INDUCE PRODUCTION OF INTERFERONS IN MELANOMA, HEPATOMA AND LUNG CANCER CELLS WITHOUT SUPPRESSING THEIR GROWTH. APOPTOSIS OF CLL, B CELL LYMPHOMA AND MULTIPLE MYELOMA REQUIRES STING, BECAUSE GENETIC DELETION OF STING RESULTS IN RESISTANCE OF ALL THESE CELLS TO STING AGONIST-MEDIATED APOPTOSIS. APOPTOSIS OF THESE CELLS IS NOT DUE TO THE PRODUCTION OF INFLAMMATORY CYTOKINES BUT MAY INVOLVE THE PROLONGED PRESENCE OF AGONIST-BOUND STING. TOGETHER WITH OUR NEW PRELIMINARY RESULTS SHOWING THAT MUTATION-MEDIATED ACTIVATION OF STING CAUSES RAPID DEGRADATION OF THE B CELL RECEPTOR (BCR) AND SIGNIFICANTLY REDUCED BCR SIGNALING (DISADVANTAGEOUS FOR CLL SURVIVAL), AND THAT NOVEL SERINE RESIDUES ON STING ARE PHOSPHORYLATED IN AGONIST-STIMULATED MALIGNANT B CELLS, WE PROPOSE TO IDENTIFY AND CHARACTERIZE DIFFERENTIAL PHOSPHORYLATION AND INTERACTING PARTNERS OF ACTIVATED STING TO FURTHER UNDERSTAND THE MECHANISMS BY WHICH ACTIVATED STING CAUSES BCR DEGRADATION AND APOPTOSIS IN CLL. SINCE OUR PRELIMINARY DATA SHOW THAT STING DEFICIENCY CAN LEAD TO INCREASED LEVELS OF THE BCR AND BCR SIGNALING IN MOUSE CLL, AND THAT MOUSE AND HUMAN MALIGNANT CLL CELLS SIGNIFICANTLY DOWNREGULATE THEIR EXPRESSION LEVELS OF STING, WE WILL TEST WHETHER STING- DOWNREGULATED OR STING-DEFICIENT CLL CELLS ARE LESS SENSITIVE TO BTKI. WE WILL ALSO EXAMINE HOW SUCH ALTERED STING EXPRESSION AND PHOSPHORYLATION CAN REGULATE THE SURVIVAL, PROGRESSION AND CHEMORESISTANCE OF CLL. BASED ON OUR HYPOTHESIS THAT ACTIVATION OF STING CAN LEAD TO THE DEGRADATION OF THE BCR AND RENDER CLL CELLS MORE SENSITIVE TO BTKI, WE PROPOSE TO COMBINE STING AGONISTS WITH BTKI TO TREAT CLL.

TARGETING THE HIV-1 RESERVOIR IN MYELOID CELLS - ABSTRACT HIV IS REVERSE-TRANSCRIBED AND INTEGRATED INTO THE DNA OF HOST CELLS, RESULTING IN PERSISTENT INFECTIONS THAT ARE DIFFICULT TO CLEAR. TO DATE, THERE IS NO EFFECTIVE CURE TO CLEAR THE VIRUS FROM HIV- INFECTED PATIENTS. WE HAVE SHOWN AN APPROACH TO ERADICATE HIV INFECTIONS BY SELECTIVE ELIMINATION OF HOST CELLS HARBORING REPLICATION-COMPETENT HIV (SECH). THE SECH TREATMENT CAN CLEAR HIV-1 IN OVER 50% MICE RECONSTITUTED WITH A HUMAN IMMUNE SYSTEM, AND IN BLOOD SAMPLES FROM HIV-1- INFECTED PATIENTS. SECH CAN CLEAR HIV-INFECTION NOT ONLY IN CD4 T CELLS, BUT ALSO IN MACROPHAGES AND MICROGLIA CELLS. EXPERIMENTS ARE PROPOSED TO DETERMINE HOW TO IMPROVE THE EFFICACY AND SAFETY FOR SECH IN CLEARING HIV-INFECTION IN THE MACROPHAGE LINEAGE: 1) TO IMPROVE THE EFFICACY AND SPECIFICITY IN THE KILLING OF HIV-1-INFECTED MACROPHAGES. WE HAVE FOUND THAT INCREASED VIRAL REACTIVATION LEADS TO IMPROVED VIRAL CLEARANCE IN HUMANIZED MICE. WE WILL IDENTIFY DRUGS THAT SHOW GREATER POTENTIALS IN VIRUS REACTIVATION THAT COULD LEAD TO BETTER HIV CLEARANCE. WE WILL EXAMINE THE INFLAMMATORY POTENTIALS FOR IDB AND ALTERNATIVE LRAS IN MACROPHAGES, AND DETERMINE WHAT INFLAMMATORY CYTOKINES MAY NEED TO BE TARGETED; 2) TO DETERMINE HOW TO IMPROVE THE EFFICACY AND SAFETY IN THE CLEARANCE OF HIV RESERVOIR IN THE MACROPHAGE LINEAGE IN LYMPHOID ORGANS AND IN THE BRAIN OF HUMANIZED MICE. WE WILL EVALUATE WHETHER ALTERNATIVE AGENTS CAN IMPROVE THE CLEARANCE OF HIV-1 IN HUMANIZED MICE IN VIVO. WE WILL ALSO DETERMINE WHETHER OPTIMUM SECH REGIMENS CAN INDUCE LIMITED AND CONTROLLABLE INFLAMMATORY RESPONSES. OUR PROPOSED WORK WILL PROVIDE NOVEL INSIGHTS INTO HOW TARGETING APOPTOSIS AND AUTOPHAGY REGULATES THE CLEARANCE OF HIV INFECTION IN THE MACROPHAGE LINEAGE. THE STUDIES WILL FACILITATE THE DEVELOPMENT OF SECH CLEAR HIV-1 INFECTION IN LYMPHOID ORGANS AND IN THE BRAIN TO ACHIEVE COMPLETE ERADICATION OF HIV-1.

TARGETING PHENETHYL ISOTHIOCYANATE TO MITOCHONDRIA IN LUNG CARCINOGENESIS - PROJECT SUMMARY DISRUPTION OF NORMAL MITOCHONDRIAL BIOENERGETICS AND OXIDATIVE PHOSPHORYLATION REPRESENTS AN EARLY EVENT DURING ONCOGENESIS BY CHANGING THE ENERGY METABOLISM OF PRECANCEROUS AND CANCEROUS CELLS. PHENETHYL ISOTHIOCYANATE (PEITC), A NATURAL COMPOUND PRESENT IN CRUCIFEROUS VEGETABLES, HAS BEEN SHOWN TO INHIBIT THE DEVELOPMENT OF SEVERAL TYPES OF CANCER IN ANIMAL MODELS. PEITC HAS BEEN SHOWN TO INHIBIT OXIDATIVE PHOSPHORYLATION AND TO INDUCE CANCER CELL APOPTOSIS THROUGH A MITOCHONDRIA-DEPENDENT MECHANISM AND ROS FORMATION, SUGGESTING THE ROLE OF MITOCHONDRIAL BIOENERGETIC FUNCTION AND REDOX HOMEOSTASIS IN ONCOGENESIS. THIS PROVIDES THE RATIONALE FOR CONJUGATING PEITC TO A TARGETING AGENT THAT DRIVES IT INTO MITOCHONDRIA TO SPECIFICALLY STUDY THE ROLE OF MITOCHONDRIAL FUNCTION AND ROS FORMATION IN LUNG CANCER DEVELOPMENT AND TO INCREASE THE EFFICACY OF PEITC. PRELIMINARY STUDIES DEMONSTRATE THAT MITOCHONDRIA-TARGETED PEITC (MITO-PEITC) IS A SIGNIFICANTLY MORE POTENT INHIBITOR OF LUNG CARCINOGENESIS IN MICE. WE HYPOTHESIZE THAT LUNG ONCOGENESIS AND METASTASIS DEPEND ON MITOCHONDRIAL RESPIRATION AND THAT MITO-PEITC IS A NOVEL, POTENT INHIBITOR OF LUNG CARCINOGENESIS AND METASTASIS ACTING PRIMARILY THROUGH MITOCHONDRIAL MECHANISMS. THIS HYPOTHESIS WILL BE TESTED USING THREE SPECIFIC AIMS. AIM 1 WILL EVALUATE THE PREVENTIVE POTENTIAL AND MECHANISMS OF ACTION OF MITO-PEITC IN VITRO. WE WILL USE MITO-PEITC AND LUNG CANCER CELLS WITH GENETICALLY MODIFIED EXPRESSION OF THE MEMBERS OF THE MITOCHONDRIAL ELECTRON TRANSPORT CHAIN TO DECIPHER THE ONCOGENIC MECHANISM AND EVALUATE THE MECHANISMS OF ACTION OF MITO-PEITC. AIM 2 WILL DETERMINE PREVENTIVE EFFICACY OF MITO-PEITC ON LUNG TUMOR INITIATION AND PROGRESSION IN A/J MICE. AIM 3 WILL DETERMINE THE IN VIVO MECHANISM AND THE EFFICACY OF MITO-PEITC TO INHIBIT LUNG CANCER BRAIN METASTASIS. WE WILL USE STATE-OF-THE-ART IN VITRO AND IN VIVO ASSAYS, INCLUDING SMALL ANIMAL IMAGING TECHNOLOGY TO MONITOR THE GROWTH OF PRIMARY TUMORS (MAGNETIC RESONANCE IMAGING) AND ENGRAFTMENT OF METASTATIC CELLS. INNOVATIVE APPROACHES FOR IN VIVO MONITORING OF CHANGES IN CANCER CELL BIOENERGETICS AND CELLULAR OXIDANT PRODUCTION (BIOLUMINESCENCE IMAGING) WILL BE EMPLOYED. DEFINING MITOCHONDRIAL MECHANISMS OF LUNG ONCOGENESIS AND BRAIN METASTASIS AND DEVELOPING A NOVEL, WELL-TOLERATED EFFICACIOUS AGENT FOR PREVENTION AND TREATMENT OF LUNG CANCER WILL BE HIGHLY SIGNIFICANT.

ETHICAL & METHODOLOGICAL STANDARDS FOR TRIALS OF INVASIVE THERAPEUTIC PROCEDURES

CORTICAL ENCODING OF UNCONSCIOUS VISUAL INFORMATION AND ITS IMPACT ON BEHAVIOR

ERADICATION OF ESCAPED VARIANT TUMOR CELLS FOR CANCER IMMUNOTHERAPY - PROJECT SUMMARY RECENTLY, WE DISCOVERED THAT ADOPTIVE TRANSFER OF CD39KO TUMOR-SPECIFIC (MIXED CD4+ AND CD8+) T CELLS, RESULTED IN LONG-TERM SURVIVAL OF MICE BEARING LARGE ESTABLISHED TUMORS. UNEXPECTEDLY, WE FOUND THAT THESE T CELLS PROMOTED KILLING OF ANTIGEN-LOSS-VARIANTS (ALVS) IN VIVO AND PREVENTED TUMOR RECURRENCE. MOREOVER, TRANSFER OF CD39KO, BUT NOT CONTROL KO, TUMOR-SPECIFIC T CELLS ERADICATED LARGE CHIMERIC TUMORS THAT CONTAINED 10% OF ALVS AND RESULTED IN LONG-TERM TUMOR-FREE SURVIVAL AND PROTECTION AGAINST RECHALLENGE WITH ALV TUMOR CELLS. BASED ON THESE NOVEL FINDINGS, WE HYPOTHESIZE THAT TRANSFER OF TUMOR-SPECIFIC CD39KO T CELLS WILL ERADICATE LARGE ESTABLISHED TUMORS AND PREVENT RECURRENCE OF ALV TUMORS, DUE TO THEIR ABILITY TO DIRECTLY KILL THE TUMOR CELLS AND INDUCE ANTI-ALV RESPONSES. AIM 1 WILL DETERMINE THE CONTRIBUTION OF TYPE I IFN PRODUCTION AT THE TUMOR SITE IN PREVENTING RECURRENCE OF ALV TUMORS. AIM 2 WILL DETERMINE THE ROLE OF CD39KO T CELLS IN THE RECRUITMENT OF INFLAMMATORY MYELOID CELLS AND THE INDUCTION OF TYPE I IFN PRODUCTION FOR TUMOR CLEARANCE. AIM 3 WILL DETERMINE WHETHER HUMAN TUMOR-SPECIFIC CD39KO T CELLS ARE ALSO ENDOWED WITH THESE ABILITIES TO EFFECTIVELY ERADICATE HUMAN TUMORS IN HUMANIZED MICE. THESE INNOVATIVE AND MECHANISTIC STUDIES WILL SHED LIGHT ON THE MECHANISMS UNDERLYING CD39KO T CELL-MEDIATED ANTITUMOR IMMUNITY AND WILL THUS ESTABLISH A FOUNDATION FOR TRANSLATING THIS DISCOVERY INTO MORE EFFECTIVE IMMUNOTHERAPIES USING TUMOR-SPECIFIC T-CELL SUBSETS IN HUMAN CANCERS.

MODULATION OF CHROMATIN DYNAMICS TO PROMOTE TRANSPLANT SURVIVAL - PROJECT SUMMARY REJECTION REMAINS A MAJOR HURDLE TO LONG-TERM TRANSPLANT SURVIVAL, IN WHICH TEFF CELLS ARE PROMINENTLY INVOLVED. WE SURMISE THAT BESIDES SIGNALS FROM THE TCR, COSTIMULATORY AND CYTOKINE RECEPTORS, TARGETING THE “T CELL EPIGENOME” DOWNSTREAM OF T CELL ACTIVATION MAY REPRESENT NOVEL THERAPEUTIC OPPORTUNITIES IN THE INDUCTION OF TRANSPLANT TOLERANCE. NOW WE HAVE NEW PRELIMINARY DATA SHOWING THAT THE CHROMATIN IN TEFF CELLS ARE HIGHLY COMPARTMENTALIZED, IN THAT “SUPER ENHANCERS” AT ACCESSIBLE CHROMATIN REGIONS APPEAR TO CONTROL A TEFF CELL FATE BY RECRUITING A CHROMATIN READER CALLED BRD4. WE THEN DEVELOPED A BRD4-FLOXED MICE AND SHOWED THAT CONDITIONAL DELETION OF BRD4 IN T CELLS COMPLETELY ABROGATED THE LETHAL AUTOIMMUNE PHENOTYPE IN SCURFY MICE AND PRODUCED LONG-TERM ALLOGRAFT SURVIVAL IN BRD4F/FCD4-CRE MICE. THUS, DISSECTING MECHANISTICALLY HOW BRD4 EPIGENETICALLY REGULATES TEFF CELLS AND TRANSPLANT SURVIVAL IS THE CENTRAL GOAL OF THIS PROPOSAL. OUR WORKING HYPOTHESIS IS THAT BRD4 OCCUPANCY AT THE ACCESSIBLE CHROMATIN REGIONS LOCKS ACTIVE CHROMATIN MODULES IN AN ACCESSIBLE STATE OR TRIGGERS REORGANIZATION OF “SUPER ENHANCERS” INTO TRANSCRIPTIONALLY HYPER-ACTIVE “HOTSPOTS” TO DRIVE A TEFF CELL FATE. WE PROPOSED 3 AIMS IN THIS PROPOSAL TO TEST THIS HYPOTHESIS. AIM 1 IS TO DETERMINE WHETHER BRD4 LOCKS “ACTIVE” CHROMATIN MODULES IN AN ACCESSIBLE STATE, ALLOWING THE FORMATION OF 3D CHROMATIN CONFIGURATIONS TO SPECIFY A TEFF CELL FATE, AND AIM 2 IS TO EXAMINE WHETHER BRD4 ENABLES STABLE SE “HOTSPOTS”, ALLOWING TEFF CELLS TO ESTABLISH ESSENTIAL FEATURES OF TEFF PROFILES. AIM 3 IS TO TEST WHETHER THERAPEUTICALLY TARGETING BRD4 ENABLES LONG-TERM ALLOGRAFT SURVIVAL IN A HEART TRANSPLANT MODEL. WE BELIEVE THAT THE PROPOSED STUDIES WILL UNCOVER NEW INSIGHTS AND OPEN NEW THERAPEUTIC OPPORTUNITIES IN TRANSPLANTATION. THE ANIMAL MODELS, TOOLS, AND CUTTING-EDGE TECHNOLOGIES WE HAVE DEVELOPED IN THE LAB PUT US IN A UNIQUE POSITION TO CARRY OUT THE PROPOSED STUDIES.

MYELIN JUNCTION THERAPY IN PERIPHERAL NEUROPATHIES

THE UNIQUE ROLES OF TUMOR-SPECIFIC TH9 CELLS FOR SOLID TUMOR ERADICATION - PROJECT SUMMARY OUR EARLY STUDIES HAVE CHARACTERIZED IL-9-PRODUCING CD4+ T HELPER (TH9) CELLS AS AN ANTITUMOR T CELL SUBSET. RECENTLY, WE ALSO REVEALED TH9 CELLS AS A NOVEL T CELL PARADIGM FOR ACT – THEY ARE LESS EXHAUSTED, FULLY CYTOLYTIC, AND HYPERPROLIFERATIVE; AND EFFICIENTLY KILLS TARGETED ANTIGEN-POSITIVE TUMOR CELLS. IN THE CURRENT PROJECT, WE WILL USE TUMOR MODELS THAT FAITHFULLY RECAPITULATE THE CLINICAL SCENARIO OF ACT IN SOLID TUMORS, TO UNCOVER UNIQUE FEATURES OF TUMOR-SPECIFIC TH9 CELLS THAT ENABLE THEM TO EFFICIENTLY ERADICATE ADVANCED SOLID TUMORS. WE HYPOTHESIZE THAT ANTI-SOLID TUMOR ACTIVITY OF TH9 CELLS ARE MAINLY ATTRIBUTED TO BOTH ENHANCED TH9 CELL PENETRATION INTO STROMA-RICH SOLID TUMORS AND RESTRAIN RELAPSE CAUSED BY ACQUIRED RESISTANCE DUE TO HIGH HETEROGENEITY IN TUMOR ANTIGEN EXPRESSION. AIM 1 WILL DETERMINE THE ROLE OF PU.1-DEPENDENT MMP12 PRODUCTION IN TH9 CELLS FOR THEIR EFFICIENT PENETRATION INTO SOLID TUMORS TO EXERT ANTITUMOR FUNCTIONS. AIM 2 WILL DETERMINE THE ROLE OF TH9 CELLS IN PROMOTING AN EATP-ENRICHED MILIEU TO ERADICATE THE TANTS IN SOLID TUMORS AND PREVENT ACQUIRED RESISTANCE. OUR PROPOSED STUDIES WILL IDENTIFY TUMOR-SPECIFIC TH9 CELLS AS THE FIRST ANTITUMOR T CELL SUBSET THAT ARE ENDOWED WITH THE CAPACITY TO ELIMINATE SOLID TUMORS WITH THE HETEROGENEITY IN TUMOR ANTIGEN EXPRESSION. THIS TRANSLATIONALLY RELEVANT WORK HOLDS PROMISE TO SIGNIFICANTLY ADVANCE THE THERAPEUTIC INDEX OF ACT IN SOLID TUMORS AND COULD THEN LAY THE FOUNDATION FOR FUTURE CLINICAL TRIALS.

EXPLORING THE ROLE OF ESTROGEN RECEPTOR BETA IN PROGRESSION AND METASTASIS OF INFLAMMATORY BREAST CANCER

ROLE OF INTEGRIN VLA-6 IN SUPPRESSION OF BONE FORMATION IN MYELOMA

NOVEL GENE DELIVERY TO MODULATE THE TUMOR MICROENVIRONMENT AND ANTIGEN-SPECIFIC ANTITUMOR IMMUNITY - ABSTRACT TECHNOLOGICAL ADVANCEMENTS HAVE YIELDED NOVEL DRUG TARGETS AND IMPROVED CANCER DETECTION, BUT TRANSFORMATIVE INNOVATIONS IN TREATMENT MODALITIES ARE LACKING. LEVERAGING OUR TEAM'S EXPERTISE IN GENE THERAPY, BREAST CANCER, CANCER IMMUNOLOGY, AND BREAST CANCER ANIMAL AND PATIENT-DERIVED XENOGRAFT MODELS, WE PROPOSE TO USE RECOMBINANT ADENOVIRUS WITH A HIGHER TUMOR-SPECIFIC EXPRESSION OF HERPES SIMPLEX VIRUS-THYMIDINE KINASE (HSV-TK), WHICH IS A SUICIDE GENE THAT CAUSES TUMOR CELL DEATH. WE WILL COMBINE HSV-TK WITH OUR IMMUNE CHECKPOINT ANTIBODY (I.E. HMR-101) THAT WILL SPECIFICALLY TARGET TUMOR CELLS, RELEASE TUMOR ANTIGENS AND CHANGE THE TUMOR MICROENVIRONMENT TO BE MORE FAVORABLE FOR IMMUNE THERAPY. THE IMMUNOTHERAPY WE ARE DEVELOPING WILL MODIFY CANCER-PROMOTING IMMUNE CELLS TO BECOME ANTI-CANCER IMMUNE CELLS. THEREFORE, AN ARMY OF ANTI-CANCER IMMUNE CELLS TRAINED TO RECOGNIZE CANCEROUS CELLS WILL MOBILIZE AND CIRCULATE AROUND THE BODY TO DESTROY METASTASES. GIVEN THE PREDOMINANCE OF TUMOR-ASSOCIATED IMMUNOSUPPRESSIVE MYELOID CELLS IN TRIPLE-NEGATIVE BREAST CANCER, LOCAL DELIVERY OF HMR-101 COULD EFFECTIVELY REPROGRAM THE TUMOR MICROENVIRONMENT TO AN ANTI-TUMOR IMMUNE PHENOTYPE. THE VIRAL TRANSDUCTION EFFICIENCY OF OUR VIRAL VECTOR IS VERY HIGH AT THE TUMOR SITE WITH CHIMERIC VIRAL VECTOR (LOW NEUTRALIZATION ANTIBODY), AND IT WILL BE LOW SPREADING TO OTHER ORGANS. THIS VIRAL VECTOR IS SAFE AND APPROVED BY THE FDA AND AT LOW COST FOR HUMAN USE. AS SUCH, OUR INTRATUMORAL STRATEGY WOULD ALLOW FOR MAXIMIZING THE FULL POTENCY OF THE IMMUNOTHERAPEUTIC ANTIBODY, THUS ENHANCING THERAPEUTIC INDEX. WE ENVISION THIS TECHNOLOGY TO MOVE FORWARD TO CLINICAL TRIALS, STRONGLY SUPPORTED BY THE SUCCESS OF OUR RECENTLY COMPLETED STOMP TRIAL FOR METASTATIC TRIPLE-NEGATIVE BREAST CANCER OR TNBC (NCT03004183, HSV-TK, RADIATION, AND IMMUNE CHECKPOINT THERAPY ON METASTATIC TNBC). CLINICALLY, WE ENVISION THAT OUR INTRATUMOR GENE DELIVERY OF HMR-101 WILL REPROGRAM THE IMMUNE SYSTEM, PREVENT PATIENTS FROM RELAPSE, AND PROLONG ANTI-TUMOR IMMUNITY, THUS BENEFITING PATIENTS RECEIVING IMMUNOTHERAPY. FOR TRIPLE-NEGATIVE BREAST CANCER PATIENTS WHO TYPICALLY HAVE DISMAL CLINICAL RESPONSE DUE TO HIGH METASTATIC RATE AND RECURRENCE, THE POTENTIAL CLINICAL IMPACT OF OUR INTRATUMORAL VIRAL DELIVERY CONTAINING NOVEL IMMUNE CHECKPOINT ANTIBODY IS PROFOUND. OVERALL, THE THERAPEUTIC REACH OF OUR NOVEL THERAPEUTIC REGIMEN IS EXPANSIVE, OFFERING A TRULY TRANSFORMATIVE APPROACH TO CANCER TREATMENT.

TARGETING ADIPOCYTE-SECRETED CHEMERIN FOR CHEMOTHERAPY RESISTANCE IN MYELOMA

NOVEL TARGETED THERAPEUTICS FOR BREAST CANCER

IMPROVING BETA-CELL FUNCTION IN MEXICAN AMERICAN WOMEN WITH PREDIABETES

TUMOR VASCULATURE-TARGETED NANOTHERAPEUTICS FOR DNA DAMAGE RESPONSE

THERAPEUTICALLY RELEVANT TARGETS OF TWIST1 DIMERS IN GLIOMA

DECOUPLING ACUTE TOXICITIES AND ANTITUMOR EFFICACY IN ADOPTIVE CELL THERAPY - PROJECT SUMMARY ADOPTIVE CELL THERAPY (ACT) WITH CHIMERIC ANTIGEN RECEPTOR (CAR) T CELLS HAS DEMONSTRATED IMPRESSIVE RESPONSE RATES IN B CELL MALIGNANCIES, BUT ACT HAS NOT MEDIATED SUSTAINED RESPONSES IN SOLID TUMORS. CD19 CAR T CELL THERAPY HAS REACHED UP TO 80% RESPONSE RATE IN THE CLINIC; HOWEVER, THE MAIN SIDE EFFECTS ARE CYTOKINE RELEASE SYNDROME (CRS) AND IMMUNE EFFECTOR CELL-ASSOCIATED NEUROTOXICITY SYNDROME (ICANS), WHICH OCCUR IN 37–83% AND ABOUT 25-35% OF PATIENTS, RESPECTIVELY. FURTHERMORE, ONE OF THE MAJOR OBSTACLES IN ACT IS THE HETEROGENEITY OF TARGETED ANTIGENS AND RELAPSE DUE TO ANTIGEN ESCAPE. RECENTLY, WE SCREENED A COHORT OF 16 FDA-APPROVED ANTI-INFLAMMATORY DRUGS AND IDENTIFIED CLOFAZIMINE (CLF) AS THE TOP CANDIDATE FOR ITS DESIRED BIFUNCTIONAL EFFECT FOR ANTI-CRS/ICANS AND ANTI-ANTIGEN ESCAPE ROLES. AIM 1 WILL DETERMINE THE ROLE OF CLF IN REDUCING MACROPHAGE-DERIVED ROS TO CURTAIL CRS/ICANS. AIM 2 WILL DETERMINE THE ROLE OF CLF IN DRIVING DSRNA/DSDNA SIGNALS IN MACROPHAGES FOR THE ERADICATION OF TUMORS. WE EXPECT THIS STUDY TO DEMONSTRATE THE ABILITY OF CLF IN POTENTIATING THE ANTI-ANTIGEN ESCAPE CAPACITY IN ACT, CURBING INTRACTABLE CRS, AND MAY ALSO FILL A DESPERATE CLINICAL NEED TO IMPROVE THE DISMAL PATIENT SURVIVAL WITH ICANS. THIS STRATEGY OF REPURPOSING THE CLINICALLY APPROVED CLF MAY HOLD GREAT PROMISE TO OVERCOME A CRITICAL OBSTACLE IN REALIZING THE FULL POTENTIAL OF ACT WITH CAR-T CELLS. THIS TRANSLATIONALLY RELEVANT WORK COULD THEN LAY THE FOUNDATION FOR FUTURE CLINICAL TRIALS.

UNDERSTANDING THE MECHANISMS THAT REGULATE CYTOPLASMIC CAPPING AND DEFINING ITS CONTRIBUTIONS TO POST-TRANSCRIPTIONAL GENE REGULATION

NOVEL THERAPY TARGETING REFRACTORY COLON CANCER

A NOVEL T-CELL SUBSET ABLE TO KILL RELAPSED CANCERS

RELATIONSHIP OF THE HUMAN ASTROCYTE MATRISOME WITH SYNAPTIC NETWORKS - PROJECT SUMMARY-ABSTRACT ASTROCYTES ARE HIGHLY-ABUNDANT CELLS IN THE NERVOUS SYSTEM AND THEY PLAY A CRITICAL ROLE IN THE ORCHESTRATION OF NEURONAL SYNAPTIC NETWORKS. ONE OF THE PRIMARY MECHANISMS THROUGH WHICH THEY INFLUENCE NEURONAL SYNAPSES IS THOUGHT TO INVOLVE SIGNALING VIA A DIVERSE MILIEU OF EXTRACELLULAR PROTEINS (I.E., THE ASTROCYTE MATRISOME). HOWEVER, IT REMAINS UNCLEAR WHICH COMPONENTS OF THE ASTROCYTE MATRISOME ARE NECESSARY AND SUFFICIENT FOR THE FORMATION AND STRENGTHENING OF NEURONAL EXCITATORY SYNAPSES, ESPECIALLY IN HUMAN-SPECIFIC CELLS DUE TO CURRENT LIMITATIONS IN EXPERIMENTALLY INVESTIGATING HUMAN NEURAL NETWORKS USING TRADITIONAL MONOLAYER CULTURES AND IMMATURE ORGANOIDS. TO OVERCOME THESE TECHNICAL LIMITATIONS, WE WILL UTILIZE OUR RECENTLY OPTIMIZED APPROACH WHICH GENERATES AND ANALYZES BIOENGINEERED NEURAL ORGANOIDS THAT ARE COMPOSED OF SPECIFIC NUMBERS OF POST-MITOTIC ASTROCYTES AS WELL AS NEURONS THAT ARE DIRECTLY TRANSDIFFERENTIATED FROM HUMAN PLURIPOTENT STEM CELLS. SPECIFICALLY, WE WILL USE THESE BIOENGINEERED NEURAL ORGANOIDS TO TEST THE HYPOTHESIS THAT MATURE HUMAN ASTROCYTES PRODUCE A CELL TYPE-RESTRICTED, MULTI-COMPONENT, ACTIVITY-DEPENDENT MATRISOME THAT ACCELERATES THE FORMATION AND FUNCTION OF NEURONAL SYNAPTIC NETWORKS. OUR PRELIMINARY DATA CONFIRMS FEASIBILITY OF OUR ORGANOID- BASED APPROACH TO TEST THIS HYPOTHESIS AND HAS IDENTIFIED TOP CANDIDATE PROTEINS POTENTIALLY UNDERLYING THE ASTROCYTE-TO-NEURON INFLUENCE ON SYNAPSES. IN AIM 1, WE WILL DETERMINE WHETHER HUMAN ASTROCYTE THROMBOSPONDIN 1 PROTEIN IS A SUFFICIENT AND NECESSARY INDUCER OF HUMAN NEURONAL SYNAPSES. WE WILL USE A COMBINATION OF GENETIC ENGINEERING AND DRUG TREATMENT TO CONCLUDE WHETHER THROMBOSPONDIN 1 PROMOTES STRUCTURAL AND FUNCTION EXCITATORY SYNAPTIC NETWORKS AND IF IT ACTS THROUGH SIGNALING TO THE NEURONAL ALPHA2DELTA- 1 RECEPTOR. IN AIM 2, WE WILL DEFINE THE EXTRACELLULAR HUMAN ASTROCYTE MATRISOME AND TEST ITS INFLUENCE UPON NEURONAL SYNAPSE FORMATION USING A NOVEL INDIRECT COCULTURE APPROACH. COCULTURES WILL BE ENABLED BY CELLULAR ENCAPSULATION WITHIN ALGINATE HYDROGEL CAPSULES TO ELUCIDATE THE EXTRACELLULAR ASTROCYTE MATRISOME COMPONENTS, TEST THEIR EFFECT UPON NEURONAL ORGANOIDS, AND IDENTIFY RELEVANT RECEPTOR-LIGAND PAIRS. WE WILL INVESTIGATE WHETHER ASTROCYTE-DERIVED EXTRACELLULAR THROMBOSPONDIN 1 AND/OR CHONDROITIN SULFATE PROTEOGLYCANS INFLUENCES SYNAPSE FORMATION AND FUNCTION. FINALLY, IN AIM 3, WE WILL TEST HOW NEURONAL ACTIVITY INFLUENCES THE SYNAPSE-PROMOTING CHARACTERISTICS OF THE HUMAN ASTROCYTE MATRISOME. NEURONS WILL BE ACTIVATED AND PACED USING OPTOGENETIC TOOLS, AND THE RESULTANT EFFECT ON COCULTURED ASTROCYTES WILL BE DETERMINED USING A COMBINATION OF SINGLE CELL RNASEQUENCING, PHARMACOLOGICAL TREATMENT, PROTEIN ASSAYS, AND CALCIUM IMAGING. THE COMPLETION OF OUR AIMS WILL DELIVER NOVEL EXPERIMENTAL CELL LINES AND PROTOCOLS TO THE SCIENTIFIC COMMUNITY, AND MAY IDENTIFY NOVEL APPROACHES TO ACCELERATE NEURONAL SYNAPTIC NETWORK FORMATION IN ORGANOID-BASED MODEL SYSTEMS. BROADLY, WE EXPECT OUR STUDIES TO MAKE SIGNIFICANT CONTRIBUTIONS TO THE NEUROBIOLOGY FIELD BY IDENTIFYING AND DEFINING INTERCELLULAR SIGNALING MECHANISMS BETWEEN HUMAN ASTROCYTES AND NEURONAL SYNAPSES.

ETIOLOGICAL LINKAGE OF DNA DAMAGE/REPAIR DEFICIENCY IN NEURODEGENERATIVE DISEASES

P38 MAPK IS A MOLECULAR SWITCH THAT CONTROLS THE ACQUIRED RESISTANCE IN ADOPTIVE CELL THERAPY

S100A4 MEDIATED IMMUNE SUPPRESSION IN GBM - PROJECT SUMMARY GLIOBLASTOMA (GBM) IS THE MOST COMMON AND AGGRESSIVE MALIGNANT BRAIN TUMOR IN ADULTS AND IS VIRTUALLY INCURABLE. DESPITE AGGRESSIVE TREATMENTS THAT INCLUDE SURGERY, RADIATION THERAPY, AND CHEMOTHERAPY, THE AVERAGE SURVIVAL IS 15.4 MONTHS, WITH LESS THAN 5% OF PATIENTS SURVIVING > 5 YEARS. IMMUNOTHERAPY IS A PROMISING NEW APPROACH TO TREAT GBM AS IT HARNESSES ONE’S OWN IMMUNE SYSTEM TO RECOGNIZE AND KILL ABERRANT CANCER CELLS. UNFORTUNATELY, ONGOING TRIALS WITH IMMUNOTHERAPIES SHOW DISAPPOINTING RESULTS IN MOST GBM PATIENTS, INDICATING THAT WE NEED TO BETTER UNDERSTAND THE INTRICATE AND DYNAMIC INTERACTIONS BETWEEN GLIOMA AND IMMUNE CELLS THAT GENERATE AND MAINTAIN HIGHLY IMMUNE SUPPRESSIVE MICROENVIRONMENT IN GBM. THE MAJOR GOAL OF THIS APPLICATION IS TO TEST OUR HYPOTHESIS THAT S100A4 IS A CRITICAL REGULATOR OF THE GBM IMMUNE LANDSCAPE. S100A4 IS A SMALL CALCIUM BINDING PROTEIN THAT FUNCTIONS INTRACELLULARLY AS WELL AS EXTRACELLULARLY. WE RECENTLY REPORTED THAT S100A4 IS NECESSARY FOR HUMAN AND MOUSE GLIOMA STEM CELL SELF- RENEWAL. OUR SINGLE CELL RNA-SEQUENCING ANALYSES OF HUMAN GBM SHOWS THAT S100A4 IS EXPRESSED IN BOTH GLIOMA CELLS AND IMMUNE SUPPRESSIVE LEUKOCYTES. CONSISTENTLY, TCGA DATA ANALYSES INDICATE THAT S100A4 EXPRESSION IS STRONGLY CORRELATED WITH PRIMARY AND RECURRENT GBM PATIENT SURVIVAL, THE MESENCHYMAL SUBTYPE, AND IMMUNE-SUPPRESSIVE MACROPHAGE MARKERS (SUCH AS CD163, CD206, AND IL10). IMPORTANTLY, OUR UNPUBLISHED RESULTS SHOW THAT S100A4 IN HUMAN AND MOUSE GLIOMA CELLS REGULATES EXPRESSION OF CHEMOKINES AND CYTOKINES THAT CONTROL IMMUNE CELL INFILTRATION AND POLARIZATION TOWARDS IMMUNE SUPPRESSIVE PHENOTYPES. NOTABLY, KNOCKING DOWN S100A4 IN GLIOMA CELLS IS SUFFICIENT TO REPROGRAM THE IMMUNE ENVIRONMENT AND ALLOW DRAMATIC INCREASE IN T-CELL INFILTRATION AND ACTIVATION IN MOUSE GLIOMAS. FURTHERMORE, DELETION OF S100A4 IN THE HOST MOUSE IS ALSO SUFFICIENT TO REPROGRAM THE GLIOMA IMMUNE LANDSCAPE AND EXTEND SURVIVAL (MANUSCRIPTS IN PREPARATION). THEREFORE, WE HYPOTHESIZE THAT S100A4 FUNCTIONS IN BOTH GLIOMA AND IMMUNE TO PROMOTE IMMUNE-SUPPRESSIVE ENVIRONMENT IN GBM THROUGH MULTIPLE MECHANISMS. TO TEST THIS HYPOTHESIS, WE WILL: 1) ELUCIDATE MECHANISMS THROUGH WHICH S100A4 EXPRESSION IN GLIOMA CELLS PROMOTES LOCAL IMMUNE-SUPPRESSIVE MICROENVIRONMENT IN GBM (AIM 1); AND 2) DETERMINE MECHANISMS THROUGH WHICH S100A4 EXPRESSION IN IMMUNE CELLS CONTROLS MYELOID AND T CELL TRAFFICKING, POLARIZATION, AND FUNCTION (AIM2). SUCCESSFUL COMPLETION OF THIS STUDY WILL REVEAL HOW S100A4 REGULATES GLIOMA ASSOCIATED IMMUNE CELL INFILTRATION/POLARIZATION AND IDENTIFY A POTENTIAL NOVEL IMMUNOTHERAPY TARGET TO TREAT GBM.

RNA-GAIN-OF-FUNCTION PATHOGENESIS IN SCA10

VENOUS ETHANOL ABLATION IN ISCHEMIC VENTRICULAR TACHYCARDIA- VELVET TRIAL - ABSTRACT RADIOFREQUENCY (RF) ABLATION OF VENTRICULAR TACHYCARDIA (VT) IN ISCHEMIC CARDIOMYOPATHY IS FRAUGHT WITH LIMITATIONS DUE TO SUBOPTIMAL EFFICACY, RISK OF COMPLICATIONS, AND FREQUENT NEED FOR REPEAT PROCEDURES. ISCHEMIC VT ARISES AS A RESULT OF REENTRANT CIRCUITS WITHIN OR AROUND THE MYOCARDIAL SCAR OF AN INFARCT. THE CO-LOCALIZATION OF VENTRICULAR ARTERIES, VEINS, AND NERVES, IS AN ANATOMICAL FACT, DETERMINED BY THE EMBRYOLOGY OF CORONARY VESSELS. JUST AS MYOCARDIAL INFARCTIONS HAVE A “CULPRIT” OR “INFARCT-RELATED ARTERY”, THERE COMMONLY EXISTS AN “INFARCT-RELATED VEIN” OR VEINS IN VT SUBSTRATE. ADDITIONALLY, IT IS WELL KNOWN THAT AUTONOMIC INNERVATION -IN ANATOMICAL PROXIMITY TO THE VEINS- PLAYS AN IMPORTANT ROLE IN POST-MI ARRHYTHMOGENESIS. WE HAVE DEVELOPED AN APPROACH TO TARGET ABLATION-REFRACTORY VTS VIA ETHANOL DELIVERY IN THE CORONARY VEINS THAT PROVIDE VENOUS RETURN FROM ARRHYTHMOGENIC SITES (VENOUS ETHANOL, VE). BEYOND AN INITIAL SET OF CASE REPORTS, WE HAVE VALIDATED THE UTILITY OF VE IN A LARGE, MULTINATIONAL REGISTRY, IN WHICH WE ESTABLISH THE SAFETY AND EFFICACY OF VE IN RF-REFRACTORY VT. GIVEN THE CO-LOCALIZATION OF EPICARDIAL ARTERIES, VEINS AND NERVES, VE MAY BE PARTICULARLY SUITED TO IMPACT INFARCT INNERVATION. THUS, A CENTRAL GOAL OF THIS PROPOSAL IS TO CAPITALIZE ON THE PRESENCE OF CORONARY VEINS ON THE EPICARDIAL ASPECT OF A MYOCARDIAL SCAR AS A THERAPEUTIC OPPORTUNITY OF UNIQUE MECHANISMS. WE HYPOTHESIZE THAT VE ADDED TO CONVENTIONAL CATHETER ABLATION IMPROVES THE RESULTS OF VT ABLATION. WE PROPOSE A SINGLE-SITE, INVESTIGATOR-INITIATED CLINICAL TRIAL ON VE. IN AIM 1-R61 PHASE-, WE PROPOSE TO FINALIZE THE DESIGN OF RANDOMIZED CLINICAL TRIAL TO ASSESS THE CLINICAL EFFICACY, AND SAFETY OF VE WHEN USED IN COMBINATION WITH RF ABLATION COMPARED WITH RF ABLATION ALONE -VENOUS ETHANOL FOR LEFT VENTRICULAR ISCHEMIC VENTRICULAR TACHYCARDIA -VELVET CLINICAL TRIAL. THE TRIAL WILL INCLUDE AN INVESTIGATIONAL NEW DRUG (IND) AUTHORIZATION BY THE FDA. PATIENTS WITH ISCHEMIC VT WILL BE RANDOMIZED TO CONVENTIONAL ENDOCARDIAL ABLATION ALONE, VS COMBINED WITH VE IN THE INFARCT-RELATED VEIN. IN AIM 2 -R33 PHASE- WE WILL ENROLL A TOTAL OF 156 PATIENTS, AND COLLECT EFFICACY, SAFETY AND PROCEDURAL DATA ON THE IMPACT OF VE ADDED TO CATHETER ABLATION. THIS TRIAL WILL ALLOW FOR A WEALTH OF NEW IMAGING DATA TO BE COLLECTED THAT WILL CHARACTERIZE THE EXTENT OF MYOCARDIAL SCAR AND INNERVATION BEFORE AND AFTER VE -COMPARED TO ENDOCARDIAL RF ALONE. IN AIM 3 -R33 PHASE- WE WILL COLLECT MULTI-MODALITY IMAGING DATA CHARACTERIZING THE VT SUBSTRATE BEFORE AND AFTER ABLATION -WITH CATHETER ABLATION ALONE VS COMBINED WITH VE. CARDIAC MAGNETIC RESONANCE, VENOUS CT ANGIOGRAMS AND REGIONAL ADRENERGIC INNERVATION MAPS WITH POSITRON EMISSION TOMOGRAPHY (PET) SCANS OF INNERVATION TRACERS (11C HYDROXYEPHEDRINE, 11C-HED) WILL PROVIDE A COMPLETE STRUCTURAL ASSESSMENT OF THE VT SUBSTRATE, BEFORE AND AFTER ABLATION. IF COMPLETED, THE PROJECT WILL VALIDATE A NEW PROCEDURAL STRATEGY AND WILL PROVIDE KEY NEW INSIGHTS INTO THE STRUCTURAL DETERMINANTS OF VT ABLATION SUCCESS.

SYSTEMS-WIDE ANALYSIS OF OXIDATIVE STRESS-RESPONSIVE M6A EPITRANSCRIPTOME

THE GERM CELL GENE TDRD1 AS A NOVEL PROSTATE CANCER BIOMARKER

STUDIES OF A NEW CHECKPOINT REGULATOR IN THE CONTROL OF INTESTINAL INFLAMMATION - PROJECT SUMMARY THE INNATE IMMUNE SYSTEM IS THE FIRST LINE OF HOST DEFENSE AGAINST INVADING PATHOGENIC MICROORGANISMS. INNATE IMMUNE CELLS CAN RECOGNIZE THESE PATHOGENS TO INDUCE CYTOKINES AGAINST INFECTIONS. HOWEVER, THIS IMMUNE RESPONSE MUST BE TIGHTLY REGULATED TO EFFECTIVELY ELIMINATE INVADING MICROORGANISMS WHILE MINIMIZING TISSUE INFLAMMATION. PROTEIN UBIQUITINATION IS IMPORTANT BOTH IN SIGNAL TRANSDUCTION AND IN PROTEASOME-MEDIATED PROTEIN DEGRADATION AND IS A PIVOTAL REGULATORY MECHANISM FOR THE INNATE IMMUNE SYSTEM. A KEY COMPONENT OF THE UBIQUITINATION SYSTEM IS E3 LIGASE TO SPECIFICALLY RECOGNIZE THE SUBSTRATE FOR MODIFICATION. WE RECENTLY FOUND THAT A TRIM FAMILY MEMBER, E3 LIGASE TRIM29, IS HIGHLY EXPRESSED IN INTESTINAL EPITHELIAL CELLS (IECS). IECS ACT AS A PHYSICAL BARRIER BETWEEN THE EXTERNAL ENVIRONMENT AND THE MUCOSAL IMMUNE SYSTEM. IMPORTANTLY, AFTER ACTIVATION, IECS BECOME INNATE IMMUNE-LIKE CELLS TO PRODUCE ROBUST PROINFLAMMATORY CYTOKINES. WE POSIT THAT TRIM29 IS A KEY IMMUNE REGULATOR TO SUPPRESS INNATE IMMUNE RESPONSE IN A CELL/TISSUE-SPECIFIC MANNER TO REDUCE INTESTINAL INFLAMMATION. NOTABLY, IN THE GUT, GENES ENCODING INFLAMMASOME COMPONENTS SUCH AS CARD9, NLRP3 AND CASPASES ARE ASSOCIATED WITH INTESTINAL INFLAMMATION, AND MICE DEFICIENT IN THESE GENES ARE SUSCEPTIBLE TO COLITIS AND FUNGAL INFECTION. HOWEVER, HYPERACTIVE INFLAMMASOME SIGNALING AND UNCONTROLLED IL-1 FAMILY CYTOKINES LEAD TO IMMUNE DISORDERS AND INFLAMMATORY DISEASE. FOR INSTANCE, OVERPRODUCTION OF INFLAMMASOME CYTOKINE IL-18, EXCESSIVE ACTIVATION OF NLRP3-INFLAMMASOME, OR ABERRANT IMMUNE RESPONSE TO FUNGUS PLAY A MAJOR PATHOGENIC ROLE IN COLITIS. THUS, THIS IMMUNE RESPONSE MUST BE FINELY TUNED DURING IMMUNE ACTIVATION FOR OPTIMAL PROTECTION AGAINST INFECTIONS WHILE AVOIDING UNWANTED INFLAMMATION. HOWEVER, LITTLE IS KNOWN ABOUT HOW SUCH ACTIVATION IS TERMINATED. OUR PRELIMINARY STUDIES SHOW THAT TRIM29 NEGATIVELY REGULATES IL-1 FAMILY CYTOKINES PRODUCED BY IECS DURING COLITIS OR FUNGAL INFECTION. MOREOVER, EXPRESSION OF TRIM29 ITSELF IS SIGNIFICANTLY DOWNREGULATED IN INFLAMED COLONS, WHEREAS PRODUCTION OF IL-1 FAMILY CYTOKINES IS ELEVATED. HOWEVER, IT IS UNKNOWN WHETHER LOST EXPRESSION OF TRIM29 IS REGULATED BY IL-1 FAMILY CYTOKINES AND/OR EPIGENETIC MODIFICATIONS TO CONTROL RESPONSES TO ENVIRONMENT STIMULI IN THE INTESTINES. WE HYPOTHESIZE THAT TRIM29 EXPRESSION IN IECS IS A CHECKPOINT REGULATOR OF IL-1 FAMILY CYTOKINES CRUCIAL TO REDUCE INTESTINAL INFLAMMATION IN COLITIS AND FUNGAL INFECTION. WE PROPOSE THE FOLLOWING THREE AIMS TO TEST THIS HYPOTHESIS: (1) TO DISSECT MOLECULAR PATHWAYS THAT CONTROL TRIM29 EXPRESSION IN EPITHELIAL CELLS UNDER NORMAL AND INFLAMMATORY CONDITIONS; (2) TO DEFINE THE MECHANISM BY WHICH TRIM29 INHIBITS IL-1 INFLAMMATION; (3) TO INVESTIGATE IN VIVO FUNCTIONS OF TRIM29 IN REGULATING IECS FUNCTION IN COLITIS AND ANTI-FUNGAL IMMUNITY.

IRF4-DEPENDENT T-CELL EFFECTOR PROGRAMS IN GOVERNING TRANSPLANT OUTCOMES

MOLECULAR MECHANISM OF STREPTOCOCCAL ADAPTATION TO HOST NUTRITIONAL DEFENSES

DEVELOPMENT OF NOVEL VACCINES FOR COCAINE ABUSE

THE ROLE OF THE NICOTINIC CHOLINERGIC PATHWAY IN RETINOPATHY OF PREMATURITY

"REPAIR CO-ORDINATION OF RADIATION-INDUCED CLUSTERED DAMAGE IN MAMMALIAN GENOMES"

TUMOR-CELL-SPECIFIC TARGETS FOR COMBINED HYPERTHERMIA AND RADIATION EFFECTS

PATHOGENESIS IN SEGMENTAL DEMYELINATION - PROJECT SUMMARY ABSTRACT: SEGMENTAL DEMYELINATION IS A PATHOLOGIC PROCESS OF STRIPPING OFF THE MYELIN SHEATH, WHICH SHUNTS CURRENT OUT OF AXONS AND RESULTS IN THE FAILURE OF ACTION POTENTIAL PROPAGATION IN A VARIETY OF PERIPHERAL NERVE DISEASES. HOWEVER, THE MOLECULAR PROGRAM THAT LEADS TO DEMYELINATION REMAINS UNCLEAR, WHICH HAMPERS THE THERAPEUTIC DEVELOPMENT FOR DEMYELINATING NEUROPATHIES. HUMANS WITH AUTOSOMAL RECESSIVE MUTATIONS IN THE FIG4 GENE DEVELOP CHARCOT-MARIE-TOOTH DISEASE TYPE-4J (CMT4J), A DISEASE CHARACTERIZED BY SEGMENTAL DEMYELINATION. OUR PRELIMINARY STUDIES IN A CMT4J MOUSE MODEL (FIG4-/-) HAVE DEMONSTRATED THAT SEGMENTAL DEMYELINATION IS ASSOCIATED WITH INCREASED INTRACELLULAR CA2+ IN MYELINATING SCHWANN CELLS AND ACCUMULATION OF MACROPHAGES IN THE SPINAL ROOTS. ADMINISTRATION OF A CA2+ CHELATOR SUPPRESSES THE DEMYELINATION IN FIG4-/- MICE. THEREFORE, THIS MOUSE IS AN APPROPRIATE MODEL TO INVESTIGATE THE MOLECULAR EVENTS UNDERLYING DEMYELINATION. TOWARD THIS END, WE WILL TEST OUR CENTRAL HYPOTHESIS THAT SIGNALS FROM FIG4-DEFICIENT AXONS AND/OR MACROPHAGES EXACERBATE THE OVERLOAD OF INTRACELLULAR CA2+ IN FIG4-DEFICIENT SCHWANN CELLS WHICH, IN TURN, LEADS TO SEGMENTAL DEMYELINATION. WE PROPOSE THREE SPECIFIC AIMS TO TEST OUR HYPOTHESIS BY DETERMINING IF: (1) FIG4-DEFICIENT SCHWANN CELLS ARE SENSITIZED TO DEMYELINATE UPON CHALLENGE WITH CA2+; (2) FIG4-DEFICIENT MACROPHAGES RELEASE CYTOKINE IL12B THAT TRIGGERS A FURTHER INCREASE IN INTRACELLULAR CA2+ IN FIG4-DEFICIENT SCHWANN CELLS, LEADING TO DEMYELINATION; AND (3) INDIVIDUAL PROTEINS IN THE PAS COMPLEX PLAY DISTINCT ROLES IN THE DEVELOPMENT AND MAINTENANCE OF MYELIN AND AXONS. THESE STUDIES HAVE THE POTENTIAL TO UNCOVER NOVEL MOLECULAR MECHANISMS UNDERLYING DEMYELINATING PERIPHERAL NEUROPATHIES AND IDENTIFY TARGETS FOR THERAPEUTIC DEVELOPMENT.

ATHEROPROTECTION VIA REDUCED PLASMA HIGH DENSITY LIPOPROTEIN-FREE CHOLESTEROL BIOAVAILABILITY

?PHARMACOLOGIC TARGETING OF NR4A1 AND NR4A2 TO ACTIVATE GLIOBLASTOMA TREATMENT RESPONSE? - RESISTANCE TO STANDARD OF CARE (SOC) TEMOZOLOMIDE (TMZ) AND RADIATION TREATMENT (XRT) LIMITS SURVIVAL OF THE MOST COMMON ADULT BRAIN CANCER, GLIOBLASTOMA (GBM), TO 12-18 MONTHS. DESPITE PROMISING RESULTS IN OTHER CANCERS, ROBUST IMMUNOSUPPRESSION IN GBM HAS ALSO LIMITED THE RESPONSES TO IMMUNE CHECKPOINT INHIBITORS (ICIS). RECIPROCAL INTERACTIONS IN THE GBM TUMOR MICROENVIRONMENT (TME) BETWEEN MESENCHYMAL CHANGES AND IMMUNOSUPPRESSION ENHANCE RESISTANCE TO CHEMORADIATION AND IMMUNOTHERAPY, RESPECTIVELY. THEREFORE, A COMPELLING THERAPEUTIC STRATEGY FOR GBM IS TO CONCURRENTLY REPROGRAM THE MESENCHYMAL AND IMMUNE SUPPRESSIVE TMES AND POTENTIATE ICI AND SOC THERAPY. THE NR4A1 AND NR4A2 ORPHAN NUCLEAR RECEPTORS ARE COMPELLING TARGETS TO ACHIEVE THIS GOAL. WE SYNTHESIZED A SERIES OF NOVEL BIS-INDOLE–DERIVED LIGANDS (CDIMS) WITH POTENT DUAL ANTAGONISM OF NR4A1 AND NR4A2 AND NEGLIGIBLE IN VIVO TOXICITY, OF WHICH SIX DEMONSTRATED NM RANGE KDS FOR BOTH RECEPTORS. GENETIC BASED NR4A 1 AND 2 LOSS OF FUNCTION STRONGLY ACTIVATES ANTI-CANCER IMMUNE RESPONSES BY REVERSING T-CELL EXHAUSTION WHICH UNDERLIES POOR ICI RESPONSES IN GBM. IN ADDITION, NR4A1/2 PROMOTE EMT IN OTHER CANCERS WHICH CONTRIBUTES TO CHEMORADIATION RESISTANCE. FINALLY, HIGH LEVELS OF NR4A1/2 EXPRESSION IN GBM ARE STRONGLY ASSOCIATED WITH DECREASED PATIENT SURVIVAL AFTER CONVENTIONAL TREATMENTS. THEREFORE, WE PROPOSE TO TEST THE HYPOTHESIS THAT CDIM INHIBITION OF NR4A1/2 REPROGRAMS THE GBM TME AND POTENTIATES ICI AND TMZ/XRT RESPONSES. FURTHER NR4A1/2 UPREGULATES PD-L1 AND TWIST1, KEY REGULATORS OF GBM IMMUNE SUPPRESSION AND MESENCHYMAL PHENOTYPES, RESPECTIVELY. PRELIMINARY STUDIES DEMONSTRATED THAT CDIMS INHIBIT PD-L1 AND TWIST1 EXPRESSION, REVERSE IMMUNE SUPPRESSIVE MYELOID AND T CELL PHENOTYPES AND PROLONG SURVIVAL OF EXPERIMENTAL GBMS. WE WILL TEST OUR HYPOTHESIS BY I) IDENTIFYING LEAD DUAL NR4A1/2 CDIM COMPOUNDS BASED ON INHIBITION OF MALIGNANT AND MESENCHYMAL GBM CELL PROPERTIES (PROLIFERATION, SELF-RENEWAL, INVASION) AND REVERSAL OF DYSFUNCTIONAL T CELL PHENOTYPES IN VIVO (AIM 1), II) ESTABLISHING THE FUNCTIONAL IMPACT OF NR4A1/2 CDIMS TO REPROGRAM THE IMMUNE SUPPRESSIVE AND MESENCHYMAL TME (AIM 2) AND III) QUANTIFYING THE EFFECTS OF NR4A1/2 CDIMS TO POTENTIATE ICI AND SOC RESPONSES IN MOUSE GBM SYNGENEIC MODELS AND DEFINE THE ROLE OF PD-L1 AND TWIST1 IN THEIR MECHANISMS OF ACTION (AIM 3). QUANTIFICATION OF POTENCY, PHARMACOKINETICS AND PRECLINICAL ANTI-TUMOR ACTIVITY ALONE AND IN COMBINATION WITH TO STANDARD OF CARE TMZ-XRT AND ICIS IS EXPECTED TO IDENTIFY CANDIDATE CDIMS FOR CONSIDERATION IN FUTURE CLINICAL TRIALS. THROUGH APPLICATION OF POWERFUL BIOINFORMATIC TOOLS THAT MODEL THE CANCER TME THROUGH INTEGRATION OF SCRNASEQ, CYTOF AND IMC DATA, THESE STUDIES ARE EXPECTED TO SHED NEW LIGHT ON NOVEL MECHANISMS BY WHICH MESENCHYMAL AND IMMUNE SUPPRESSIVE TMES INTERACT TO COORDINATELY DRIVE TREATMENT RESISTANCE IN GBM THROUGH NR4A1/2. TO ACHIEVE OUR GOALS, WE ASSEMBLED A UNIQUE MULTIDISCIPLINARY TEAM WITH EXPERTISE IN MEDICINAL CHEMISTRY, NEURO-ONCOLOGY, GBM BIOLOGY AND PRECLINICAL MODELING, AND IMMUNO-ONCOLOGY.

ROLE OF S-NITROSYLATION IN TRANSDIFFERENTIATION

NANOTECHNOLOGY-BASED TARGETING OF BREAST CANCER LIVER METASTASES

NANOPARTICLE DELIVERY OF MIRNA-BASED THERAPEUTICS TO OVERCOME CLINICAL CHALLENGES IN TRIPLE NEGATIVE BREAST CANCER - PROJECT SUMMARY TRIPLE-NEGATIVE BREAST CANCER (TNBC) HAS THE HIGHEST PATIENT DEATH RATE OF ALL BREAST CANCER SUBTYPES. SEVERAL MOLECULAR TARGETS HAVE BEEN IDENTIFIED FOR BREAST CANCER TREATMENT, BUT CURRENTLY, THERE IS NO APPROVED, BROADLY APPLICABLE TARGETED THERAPY FOR TNBC. THROUGH 10 YEARS OF RESEARCH, WE FOUND THAT ELONGATION FACTOR 2-KINASE (EF2K) EXPRESSION IS A CRITICAL DRIVER OF TNBC TUMORIGENESIS AND PROGRESSION. WE ALSO FOUND THAT MICRORNA-22 (MIR-22) EXPRESSION IS BROADLY REPRESSED IN TNBC PATIENTS, AND IS INVERSELY CORRELATED WITH EF2K EXPRESSION. FURTHER ANALYSIS REVEALED THAT MIR-22 SUPPRESSES TUMORS BY SPECIFICALLY BINDING TO EF2K, WHICH INHIBITS EF2K EXPRESSION AND REDUCES TUMOR GROWTH IN MULTIPLE TNBC MODELS. CONSIDERING THE CLINICAL SIGNIFICANCE AND POTENTIAL THERAPEUTIC VALUE OF EF2K IN TNBC, WE HAVE THUS DEVELOPED AN AXL RECEPTOR-TARGETED AXL APTAMER-COATED SLNP-MIR-22 NANOPARTICLE SYSTEM THAT CAN SPECIFICALLY DELIVER MIR-22 TO TNBC TUMORS IN VIVO (BUT DOES NOT LEAD TO MIR-22 ACCUMULATION IN NORMAL TISSUES). ON THE BASIS OF THIS PRELIMINARY WORK, WE HYPOTHESIZE THAT EF2K IS AN EFFECTIVE THERAPEUTIC TARGET IN TNBC, AND THAT TARGETING EF2K USING OUR AXL-APTAMER-SLNP-MIR-22 NANOTHERAPEUTICS CAN PROVIDE SIGNIFICANT THERAPEUTIC EFFICACY IN TNBC TREATMENT. HOWEVER, UNDERSTANDABLY, THIS THERAPEUTIC SYSTEM IS COMPLEX, AND IT HAS BEEN DIFFICULT TO FURTHER UNDERSTAND THE UNDERLYING BIOLOGICAL AND PHYSICAL PROCESSES THAT SIGNIFICANTLY IMPACT TREATMENT OUTCOME, AND TO IDENTIFY THE OPTIMAL DOSES AND DOSING SCHEDULES FOR MAXIMIZING TREATMENT EFFICACY. THEREFORE, IN THIS PROJECT, WE PROPOSE TO OVERCOME THIS CHALLENGE BY INTEGRATING BIOLOGICAL EXPERIMENTS WITH MATHEMATICAL MODELING BASED ON THE UNDERLYING BIOLOGICAL AND PHYSICAL MECHANISMS THAT ARE INVOLVED IN CANCER INVASION, DRUG PENETRATION, AND DRUG-CANCER CELL INTERACTIONS IN THE EF2K-TARGETED MIR-22 NANOTHERAPEUTICS FOR TNBC TREATMENT. OUR HYPOTHESIS WILL BE TESTED BY ACHIEVING THE FOLLOWING TWO SPECIFIC AIMS: 1) EXPERIMENTAL TESTING OF THE EF2K-TARGETED MIR-22 NANOTHERAPY (AIM 1), AND 2) MATHEMATICAL MODELING (AIM 2). IN AIM 1, WE WILL FOCUS ON CHARACTERIZING AND DETERMINING THE IN VIVO THERAPEUTIC EFFICACY OF EF2K-TARGETED MIR-22 MEDIATED THERAPIES IN ORTHOTOPIC MOUSE MODELS. IN AIM 2, WE WILL FOCUS ON DEVELOPING, TESTING, AND VALIDATING A MATHEMATICAL MODEL OF EF2K-TARGETED, MIR-22 BASED NANOTHERAPY, USING A LOGICALLY INTEGRATED STATISTICAL AND MULTISCALE MECHANISTIC MODELING APPROACH. EXPERIMENTAL DATA FROM AIM 1 WILL BE SUPPLIED TO AIM 2 FOR DEVELOPING AND VALIDATING THE MATHEMATICAL MODEL, AND EXPERIMENTS IN AIM 1 WILL BE GUIDED BY DISCOVERIES OBTAINED FROM COMPUTATIONAL INVESTIGATIONS IN AIM 2. THROUGH THIS ITERATION-BASED FEEDBACK APPROACH, THE MATHEMATICAL MODEL WILL BE USED TO PREDICT AND DETERMINE THE EFFECTS OF VARIOUS PARAMETERS, INCLUDING SIRNA DOSE AND DOSING SCHEDULES, ON TUMOR RESPONSE TO EF2K-TARGETED MIR-22 MEDIATED THERAPIES (WITH OR WITHOUT CHEMOTHERAPY), AND TO DETERMINE THE OPTIMAL DRUG DOSES AND DOSING SCHEDULES FOR OPTIMIZING THERAPEUTIC EFFICACY. THE LONG-TERM GOAL OF THIS PROJECT IS TO DEMONSTRATE THAT THIS MIR-22-BASED NANOTHERAPY IS SAFE AND EFFECTIVE, BOTH ALONE AND IN COMBINATION WITH STANDARD CHEMOTHERAPEUTIC AGENTS AS A CO-ADJUVANT THERAPY, AND TO COMPLETE PRECLINICAL DEVELOPMENT FOR POTENTIAL FUTURE CLINICAL TRANSLATION FOR TNBC PATIENTS.

DENDRITIC NANOMEDICINE FOR CANCER IMAGING AND TREATMENT

A BIFUNCTIONAL NANOMEDICINE FOR BOTH SPECIFIC IMAGING AND TARGETING THERAPY OF AN

CHEMOPREVENTION OF LUNG CANCER BY TARGETING LONIDAMINE TO MITOCHONDRIA

THE ROLE OF CD4+ T CELLS IN ANTITUMOR IMMUNITY

"PHOSPHOLIPIDS, METABOLIC SYNDROME AND RESPONSE TO INJURY"

INHIBITION OF OLA1: A NOVEL STRATEGY FOR ANTI-METASTASIS THERAPY

POR PROGRAM ON GENOMIC PREDICTION OF ANTIMICROBIAL RESISTANCE IN VRE

TUMOR-SPECIFIC CD8+ TC9 CELLS ACTIVATE HOST CD4+ T CELLS TO CONTROL ANTIGEN-LOST TUMORS - PROJECT SUMMARY ADOPTIVE CELL TRANSFER USING TUMOR-SPECIFIC T CELLS IS A PROMISING OPTION FOR CANCER TREATMENT AND THE MOST POWERFUL TUMOR-KILLING EFFECTOR T CELLS ARE CD8+ TYPE-1 CYTOTOXIC T CELLS (TC1). WE HAVE SHOWN THAT IL-9- SECRETING CD8+ TC9 CELLS MEDIATE STRONGER AND LONG-LASTING ANTITUMOR EFFECTS IN VIVO COMPARED TO THE CLASSICAL IFN-Γ-SECRETING TC1. HOWEVER, THE UNDERLYING MECHANISMS REMAIN UNCLEAR. RECENTLY, WE DISCOVERED THAT, WHEN WE RECHALLENGED PMEL-1 TC9- OR TC1-TREATED MICE ON THE CONTRALATERAL FLANK WITH GP100-KNOCKOUT (KO) B16 TUMOR CELLS, THESE CELLS RAPIDLY GREW AND ESTABLISHED TUMORS IN TC1 BUT NOT TC9-TREATED MICE. SIMILARLY, RELAPSED GP100– B16 CELLS RECOVERED FROM PMEL-1 TC1-TREATED MICE RAPIDLY GREW AND ESTABLISHED TUMORS AFTER RE-IMPLANTING TO PMEL-1 TC1- BUT NOT TC9-TREATED MICE, SUGGESTING THAT TC9- BUT NOT TC1-TREATED MICE DEVELOP A HOST IMMUNITY AGAINST OTHER (THAN THE COGNATE) ANTIGENS EXPRESSED BY RELAPSED TUMORS. IMPORTANTLY, SIGNIFICANTLY LARGER NUMBERS OF CD4+ BUT NOT CD8+ T CELLS WERE DETECTED IN LATE-STAGE TUMORS, RECHALLENGED TUMORS AND TUMOR-DRAINING LYMPH NODES (TDLNS) OF TC9-TREATED MICE COMPARED TO TC1-TREATED MICE. THESE CD4+ T CELLS EXPRESSED HIGH LEVELS OF IFNΓ AND GRANZYME-B AND EFFECTIVELY KILLED WILD-TYPE B16, GP100– RELAPSED AND GP100-KO B16 TUMOR CELLS. HENCE, THESE NOVEL FINDINGS STRONGLY INDICATE THAT ADOPTIVELY TRANSFERRED TC9 BUT NOT TC1 CELLS EFFECTIVELY INDUCE A HOST CD4+ T CELL RESPONSE AGAINST RELAPSED TUMORS. TO DETERMINE THE MECHANISM UNDERLYING TC9 CELL-INDUCED HOST CD4+ T CELL RESPONSE, WE PERFORMED PRELIMINARY STUDIES TO EXAMINE IMMUNE CELLS IN TUMOR MICROENVIRONMENT (TME). WE OBSERVED THAT DENDRITIC CELLS (DCS) WERE ENRICHED IN TC9-TREATED PRIMARY TUMORS, RECHALLENGED TUMORS, AND TDLNS. MORE IMPORTANTLY, TC9 CELLS SECRETED A HIGH LEVEL OF IL-24, AND TC9-TREATED TUMOR SUPERNATANTS CONTAINED ABUNDANT IL-24. AS OUR PRELIMINARY STUDIES SHOWED THAT IL-24 ATTRACTED DC MIGRATION IN VITRO, WE SPECULATE THAT TC9 CELLS INDUCE A HOST CD4+ T CELL RESPONSE THROUGH IL-24-DC CIRCUIT. WE HYPOTHESIZE THAT TUMOR-SPECIFIC TC9 CELLS MAY BE A SUPERB T-CELL SUBSET FOR CANCER IMMUNOTHERAPY DUE TO THEIR CAPACITY TO ELICIT A HOST CD4+ T CELL RESPONSE TO SUPPRESS THE GROWTH OF RELAPSED TUMORS. TO TEST OUR HYPOTHESIS, AIM 1 WILL DETERMINE THE ROLE AND IMPORTANCE OF TC9 CELL- INDUCED HOST CD4+ T CELL RESPONSES IN RECOGNIZING AND ELIMINATING RELAPSED TUMOR CELLS, AND AIM 2 WILL DETERMINE THE ROLE AND MECHANISM OF IL-24-DC CIRCUIT IN TME AND TDLNS IN ELICITING TUMOR-SPECIFIC CD4+ T CELL RESPONSES AND PREVENTING THE RECURRENCE OF TUMORS. COMPLETING THIS PROJECT WILL UNCOVER A NOVEL MECHANISM OF TC9 CELLS IN MEDIATING LONG-LASTING ANTITUMOR ACTIVITY IN TME BY INDUCING HOST CD4+ T CELLS TO RECOGNIZE TUMOR- ASSOCIATED ANTIGENS OR NEOANTIGENS VIA IL-24-DC CIRCUIT AND SUPPRESSING OR PREVENTING TUMOR RECURRENCE IN TC9-TREATED MICE.

SELF-ASSEMBLED MULTIFUNCTIONAL APTAMER-COMPLEX BIOMATERIAL FOR PRECISION MEDICINE

ROLE OF MIF IN MYELOMA BONE HOMING AND DRUG RESPONSE

ROLE OF LIPID METABOLISM IN CD8+ T CELL FERROPTOSIS - PROJECT SUMMARY RECENTLY, WE MADE A NOVEL AND EXCITING DISCOVERY THAT TUMORS OR THE TUMOR MICROENVIRONMENT (TME) CAUSE T- CELL DYSFUNCTION AND DEATH BY INDUCING FERROPTOSIS IN T CELLS. WE ANALYZED THE SC-RNA-SEQ DATA OF TUMOR- INFILTRATING T CELLS FROM MELANOMA PATIENTS AND DISCOVERED THAT TUMOR-INFILTRATING CD8+ T CELLS HAD SIGNIFICANTLY INCREASED EXPRESSIONS OF GENES ASSOCIATED WITH LIPID PEROXIDATION AND FERROPTOSIS COMPARED TO BLOOD CD8+ T CELLS FROM HEALTHY INDIVIDUALS. MORE IMPORTANTLY, OUR UNPUBLISHED STUDIES FURTHER REVEALED THAT AMONG DIFFERENT CD8+ T CELL SUBPOPULATIONS, EFFECTOR MEMORY (TEM) AND TERMINALLY DIFFERENTIATED EFFECTOR (TTE) CD8+ T CELLS ARE MORE SENSITIVE TO TUMOR-INDUCED FERROPTOSIS. WE EXAMINED TUMOR-INFILTRATING CD8+ T CELLS FROM THE BONE MARROW (BM; TUMOR BED) OF PATIENTS WITH MULTIPLE MYELOMA. BY SEPARATING THE T CELLS INTO NAÏVE, TTE OR TEM CELLS BASED ON THEIR EXPRESSION OF CCR7 AND CD45RA, WE FOUND THAT TEM AND TTE CD8+ T CELLS EXPRESSED HIGHER LEVELS OF LIPID PEROXIDATION- AND FERROPTOSIS-ASSOCIATED GENES AND WERE MORE SENSITIVE TO TUMOR-INDUCED FERROPTOSIS COMPARED TO NAÏVE CD8+ T CELLS ALTHOUGH THEY EXPRESSED SIMILAR LEVELS OF CD36. SIMILARLY, IN MOUSE MELANOMA AND MM MODELS, INCREASED FERROPTOSIS MAINLY OCCURRED IN TUMOR-INFILTRATING CD8+ TEM AND TTE CELLS BUT NOT IN NAÏVE CD8+ T CELLS FROM MICE WITH LARGE TUMOR BURDENS COMPARED TO THOSE WITH SMALL TUMOR BURDENS. OUR EX VIVO STUDIES CONFIRMED THAT CD8+ TEM AND TTE CELLS WERE MORE SENSITIVE TO TUMOR- OR FA- INDUCED FERROPTOSIS THAN NAÏVE T CELLS AND THEIR PRODUCTION OF CYTOTOXIC CYTOKINES SUCH AS IFN AND TNFA WAS INHIBITED. TO ELUCIDATE THE UNDERLYING MECHANISMS, RNA-SEQ WAS USED AND SHOWED THAT CD8+ TEM AND TTE CELLS EXPRESSED A SIGNIFICANTLY LOWER LEVEL OF 2,4-DIENOYL-COA REDUCTASE 1 (DECR1), A RATE-LIMITING ENZYME FOR POLYUNSATURATED FATTY ACID (PUFA) SS-OXIDATION, COMPARED TO NAÏVE CD8+ T CELLS. KNOCKDOWN (KD) OF DECR1 IN NAÏVE CD8+ T CELLS RESULTED IN AN INCREASED PUFA EXPRESSION AND PEROXISOMAL DYSFUNCTION AND SENSITIZED THEM TO TUMOR- OR FA-INDUCED FERROPTOSIS THAN CONTROL T CELLS. BASED ON THESE NOVEL FINDINGS, WE HYPOTHESIZE THAT CD8+ TEM AND TTE CELLS, DUE TO UPTAKE OF MORE FAS AND REDUCED EXPRESSION OF DECR1 AND PUFA OXIDATION, ARE SENSITIVE TO TUMOR/TME-INDUCED FERROPTOSIS, AND INHIBITING TEM AND TTE CELL FERROPTOSIS MAY EFFECTIVELY ENHANCE THE THERAPEUTIC EFFICACY OF IMMUNOTHERAPY IN CANCER PATIENTS. AIM 1 WILL DETERMINE THE MECHANISM UNDERLYING TUMOR- OR FA-INDUCED FERROPTOSIS IN CD8+ TEM AND TTE CELLS IN TME. AIM 2 WILL ELUCIDATE THE ROLE AND MECHANISMS OF TUMOR AND TME ACCUMULATION OF FAS AND INDUCTION OF LIPID PEROXIDATION IN CD8+ TEM AND TTE T CELLS AND AIM 3 WILL INHIBIT CD8+ TEM AND TTE CELL FERROPTOSIS TO ENHANCE THE EFFICACY OF CANCER IMMUNOTHERAPIES. ACCOMPLISHING THESE AIMS WILL PROVIDE US WITH IN-DEPTH UNDERSTANDING OF THE MECHANISMS UNDERLYING HOW TUMORS AND THE TME INDUCE T CELL LIPID PEROXIDATION AND HOW FERROPTOSIS MEDIATES T CELL METABOLIC MALFUNCTION AND DEATH. UNDERSTANDING THESE MECHANISMS IS EXTREMELY IMPORTANT AND WILL GREATLY ASSIST US IN DEVELOPING NOVEL THERAPEUTIC APPROACHES TO TARGET T CELL LIPID METABOLISM AND TME TO SIGNIFICANTLY IMPROVE THE EFFICACY OF CANCER IMMUNOTHERAPY.

EXPLORING THE ROLE AND POTENTIAL OF ANTI-VIRAL DRUGS TO SENSITIZE CANCER CELLS TO CHEMOTHERAPY - PROJECT SUMMARY CHEMOTHERAPY REMAINS THE MOST USED SYSTEMIC TREATMENT FOR CANCERS. HOWEVER, DESPITE SIGNIFICANT IMPROVEMENT IN CHEMOTHERAPY AGENTS, CHEMORESISTANCE REMAINS THE MAJOR PROBLEM IN CANCER MANAGEMENT. RECENTLY, WE DISCOVERED THAT THE SELECTIVE INHIBITORS OF HEPATITIS C VIRUS (HCV) NS5A REPLICATION COMPLEX ELBASVIR (ELB) AND DACLATASVIR (DAC), AN ANALOG OF ELB, MAY BE USED TO SENSITIZE AND RE-SENSITIZE SOLID AND HEMATOLOGIC CANCER CELLS TO CHEMO-DRUGS. WE SCREENED 1855 FDA-APPROVED DRUGS AND FOUND THAT ELB WAS AMONG THE TOP DRUGS THAT SIGNIFICANTLY SENSITIZED MULTIPLE MYELOMA (MM) CELLS TO CARFILZOMIB (CFZ, PROTEASOME INHIBITOR), DEXAMETHASONE (DEX, CORTICOSTEROID), AND MELPHALAN (MEL, ALKYLATING AGENT). ELB AND DAC COULD ALSO RE-SENSITIZE CFZ- OR DEX-RESISTANT MM CELLS TO CFZ OR DEX RESPECTIVELY. IN ADDITION, WE OBSERVED THAT ELB AND DAC ENHANCED CHEMOSENSITIVITY AND RE-SENSITIZED DIFFERENT TYPES OF CANCERS SUCH AS PANCREATIC DUCTAL ADENOCARCINOMA TO 5-FLUOROURACIL AND GEMCITABINE (GEM), ESTROGEN RECEPTOR‑POSITIVE BREAST CANCER (BC) CELLS TO TAMOXIFEN, AND TRIPLE NEGATIVE BC CELLS TO GEM. MOREOVER, IN THE PRESENCE OF ELB OR DAC, LOWER DOSES OF CHEMO-DRUGS WERE REQUIRED TO INDUCE SIMILAR CANCER CELL DEATH COMPARED TO CHEMOTHERAPY DRUGS ALONE. WE DISCOVERED THAT ELB AND DAC SIGNIFICANTLY ENHANCED DRUG RETENTION IN CANCER CELLS BY INHIBITING DRUG EFFLUX THROUGH ATPASE PHOSPHOLIPID TRANSPORTING 9B (ATP9B). IMPORTANTLY, ALTHOUGH TUMOR MICROENVIRONMENT (TME) COMPONENTS SUCH AS TUMOR-ASSOCIATED STROMAL CELLS (TASCS) AND TUMOR-ASSOCIATED MACROPHAGES (TAMS) CAN PROTECT CANCER CELLS FROM CHEMOTHERAPY-INDUCED CELL DEATH, ELB AND DAC ABROGATED TASC AND TAM PROTECTIVE EFFECTS BY SUPPRESSING PRO-TUMOR LIPID SECRETION, REPROGRAMMING THEM TO SECRETE TYPE-I-IFNS FOR SENSITIZING MM CELLS TO CFZ, AND INHIBITING TAM RELEASE OF DEOXYCYTIDINE FOR OVERCOMING GEM RESISTANCE IN SOLID TUMORS. FINALLY, ELB AND DAC SIGNIFICANTLY IMPROVED THE THERAPEUTIC EFFICACY OF CHEMOTHERAPIES IN MM IN VIVO WITHOUT INCREASED TOXICITY TO NORMAL TISSUES. THEREFORE, WE HYPOTHESIZE THAT HCV NS5A INHIBITORS ELB AND DAC CAN BE DEVELOPED INTO CANCER THERAPEUTIC AGENTS DUE TO THEIR ABILITY TO (RE)SENSITIZE CANCER CELLS TO CHEMOTHERAPIES BY ENHANCING CHEMO-DRUG RETENTION IN TUMOR CELLS AND REMOVING TME-PROVIDED PROTECTION. TO MINIMIZE THE SCOPE OF THIS APPLICATION, WE WILL FOCUS ON HUMAN MM (HEMATOLOGICAL MALIGNANCY) AND USE HUMAN MM CELL LINES AND PRIMARY MM CELLS FROM PATIENTS AND MM PDX MOUSE MODELS TO TEST THE HYPOTHESIS. AIM 1 WILL ELUCIDATE THE ROLE AND MECHANISM OF ELB AND DAC IN SENSITIZING TUMOR CELLS TO CHEMOTHERAPY BY INHIBITING CHEMOTHERAPEUTIC DRUGS EFFLUX THROUGH ATP9B AND AIM 2 WILL ELUCIDATE THE ROLE AND MECHANISMS OF ELB AND DAC IN OVERCOMING TME-MEDIATED EXTRINSIC RESISTANCE BY INHIBITING ATP9B-MEDIATED CHOLESTEROL UPTAKE AND DEOXYCYTIDINE EFFLUX. ACCOMPLISHING THESE AIMS WILL PROVIDE THE JUSTIFICATION AND TOOLS FOR DEVELOPING NOVEL AND EFFECTIVE STRATEGIES FOR TARGETING BOTH CANCER DRUG EFFLUX AND TME TO IMPROVE THE THERAPEUTIC EFFICACY OF CHEMOTHERAPY.

HEPARANASE MECHANISMS IN BRAIN-METASTATIC BREAST CANCER

TRANSFORMING TRIPLE-NEGATIVE BREAST CANCER TREATMENT THROUGH INTRATUMORAL IMMUNOTHERAPY VIA NANOFLUIDIC DRUG-ELUTING SEED

TRAINING IN NEURAL CONTROL OF ORGAN DEGENERATION AND REGENERATION (NEURALCODR) - NEURAL ACTIVITY DIRECTLY CONTROLS THE DEVELOPMENT AND HOMEOSTASIS OF ORGAN FUNCTION. YET, A SURPRISING GAP EXISTS BETWEEN LABORATORIES FOCUSED ON VASCULAR, GASTROINTESTINAL, IMMUNE, AND MUSCULOSKELETAL SYSTEMS AND THOSE FOCUSED ON NEUROPHYSIOLOGY, NEUROMODULATION, AND NEURAL INJURY. TRADITIONALLY, DEPARTMENTS ARE ORGANIZED BY ORGAN SYSTEM AND THIS SILOED STRUCTURE LEADS TO A PHYSICAL AND SCIENTIFIC JARGON SEPARATION, IMPEDING COLLABORATION AND TRAINING. HENCE, OUR UNDERSTANDING OF HOW THE CENTRAL NERVOUS SYSTEM COMMUNICATES WITH END ORGANS THROUGHOUT THE BODY IS IN ITS INFANCY. NEURAL CONTROL OF ORGAN DEGENERATION AND REGENERATION (NEURALCODR), A CROSS-DISCIPLINARY TRAINING PROGRAM UNITES 30 FACULTY MEMBERS AND 12 CLINICIAN RESEARCHERS TO FORM MENTORSHIP TEAMS WITHIN THE LARGEST MEDICAL RESEARCH CENTER IN THE WORLD (TEXAS MEDICAL CENTER). MENTORSHIP TEAMS INCLUDE A PRIMARY AND SECONDARY MENTOR (EACH FROM ONE OF OUR THREE MAIN AREAS OF RESEARCH: 1) NEURAL DEVELOPMENT AND TOOLS, 2) NEURAL INNERVATION AND ORGAN ENGINEERING, AND 3) NERVOUS SYSTEM AND PERIPHERAL ORGAN DISORDERS) AND A CLINICAL MENTOR WHO PROVIDES MUCH NEEDED AND RARE EXPOSURE TO REAL-WORLD DISEASES, CLINICAL CHALLENGES, AND HUMAN SAMPLES. NEURALCODR AIMS TO: 1) CATALYZE THE COLLISION OF TALENT AND IDEAS THAT SPAWN RESEARCH PROJECTS BRIDGING NEUROSCIENCE WITH ORGAN SYSTEMS THROUGH FACILITATED INTERACTIONS, 2) BUILD CO-MENTOR TEAMS THAT INCLUDE NEUROSCIENCE, ORGAN SYSTEMS, AND CLINICAL PERSPECTIVES, ENSURING TRAINEES ARE GUIDED TOWARD A UNIQUE RESEARCH NICHE, AND 3) TRAIN FELLOWS IN RESEARCH RIGOR, ANALYSIS, AND CAREER SKILLS THAT SUPPORT THEIR DEVELOPMENT AS SCIENTIFIC LEADERS. NEURALCODR LEVERAGES A NETWORK OF FACULTY WHO COLLABORATE ON PROJECTS THAT BRIDGE THE GAP BETWEEN ORGAN SYSTEM BIOLOGY AND ENGINEERING AND NEURAL FUNCTION, DEGENERATION AND REGENERATION, AS EVIDENCED BY AN IMPRESSIVE LIST OF MULTI-LABORATORY SHARED GRANTS, PUBLICATIONS, AND NASCENT COLLABORATIONS. THE TRAINING STRUCTURE EMPHASIZES EXPERIENCES IN ORGAN ENGINEERING AND ORGAN PHYSIOLOGY LABORATORIES IN PARALLEL WITH EDUCATION IN NEUROPHYSIOLOGY AND NEURAL ENGINEERING, TRANSLATIONAL THEORY, AND PRACTICE. CURRENTLY, TWO NEURALCODR HOUSTON METHODIST-SPECIFIC POSTDOCTORAL POSITIONS EXIST, THANKS TO A PHILANTHROPIC ENDOWMENT THAT HAS ALLOWED US TO BUILD THE PROGRAM STRUCTURE AND TO TEST BOTH THE COURSEWORK AND MENTORSHIP TEAM CONCEPT. TRAINEES ARE FUNDED FOR TWO YEARS, DURING WHICH TIME THEY HAVE ACCESS TO A CORE CURRICULUM, CAREER DEVELOPMENT AND PROGRAM ENRICHMENT OPPORTUNITIES, SYMPOSIUMS/RETREATS, AND ELECTIVE COURSES. YEAR ONE CULMINATES WITH THE SUBMISSION OF AN NRSA AND A MANUSCRIPT BASED ON TRAINEES’ DEVELOPMENT OF A TRANSFORMATIVE RESEARCH PROJECT THAT INCORPORATES NON-NEUROSCIENCE EXPERTISE AND TOOLS INTO A NEUROSCIENCE PROBLEM. YEAR TWO SOLIDIFIES THE RELATIONSHIPS BETWEEN THE MENTORSHIP TEAM AND TRAINEE AND PROVIDES TRAINING IN LABORATORY MANAGEMENT, DIDACTIC TEACHING, AND MENTORSHIP SKILLS. T32 FUNDING WILL ALLOW US TO INCREASE THE CURRENT MODEL TO SIX POSTDOCTORAL TRAINING SLOTS. THIS TRAINING PROGRAM WILL FOSTER A NEW GENERATION OF SCIENTIFIC LEADERS WHO PIONEER RESEARCH ON THE CONNECTED PATHWAYS BETWEEN BRAIN AND ORGAN SYSTEMS TO SOLVE FUNDAMENTAL CHALLENGES IN NEUROSCIENCE.

NANO-IMAGING AGENTS FOR EARLY DISEASE DETECTION

DEVELOPMENT OF A NOVEL THERAPEUTIC TO TREAT DNA DAMAGE ASSOCIATED WITH ATYPICAL BURNS

NOVEL MECHANISM OF INDUCTION OF TUMOR PYROPTOSIS BY IL-9-SECRETING TC9 CELLS - PROJECT SUMMARY BLOOD CD8+ T CELLS CAN BE SUBDIVIDED INTO DIFFERENT SUBSETS BASED ON THEIR CYTOKINE SECTION PROFILES AND FUNCTIONS. WE HAVE SHOWN THAT IL-9-SECRETING CD8+ TC9 CELLS MEDIATE A STRONGER ANTITUMOR EFFECT IN VIVO COMPARED TO THE CLASSICAL IFN--SECRETING TC1 OR CTLS. HOWEVER, THE UNDERLYING MECHANISMS REMAIN UNCLEAR. RECENTLY, WE DISCOVERED THAT TC9 CELLS COULD ERADICATE LARGE-ESTABLISHED TUMORS BY INDUCING AN ENHANCED TUMOR PYROPTOSIS, A FORM OF PROGRAMMED CELL DEATH DEPENDENT ON CASPASE-1 ACTIVATION. OUR PRELIMINARY STUDIES SHOWED THAT TC9 CELL-TREATED TUMORS HAD INCREASED EXPRESSIONS OF IL-1 AND IL-18 AND ACTIVATED CASPASE-1, AND MORE TUMOR CELL DEATH COMPARED TO TC1-TREATED TUMORS. NEUTRALIZING IL-1 AND IL-18 IN VIVO COMPLETELY ABROGATED THE THERAPEUTIC ADVANTAGE OF TC9 CELLS OVER TC1 CELLS IN TUMOR CONTROLS. WE FURTHER SHOWED THAT TUMOR-SPECIFIC TC9 CELLS KILLED TUMOR CELLS BY INDUCING BOTH CASPASE-1-DEPEDENT PYROPTOSIS AND CASPASE-3- DEPENDENT APOPTOSIS WHILE TC1 CELLS MAINLY INDUCED APOPTOSIS IN TUMOR CELLS. SIMILARLY, HUMAN TUMOR-SPECIFIC TC9 CELLS ALSO DISPLAYED STRONGER ANTITUMOR EFFECTS IN VIVO COMPARED TO TC1 CELLS, WHICH WAS IL-1- AND IL-18- DEPENDENT. INTERESTINGLY, WE OBSERVED THAT IL-1 PLUS IL-18 INDUCED APOPTOSIS IN CULTURED TC1 BUT NOT TC9 CELLS, TC9 CELL-DERIVED IL-9 IS CRITICAL FOR TC9 CELL ACTIVATION OF STAT3 AND PRO-GROWTH AND SURVIVAL SIGNALING AND FUNCTION, AND IL-1 PLUS IL-18 CAN RE-ACTIVATE STAT3 AND PRO-GROWTH AND SURVIVAL SIGNALING IN AGED TC9 CELLS WITH REDUCED IL-9 PRODUCTION. THUS, THESE FINDINGS REVEAL NOVEL MECHANISMS UNDERLYING HOW TC9 CELLS OVERCOME THE SUPPRESSIVE TUMOR MICROENVIRONMENT AFTER TRANSFER, SURVIVE AND PERSIST LONGER, AND EXERT A GREATER ANTITUMOR ACTIVITY AS COMPARED TO THE TRADITIONAL TC1 CELLS. WE HYPOTHESIZE THAT TUMOR-SPECIFIC TC9 SUBSET MAY BE SUPERB EFFECTOR T CELLS FOR CANCER IMMUNOTHERAPY DUE TO THEIR CAPACITY TO INDUCE BOTH PYROPTOSIS AND APOPTOSIS IN TUMOR CELLS AND UTILIZE TC9 CELL-SECRETED IL-9 AND TUMOR-PRODUCED IL-1 AND IL-18 FOR THEIR FITNESS, LONGEVITY, AND FUNCTION IN VIVO TO EFFECTIVELY ERADICATE LARGE ESTABLISHED TUMORS. TO TEST OUR HYPOTHESIS, AIM 1 WILL DETERMINE THE IMPORTANCE AND MECHANISMS OF TC9 CELL-INDUCED TUMOR CELL PYROPTOSIS IN TUMOR CLEARANCE VIA IL-9- AND GRZB-INDUCED NFB-NLRP3-CASPASE-1-GASDERMIN ACTIVATION, AND AIM 2 WILL DETERMINE THE ROLE AND MECHANISMS UNDERLYING TC9 CELL PERSISTENCE AND FUNCTION MEDIATED BY TC9 CELL- SECRETED IL-9 AND TUMOR-PRODUCED IL-1 AND IL-18. COMPLETING THIS PROJECT WILL REVEAL THE IMPORTANCE AND MECHANISM IN INDUCTION OF TUMOR PYROPTOSIS BY TUMOR-SPECIFIC TC9 CELLS AND ELUCIDATE THE MECHANISMS UNDERLYING THE LONGEVITY AND FUNCTION OF TC9 CELLS IN TUMOR MICROENVIRONMENT.

ROLE OF CDK9 IN THE PATHOGENESIS OF PULMONARY FIBROSIS - PROJECT SUMMARY/ABSTRACT: THE LONG-TERM OBJECTIVES OF THIS PROPOSAL ARE TO REVEAL NEW AND NOVEL DRUGGABLE TARGETS AND/OR ELEMENTS OF FIBROGENIC PATHWAYS, THEREBY YIELDING NEW INSIGHTS INTO BETTER TREATMENT STRATEGIES FOR IDIOPATHIC PULMONARY FIBROSIS (IPF). IPF IS ONE OF THE MOST FREQUENT INTERSTITIAL LUNG DISEASES AND HAS A POOR PROGNOSIS WITH WORSE OUTCOMES THAN MANY MALIGNANT CANCERS. THE TWO FDA-APPROVED THERAPEUTIC AGENTS (NINTEDANIB AND PIRFENIDONE) HAVE ENTERED CLINICAL USE RECENTLY. OF MOST CONCERN, WHILE THESE DRUGS MAY SLOW THE DECLINE IN LUNG FUNCTION TO A CERTAIN EXTENT, BOTH DRUGS FAIL TO EFFECTIVELY HALT LUNG FIBROSIS AND IMPROVE PATIENT SURVIVAL. THEREFORE, THERE IS AN URGENT NEED FOR NEW AND IMPROVED THERAPEUTICS. IN ORDER TO ACHIEVE HIGHER CLINICAL EFFICACY, CURRENT THERAPEUTIC APPROACHES SEEK DRUGS THAT HAVE DUAL ACTIVITIES AGAINST INFLAMMATION AND FIBROSIS. TO ACHIEVE DUAL ACTIVITIES EFFECTIVELY, WE IDENTIFIED A NOVEL CENTRAL REGULATOR, CYCLIN- DEPENDENT KINASE 9 (CDK9), THAT CONTROLS A COMMON CHECKPOINT FOR TRANSCRIPTIONAL ACTIVATION OF GENES INVOLVED IN MAJOR INFLAMMATORY AND FIBROTIC SIGNALING PATHWAYS. PRELIMINARY DATA SHOW THAT CDK9 IS ELEVATED IN IPF FIBROBLASTS, WHICH DRIVES THE AGGRESSIVE NATURE OF THESE CELLS. TREATMENT WITH CDK9 INHIBITOR REDUCED THE AGGRESSIVENESS OF THE CELLS, AND IN MOUSE MODELS OF IPF, CDK9 INHIBITOR INCREASED SURVIVAL AND REDUCED FIBROSIS. THESE DATA SUPPORT THE HYPOTHESIS THAT CDK9 INHIBITORS CAN BE EFFECTIVE THERAPEUTICS TO HALT OR REVERSE THE PROGRESSION OF PULMONARY FIBROSIS. THIS PROPOSAL AIMS TO INVESTIGATE THE MECHANISTIC ROLES OF CDK9 IN IPF AND VALIDATE CDK9 AS A NEW ANTI-INFLAMMATORY AND ANTI-FIBROTIC PHARMACOTHERAPEUTIC TARGET FOR TREATING IPF. THE APPROACH IS TO FIRMLY ESTABLISH A CENTRAL ROLE OF CDK9 IN GOVERNING BOTH INFLAMMATORY AND FIBROGENIC PROCESSES. OUR SCIENTIFIC PREMISE RESTS UPON THE PRINCIPLE THAT A POTENT THERAPEUTIC APPROACH TO COUNTERACT INFLAMMATION AND FIBROSIS CONCURRENTLY CAN BE ATTAINED BY TARGETING CDK9. THREE SPECIFIC AIMS ARE PROPOSED, SA1) USE GAIN-AND-LOSS OF FUNCTION ANALYSES TO ELUCIDATE THE MOLECULAR MECHANISM BY WHICH CDK9 REGULATES IPF PATHOGENESIS AND PROGRESSION, AND SCREEN A PANEL OF CLINICAL STAGE CDK9 INHIBITORS FOR THEIR EFFECTIVENESS AGAINST FIBROSIS; SA2) TEST THE THERAPEUTIC POTENTIAL OF THE TOP PERFORMING CDK9 INHIBITOR IN A BLEOMYCIN-INDUCED LUNG INJURY MOUSE MODEL; AND SA3) ESTABLISH THERAPEUTIC PROOF-OF-CONCEPT BY DEVELOPING AN INHALABLE FORMULATION OF CDK9 INHIBITOR AND DEMONSTRATING EFFICACY IN THE BLEOMYCIN MOUSE MODEL. OUR VISION IS TO GENERATE PRE-CLINICAL MECHANISM-OF-ACTION DATA TO SUPPORT FUTURE DEVELOPMENT OF A CDK9 INHIBITOR-BASED IPF THERAPEUTICS.

A RANDOMIZED, SHAM-CONTROLLED CLINICAL TRIAL EVALUATING INDIVIDUALIZED NEUROMODULATION OF CORTICAL REGIONS INVOLVED IN NEUROGENIC OVERACTIVE BLADDER IN MULTIPLE SCLEROSIS - NEUROGENIC OVERACTIVE BLADDER (NOAB), CHARACTERIZED BY URINARY FREQUENCY, URGENCY OR URGENCY INCONTINENCE SYMPTOMS OCCURRING DURING THE STORAGE PHASE OF THE BLADDER, IS THE MOST COMMON URINARY COMPLAINT IN MULTIPLE SCLEROSIS (MS). CURRENT MANAGEMENT OPTIONS FOR NOAB IN MS HAVE LIMITED EFFICACY AND CONSIDERABLE ADVERSE EFFECTS, WHICH UNDERSCORES THE SIGNIFICANCE OF OUR STUDY AND HIGHLIGHTS THE NEED FOR BETTER, LESS INVASIVE THERAPIES. OUR NOVEL STUDY INVESTIGATES BRAIN THERAPEUTIC TARGETS THAT COULD SHIFT THE FOCUS OF NOAB MANAGEMENT IN MS FROM A BLADDER-CENTRIC FOCUS TO BRAIN RESTORATION; SPECIFICALLY MODULATING THE BRAIN REGIONS IDENTIFIED IN OUR PRIOR FUNCTIONAL MAGNETIC RESONANCE IMAGINING STUDIES. REPETITIVE TRANSCRANIAL MAGNETIC STIMULATION (RTMS) IS A NONINVASIVE BRAIN STIMULATION THAT CAN MODULATE NEURONS (EXCITE OR INHIBIT) TO IMPROVE THE CONNECTIVITY OF THE REGIONS OF INTEREST (ROI). OUR PRELIMINARY DATA DEMONSTRATE, FOR THE FIRST TIME, SIGNIFICANT IMPROVEMENT IN BLADDER SYMPTOMS IN TEN WOMEN WITH MS WHO HAVE VOIDING DYSFUNCTION FOLLOWING MULTIFOCAL TRANSCRANIAL MAGNETIC STIMULATION WITHOUT ANY TREATMENT-RELATED ADVERSE EFFECTS. THIS RANDOMIZED DOUBLE-BLIND, SHAM-CONTROLLED SINGLE CENTER CLINICAL TRIAL WITH AN OPTIONAL OPEN-LABEL EXTENSION (OLE) PHASE IS DESIGNED TO EVALUATE THE EFFECTS OF TARGETED RTMS IN WOMEN WITH MS AND NOAB BY INVESTIGATING RESTORATIVE REORGANIZATION OF BRAIN FUNCTION AND IMPROVEMENT OF URINARY FREQUENCY, URGENCY AND INCONTINENCE. WE HYPOTHESIZE THAT CORTICAL ALTERATIONS IN BLADDER VOLUME SENSING AND THEIR RESPONSE TO STIMULATION CONTRIBUTE TO NOAB SYMPTOMS IN MS, AND THAT IMPROVING THE RESPONSE TO BLADDER DISTENTION (ROI WITHIN CIRCUITS 1 AND 2) WITH NEURONAVIGATED RTMS CAN RESTORE BRAIN ACTIVITY AND IMPROVE SYMPTOMS (FREQUENCY, URGENCY, AND INCONTINENCE). WE WILL TEST OUR HYPOTHESIS WITH THSES SPECIFIC AIMS: AIM 1: TO DETERMINE THE CLINICAL EFFECTS OF NEURONAVIGATED AND MULTIFOCAL ACTIVE/SHAM RTMS IN WOMEN WITH MS AND NOAB; AIM 2: TO ASSESS THE NEUROIMAGING RESTORATIVE EFFECTS OF NEURONAVIGATED ACTIVE/SHAM RTMS IN WOMEN WITH MS AND NOAB; AIM 3: TO ASSESS THE LONG-TERM SAFETY AND THERAPEUTIC EFFECTS OF REPEATED RTMS IN WOMEN WITH MS AND NOAB WHO PARTICIPATE IN THE OLE PHASE (WHICH SUBJECTS FROM BOTH GROUPS WILL BE INVITED TO ENTER AT THE 3-MONTH FOLLOW UP). EFFORTS TO IMPROVE OUR CURRENT KNOWLEDGE OF BRAIN CONTRIBUTION TO LOWER URINARY TRACT FUNCTION AND THE DEVELOPMENT OF AN INDIVIDUALIZED, NONINVASIVE, AND EFFECTIVE TREATMENT MODALITY AT THE LEVEL OF THE BRAIN WILL GREATLY IMPACT THE QUALITY OF LIFE FOR INDIVIDUALS WITH MS AND SUBSEQUENTLY OTHERS WITH OAB, WHETHER NEUROGENIC OR NON-NEUROGENIC.

MODULATING B AND T CELL FUNCTIONS IN EAE THROUGH GAMMA SECRETASE AND NOTCH

SPINAL NEUROMODULATION TO PROMOTE PHYSIOLOGIC AND MOLECULAR PLASTICITY IN THEINJURED SPINAL CORD - ABSTRACT THERE IS GROWING INTEREST IN THE USE OF ELECTRICAL STIMULATION TO PROMOTE THE RECOVERY OF SENSORIMOTOR AND AUTONOMIC FUNCTION AFTER NEURAL INJURY. PREVIOUS RESEARCH FROM OUR LAB AND OTHERS HAS DEMONSTRATED THAT STIMULATION OF SPINAL LUMBAR SEGMENTS ACTIVATES CENTRAL PATTERN GENERATORS, WHICH, IN TURN, FACILITATES STANDING AND WALKING. HOWEVER, WHILE THERE IS EXTENSIVE ANIMAL, PRE-CLINICAL, AND CLINICAL DATA EXAMINING THE IMPACT OF LUMBAR STIMULATION, STUDIES THAT APPLY NEUROMODULATION TO THE CERVICAL SPINAL CORD FOR UPPER LIMB ARE VERY LIMITED. HERE, WE PROPOSE THAT FUNDAMENTAL BLIND SPOTS EXIST IN THE FIELD OF NEUROMODULATION FOR UPPER LIMB, INCLUDING WHERE TO STIMULATE ANATOMICALLY, HOW TO DOSE AND, ESPECIALLY, MECHANISMS OF ACTION. THE HORNER LAB HAS DEVELOPED A CLINICALLY RELEVANT RAT MODEL OF CERVICAL SPINAL CORD INJURY AND ENGINEERED A SELF-CONTAINED EPIDURAL STIMULATION DEVICE THAT CAN BE DEPLOYED IN FREELY BEHAVING RATS TO STIMULATE SENSORIMOTOR CIRCUITRY FROM MULTIPLE SURFACES OF THE SPINAL CORD. HENCE, WE ARE WELL POSITIONED TO TEST CRITICALLY IMPORTANT HYPOTHESES ON THE ROLE OF CERVICAL STIMULATION IN THE RESTORATION OF UPPER LIMB FUNCTION. WE PRESENT EXCITING PRELIMINARY DATA DEMONSTRATING THAT EPIDURAL STIMULATION OF THE CERVICAL SPINAL CORD IMPROVES FORELIMB FUNCTION. THE RATIONALE FOR THE PROPOSED RESEARCH IS THAT THE SITE OF EPIDURAL STIMULATION PROVIDES UNIQUE ACCESS TO MOTOR CIRCUITRY. WE HYPOTHESIZE THAT VENTRAL POSITIONING OF ELECTRODES (VSS) WILL PROVIDE ACCESS TO STIMULATE MOTOR CIRCUITRY AT THE SITE OF LESION THAT ARE INACCESSIBLE FROM THE MORE COMMON DORSAL APPROACH (DSS). FURTHER, WE PROPOSE THAT VSS WILL PRODUCE NOVEL MECHANISMS OF FUNCTION PLASTICITY THAT CAN AMPLIFY RECOVERY WHEN COMBINED WITH DSS. TO TEST THIS HYPOTHESIS, WE PROPOSE THE FOLLOWING AIMS: AIM 1: DETERMINE ACUTE MOLECULAR AND PHYSIOLOGICAL MECHANISMS OF VSS WHEN APPLIED TO SUBACUTE CERVICAL SPINAL CORD INJURY. AIM 2: ESTABLISH THE FUNCTIONAL IMPACT OF SITE OF STIMULATION AND REHABILITATIVE TRAINING ON RECOVERY FROM EARLY CHRONIC CERVICAL SPINAL CORD INJURY. AIM 3: ESTABLISH THE SYNERGISTIC EFFECTS OF COMBINED VSS AND DSS AFTER CERVICAL SPINAL CORD INJURY. THESE STUDIES WILL EXPLORE AN EXCITING NEW APPROACH TO PROMOTE NEURAL RECOVERY OF THE UPPER LIMB, AN AREA OF RESEARCH THAT HAS HAD LIMITED INVESTIGATION, BUT REMAINS A PRIMARY CONCERN FOR THE PATIENT. OUR APPROACH WILL RIGOROUSLY ESTABLISH THE PHYSIOLOGICAL AND FUNCTIONAL EFFECTS OF THE SITE OF STIMULATION ON THE MOLECULAR AND PHYSIOLOGICAL MECHANISMS OF UPPER LIMB PLASTICITY.

SYSTEM BIOLOGY APPROACH FOR SIGNALING TRANSDUCTION STUDY OF COMPLEX PHENOTYPES

TARGETING MACROPHAGES TO SENSITIZE MYELOMA TO IMMUNE CHECKPOINT BLOCKADE

MICROBIOTA-TARGETED APPROACHES TO RESOLVE DYSBIOSIS-INDUCED AD NEUROPATHOLOGY FOLLOWING BRAIN INJURY. - PROJECT SUMMARY / ABSTRACT ALZHEIMER’S DISEASE (AD) IS A PROGRESSIVE NEURODEGENERATIVE DISEASE THAT RESULTS IN COGNITIVE DECLINE. THE NEUROINFLAMMATORY EVENTS THAT ARE ASSOCIATED WITH THE PATHOLOGY OF AD EXACERBATE NEURODEGENERATION. ELDERLY PEOPLE WITH DEMENTIA OR AD HAVE A HIGHER RISK OF FALLING BECAUSE THEIR COGNITIVE FUNCTION IS AFFECTED. THEY LOSE THEIR ORIENTATION AND ARE VERY PRONE TO BREAKING THEIR HIPS, OTHER BONES, OR EVEN MORE SEVERE FALLS THAT CAN CAUSE BRAIN TRAUMA. ALTHOUGH THEY ARE TREATED IN THE HOSPITALS WITH SUBSEQUENT REHABILITATION, THEIR WEAKENED IMMUNE STATE TYPICALLY AFFECTS THEIR RECOVERY PHASES. WE NEED TO DEVELOP NEW TREATMENTS FOR AD PATIENTS WHO HAVE HAD A TRAUMATIC BRAIN INJURY (TBI). THIS VULNERABLE POPULATION COULD BE PARTICULARLY AMENABLE TO PRECISION-MICROBIOTA THERAPY. ALTHOUGH NOT FULLY UNDERSTOOD, THE BRAIN-GUT-MICROBIOTA AXIS PLAYS A MAJOR ROLE IN THE ONSET AND SEVERITY OF MANY NEUROLOGICAL DISEASES. WE PLAN TO DEVELOP PRECISION-MICROBIOTA THERAPY TO PROTECT AGAINST NEURODEGENERATION AND FUNCTIONAL BEHAVIOR IN AD MICE AFTER TBI. THE GUT MICROBIOME IS EMERGING AS AN ESSENTIAL NEUROMODULATOR OF BRAIN-GUT AXIS SIGNALING. IT CAN SIGNIFICANTLY IMPACT BRAIN INFLAMMATION AND OUTCOME AFTER CNS TRAUMA AND ALTER ANXIETY- AND DEPRESSION-LIKE BEHAVIORS. THIS RESEARCH PROPOSAL IS BASED ON THE SCIENTIFIC PREMISE THAT GUT BACTERIA CAN IMPACT BEHAVIOR AND THAT BRAIN INJURY CAN DISRUPT DIVERSITY IN A HEALTHY GUT MICROBIOME. THE OVERARCHING HYPOTHESIS IS THAT THE GUT MICROBIOTA DYSFUNCTION CAUSED BY BRAIN TRAUMA CONTRIBUTES DIRECTLY TO NEUROINFLAMMATION AND NEURODEGENERATION RESPONSES IN AD MICE, AND THE MICROBIOTA MODULATION WILL DELAY THE PROGRESSION OF AD NEUROPATHOLOGY. OUR PRELIMINARY DATA INDICATE HOW THE MICROBIOTA OF AD MICE AFFECTED ADVERSELY TO TBI OUTCOMES AND HOW PROBIOTICS HELP BRAIN RECOVERY IN C57B6/J YOUNG MICE. THIS PROPOSAL INVESTIGATES THE MECHANISTIC LINKAGE BETWEEN GUT MICROBIOTA AND AD PROGRESSION FOLLOWING TBI AND EXPLORES POTENTIAL INTERVENTION STRATEGIES. WE WILL TEST THIS PREMISE WITH THE FOLLOWING THREE SPECIFIC AIMS: WE 1) WILL DETERMINE IF THE GUT MICROBIOME PLAYS A ROLE IN ACCELERATING AD PATHOLOGY AND COGNITIVE DECLINE INDUCED BY TBI, AND 2) WILL INVESTIGATE THE IMPACT OF FECAL TRANSPLANTS ON AD PATHOLOGY AND COGNITIVE DECLINE AFTER TBI, AND 3) WILL ASSESS THE ABILITY OF MULTI-STRAIN PROBIOTICS TO REDUCE AD PATHOLOGY AFTER TBI. WE EXPECT TO IDENTIFY A PARTICULARLY VULNERABLE PATIENT POPULATION BY DEFINING WHICH INFLAMMATORY RESPONSES ASSOCIATED WITH BRAIN TRAUMA ACCELERATE AD PATHOGENESIS AND ARE REGULATED BY BRAIN-GUT-MICROBIOME SIGNALS. OUR APPROACH IS SIGNIFICANT BECAUSE WE WILL ESTABLISH A “TRANSLATABLE” FOUNDATION FOR THE POTENTIAL OF THERAPEUTIC APPROACHES FOR AD AFTER TBI BY USING THE GUT AS A NON-CNS TARGET OF PRECISION-MICROBIOTA THERAPY.

A NANOFLUIDIC PLATFORM FOR TUNABLE DRUG DELIVERY

HIGH-CONTENT IMAGE ANALYSIS AND MODELING FOR RANIGENOME-WIDE SCREENING

NATURAL PHENOLIC COMPOUNDS AGAINST ORAL CANDIDIASIS (OC) - CONTACT PD/PI: MYLONAKIS, ELEFTHERIOS PROJECT SUMMARY DESPITE THE AVAILABILITY OF ANTI-RETROVIRAL THERAPY, ORAL CANDIDIASIS CAUSED BY THE OVERGROWTH OF CANDIDA SPP. IS THE MOST FREQUENT OPPORTUNISTIC INFECTION AMONG INDIVIDUALS LIVING WITH HIV. THE WIDESPREAD USE OF AZOLES TO TREAT ORAL CANDIDIASIS IN INDIVIDUALS LIVING WITH HIV OFTEN RESULTS IN TREATMENT FAILURE. IN ADDITION TO ANTIFUNGAL RESISTANCE, THE CURRENT THERAPEUTIC OPTIONS FOR ORAL CANDIDIASIS ARE LIMITED BY SIDE EFFECTS THE CHALLENGES POSED BY BIOFILM DRUG RECALCITRANCE. MOREOVER, CANDIDA AURIS HAS RECENTLY EMERGED AS A MULTI-DRUG–RESISTANT FUNGAL PATHOGEN AND IS CLASSIFIED AS 1 OF THE 5 PATHOGENS IN THE HIGHEST CATEGORY—URGENT THREATS—IN THE ANTIBIOTIC RESISTANCE THREATS IN THE UNITED STATES. POLYPHENOLS ARE SECONDARY PLANT METABOLITES THAT HAVE PROTECTIVE PROPERTIES FOR HUMAN HEALTH, INCLUDING ANTIOXIDANT, ANTI-INFLAMMATORY, ANTICANCER, AND ANTIMICROBIAL ACTIVITY. WE FOUND THAT CAFFEIC ACID PHENETHYL ESTER (CAPE) AND ELLAGIC ACID (EA) ARE BOTH ACTIVE AGAINST CANDIDA SPP., INCLUDING C. AURIS RESISTANT STRAINS. THE MINIMUM INHIBITORY CONCENTRATION FOR EA RANGED FROM 0.125 TO 0.25 ΜG/ML AND, AT SUBINHIBITORY CONCENTRATIONS, EA INHIBITED PHOSPHOLIPASE PRODUCTION BY C. AURIS. CAPE INHIBITED FUNGAL FILAMENTATION AND BIOFILM FORMATION. BOTH COMPOUNDS WERE ACTIVE IN THE INVERTEBRATE MODEL HOSTS CAENORHABDITIS ELEGANS AND GALLERIA MELLONELLA. WE FURTHER DEMONSTRATED THAT NEITHER CAPE NOR EA IS TOXIC TO HUMAN ERYTHROCYTES. IMPORTANTLY, IN A MOUSE MODEL OF ORAL CANDIDIASIS, CAPE SIGNIFICANTLY INCREASED THE EXPRESSION OF THE MURINE ANTIFUNGAL DEFENSIN Β- DEFENSIN 3 AND REDUCED PSEUDOMEMBRANOUS LESIONS, INVASION OF HYPHAE ON EPITHELIUM SURFACES, TISSUE DAMAGE, AND INFLAMMATORY INFILTRATES. THE PURPOSE OF THIS PROPOSAL IS TO ADVANCE THE DEVELOPMENT OF THESE NATURAL PHENOLIC COMPOUNDS FOR THE MANAGEMENT OF ORAL CANDIDIASIS. WE HAVE PUBLISHED “PROOF OF CONCEPT” STUDIES THAT GELLAN CAN BE IMPREGNATED WITH CAPE, AND THAT THE HYDROGEL IS ACTIVE IN A MOUSE MODEL OF ORAL CANDIDIASIS. WE WILL FURTHER ADVANCE THESE GOALS WITH THE FOLLOWING AIMS: 1. TO OPTIMIZE HYDROGELS LOADED WITH EA AND/OR CAPE FOR MAXIMAL RELEASE OF COMPOUNDS AND TEST THEIR ANTIFUNGAL ACTIVITY AND CYTOTOXICITY IN VITRO. 2. TO TEST THE OPTIMIZED HYDROGELS LOADED WITH EA AND/OR CAPE IN AN ESTABLISHED MOUSE MODEL OF ORAL CANDIDIASIS. 3. TO TEST THE ANTIFUNGAL IMMUNE RESPONSE TO HYDROGELS LOADED WITH EA AND/OR CAPE IN THE ORAL MUCOSA OF VOLUNTEERS LIVING WITH HIV. OUR OVERALL HYPOTHESIS IS THAT THE COMBINATION OF CAPE AND EA COULD PROVIDE AN IDEAL NATURAL TREATMENT AGAINST ORAL CANDIDIASIS WITH DIRECT ANTIFUNGAL ACTIVITY AND ACTIVITY AGAINST RESISTANT STRAINS (INCLUDING C. AURIS), EFFICACY AGAINST FILAMENTS AND BIOFILMS, AND IMMUNOMODULATORY ACTIVITY THROUGH THE PRODUCTION OF ANTIMICROBIAL PEPTIDES AND CHEMOKINES. IN THIS CONTEXT, THE PROPOSED STUDIES COULD REPRESENT A PIVOTAL STAGE IN THE DEVELOPMENT AND TRANSLATION OF THESE NATURAL COMPOUNDS AS CLINICAL ANTIFUNGAL AGENTS. REFERENCES CITED PAGE 1

DEEPSTROKE+: AN ADVANCED MOBILE AI DIAGNOSTIC TOOL FOR FAST AND PRECISE DETECTION OF ACUTE STROKES IN MOBILE STROKE UNITS, EMERGENCY ROOMS, AND TELESTROKE TRIAGE - PROJECT SUMMARY/ABSTRACT NEARLY ONE MILLION AMERICANS SUFFER FROM A STROKE EVERY YEAR, OFTEN RESULTING IN SERIOUS LONG-TERM DISABILITY OR DEATH. EARLY DETECTION AND TREATMENT OF STROKE IS CRITICAL TO IMPROVING PATIENT OUTCOMES. SEVERAL TOOLS HAVE PREVIOUSLY BEEN DEVELOPED THAT ATTEMPT TO SCREEN AND HELP IDENTIFY PATIENTS HAVING A STROKE, BUT THERE ARE STILL MANY STROKE CASES THAT ARE MISSED, EVEN AFTER THE PATIENT ARRIVES TO THE EMERGENCY ROOM. THIS STUDY AIMS TO IMPROVE THE IDENTIFICATION AND TRIAGE OF STROKE PATIENTS IN EMERGENCY DEPARTMENTS USING ARTIFICIAL INTELLIGENCE (AI) ON MULTIMEDIA PATIENT DATA. OUR HYPOTHESIS IS THAT REAL-TIME, STANDARDIZED, AND REPRODUCIBLE STROKE ASSESSMENT TOOLS USING AI CAN IMPROVE STROKE TRIAGE AND DECREASE MISSED DIAGNOSES IN PATIENTS WITH MINOR-TO-MODERATE NEUROLOGICAL SYMPTOMS. THE STUDY PLANS TO DEVELOP A DEEPSTROKE+ AUGMENTED-INTELLIGENCE FRAMEWORK TO TRIAGE ANY KIND OF STROKES (ISCHEMIC STROKE, HEMORRHAGIC STROKE, TRANSIENT ISCHEMIC ATTACK, ETC) VERSUS STROKE MIMICS COMMONLY SEEN IN EMERGENCY SETTINGS. WE DEVELOPED A DEEPSTROKE+ FRAMEWORK FOR STROKE TRIAGE USING FACIAL VIDEOS OF ENGLISH-SPEAKING PATIENTS WHO ARE DESCRIBING THE “COOKIE THEFT” PICTURE FROM THE BOSTON DIAGNOSTIC APHASIA EXAMINATION. BASED ON THIS TECHNOLOGY, WE WILL DEVELOP A STROKE TRIAGE APP AND VALIDATE IT IN DIFFERENT TRIAGE SCENARIOS FROM MOBILE STROKE UNITS TO EMERGENCY ROOM TRIAGE IN LOCAL AND TELESTROKE SCENARIOS.

METABOLISM AND EPIGENETIC REGULATION ARE COUPLES IN TRANSDIFFERENTIATION AND VASCULAR REGENERATION - ABSTRACT WE DISCOVERED THAT NUCLEAR REPROGRAMMING OF SOMATIC CELLS TO A DIFFERENT SOMATIC CELL LINEAGE, OR INDUCED PLURIPOTENT STEM CELLS, REQUIRES ACTIVATION OF INFLAMMATORY SIGNALING WITHIN THE CELL. SPECIFICALLY, PATTERN RECOGNITION RECEPTORS (PRRS) SUCH AS TOLL-LIKE RECEPTORS (TLRS) MEDIATE A CELL-AUTONOMOUS INNATE IMMUNE RESPONSE VIA NFKB AND IRF3. WE FOUND THAT THIS INFLAMMATORY SIGNALING CAUSES GLOBAL CHANGES IN THE EXPRESSION AND/OR ACTIVITY OF EPIGENETIC MODIFIERS TO INCREASE DNA ACCESSIBILITY AND FLUIDITY OF CELL PHENOTYPE. SUBSEQUENT WORK HAS SUGGESTED THAT THIS PROCESS OF “TRANSFLAMMATION” MAY BE INVOLVED IN VASCULAR REGENERATION. SPECIFICALLY, WE HAVE SHOWN THAT FIBROBLASTS IN AN ISCHEMIC REGION CAN BE TRANSFORMED INTO ENDOTHELIAL CELLS (ECS) THROUGH AN “ANGIOGENIC TRANSDIFFERENTIATION” PROCESS. THIS PROCESS CONTRIBUTES TO THE RECOVERY OF PERFUSION IN THE ISCHEMIC REGION, AS THE RECOVERY OF THE MICROVASCULATURE, AND THE RESTORATION OF BLOOD FLOW IN AN ISCHEMIC REGION IS ANTAGONIZED BY FACTORS REQUIRED FOR ANGIOGENIC TRANSDIFFERENTIATION (E.G., INFLAMMATORY SIGNALING). MY RECENT WORK INDICATES THAT CELL METABOLISM MAY BE AN IMPORTANT CONTRIBUTOR TO THIS PROCESS. SPECIFICALLY, A GLYCOLYTIC SHIFT IS INDUCED BY INFLAMMATORY SIGNALING WHICH IS REQUIRED FOR THE TRANSDIFFERENTIATION OF FIBROBLASTS TO ECS. THUS, REGULATING CELL METABOLISM WITHIN FIBROBLASTS TO FACILITATE THEIR TRANSDIFFERENTIATION INTO REPARATIVE ECS MAY BE A NOVEL STRATEGY FOR TREATING ISCHEMIA. TO DETERMINE THE MOLECULAR METABOLIC PATHWAY THAT LEADS TO TRANSDIFFERENTIATION, WE WILL ALTER THE FUNCTION OF KEY METABOLIC ENZYMES IN MICE PHARMACOLOGICALLY AND GENETICALLY IN VIVO TO CONFIRM OUR PROPOSED PATHWAY AND DEMONSTRATE THE METABOLIC REGULATION OF TRANSDIFFERENTIATION IN A MOUSE MODEL OF PERIPHERAL ARTERY DISEASE (PAD). WE WILL PURSUE EXPERIMENTS TO TRACE KEY METABOLITES AND DEMONSTRATE THEIR IMPORTANCE IN MEDIATING DNA ACCESSIBILITY AND TRANSDIFFERENTIATION TO IDENTIFY THEIR ROLE IN EPIGENETIC REGULATION IN CELL FATE TRANSITION. COMPLETION OF THESE STUDIES WILL DEMONSTRATE THE NOVEL CONCEPT THAT METABOLIC REGULATION WITHIN CELLS CONTRIBUTES TO THEIR FATE AND PROVIDE NOVEL TARGETS TO ENHANCE THIS PROCESS FOR THE TREATMENT OF PAD.

LANDSCAPES FOR CELL STATE TRANSITION LEVERAGING BY SINGLE-CELL MULTI-OMICS - THE OVERALL GOAL OF THIS PROJECT IS TO DEVELOP NOVEL MATHEMATIC METHODS AND TOOLKITS TO CONNECT CELL FATE TRANSITION AND EPIGENETIC REGULATION ACROSS TISSUES AND DISEASES. CELL FATE TRANSITION OFTEN OCCURS IN ORGAN DEVELOPMENT, TISSUE REGENERATION, AND PATHOGENESIS. DYSREGULATION OF THE CELL FATE TRANSITION CAN LEAD TO ABNORMAL DEVELOPMENT OR DISEASES, SUCH AS TYPE 2 DIABETES, OBESITY, HEART FAILURE, AND ALZHEIMER’S DISEASE. QUANTITIVELY DECODING HOW CELL FATE CHANGES CAN PROVIDE NOVEL MECHANISTIC INSIGHT INTO ORGANOGENESIS AND TISSUE REGENERATION, AND HELP IDENTIFY NEW STRATEGIES FOR THE TREATMENT OF HUMAN DISEASES. HOWEVER, OUR KNOWLEDGE OF CELL FATE TRANSITION AND ITS REGULATION IS ONLY THE TIP OF THE ICEBERG DUE TO THE IMPRACTICALITY OF LONG- TERM TRACING OF CELL TRANSCRIPTOMES. IN THE PAST DECADE, NUMEROUS SINGLE-CELL ATLASES CONTAINING MILLIONS OF CELLS IN DIFFERENT TISSUES, ORGANS, DEVELOPMENTAL STAGES, AND BIOLOGICAL CONDITIONS ARE ROUTINELY DEVELOPED BY CONSORTIA SUCH AS THE HUMAN CELL ATLAS (HCA). THESE ATLASES PROVIDE AN OPPORTUNITY FOR THE UNBIASED STUDY OF CELLULAR DYNAMICS AND THE REGULATION MECHANISM. THE LACK OF COMPUTATIONAL METHODS PRESENTS A MAJOR KNOWLEDGE GAP IN THE UNDERSTANDING OF CELL DYNAMICS AND THE REGULATION LEVERAGING BY THOSE LARGE REFERENCE ATLASES. TO ADDRESS THIS KNOWLEDGE GAP, WE PROPOSED A NEW CONCEPT OF “REFERENCE-BASED CELLULAR DYNAMIC INFERENCE”, WHICH IS A NOVEL STRATEGY TO AUTOMATICALLY ANNOTATE THE CELL STATE TRANSITION IN NEW DATASETS BY LEARNING FROM THE APPROPRIATE REFERENCE, ALLOWING US TO EASILY PERFORM COMPARATIVE ANALYSIS AMONG DIFFERENT TISSUES AND DISEASE CONDITIONS. IN THIS PROJECT, WE WILL PURSUE THREE PARALLEL BUT COMPLEMENTARY RESEARCH DIRECTIONS: 1) TO DEVELOP THE FIRST COMPUTATIONAL METHODS AND TOOLKITS FOR GENERATING CELL DYNAMICS ATLASES AND ANALYZING CELL STATE TRANSITION BASED ON THE APPROPRIATE REFERENCE ATLASES; 2) TO DEVELOP NOVEL STATISTICAL MODELS FOR STUDYING EPIGENETIC REGULATION OF CELL FATE FROM SINGLE-CELL MULTIOMICS DATA; 3) TO GENERATE THE FIRST DYNAMIC REFERENCE LANDSCAPES OF CELL DIFFERENTIATION, SUCH AS CARDIOGENESIS, HEMATOPOIESIS, AND NEUROGENESIS, AND IN- HOUSE LANDSCAPES OF TRANSDIFFERENTIATION. THIS PROJECT WILL BE BUILT ON THE FOUNDATION OF OUR RECENT STUDIES FOR THE DEVELOPMENT OF COMPUTATIONAL APPROACHES TO UNCOVER CELL STATE TRANSITION FROM SINGLE-CELL TRANSCRIPTOMES IN BOTH HOMOGENEOUS AND HETEROGENEOUS CELL POPULATIONS AND THE STUDIES FOR INVESTIGATING THE ROLE OF EPIGENETIC REGULATION ON CELL FATE TRANSITION. THE PROPOSED STUDIES WILL GENERATE ADVANCED COMPUTATIONAL TOOLKITS AND BROADLY APPLICABLE DYNAMIC REFERENCE ATLASES, WHICH ARE EXPECTED TO REVEAL PROFOUND MECHANISMS CONTROLLING CELL STATE TRANSITION IN HEALTH AND DISEASE. IN THE LONG TERM, THE ABILITY TO BUILD CELL DYNAMICS REFERENCE LANDSCAPES WILL OPEN A NEW HORIZON TO UNDERSTAND THE DIVERSITY OF CELL FATE THROUGH COMPARATIVE ANALYSES ACROSS TISSUES AND DISEASES AND ENHANCE REGENERATIVE MEDICINE.

FUNCTIONAL ROLES OF MTOR IN TUMOR PERSISTENCE - PROJECT SUMMARY THE MTOR PATHWAY IS A MASTER REGULATOR OF NUTRIENT METABOLISM AND CELL PROLIFERATION IN RESPONSE TO ENVIRONMENTAL CUES. IT HAS LONG BEEN VIEWED AS AN IMPORTANT DRIVER OF TUMORIGENESIS AND AN IDEAL CANCER THERAPEUTIC TARGET. MULTIPLE MTOR INHIBITORS HAVE BEEN TESTED IN MORE THAN 500 CLINICAL TRIALS, EITHER AS SINGLE AGENTS OR IN COMBINATION WITH CHEMOTHERAPY FOR CANCER TREATMENT. HOWEVER, AMONG NEARLY 200 COMPLETED TRIALS, FEW SHOW POSITIVE OUTCOMES, CHALLENGING THE UTILITY OF MTOR AS AN EFFECTIVE CANCER THERAPEUTIC TARGET. AS AN INTEGRATOR OF EXTRACELLULAR SIGNALS AND CELLULAR RESPONSES, THE MTOR PATHWAY IS CRITICAL FOR CELLULAR ADAPTATION TO ENVIRONMENTAL CHANGES. IN PARTICULAR, MTOR INHIBITION INDUCES EMBRYONIC DIAPAUSE, A REVERSIBLE DORMANCY STATE IN DEVELOPMENT IN RESPONSE TO UNFAVORABLE ENVIRONMENT CUES. WE REASON THAT, ANALOGOUS TO ITS ROLE IN EMBRYONIC DIAPAUSE, MTOR INHIBITION MAY INDUCE AN ADAPTIVE DIAPAUSE-LIKE DORMANT STATE IN CANCERS, LEADING TO THE PERSISTENCE OF RESIDUAL TUMORS FOLLOWING CHEMOTHERAPY. IN LINE WITH THIS NOTION, RECENT STUDIES HAVE INDEED REPORTED MARKEDLY REDUCED MTOR ACTIVITY IN RESIDUAL TUMORS. HOWEVER, A CAUSAL RELATIONSHIP BETWEEN MTOR INHIBITION AND TUMOR PERSISTENCE HAS NOT BEEN ESTABLISHED OR INVESTIGATED. INTEGRATING MULTIPLE STATE-OF-THE-ART APPROACHES, OUR RECENT STUDIES SHOW THAT MTOR INHIBITION INDEED INDUCES A DIAPAUSE-LIKE PERSISTER STATE IN TUMOR CELLS. WE CONFIRM THAT THIS IS A PAN-CANCER PHENOMENON USING TUMORS FROM DIVERSE TISSUE ORIGINS AND DEMONSTRATE THAT THE PERSISTER STATE RECAPITULATES THE RESIDUAL TUMORS IN PATIENTS FOLLOWING CHEMOTHERAPY. IMPORTANTLY, OUR STUDIES IDENTIFY FERROPTOSIS AS A KEY DRUGGABLE VULNERABILITY OF PERSISTERS. BUILT ON THESE COMPELLING FINDINGS, WE HYPOTHESIZE THAT MTOR IS A MASTER REGULATOR OF A DIAPAUSE-LIKE PERSISTER STATE IN TUMORS BY MODULATING SURVIVAL AND DORMANCY. THE CURRENT PROPOSAL AIMS TO INVESTIGATE THE MECHANISTIC REGULATION OF THE PERSISTER STATE, AND TEST THERAPEUTIC STRATEGIES TO TARGET RESIDUAL TUMORS IN PRECLINICAL MODELS, WITH THE ULTIMATE GOAL TO PREVENT TUMOR RECURRENCE IN PATIENTS.

MULTILEVEL INTERVENTION STRATEGIES TO TRANSFORM KIDNEY CARE AND IMPROVE PURSUIT OF TRANSPLANT IN AN INTEGRATED HEALTHCARE DELIVERY SYSTEM - PROJECT SUMMARY APPROXIMATELY 15% OF THE U.S. POPULATION HAS CHRONIC KIDNEY DISEASE, AND ~700,000 PATIENTS ARE IN FULL KIDNEY FAILURE ALSO CALLED END-STAGE KIDNEY DISEASE (ESKD). THE OPTIMAL TREATMENT FOR ESKD IS LIVING DONOR KIDNEY TRANSPLANTATION (LDKT), FOLLOWED BY DECEASED DONOR KIDNEY TRANSPLANTATION (DDKT); HOWEVER, THE STANDARD OF CARE CONTINUES TO BE ONGOING DIALYSIS, WHICH HAS POOR CLINICAL OUTCOMES IN COMPARISON TO LDKT AND DDKT. BEST PRACTICES TO TRANSFORM KIDNEY CARE RECOMMENDED BY THE CENTERS FOR MEDICARE AND MEDICAID SERVICES (CMS), THE AMERICAN SOCIETY OF NEPHROLOGY (ASN) AND THE 2019 EXECUTIVE ORDER ADVANCING AMERICAN KIDNEY HEALTH INITIATIVE INCLUDE EARLIER DETECTION OF PATIENTS WHOSE KIDNEYS ARE DETERIORATING RAPIDLY, INTRODUCING TRANSPLANT AS A POTENTIAL TREATMENT OPTION EARLIER, OPTIMALLY BEFORE THEIR KIDNEYS FAIL, IMPROVED DISSEMINATION OF HEALTH LITERATE TRANSPLANT EDUCATION TOOLS, OFTEN THROUGH DIGITAL TECHNOLOGY OR MHEALTH, AND INCREASING LDKT RATES BY HELPING PATIENTS LOCATE LIVING DONORS OR MOTIVATING OTHERS TO DONATE. BARRIERS AT THE PATIENT-, SUPPORT NETWORK-, CLINICIAN- AND SYSTEM-LEVELS OF THE SOCIO-ECOLOGICAL MODEL PERSIST, INCLUDING POOR IDENTIFICATION OF HIGH-RISK PATIENTS, INSUFFICIENT CLINICIAN TIME TO DISCUSS TRANSPLANT, POOR TRANSPLANT KNOWLEDGE, RELUCTANCE OR INSUFFICIENT SUPPORT TO ASK LIVING DONORS TO DONATE, AND DISENGAGED FRIENDS AND RELATIVES, SOME OF WHOM WHO MIGHT BECOME LIVING DONORS. WHILE EXTENSIVE POLICY AND INTERVENTION EFFORTS ARE UNDERWAY, NONE HAVE ACHIEVED SIGNIFICANT INCREASES IN PURSUIT AND RECEIPT OF TRANSPLANT, ESPECIALLY LDKT RATES. IN 2017, KAISER PERMANENTE SOUTHERN CALIFORNIA (KPSC), AN INTEGRATED CARE SYSTEM SERVING 24,000 CKD PATIENTS, PARTNERED WITH THE TRANSPLANT RESEARCH AND EDUCATION CENTER (TREC) AT HOUSTON METHODIST RESEARCH INSTITUTE (HMRI) AND J.C. WALTER JR. TRANSPLANT CENTER HOUSTON METHODIST HOSPITAL (HMH) TO LAUNCH A MULTI-YEAR PLAN FOR TRANSFORMING CKD AND ESKD CARE. WE NOW PROPOSE TO CONDUCT A PRAGMATIC STEPPED WEDGE CLUSTER RANDOMIZED TRIAL OF A NOVEL MULTILEVEL INTERVENTION TO IMPROVE CKD AND ESKD CARE, IMPROVE TRANSPLANT RATES AND REDUCE DISPARITIES. ONE INNOVATIVE COMPONENT OF THE MULTILEVEL INTERVENTION IS A STATE-OF-THE-ART TECHNOLOGY-SUPPORTED GROVE KIDNEY HEALTH MHEALTH APPLICATION, DEVELOPED IN PARTNERSHIP WITH PATIENTS, TO ENGAGE PATIENTS, FAMILY MEMBERS, AND POTENTIAL LIVING DONORS TO IMPROVE THEIR CKD KNOWLEDGE, VIEW TRANSPLANT SUCCESS STORIES, AND SEEK KIDNEY-RELATED SUPPORT TO PURSUE TRANSPLANT, INCLUDING LEARNING HOW TO FIND LIVING DONORS. WE ALSO SEEK TO IDENTIFY MODERATORS AT VARIOUS SOCIO- ECOLOGICAL LEVELS, ESPECIALLY FACTORS INFLUENCING VARIATIONS IN EFFECTIVENESS ACROSS DIFFERENT SETTINGS AND AMONG UNDERSERVED PATIENT SUBGROUPS KNOWN TO HAVE REDUCED ACCESS TO TRANSPLANT AND BUILD IMPLEMENTATION TOOLS TO INCREASE ACCESS TO AND PURSUIT OF TRANSPLANT WITHIN LARGE INTEGRATED HEALTH SYSTEMS INCLUDING COMPARABLE SYSTEMS (COMMERCIAL, ACADEMIC, SAFETY NET) ACROSS THE U.S.

NANOPARTICLES FOR HARVESTING AND TARGETING ANGIOGENIC PROTEINS

A NOVEL PATHWAY INVOLVING ATM PP1 AND I-2

4/4-AMERICAN CONSORTIUM OF EARLY LIVER TRANSPLANTATION-PROSPECTIVE ALCOHOL-ASSOCIATED LIVER DISEASE COHORT EVALUATION (ACCELERATE-PACE) - PROJECT SUMMARY THE AMERICAN CONSORTIUM OF EARLY LIVER TRANSPLANTATION-PROSPECTIVE ALCOHOL-ASSOCIATED LIVER DISEASE COHORT EVALUATION (ACCELERATE-PACE) STUDY IS A PROSPECTIVE LONGITUDINAL COHORT OF PATIENTS WITH SEVERE ALCOHOL- ASSOCIATED LIVER DISEASE (ALD) EVALUATED FOR EARLY LIVER TRANSPLANTATION (ELT). THE COHORT LEVERAGES THE ACCELERATE CONSORTIUM WITH 4-LINKED R01S AND 5 ADDITIONAL RECRUITMENT SITES IN THE SOUTH/SOUTHEAST, MID- ATLANTIC, MIDWEST, AND WEST, AND WILL REFINE AND IDENTIFY BEST PRACTICES IN THE SELECTION AND MANAGEMENT OF PATIENTS WITH SEVERE ALD CONSIDERED FOR ELT ACROSS THEIR CONTINUUM OF CARE. ALD IS NOW THE MOST COMMON INDICATION FOR LIVER TRANSPLANTATION (LT) IN THE U.S. HISTORICALLY, LT CENTERS REQUIRED AT LEAST 6 MONTHS OF ALCOHOL ABSTINENCE BEFORE LT REFERRAL AND EVALUATION, THOUGH EMPIRIC EVIDENCE TO SUPPORT MINIMUM SOBRIETY PERIODS WAS LIMITED. ELT, DEFINED AS LT BEFORE 6 MONTHS OF ABSTINENCE, IS INCREASINGLY PERFORMED BUT WITH SIGNIFICANT PRACTICE VARIABILITY. THERE IS NO CONSENSUS ON OPTIMAL ELT CANDIDATE SELECTION, AND SELECTION CRITERIA VARY WIDELY, CONTRIBUTING TO DISPARITIES IN ACCESS TO LIFESAVING CARE. ELT IS ALSO CONTROVERSIAL DUE TO THE POTENTIAL FOR LIVER RECOMPENSATION WITH ABSTINENCE, WHICH WOULD OBVIATE THE NEED FOR LT—ACCURATE PREDICTION OF RECOMPENSATION HAS THE POTENTIAL TO INCREASE ORGAN UTILITY AND STEWARDSHIP. DETAILED EVALUATION OF THE EFFICACY OF ALCOHOL USE DISORDER TREATMENTS AND IMPROVED RISK SCORES BASED ON PRE-LT PSYCHOSOCIAL FACTORS TO PREDICT RETURN TO ALCOHOL USE ARE NEEDED TO REFINE SELECTION CRITERIA, OPTIMIZE POST-LT CARE, AND EFFECTIVELY TREAT AUD. SHORT- AND INTERMEDIATE-TERM SURVIVAL AFTER ELT IS EXCELLENT, BUT THE INCIDENCE AND PREDICTORS OF POST-LT COMPLICATIONS ARE POORLY DEFINED. TO FILL THESE KEY KNOWLEDGE GAPS, WE WILL ENROLL AND PROSPECTIVELY FOLLOW 770 ELT CANDIDATES AND 270 ELT RECIPIENTS FOR 3 YEARS AT 9 SOCIO-DEMOGRAPHICALLY DIVERSE CENTERS. THE PROPOSED AIMS WILL: (I) INFORM ELT SELECTION CRITERIA AND INVESTIGATE POTENTIAL SOURCES OF BIAS IN ELT EVALUATION AND HEALTHCARE DISPARITIES IN ELT ACCESS; (II) DEVELOP RISK PREDICTION SCORES FOR LT-FREE SURVIVAL AND RECOMPENSATION; (III) IDENTIFY EFFECTIVE TREATMENTS (MEDICAL, BEHAVIORAL) FOR ALCOHOL USE DISORDER AMONG PATIENTS WITH SEVERE ALD AND POST-ELT; (IV) EVALUATE CLINICAL OUTCOMES AMONG ELT CANDIDATES AND RECIPIENTS, INCLUDING MORTALITY, TRANSPLANTATION, POST-LT COMPLICATIONS (E.G. CANCER, CARDIOVASCULAR EVENTS, GRAFT REJECTION/FAILURE), AND QUALITY OF LIFE. A COMPREHENSIVE DATA REPOSITORY WILL INCLUDE SOCIODEMOGRAPHIC, CLINICAL, GEOSPATIAL, PSYCHOSOCIAL, BEHAVIORAL, AND PATIENT-REPORTED OUTCOME VARIABLES. LT DOCUMENTS, CHECKLISTS, RECORDINGS OF SELECTION MEETINGS, DIRECT OBSERVATIONS OF LT PROCEDURES, AND CLINICIAN INTERVIEWS WILL IDENTIFY BEST PRACTICES AND PITFALLS IN CANDIDATE SELECTION. A BIOREPOSITORY OF BLOOD, URINE, EXPLANT/DONOR TISSUE, PRE- AND POST-LT LIVER TISSUE, PERIPHERAL BLOOD MONONUCLEAR CELLS, AND CROSS-SECTIONAL RADIOLOGIC IMAGING WILL INFORM FUTURE ANCILLARY STUDIES.

RFA-CA -12-003 - ACTIVATABLE ONE-DROP AND ONE-STEP ASSAY FOR CIRCULATING TUMOR CE

QUANTITATION OF NUTRIENT METABOLISM IN BRAIN TUMOR PATIENTS USING ADVANCED 13C ISOTOPOMER TECHNOLOGY - PROJECT SUMMARY/ABSTRACT MALIGNANT BRAIN TUMORS ARE AGGRESSIVE CANCERS THAT HAVE HIGH PROLIFERATIVE RATES WITH MUCH HIGHER ENERGY REQUIREMENTS AND HIGH MORTALITY RATES. DESPITE INTENSE CLINICAL AND DRUG DEVELOPMENT EFFORTS IN THE LAST TWO DECADES, THERE HAS BEEN NO IMPROVEMENT IN SURVIVAL. TO SUPPORT THE ABNORMAL GROWTH COMMONLY SEEN IN TUMORS, THE CANCER CELLS HAVE ALTERED THEIR METABOLISM COMPARED TO NORMAL CELLS IN HEALTHY TISSUES. MOST OF THE KNOWLEDGE TO DATE ON CANCER METABOLISM IS DERIVED FROM CULTURED CELL LINES. PROBING METABOLISM IN INTACT TUMORS WILL BE CRITICAL TO UNDERSTAND HOW THE TUMOR CELLS GROW IN A PATIENT UNDER THE COMPLEX BIOLOGICAL TUMOR ENVIRONMENT. FROM OUR PILOT STUDY INVOLVING A SMALL NUMBER OF PATIENTS, WE HAVE DEMONSTRATED THAT GLIOMAS AND BRAIN METASTASES HAVE THE CAPACITY TO OXIDIZE ACETATE IN THE CITRIC ACID CYCLE TO MEET THEIR BIOENERGETIC REQUIREMENTS, AND GLUCOSE AND ACETATE TOGETHER CONTRIBUTE UP TO 63.0% OF THE TOTAL ACETYL-COA POOL IN THESE TUMORS. THE REMAINING ACETYL-COA THAT PROVIDES CARBON SOURCES FOR BIOMOLECULAR SYNTHESIS, MUST BE DERIVED FROM OTHER NUTRIENTS. THE FOLLOWING ARE THE GOALS OF THIS PROPOSAL: (1) DETERMINE IF ACETATE AND KETONE BODY (BETA HYDROXYBUTYRATE, BHB) UTILIZATION IS A COMMON PROPERTY OF ALL GLIOMAS OR SPECIFICALLY LINKED TO HIGH GRADE GBMS (2) EXAMINE WHETHER ACETATE AND BHB PROVIDE CARBONS FOR 2-HYDROXYGLUTARATE (2-HG) SYNTHESIS IN IDH MUTATED GLIOMA PATIENTS (3) PRECLINICAL TESTING OF THE EFFECTS OF SMALL MOLECULE INHIBITORS OF ACETATE AND BHB, IN FRESHLY RESECTED TUMOR TISSUE SLICES. WE HAVE INSTITUTIONAL REVIEW BOARD (IRB) APPROVED CLINICAL PROTOCOL TO INFUSE NON-TOXIC AND NON-RADIOACTIVE 13C-ENRICHED ACETATE IN PATIENTS WHO WILL BE UNDERGOING SURGICAL REMOVAL OF A BRAIN TUMOR. USING NUCLEAR MAGNETIC RESONANCE (NMR) SPECTROSCOPY AND MASS SPECTROMETRY OF THESE SURGICALLY RESECTED TUMOR TISSUES, WE WILL INVESTIGATE THE ABOVE DESCRIBED AIMS ON ENERGY METABOLISM OF PRIMARY BRAIN TUMORS. THE ATTRACTIVENESS OF THIS TECHNOLOGY IS THAT NO RADIOACTIVITY IS INVOLVED. WE ANTICIPATE THAT THE OUTCOME OF THIS STUDY WILL GENERATE A DETAILED UNDERSTANDING OF IN VIVO UTILIZATION OF ACETATE AND KETONE BODY IN BRAIN TUMOR PATIENTS. THIS KNOWLEDGE WILL LEAD TO IDENTIFICATION OF KEY METABOLIC TARGETS THAT MAY BE FURTHER EXPLOITED FOR THE DEVELOPMENT OF NEW THERAPIES. ADDITIONALLY, IT MAY IDENTIFY NOVEL BIOMARKERS WHICH MAY BE HELPFUL IN DESIGNING NON-INVASIVE IN VIVO MRI METHODS TO TRACK ACETATE UTILIZATION BY TUMORS FOR DIAGNOSTIC PURPOSES.

ADVANCING THYROID CANCER DIAGNOSTICS WITH AI-ENHANCED MULTIMODAL OPTICAL HISTOPATHOLOGY - PROJECT SUMMARY/ ABSTRACT THE INCREASING INCIDENCE OF THYROID CANCER WORLDWIDE IS PARTLY FUELED BY ENHANCED DETECTION OF SMALLER NODULES THROUGH IMAGING TECHNIQUES LIKE ULTRASOUND, COMPUTED TOMOGRAPHY (CT), AND MAGNETIC RESONANCE IMAGING (MRI), LEADING TO OVERDIAGNOSIS OF LOW-RISK CANCERS AND BENIGN THYROID CONDITIONS. PAPILLARY THYROID CARCINOMA, THE MOST PREVALENT TYPE OF THYROID CANCER, PRESENTS LYMPH NODE METASTASIS (LNM) IN 60 TO 70% OF CASES, INCREASING THE RISK OF LOCAL RECURRENCE, DISTANT METASTASIS, AND MORTALITY. FINE NEEDLE ASPIRATION (FNA), THE PRIMARY DIAGNOSTIC METHOD, YIELDS INDETERMINATE RESULTS FOR 10 TO 20% OF NODULES, OF WHICH 20 TO 30% OF NODULES MAY BE MALIGNANT. SUCH UNCERTAINTY RESULTS IN REPEAT BIOPSIES, COSTLY MOLECULAR TESTS, OR UNNECESSARY SURGERIES, CAUSING INCREASED HEALTHCARE COSTS, REDUCED QUALITY OF LIFE, AND EMOTIONAL DISTRESS. CORE NEEDLE BIOPSIES (CNB) ARE SUGGESTED FOR INDETERMINATE FNA FINDINGS. HOWEVER, CNB IS ALSO PRONE TO SAMPLING ERRORS, NOT SUITABLE NEAR CRITICAL STRUCTURES, AND BOTH FNA AND CNB ARE TIME-CONSUMING PROCEDURES AND CAN ALTER TISSUE MORPHOLOGY. GIVEN THE PREVALENCE OF FALSE POSITIVES, COMPLICATIONS ARISING FROM INDETERMINATE DIAGNOSES, PROLONGED ANALYSIS PERIODS, AND SAMPLING CHALLENGES IN THYROID CANCER DETECTION, THERE IS A PRESSING DEMAND FOR A MORE EFFECTIVE DIAGNOSTIC SOLUTION. OUR PROPOSAL INTRODUCES AN AI-AUGMENTED, MULTIMODAL, LABEL-FREE NONLINEAR OPTICAL MICROSCOPY SYSTEM THAT INCORPORATES COHERENT ANTI-STOKES RAMAN SCATTERING MICROSCOPY (CARS), SECOND HARMONIC GENERATION MICROSCOPY (SHG), AND TWO-PHOTON AUTOFLUORESCENCE MICROSCOPY (TPAF). THIS INNOVATIVE SYSTEM AIMS FOR RAPID AND PRECISE DIAGNOSIS OF THYROID CANCER AND LNM. BY ELIMINATING THE NEED FOR EXTERNAL DYES, THE LABEL-FREE TECHNIQUE STREAMLINES THE DIAGNOSTIC PROCESS AND MINIMIZES COMPLICATIONS. THIS COMBINATION OF IMAGING MODALITIES WITH AI AUGMENTATION OFFERS DETAILED AND COMPLIMENTARY SUBCELLULAR MORPHOLOGICAL AND BIOCHEMICAL INFORMATION, ENHANCING OUR UNDERSTANDING OF BIOCHEMICAL COMPOSITION OF THYROID NODULES AND LYMPH NODES, FACILITATING IDENTIFICATION OF MALIGNANCY MARKERS MORE ACCURATELY, AND SIGNIFICANTLY IMPROVING DIAGNOSTIC ACCURACY. IN ADDITION, WE PLAN TO DEVELOP A COMPACT, PORTABLE, AI INTEGRATED LABEL-FREE MICROENDOSCOPE FOR CLINICAL TRANSLATION, DESIGNED TO FIT WITHIN A CORE BIOPSY NEEDLE, FOR DIFFERENTIATING BETWEEN CANCEROUS AND NORMAL TISSUES AND DETECTING LNM WITHOUT TISSUE EXCISION. VALIDATED THROUGH STUDIES ON EX VIVO HUMAN TISSUES AND IN VIVO PORCINE MODELS, THIS DEVICE PROMISES TO TRANSFORM THYROID CANCER DIAGNOSIS BY SERVING AS A VIRTUAL HISTOPATHOLOGY TOOL, ENABLING REAL-TIME, LABEL-FREE IMAGING AND ADDRESSING THE DRAWBACKS OF FNA AND CNB, SUCH AS OVERDIAGNOSES, INDETERMINATE DIAGNOSES, AND FALSE POSITIVES, WITHOUT TISSUE EXCISION OR HARM TO CRITICAL ADJACENT STRUCTURES. UPON PROJECT COMPLETION, WE ANTICIPATE DELIVERING AN AI-ENHANCED, LABEL-FREE BENCHTOP SYSTEM FOR TESTING IN OUTPATIENT CLINICS, CAPABLE OF IMAGING FNA SAMPLES FOR RAPID AND PRECISE DIAGNOSIS WITHIN MINUTES, AND A COMPARABLE LABEL-FREE MICROENDOSCOPE AS A POTENTIAL OPTICAL HISTOPATHOLOGY DEVICE TO FACILITATE NEAR REAL-TIME CANCER DIAGNOSIS WITHOUT TISSUE EXCISION.

NEURONAL SPINES TRACKING AND ANALYSIS FOR TIME-LAPSE, 3D OPTICAL MICROSCOPY

MCP-1 AND MMP-1 CONTRIBUTIONS IN SUSCEPTIBILITY TO DEVELOPING ACTIVE TUBERCULOSIS

MITOCHONDRIAL FACTORIES FOR AMD THERAPY - PROJECT SUMMARY AGE-RELATED MACULAR DEGENERATION (AMD) IS THE LEADING CAUSE OF IRREVERSIBLE BLINDNESS INVOLVING GRADUAL DYSFUNCTION AND DEGENERATION OF PHOTORECEPTORS AND THE RETINAL PIGMENT EPITHELIUM (RPE). MITOCHONDRIAL DYSFUNCTION REPRESENTS A HALLMARK OF AMD. EXCESSIVE AND PERSISTENT REACTIVE OXYGEN SPECIES (ROS) DAMAGES MITOCHONDRIAL DNA (MTDNA), RESULTING IN FEWER MITOCHONDRIA. IMPAIRMENTS IN RESPIRATORY CHAIN ACTIVITY AND REDUCED OXIDATIVE PHOSPHORYLATION (OXPHOS) IMPACTS ATP SYNTHESIS AND FURTHER DRIVES ROS PRODUCTION. OUR PRIOR WORK HAS SHOWN THAT EXOGENOUS MITOCHONDRIAL TRANSPLANTATION TO CELLS CAN INDUCE A BIOENERGETIC SHIFT TOWARDS INCREASED OXPHOS AND ATP PRODUCTION, AND REDUCED ROS. MITOCHONDRIAL CELL-TO-CELL TRANSFER IN RESPONSE TO STRESS RESCUES AEROBIC RESPIRATION TO REDUCE DELETERIOUS CELL DYNAMICS. WE AIM TO CREATE A SUSTAINABLE SOURCE OF MITOCHONDRIA PROXIMAL TO THE RPE IN THE FORM OF MESENCHYMAL STEM CELLS (MSCS) TRANSFECTED WITH NUCLEAR RESPIRATORY FACTOR 1 (NRF1), A DRIVER OF MITOCHONDRIAL BIOGENESIS, FOR ENHANCED CELL-TO- CELL TRANSFER OF MITOCHONDRIA. OUR OBJECTIVES ARE TO DETERMINE HOW NRF1 TRANSFECTION CHANGES MSC TRANSCRIPTIONAL ACTIVITY, MITOCHONDRIAL PRODUCTION AND TRAFFICKING, AND EXTRACELLULAR VESICLE (EV) CONTENTS, AND TEST THE STRATEGY IN TWO RODENT MODELS OF RETINAL DEGENERATION. WE HYPOTHESIZE THAT OUR CELL THERAPY USING NRF1- OVEREXPRESSING MSCS WILL NOT ONLY PROVIDE THE NEEDED ENERGETIC BOOST VIA MITOCHONDRIAL MASS TRANSFER TO RETINAL CELLS, BUT ALSO INDUCE PROFOUND CELLULAR REPROGRAMMING THROUGH THE SECRETION OF EV-CONTAINING FACTORS, THEREBY COUNTERACTING AND POTENTIALLY REVERSING THE ONSET OF LATE-STAGE DRY AMD. WE WILL USE A COMBINATION OF MOLECULAR AND BIOCHEMICAL METHODS, INCLUDING TRANSCRIPTOMICS, TO CHARACTERIZE THE POTENCY OF MITOCHONDRIAL TRANSFER TO IMPACT AMD. WE WILL FIRST INVESTIGATE THE IMPACT OF NRF1 ON MITOCHONDRIAL TRANSFER MECHANISMS IN MSCS. WE WILL ELUCIDATE THE ROLE OF NRF1 ON MITOCHONDRIAL TRANSFER MECHANISMS, SPECIFICALLY TUNNELING NANOTUBES (TNTS) AND EVS, BETWEEN MSCS AND HUMAN RPE CELLS FROM DONOR EYES AFFLICTED WITH AMD, SHEDDING LIGHT ON NRF1- PRIMED MSCS AS MITOCHONDRIA GENERATORS, RESPONSIVE TO SIGNALS FROM STRESSED CELLS. WE WILL THEN EXAMINE THE INFLUENCE OF NRF1 ON MSC SENESCENCE AND EV COMPOSITION, PROVIDING INSIGHTS INTO THE MULTIFACETED BENEFITS OF NRF1 IN MSCS, INCLUDING METABOLIC ADAPTATION TO AMD STRESS ENVIRONMENTS AND MODULATION OF EV CONSTITUENTS. LASTLY, WE WILL EVALUATE THE IMPACT OF NRF1 PRIMED MSC TRANSPLANTATION IN VIVO. THROUGH LONGITUDINAL AND LONG- TERM EFFICACY STUDIES AND TISSUE TRANSCRIPTOMIC ANALYSIS, WE WILL IDENTIFY DISEASE STAGES MOST RESPONSIVE TO NRF1 MSCS WHEN TRANSPLANTED SUBRETINALLY IN A SODIUM IODATE (NAIO3)-INDUCED MODEL OF RETINAL DEGENERATION AND SENESCENCE ACCELERATED MOUSE (SAMP8) MODEL, HIGHLIGHTING THE POTENTIAL TO RESCUE RPE CELL DEGENERATION AND ABROGATE AMD PROGRESSION. FINDINGS FROM THIS WORK WILL PROVIDE CRUCIAL INSIGHTS INTO THE POTENTIAL OF NRF1-BASED THERAPIES FOR AMD, OFFERING A PROMISING AVENUE FOR THE TREATMENT AND MANAGEMENT OF THIS DEBILITATING DISEASE.

ANTIMICROBIAL RESISTANCE TRAINING PROGRAM IN THE TEXAS MEDICAL CENTER (AMR-TPT) - PROJECT SUMMARY/ABSTRACT THE RISE OF ANTIBIOTIC-RESISTANT MICROORGANISMS GLOBALLY HAS BECOME A CRITICAL PUBLIC HEALTH PRIORITY DUE TO THE DEVASTATING CONSEQUENCES THAT IT MAY HAVE TO THE WORLD HEALTH AND ECONOMY. ANTIBIOTIC RESISTANCE (AMR) THREATENS THE PROGRESS OF MEDICINE IN ALL AREAS, AND AS SUCH, THE ISSUE HAS REACHED THE HIGHEST LEVEL OF GOVERNMENT, INCLUDING THE OFFICE OF THE US PRESIDENT AND THE UNITED NATIONS. AMR IS A TOP PUBLIC HEALTH PRIORITY FOR BOTH CDC AND WHO AND HAS BEEN DESIGNATED AS THE “SILENT” PANDEMIC. HOUSTON IS THE HOME TO THE LARGEST CLUSTER OF HEALTHCARE INSTITUTIONS IN THE WORLD, THE TEXAS MEDICAL CENTER (TMC), WITH MORE THAN 9,200 HOSPITAL BEDS AND 10 MILLION PATIENT VISITS PER YEAR. THE HOUSTON AREA’S STRONG HISTORY OF OUTSTANDING INFECTIOUS DISEASES RESEARCH AND TRAINING INCLUDES A FOCUS ON AMR AND ANTIBIOTIC STEWARDSHIP. THE COMBINED EFFORTS OF THE RECENTLY FORMED HOUSTON METHODIST CENTER OF EXCELLENCE FOR INFECTIOUS DISEASES AND THE EXISTING GULF COAST CONSORTIUM (GCC) FOR ANTIMICROBIAL RESISTANCE HAVE RESULTED IN AN ACTIVE, MULTIDISCIPLINARY AND COMPREHENSIVE RESEARCH AND EDUCATIONAL PROGRAM AMALGAMATING THE ENDEAVORS OF AMR RESEARCHERS AND CREATING THE RESOURCES, PERSONNEL, FUNDING AND FERTILE GROUND TO DEVELOP AN AMBITIOUS, INNOVATIVE, UNPARALLELED AMR TRAINING PROGRAM. WE PROPOSE TO ESTABLISH THE AMR TRAINING PROGRAM IN THE TEXAS MEDICAL CENTER (AMR-TPT) THAT TRAINS POSTDOCTORAL SCHOLARS, CLINICAL RESIDENTS/FELLOWS AND PHARMD FELLOWS FROM EIGHT INSTITUTIONS IN THE TMC (HOUSTON METHODIST RESEARCH INSTITUTE, UNIVERSITY OF TEXAS HEALTH SCIENCE CENTER AT HOUSTON, MD ANDERSON CANCER CENTER, BAYLOR COLLEGE OF MEDICINE, UNIVERSITY OF TEXAS MEDICAL BRANCH, RICE UNIVERSITY, UNIVERSITY OF HOUSTON, AND INSTITUTE OF BIOSCIENCES AND TECHNOLOGY OF TEXAS A&M UNIVERSITY) ON ADVANCED ASPECTS OF AMR RESEARCH. WE WILL TAKE ADVANTAGE OF THE STRONG ADMINISTRATIVE EXPERTISE OF THE GCC ON SUCCESSFUL T32 PROGRAMS IN OTHER AREAS AND THE EDUCATIONAL ACTIVITIES ALREADY IN PLACE, COMBINED WITH THE EXPERTISE OF WORLD-CLASS AMR RESEARCHERS. AMR-TPT SEEKS TO LEVERAGE AND CREATE RESOURCES TO TRAIN THE NEXT GENERATION OF SCIENTISTS AND CLINICIAN-SCIENTISTS (MDS, PHDS AND PHARMDS) FOCUSED ON TACKLING THE PRESSING AMR PUBLIC HEALTH CRISIS. THE HIGHLY COLLABORATIVE ENVIRONMENT PROVIDES THE PERFECT OPPORTUNITY FOR TRAINEES TO ACQUIRE THE SKILLS, EXPERTISE AND INTELLECTUAL ABILITIES TO FOSTER INNOVATIVE RESEARCH THAT HAS A STRONG TRANSLATIONAL COMPONENT AND COULD BE DEVELOPED TO DIRECTLY INFLUENCE PATIENT CARE. ALONG WITH A HIGHLY INTERACTIVE AMR FOUNDATIONS COURSE AND MYRIAD CAREER/PROFESSIONAL DEVELOPMENT OPPORTUNITIES, THE PROPOSED TRAINING GRANT INCLUDES EXPERTISE IN AND A FOCUS ON I) MOLECULAR BASIS OF RESISTANCE, II) BACTERIAL GENOMICS AND BIOINFORMATICS, III) DIAGNOSTICS, IV) PHARMACOLOGICAL ASPECTS OF RESISTANCE, V) MICROBIOME SCIENCE, VI) CLINICAL EPIDEMIOLOGY AND BIOSTATISTICS OF AMR AND VII) ANTIBIOTIC STEWARDSHIP. WE BELIEVE WE ARE POISED TO CONTINUE DEVELOPING A UNIQUE, INNOVATIVE AND COMPREHENSIVE TRAINING PROGRAM THAT TRULY PROVIDES TRAINEES WITH EXCEPTIONAL TOOLS AND ABILITIES AND CREATES A STRONG COHORT OF NEW WORLD-CLASS AMR LEADERS AND RESEARCHERS.

THYROID HORMONE RECEPTOR

PROJECT TITLE: LARGE-SCALE MULTIOMIC APPROACH TO IDENTIFY SYSTEMIC AND CENTRAL IMMUNE-RELATED BIOMARKERS IN AMYOTROPHIC LATERAL SCLEROSIS.

EXPLORING FUNCTIONAL COMPLEXES AND DISEASE NETWORKS WITHIN HUMAN RNA-BINDING PROTEIN INTERACTOMES

HIGH-CONTENT IMAGE ANALYSIS AND MODELING FOR NEURON ASSAY BASED SCREENING

TRANSFORMING CURIOSITY INTO DONATION: VALIDATING A RISK PREDICTION INDEX TO DETECT AND PREVENT DROP-OUT IN POTENTIAL LIVING KIDNEY DONORS WHO ARE RACIAL/ETHNIC MINORITIES - PROJECT SUMMARY APPROXIMATELY 1 IN 7 US ADULTS HAVE CHRONIC KIDNEY DISEASE (CKD), AND >800,000 PATIENTS ARE IN FULL KIDNEY FAILURE ALSO CALLED END-STAGE KIDNEY DISEASE (ESKD). THE OPTIMAL TREATMENT FOR ESKD IS LIVING KIDNEY DONOR TRANSPLANTATION (LKDT); HOWEVER, THE STANDARD OF CARE CONTINUES TO BE ONGOING DIALYSIS, WHICH HAS POOR CLINICAL OUTCOMES IN COMPARISON TO LKDT. CKD CONTINUES TO INCREASE AMONG HISPANICS AND LOW-INCOME PATIENTS. BLACK AND HISPANIC/LATINO PERSONS ARE RESPECTIVELY 3.1- AND 1.3-TIMES MORE LIKELY THAN WHITE PERSONS TO DEVELOP ESKD. MANY INDIVIDUALS ARE CURIOUS ABOUT BECOMING A LIVING DONOR, BUT SOME DECIDE NOT TO DONATE FOR UNKNOWN REASONS. THIS PREVENTS TRANSPLANT CENTERS AND KIDNEY ORGANIZATIONS FROM IMPROVING THEIR KIDNEY DONATION PROCESS AND TAILORING DONOR CARE TO OVERCOME BARRIERS, PARTICULARLY FOR VULNERABLE AND UNDERSERVED POPULATIONS. DROP-OUT IS ESPECIALLY COMMON FOR POTENTIAL DONORS WHO ARE BLACK AND HISPANIC, SPECIFICALLY THOSE WHO FACE GREATER SOCIOECONOMIC CHALLENGES TO DONATING, AND THOSE WHO ARE NOT BIOLOGICALLY RELATED TO THEIR KIDNEY PATIENTS. DROP- OUT FROM THESE GROUPS OF POTENTIAL DONORS CONTRIBUTES TO KNOWN DISPARITIES IN LKDT. TO REDUCE DISPARITIES IN LIVING DONATION AND INCREASE LKDT, WE PROPOSE A MULTI-STEP STUDY THAT INCLUDES: AIM 1A: EARLY IDENTIFICATION OF PATIENTS OF ALL RACES/ETHNICITIES AT HIGHER RISK OF DROPPING OUT FROM THE DONOR EVALUATION PROCESS VIA SECONDARY DATA ANALYSES OF MULTIPLE REGIONAL AND NATIONAL DONOR DATABASES. A TAILORED RISK PREDICTION INDEX (LD-DROP) WILL BE DEVELOPED AND TAILORED TO CHARACTERIZE THE RISK OF DROP-OUT AMONG POTENTIAL DONORS AT THE TIME OF INITIAL SCREENING OR CONTACT, INCLUDING SUBGROUP ANALYSES BY RACE, ETHNICITY, AND SOCIOECONOMIC STATUS; AIM 1B: VALIDATION OF THE LD-DROP INDEX BY PROSPECTIVELY TRACKING A LARGE, DIVERSE GROUP OF INDIVIDUALS PRESENTING FOR LIVING KIDNEY DONOR EVALUATION AT TWO LARGE AND GEOGRAPHICALLY DIVERSE TRANSPLANT CENTERS; AIM 2: INDIVIDUAL INTERVIEWS OF POTENTIAL LIVING DONORS OF VARIOUS RACE, GENDER, AND LEVEL OF RISK FOR DROP- OUT WILL BE CONDUCTED TO EXPLORE THEIR DECISION-MAKING AND READINESS TO DONATE, WHAT VULNERABILITIES AND BARRIERS TO DONATION THEY FACE, AND WHAT FINANCIAL, PSYCHOSOCIAL, PEER MENTORING, AND EDUCATIONAL INTERVENTIONS MIGHT BE MOST HELPFUL FOR OVERCOMING THESE BARRIERS; AIM 3: AN INTERVENTION TO ADDRESS DONATION BARRIERS WILL BE PILOTED FOR POTENTIAL DONORS AT HIGH DROP-OUT RISK THROUGH A NATIONAL ONLINE PEER SUPPORT PROGRAM AND TRANSPLANT CENTER- BASED EDUCATION PROGRAMS FOR POTENTIAL LIVING DONORS. PROJECT DELIVERABLES WILL BE MADE FREELY AVAILABLE NATIONWIDE VIA PARTNERSHIPS WITH THE SCIENTIFIC REGISTRY OF TRANSPLANT RECIPIENTS LIVING DONOR COLLECTIVE INITIATIVE AND THE NATIONAL KIDNEY FOUNDATION.

ROLE OF YXDJK AND DAK IN THE ENTEROCOCCAL ENVELOPE STRESS RESPONSE

CENTER FOR THE DISSEMINATION OF ULTRA-LONG-ACTING ANTIRETROVIRAL RELEASE TECHNOLOGY: DART RESOURCE PROGRAM - ABSTRACT HUMAN IMMUNODEFICIENCY VIRUS (HIV) IS ONE OF THE TOP AREAS OF THE NATIONAL INSTITUTES OF HEALTH- FUNDED NONHUMAN PRIMATES (NHP) RESEARCH. WITHIN THIS CONTEXT, ANTIRETROVIRAL (ARVS) USED IN RESEARCH FOR HIV PRE-EXPOSURE PROPHYLAXIS AND TREATMENT REQUIRE FREQUENT DOSING REGIMENS. ARVS ARE OFTEN ADMINISTERED THROUGH DAILY ORAL DOSING OR REPEATED INJECTIONS, WHICH ARE COSTLY AS WELL AS BURDENSOME ON ANIMALS, VETERINARY STAFF AND OPERATIONAL RESOURCES. MOREOVER, INCONSISTENCY IN DOSING REGIMEN OR DOSAGES COULD BE DETRIMENTAL TO RESEARCH OUTCOMES. GIVEN THIS, WE PROPOSE THE DISSEMINATION OF A DRUG-AGNOSTIC ULTRA-LONG-ACTING SUBCUTANEOUS DRUG DELIVERY IMPLANT, THE NANODDI, AS A RESEARCH TOOL FOR NHP STUDIES, THROUGH THE CENTER FOR THE DISSEMINATION OF ULTRA-LONG-ACTING ANTIRETROVIRAL RELEASE TECHNOLOGY: DART RESOURCE PROGRAM. THE DART PROGRAM WILL PROVIDE THE NANODDI FOR USE IN NHP STUDIES ON HIV FOR LONG- TERM ARV RELEASE TO REPLACE CONVENTIONAL MODES OF ADMINISTRATION. SUSTAINED AND CONSTANT ARV DELIVERY FROM THE NANODDI IS ACHIEVED THROUGH CONTROLLED DRUG DIFFUSION ACROSS A NANOFLUIDIC MEMBRANE WITHOUT PUMPS OR PORTS. THE NANODDI TECHNOLOGY IS IMMEDIATELY READY FOR USE AND RAPID AND REPRODUCIBLE MANUFACTURING PERMITS WIDESPREAD FACILE DISSEMINATION. THE DRUG-AGNOSTIC TECHNOLOGY IS SUPPORTED BY 10 YEARS OF DEVELOPMENT AND DEPLOYMENT IN VARIOUS ANIMAL MODELS, WITH DEMONSTRATIONS OF LONG-TERM SUSTAINED RELEASE OF VARIOUS ARVS IN NHP FOR UP TO 29 MONTHS WITHOUT INTERRUPTION. THE CENTRAL TENET OF THE HIV-RELATED NHP STUDIES AND ALLEVIATE ANIMAL, OPERATIONAL AND FINANCIAL BURDEN ASSOCIATED WITH CONVENTIONAL ADMINIST ATION METHODS. THE AIMS OF THE PROPOSED DART PROGRAM, WHICH WILL BE PURSUED CONCURRENTLY OVER FIVE DART PROGRAM IS THAT THE NANODDI CAN STANDARDIZE ARV DELIVERY ACROSS WIDE-RANGING YEARS, ARE, AIM 1: OUTREACH AND DISSEMINATION OF NANODDI FOR HIV RESEARCH IN NHP, AIM 2: INTEGRATION AND DEPLOYMENT OF NANODDI FOR ARV DELIVERY IN INDEPENDENT USERS’ LABORATORIES, AND AIM 3: BROADEN THE SPECTRUM OF APPLICABILITY OF NANODDI WITHIN THE HIV RESEARCH COMMUNITY. OUTREACH INITIATIVES WILL CREATE AWARENESS AND PROMOTE ADOPTION OF THE NANODDI INTO CURRENT AND FUTURE NHP HIV RESEARCH. THE DART PROGRAM WILL ENSURE STANDARDIZED NANODDI DEPLOYMENT IN USERS’ NHP STUDIES FOR REPRODUCIBLE RESEARCH OUTCOMES THROUGH COMPREHENSIVE TRAINING, SUPPORTED BY OUR RESEARCH AND VETERINARY STAFF, AND VETERINARY SCIENTIFIC ADVISORY BOARD. CONCURRENTLY, USER-GENERATED DATA AND USER-INTRODUCED NOVEL ARVS COULD ENHANCE OUTREACH AS WELL AS BROADEN NANODDI DRUG PORTFOLIO. ALTOGETHER, RESEARCH OUTCOMES AND DATASET GENERATED INCLUSIVE OF ARV PHARMACOKINETIC AND USER-SPECIFIC DATA, COULD BE USED TO FILL KNOWLEDGE GAPS, IMPROVE DRUG DELIVERY AND ACCELERATE DRUG DEVELOPMENT IN HIV RESEARCH. EXPECTED OUTCOMES OF THE DART PROGRAM INCLUDE: SUCCESSFUL INTEGRATION OF THE NANODDI INTO NHP HIV STUDIES WITH SIGNIFICANT COST AND BURDEN REDUCTION, INCREASE RESEARCH OUTPUT DUE TO COST- AND TIME-EFFICIENT CHANGES IN ARV DELIVERY PROTOCOLS, HIGHER QUALITY RESEARCH DATA DUE TO LESS VARIABILITY IN ARV DELIVERY, AND MINIMIZE SCIENTIFIC AND FINANCIAL RISKS ASSOCIATED WITH NHP RESEARCH.

COMPUTER-BASED TRAINING METHODS FOR SURGICAL TESTING

TRANSFORMING TRIPLE-NEGATIVE BREAST CANCER TREATMENT THROUGH INTRATUMORAL IMMUNOTHERAPY VIA NANOFLUIDIC DRUG-ELUTING SEED

NY-ESO-1-SPECIFIC TCR-ENGINEERED T CELL IMMUNOTHERAPY FOR TRIPLE NEGATIVE BREAST CANCER

BIONANOSCAFFOLDS (BNS) FOR POST-TRAUMATIC OSTEOREGENERATION MODULE I: RESTORED MECHANICAL AND AMBULATORY FUNCTION

ARTIFICIAL INTELLIGENCE-INTEGRATED MECHANISTIC MODELING FOR RATIONAL DESIGN OF NANOPARTICLES TO IMPROVE ORGAN TARGETING AND SAFETY - PROJECT SUMMARY. NANOPARTICLES (NPS) HOLD GREAT PROMISE AS TARGETED DRUG DELIVERY SYSTEMS BUT TAILORING THEIR PHARMACOKINETICS (PK) TO SPECIFICALLY TARGET REGIONS OF INTEREST REMAINS A CHALLENGE. THIS LIMITS THE CLINICAL TRANSLATION OF NPS DUE TO POOR EFFICACY AND SAFETY CONCERNS ASSOCIATED WITH OFF-TARGET ACCUMULATION OF NP-BASED FORMULATIONS. DUE TO THE INTERACTIONS OF NPS WITH BIOLOGICAL COMPONENTS, DRIVEN BY THEIR STRUCTURAL PROPERTIES, CUSTOMIZING THE PHARMACOKINETICS (PK) OF NPS REQUIRES A QUANTITATIVE UNDERSTANDING OF THE EFFECT OF NP STRUCTURAL PROPERTIES ON THEIR WHOLE-BODY BIODISTRIBUTION, WHICH IN TURN ALSO GOVERNS THEIR SAFETY PROFILE. THEREFORE, TO ENABLE RATIONAL DESIGN OF NPS TO ACHIEVE ORGAN TARGETING AND SAFETY, WE PROPOSE TO LEVERAGE ARTIFICIAL INTELLIGENCE TO DEVELOP A TOXICOLOGY-INTEGRATED PHYSIOLOGICALLY-BASED PHARMACOKINETIC MODEL (PBPK-TOX) CAPABLE OF ACCURATELY PREDICTING THE WHOLE-BODY EXPOSURE AND SAFETY OF NOVEL NANOMATERIALS, BASED SOLELY ON THEIR STRUCTURAL PROPERTIES, DOSE, AND ROUTE OF ADMINISTRATION. FOR THIS, WE WILL (1) DEVELOP THE PBPK-TOX MODEL BASED ON DIVERSE DATASETS FROM LITERATURE, (2) ESTABLISH THE QUANTITATIVE RELATIONSHIP BETWEEN NP PROPERTIES, EXPOSURE, AND TOXICITY, AND (3) EXPERIMENTALLY TEST THE MODEL PREDICTIONS OF RATIONAL DESIGN TO TARGET ONE OR MORE ORGANS. OUR PROPOSED MODELING FRAMEWORK WILL ENABLE EFFICIENT PRECLINICAL DEVELOPMENT OF NOVEL NANOMATERIALS (AND ACCELERATE THEIR CLINICAL TRANSLATION) BY PROVIDING RATIONAL DESIGN GUIDELINES THROUGH IN-DEPTH COMPUTATIONAL INVESTIGATION OF BIOLOGICAL AND PHYSICOCHEMICAL VARIABILITY ON BIODISTRIBUTION AND SAFETY OF NPS.

ANTIGEN SPECIFICITY, SUPPRESSIVE MECHANISMS AND REGULATION OF CD4+ REGULATORY T C

REVERSAL OF REGULATORY T CELL FUNCTION IN PROSTATE CANCER

ALTERNATIVE POLYADENYLATION IN RIGHT VENTRICULAR FIBROSIS - PROJECT SUMMARY: PULMONARY HYPERTENSION (PH) IS A CARDIOVASCULAR DISORDER CHARACTERIZED BY HIGH MORTALITY, PRIMARILY DUE TO RIGHT VENTRICULAR (RV) FAILURE (RVF) CAUSED BY INCREASED PULMONARY VASCULAR RESISTANCE. RV FIBROSIS, A HALLMARK OF DECOMPENSATED RVF, LACKS TARGETED THERAPIES, HIGHLIGHTING THE NEED TO ELUCIDATE THE MOLECULAR MECHANISMS UNDERLYING RV FIBROSIS AND DYSFUNCTION. OUR RESEARCH FOCUSES ON THE ROLE OF ALTERNATIVE POLYADENYLATION (APA), A PROCESS ASSOCIATED WITH EXCESSIVE PRODUCTION OF EXTRACELLULAR MATRIX (ECM) PROTEINS, IN RV FIBROSIS. APA SHORTENS THE 3' UNTRANSLATED REGION (UTR) OF TRANSCRIPTS, LEADING TO LOSS OF MICRORNA BINDING SITES AND INCREASED TRANSCRIPT STABILITY. WE HAVE IDENTIFIED CLEAVAGE AND POLYADENYLATION SPECIFIC FACTOR 6 (CPSF6), A KEY REGULATOR OF APA, AS BEING INVOLVED IN END- STAGE RVF. IN RVF PATIENTS, CPSF6 EXHIBITS LENGTHENED 3' UTR AND DECREASED PROTEIN EXPRESSION. SILENCING CPSF6 IN CARDIAC FIBROBLASTS (CFS) RESULTS IN 3' UTR SHORTENING AND UPREGULATION OF MAJOR FIBROTIC MEDIATORS, INCLUDING TGF-Β1 AND ITS RECEPTOR, TGFΒR1. PATHWAY ANALYSIS FURTHER SUPPORTS 3' UTR SHORTENING IN MRNAS ENCODING ECM PROTEINS IN CPSF6 KNOCKDOWN CFS. ADDITIONALLY, WE HAVE DISCOVERED THE ROLE OF 4-HYDROXY-2- NONENAL (4HNE), A REACTIVE ALDEHYDE GENERATED DURING OXIDATIVE STRESS, IN RVF. INCREASED 4HNE DOWNREGULATES CPSF6, INDUCING 3' UTR SHORTENING IN PROFIBROTIC GENES AND PROMOTING RV FIBROSIS. THE PROPOSED RESEARCH AIMS TO INVESTIGATE THESE MECHANISMS AND IDENTIFY THERAPEUTIC TARGETS FOR MITIGATING RV FIBROSIS. OUR HYPOTHESIS POSITS THAT CPSF6 DEPLETION SHORTENS THE 3' UTRS OF ECM GENES, CAUSING THEIR ESCAPE FROM REGULATION, PROMOTING THEIR EXPRESSION, AND LEADING TO RV FIBROSIS. SPECIFICALLY, WE WILL: INVESTIGATE THE IMPACT OF CPSF6 LOSS ON THE 3' UTR LANDSCAPE AND PROFIBROTIC GENE EXPRESSION IN RVF (SPECIFIC AIM 1). UNCOVER THE MECHANISM UNDERLYING CPSF6 REDUCTION-DEPENDENT 3' UTR SHORTENING IN CFS AND ITS FUNCTIONAL CONSEQUENCES IN RVF (SPECIFIC AIM 2). ASSESS THE IMPACT OF ALDH2 RESTORATION ON ALLEVIATING RV FIBROSIS THROUGH CPSF6 REGULATION (SPECIFIC AIM 3). THE VALIDATION OF OUR HYPOTHESES AND THE COMPLETION OF THESE AIMS WILL HIGHLIGHT THE IMPORTANCE OF 3' UTR SHORTENING IN ECM DEPOSITION AND FIBROSIS IN RVF, POTENTIALLY GUIDING THE DEVELOPMENT OF THERAPEUTIC INTERVENTIONS. GIVEN THE LIMITED TREATMENT OPTIONS AND SEVERE CONSEQUENCES OF PH, OUR RESEARCH HOLDS SIGNIFICANT PROMISE FOR IMPROVING PUBLIC HEALTH.

MULTI-OMIC INTEGRATIVE ANALYSIS OF SHARED AND DISTINCT INFLAMMATION NETWORKS ACROSS NEURODEGENERATIVE DISORDERS - SUMMARY/ABSTRACT: ALZHEIMER'S DISEASE (AD) AND FRONTOTEMPORAL DISORDER (FTD) ARE TWO OF THE MOST COMMON NEURODEGENERATIVE DISORDERS, WITH LIMITED AVAILABLE THERAPEUTIC OPTIONS. ALTHOUGH THEY DIFFER IN TERMS OF GENES ASSOCIATIONS, UNDERLYING PROTEINOPATHY, AND AFFECTED BRAIN REGIONS, THEY SHARE PATHOLOGICAL FEATURES SUCH AS INCREASED INFLAMMATION AND NEURONAL LOSS. RECENT FINDINGS INDICATE THAT ADDRESSING INFLAMMATION MIGHT BE A POTENTIAL TREATMENT FOR AD AND FTD. DESPITE CONSIDERABLE EVIDENCE SUPPORTING THE PRESENCE OF PERIPHERAL AND CENTRAL CHRONIC LOW-GRADE INFLAMMATION IN AD AND FTD, A COMPREHENSIVE MOLECULAR UNDERSTANDING OF INFLAMMATORY CASCADES AND NETWORKS ACROSS THESE DISORDERS IS LACKING. THIS KNOWLEDGE GAP REPRESENTS A SIGNIFICANT BARRIER TO ADVANCING THE FIELD. OUR PRELIMINARY ANALYSIS OF INFLAMMATION-RELATED ANALYTES FROM AD, FTD, AND HEALTHY CONTROL SERA, IDENTIFIED THE ACTIVATION OF SPECIFIC INFLAMMATORY ANALYTES AND SIGNALING NODES IN BOTH AD AND FTD. IN ADDITION TO THE SHARED INFLAMMATORY SIGNALING NODES, SEVERAL DISEASE-SPECIFIC INFLAMMATORY ANALYTES WERE ALTERED IN EACH NEURODEGENERATIVE DISORDER. BASED ON OUR PRELIMINARY RESULTS, WE HYPOTHESIZE THAT THE IMMUNE RESPONSES AND ACTIVATED INFLAMMATORY PATHWAYS IN AD AND FTD HAVE SHARED AND DISEASE-SPECIFIC CHARACTERISTICS AND THAT IT MAY BE POSSIBLE TO IDENTIFY NOVEL INTERVENTIONAL STRATEGIES FOR PAN-NEURODEGENERATION AND DISEASE-SPECIFIC INFLAMMATORY RESPONSES. TO CONFIRM THE PRELIMINARY FINDINGS AND EXPLORE NOVEL GROUPS, WE WILL PERFORM A LARGE-SCALE PROTEOMIC ANALYSIS TO INTERROGATE MATCHED SERUM AND CSF SAMPLES FROM 250 AD, 100 FTD PATIENTS AND 150 AGE- AND GENDER- MATCHED HEALTHY CONTROLS FROM TWO WELL-CHARACTERIZED NATIONAL COHORTS (ALZHEIMER'S DISEASE NEUROIMAGING INITIATIVE (ADNI) AND ALL-FTD) (AIM 1). IN THE NEXT STEP, WE WILL EVALUATE THE CONTRIBUTION OF THE ALTERED INFLAMMATORY SIGNALS TO AD AND FTD CHANGES. IN THIS REGARD, WE WILL INVESTIGATE THE ASSOCIATION OF INFLAMMATORY ANALYTES AND SIGNALING NODES WITH 1) KNOWN GENETIC RISK FACTORS AND MUTATIONS, 2) THEIR SEX-SPECIFIC DIFFERENCES, 3) THE CORRELATION WITH RELEVANT ESTABLISHED BIOFLUID BIOMARKERS IN AD AND FTD, AND 4) THEIR IMPACT ON CLINICAL OUTCOMES (AIM-2). FINALLY, WE WILL IDENTIFY THE NEURAL AND IMMUNE CELL TYPES RESPONSIBLE FOR THE ELEVATED INFLAMMATION IN AD AND FTD THROUGH SINGLE-CELL RNA-SEQUENCING AND SPATIAL TRANSCRIPTOMICS (BLOOD AND BRAIN TISSUE) ANALYSES (AIM 3). THIS MULTI-OMICS INTEGRATIVE APPROACH WILL OFFER NOVEL INSIGHTS INTO THE MECHANISMS DRIVING THE PERIPHERAL AND CENTRAL INFLAMMATORY SIGNALS IN AD AND FTD AND SUGGEST TREATMENT OPTIONS. THE INNOVATION IN THIS PROJECT RESULTS FROM THE COLLABORATION OF MULTIPLE INVESTIGATORS WITH EXPERIENCE IN NEURODEGENERATION MOLECULAR IMMUNOLOGY (ALIREZA FARIDAR, MD.), NEUROPATHOLOGY, BIOMARKERS, AND DATA ANALYSIS (JON B. TOLEDO, MD., PHD.), AND SINGLE-CELL ANALYSIS (KYUSON YUN, PHD.).

HEALTH CARE AND OTHER FACILITIES

HIGH THROUGHPUT SINGLE-CELL PHENOTYPE ISOLATION BY PROTRUSION ANALYSIS CHIP (PAC)

AIBP-MEDIATED CHOLESTEROL EFFLUX AND ANGIOGENESIS

THE STEROID RECEPTOR COACTIVATOR SRC-2 COORDINATES AND MODULATES THE METABOLIC, S

MITIE TELEMENTORING PLATFORM: A NOVEL TRANSPORTABLE, AND MOBILE TELEMEDICINE PLATFORM FOR COMMUNICATION

NANOMEDICINE FOR EARLY DISEARS DETECTION AND TREATMENT

NEURAL CODING OF NATURAL STIMULI IN FREELY MOVING MACAQUE - DESPITE THE FACT THAT VISUAL PERCEPTION REPRESENTS SUCH A FUNDAMENTAL ASPECT OF OUR EVERYDAY LIFE, OUR KNOWLEDGE OF THE UNDERLYING NEURAL CODING OF NATURAL STIMULI IS WOEFULLY LACKING. ONE MAJOR LIMITATION PREVENTING OUR UNDERSTANDING OF THE NEURAL UNDERPINNINGS OF NATURAL VISION IS THE LACK OF VIABLE METHODOLOGIES FOR RECORDING AND SYNCHRONIZING EYE MOVEMENTS AND INCOMING VISUAL STIMULI FROM FREELY-MOVING MONKEYS DURING UNRESTRAINED EXPLORATORY BEHAVIOR. INDEED, EXAMINING THE NEURAL BASES OF VISUAL PERCEPTION HAS BEEN TRADITIONALLY PERFORMED BY STUDYING THE BRAIN OF NONHUMAN PRIMATES IN A LABORATORY ENVIRONMENT IN WHICH THE HEAD AND BODY ARE RESTRAINED WHILE SYNTHETIC STIMULI ARE PRESENTED ON A COMPUTER MONITOR. HOWEVER, IT HAS BECOME INCREASINGLY CLEAR THAT STUDYING THE BRAIN IN SPATIALLY CONFINED, ARTIFICIAL LABORATORY RIGS POSES SEVERE LIMITS ON OUR CAPACITY TO UNDERSTAND THE FUNCTION OF BRAIN CIRCUITS. TO OVERCOME THESE LIMITATIONS, WE PROPOSE A NOVEL APPROACH BY DESIGNING AN INTEGRATED WIRELESS EYE TRACKING SYSTEM TO PRECISELY RECORD NATURALLY-OCCURRING EYE MOVEMENTS AND RETINOTOPIC VISUAL INPUTS THROUGHOUT UNSTRUCTURED, FREELY-MOVING BEHAVIOR IN CONJUNCTION WITH MASSIVE, WIRELESS ELECTRICAL RECORDINGS OF POPULATION ACTIVITY ACROSS FOUR CORTICAL AREAS. OUR NOVEL SYSTEM WILL BE USED TO STUDY THE VISUAL CODING AND BRAIN STATE-DEPENDENCE OF CORTICAL DYNAMICS DURING NATURAL VISUAL EXPLORATION BY RECORDING POPULATION ACTIVITY IN MULTIPLE VISUAL, TEMPORAL, AND FRONTAL CORTICAL AREAS WHILE NONHUMAN PRIMATES ARE INTERACTING WITH THEIR ENVIRONMENT. WE WILL FOCUS ON THE SPARSENESS OF THE POPULATION RESPONSE, THE REAL-TIME ENCODING OF NATURAL SCENES, AND SPATIAL POSITION DEPENDENCY OF STIMULUS CODING. WE HYPOTHESIZE THAT THE SPARSE CODING OF NATURAL STIMULI DURING FREE VIEWING IMPROVES THE ACCURACY WITH WHICH NEURAL POPULATIONS ENCODE STIMULI ACROSS THE VISUAL CORTICAL HIERARCHY. OUR PROPOSED RESEARCH WILL CONSTITUTE A PARADIGM SHIFT BY MOVING VISUAL NEUROSCIENCE - FROM SIMPLY OBSERVING ANIMAL BEHAVIOR AND RECORDING THE RESPONSES OF SINGLE CELLS - TO A QUANTITATIVE UNDERSTANDING OF THE DISTRIBUTED NEURONAL NETWORK ENCODING DURING TASK-FREE BEHAVIOR IN FREELY MOVING NONHUMAN PRIMATES INTERACTING WITH THEIR PHYSICAL ENVIRONMENT. WE ANTICIPATE THAT THE LARGE QUANTITY OF NEURAL DATA RECORDED USING OUR APPROACH WILL BE OF GREAT INTEREST TO CLINICIANS AND COMPUTATIONAL NEUROSCIENTISTS STUDYING GENERAL PROPERTIES OF NORMAL AND DYSFUNCTIONAL NEURAL NETWORKS, POSSIBLY LEADING TO MEDICAL INSIGHTS INTO CORTICAL VISUAL IMPAIRMENTS AND INNOVATIONS IN CORTICAL PROSTHETICS FOR RESTORING NATURAL VISUAL FUNCTIONS.

NOVEL GROUP A STREPTOCOCCUS HUMAN VACCINE CANDIDATES