Thomas Jefferson University

TRANSLATIONAL RESEARCH IN CANCER

CARES ACT - HIGHER EDUCATION EMERGENCY RELIEF FUND-IHE/INSTITUTION - THOMAS JEFFERSON UNIVERSITY

CARES ACT FUNDS - THOMAS JEFFERSON UNIVERSITY

ETHANOL EFFECTS ON THE TRANSCRIPTIONAL REGULATORY NETWORK IN LIVER REGENERATION

CLINICAL PHARMACOLOGY TRAINING PROGRAM

G PROTEIN-COUPLED RECEPTOR REGULATION IN AIRWAY MYOCYTES

PARADIGMS OF WOUND HEALING AND FIBROSIS IN THE EYE

JEFFERSON ALUMNI HALL 3 WEST LAB MODERNIZATION

CONTINUATION AND EXPANSION OF THE DRUG INDUCED LIVER INJURY NETWORK(PHILLY-DILIN)

ROLE OF HIF-1 IN INTERVERTEBRAL DISC FUNCTION

RESPIRATORY MOTOR NEURON PROTECTION FOLLOWING CERVICAL SPINAL CORD INJURY

REPURPOSING CLASSICAL DEATH PATHWAYS FOR SIGNALING ROLES IN LENS DIFFERENTIATION

TRNA IN CODON USAGE

POST DPP FOLLOW-UP STUDY

NEUROHUMORAL CONTROL OF INTERNAL ANAL SPHINCTER

ENVIRONMENT AND GENE EFFECTS ON BRAIN AND BEHAVIOR

CA2+ AND ROS CROSSTALK SIGNALING IN CARDIAC MITOCHONDRIA

PRECLINICAL CHARACTERIZATION OF A MULTIVALENT KILLED FILOVIRUS/RABIES VACCINE

TJU-JNMC GLOBAL NETWORK FOR WOMEN'S & CHILDREN'S HEALTH RESEARCH UNIT

ALCOHOLIC TISSUE INJURY

VIRUS CLEARANCE FROM THE CENTRAL NERVOUS SYSTEM

BASIC AND TRANSLATIONAL RESEARCH PROGRAM

ORTHROPOXVIRUS PATHOGENESIS AND VACCINES

NONINVASIVE SUBHARMONIC AIDED PRESSURE ESTIMATION OF PORTAL HYPERTENSION

TARGETING ADIPONECTIN FOR CARDIOPROTECTION IN THE ISCHEMIC HEART

EXPLORING THE EPHB2-NMDA RECEPTOR INTERACTION IN SPINAL CORD INJURY-INDUCED NEUROPATHIC PAIN

MOLECULAR MECHANISMS OF MITOCHONDRIAL TRANSCRIPTION AND REPLICATION

ALTERED PROTEOGLYCAN GENE EXPRESSION AND CANCER

RETRIEVAL PRACTICE PRINCIPLES: A THEORY OF LEARNING FOR APHASIA REHABILITATION

CYSTEINE PROTEASES IN APOPTOSIS

TRMD-TARGETING ACTINOBACTERIAL NATURAL PRODUCTS AS NEXT GENERATION ANTIBIOTICS - PROJECT SUMMARY. DISCOVERING NEW ANTIBIOTICS FOR GRAM-NEGATIVE BACTERIA IS UNIQUELY CHALLENGING, DUE TO THEIR DOUBLE-MEMBRANE STRUCTURE THAT ACTS AS A PERMEABILITY BARRIER TO DRUGS AND AS AN ANCHOR FOR EFFLUX PUMPS. EFFORTS THAT TARGET ONE MEMBRANE PROTEIN OR ONE EFFLUX PUMP AT A TIME ARE INEFFECTIVE, DUE TO RAPID RISE OF RESISTANCE MUTATIONS. WE WILL TARGET THE TRMD-CATALYZED M1G37 METHYLATION OF TRNA TO INHIBIT BIOSYNTHESIS OF MULTIPLE CLASSES OF MEMBRANE PROTEINS, WITH THE POTENTIAL TO ACCELERATE BACTERICIDAL ACTION. TRMD IS A BACTERIA-SPECIFIC S- ADENOSYL-METHIONINE (ADOMET)-DEPENDENT METHYL TRANSFERASE THAT CONTROLS ACCURACY OF THE PROTEIN-SYNTHESIS READING FRAME. LOSS OF TRMD INCREASES +1 FRAMESHIFTING AND CAUSES CELL DEATH. WE HAVE SHOWN THAT GENES FOR MULTIPLE MEMBRANE PROTEINS AND EFFLUX PUMPS IN E. COLI AND IN OTHER GRAM-NEGATIVE BACTERIA CONTAIN TRMD- DEPENDENT CODONS NEAR THE START OF THE READING FRAME. WE HYPOTHESIZE THAT TARGETING TRMD WILL REDUCE PROTEIN SYNTHESIS OF ALL OF THESE GENES, THUS OFFERING A NOVEL SOLUTION TO AN UNMET NEED. WHILE ASTRAZENECA (AZ), GSK, AND ACADEMIC LABS HAVE ATTEMPTED TO TARGET TRMD BY SCREENING SMALL MOLECULAR COMPOUND LIBRARIES, ISOLATED HITS LACK THE CELL-PERMEABILITY NEEDED TO EXHIBIT AN ANTIBACTERIAL EFFECT. HERE, WE PROPOSE TO SCREEN A LARGE COLLECTION OF MICROBIAL EXTRACTS AND FRACTIONS FOR CELL-PERMEABLE AND TRMD-TARGETING NATURAL PRODUCTS (NPS) THAT ARE POTENT AND SELECTIVE OVER THE HUMAN COUNTERPART TRM5. WE WILL USE A CELL-BASED ASSAY, CONSISTING OF A 1:1 MIX OF AN E. COLI (EC) TRMDMCH STRAIN (DEPENDENT ON TRMD FOR SURVIVAL AND EXPRESSING MCH (MCHERRY) AS A FLUORESCENCE MARKER) AND AN EC TRM5YFP STRAIN (DEPENDENT ON TRM5 FOR SURVIVAL AND EXPRESSING YFP), IN A HIGH- THROUGHPUT SCREENING (HTS) CAMPAIGN TO ISOLATE NPS THAT SELECTIVELY INHIBIT THE TRMDMCH STRAIN. WE PERFORM THIS ASSAY IN EC TOLC+ CELLS, WHICH MAINTAIN THE ENTIRE GRAM-NEGATIVE EFFLUX MACHINERY INCLUDING THE MAJOR EFFLUX PUMP ENCODED BY TOLC, TO SCREEN FOR NPS THAT ARE CELL-PERMEABLE AND RESISTANT TO EFFLUX. A PILOT SCREEN WITH THIS TOLC+ CELL-BASED ASSAY HAS IDENTIFIED AN ATTRACTIVE HIT, DEMONSTRATING THE HTS-READINESS OF THE ASSAY. IN AIM 1, WE WILL USE THIS TOLC+ CELL-BASED ASSAY TO SCREEN 74,770 ACTINOBACTERIAL EXTRACTS AND FRACTIONS AVAILABLE AT THE SCRIPPS RESEARCH INSTITUTE (TSRI). WE WILL ASSESS HITS IN SECONDARY ASSAYS, REMOVE FALSE POSITIVES, EVALUATE THEIR ACTIVITY AT THE WHOLE-CELL LEVEL, AND TEST THEM FOR PERMEABILITY AND EFFLUX IN A PANEL OF GRAM-NEGATIVE BACTERIA. IN AIM 2, WE WILL DE-REPLICATE THE TOP 20 HITS TO ISOLATE THE ACTIVE NPS, DETERMINE THEIR STRUCTURES, AND USE A COMBINATION OF GENOME SEQUENCING AND MINING TO IDENTIFY THEIR BIOSYNTHETIC GENE CLUSTERS (BGCS) FOR DEVELOPING BIOTECHNOLOGY PLATFORMS TO SCALE UP THEIR PRODUCTION. IN AIM 3, WE WILL TEST ACTIVE NPS FOR CONFERRING TRMD- DEFICIENT PHENOTYPES IN WHOLE-CELL ASSAYS, DETERMINE THEIR POTENCY, SELECTIVITY, MECHANISM OF ACTION, AND ASSESS THEIR RISK OF RESISTANCE. THESE NPS REPRESENT NOVEL LEADS IN A NEW PARADIGM OF ANTIBIOTIC DISCOVERY THAT ADDRESSES THE MULTI-DRUG RESISTANCE PROBLEM OF GRAM-NEGATIVE BACTERIA.

MITOCHONDRIA-SR TETHERING: ITS ROLE IN CARDIAC BIOENERGETICS AND CA2+ DYNAMICS

TRANSCRIPTIONAL REGULATION BY EPIGENETIC FACTORS

RPE LACTATE TRANSPORTERS: A ROLE IN RETINAL SURVIVAL

BIOCHEMISTRY AND MORPHOLOGY OF CONNECTIVE TISSUE

ENDOGENOUS SUPPRESSION OF INTEGRIN SIGNALING

USING REPORTER HUMAN IPS CELLS TO STUDY FATE, FUNCTION AND PARKINSON'S DISEASE

CCHS-JNMC GLOBAL NETWORK FOR WOMEN'S & CHILDREN'S HEALTH RESEARCH UNIT

LOW VISION DEPRESSION PREVENTION TRIAL FOR AGE RELATED MACULAR DEGENERATION

FOXD3 UP-REGULATION AND ERBB3 SIGNALING AS AN ADAPTIVE RESPONSE TO RAF INHIBITORS

HEALTH BELIEFS, GLYCEMIC CONTROL, AND PREVENTING COGNITIVE DECLINE IN AFRICAN AMERICANS WITH DIABETES AND MILD COGNITIVE IMPAIRMENT: A RANDOMIZED CLINICAL TRIAL

ROLE OF ARRESTINS IN G PROTEIN-COUPLED RECEPTOR REGULATION

2D AND 3D CONTRAST-ENHANCED ULTRASOUND EVALUATION OF HCC CHEMOEMBOLIZATION

EXOSOME-MEDIATED PROPAGATION OF DISEASE LINKED POLY-DIPEPTIDES IN C9ORF72-FTD/ALS

REGULATION OF ENDOTHELIAL NITRIC OXIDE SYNTHASE MRNA STABILITY

SIDNEY KIMMEL CANCER CENTER LAPS INSTITUTIONAL COLLABORATION

MOLECULAR MECHANISMS OF AXON BRANCHING IN SYNAPTIC DEVELOPMENT

ASK1 A NOVEL REGULATOR OF PLATELET FUNCTION

TARGETED THERAPIES IN MUTANT BRAF MELANOMA

THE BIOLOGY OF PERLECAN IN CANCER AND ANGIOGENESIS

PLASMA-BASED THERAPIES FOR BONE INFECTION: A TRIPARTITE USA/NORTHERN IRELAND/REPUBLIC OF IRELAND CONSORTIUM

A PRAGMATIC RANDOMIZED CONTROLLED TRIAL ASSESSING THE IMPACT OF MEDICALLY TAILORED MEALS AND MEDICAL NUTRITION THERAPY VIA TELEHEALTH AMONG PATIENTS WITH POORLY CONTROLLED DIABETES

PROVIDER SUPPORT AND PATIENT OUTREACH IN LUNG CANCER SCREENING - WE PROPOSE TO CONDUCT A MULTI-LEVEL INTERVENTION RANDOMIZED TRIAL IN FOUR HEALTH SYSTEMS (CHRISTIANA CARE HEALTH SYSTEM (CCHS), JEFFERSON HEALTH (JH), LEHIGH VALLEY HEALTH NETWORK (LVHN), AND PENNSTATE HEALTH (PSH)) TO TEST THE COMBINED AND SEPARATE EFFECTS OF A PROVIDER SUPPORT INTERVENTION AND A PATIENT OUTREACH INTERVENTION ON SDM AND LCS IN PRIMARY CARE PRACTICE. USING A MIXED METHODS 2X2 FACTORIAL DESIGN, WE INITIALLY WILL RECRUIT AND RANDOMIZE 28 PRIMARY CARE PRACTICES AND PROVIDERS EITHER TO A PROVIDER SUPPORT GROUP (N=14) OR A CONTROL GROUP (N=14). OVER THREE YEARS, WE WILL ALSO IDENTIFY APPROXIMATELY 4,480 PATIENTS WHO ARE POTENTIALLY ELIGIBLE FOR LCS AND HAVE A SCHEDULED PRIMARY CARE VISIT. ON A WEEKLY BASIS, WE WILL ALLOCATE IDENTIFIED PATIENTS TO COHORTS AND INITIATE TELEPHONE CONTACT WITH EACH COHORT TO VERIFY SCREENING ELIGIBILITY, OBTAIN CONSENT, AND COMPLETE A BASELINE SURVEY. WE WILL RANDOMIZE AT LEAST 672 PATIENT PARTICIPANTS EITHER TO A PATIENT OUTREACH GROUP (N=336) OR A CONTROL GROUP (N=336). THUS, THE TRIAL WILL INVOLVE A PROVIDER SUPPORT + PATIENT OUTREACH GROUP, A PROVIDER SUPPORT GROUP, A PATIENT OUTREACH GROUP, AND A CONTROL GROUP. WE WILL COLLECT HEALTH SYSTEM ELECTRONIC MEDICAL RECORDS (EMRS) DATA, STUDY ADMINISTRATIVE DATA, PHYSICIAN AND PATIENT SURVEY DATA, AND INTERVIEW DATA FROM PHYSICIANS, PRACTICE STAFF, HEALTH SYSTEM CARE COORDINATORS, AND HEALTH SYSTEM LEADERS FOR USE IN ADDRESSING TRIAL AIMS. SPECIFIC AIMS OF THE TRIAL ARE AS FOLLOWS: AIM 1. ASSESS INTERVENTION IMPACT ON LUNG CANCER SCREENING (LCS); AIM 2. ASSESS INTERVENTION IMPACT ON SHARED DECISION MAKING (SDM); AIM 3. IDENTIFY MEDIATORS AND MODERATORS OF SCREENING; AND, AIM 4. ASSESS INTERVENTION IMPLEMENTATION. WE WILL USE HEALTH SYSTEM EMR DATA, STUDY ADMINISTRATIVE DATA, PROVIDER BASELINE AND ENDPOINT SURVEY DATA, AND PATIENT BASELINE AND ENDPOINT SURVEY DATA, FOCUS GROUP DATA, AND HEALTH SYSTEM LEADER INTERVIEW DATA TO IDENTIFY PRACTICE-, PROVIDER- AND PATIENT-LEVEL FACTORS THAT IMPACT INTERVENTION EFFECTS ON SCREENING. IN TERMS OF OVERALL IMPACT, THIS TRIAL WILL IDENTIFY EFFECTIVE STRATEGIES FOR INCREASING SDM AND LCS RATES IN PRIMARY CARE AND FOR IMPLEMENTING EFFECTIVE STRATEGIES IN HEALTH SYSTEMS. WE WILL DISSEMINATE STUDY FINDINGS THROUGH COMMUNICATION CHANNELS IN PARTICIPATING HEALTH SYSTEMS AND MORE BROADLY BY REPORTING RESULTS ON CLINICALTRIALS.GOV, AT HEALTH CARE PROFESSIONAL ORGANIZATION MEETINGS, THROUGH LOCAL, STATE AND REGIONAL CONFERENCES, AND VIA PUBLICATIONS IN PEER-REVIEWED JOURNALS.

MODEL SYSTEMS FOR PXE

GENOMIC PROFILING OF SINGLE CIRCULATING TUMOR CELLS IN THE PRECISION MEDICINE OF METASTATIC PROSTATE CANCER - PROJECT SUMMARY AND ABSTRACT METASTATIC CASTRATION-RESISTANT PROSTATE CANCER (MCRPC) IS THE MOST LETHAL STATE OF PROSTATE CANCER (PCA) AND REMAINS AS THE MOST CHALLENGING ISSUE IN PCA TREATMENT. THE DEVELOPMENT OF MORE EFFECTIVE TREATMENTS FOR MCRPC IS A SIGNIFICANT UNMET CLINICAL NEED. THERE ARE SUBSTANTIAL HETEROGENEITIES IN TREATMENT RESPONSES OF MCRPC, NECESSITATING THE USE OF MORE PERSONALIZED STRATEGIES IN GUIDING TREATMENT BASED ON THE SPECIFIC GENOMIC CHARACTERISTICS OF INDIVIDUAL PATIENTS. THERE ARE CONSIDERABLE LIMITATIONS IN USING THE TRADITIONAL TISSUE- BASED GENOMIC PROFILING TO GUIDE MCRPC TREATMENT, PARTLY BECAUSE MCRPC IS A BONE-PREDOMINANT METASTATIC DISEASE, THUS TISSUE SAMPLES ARE DIFFICULT TO OBTAIN AND YIELDS ARE GENERALLY LOW. MOREOVER, DURING TREATMENTS, THE GENOMIC PROFILES OF TUMORS MAY CHANGE QUICKLY TO EVADE THERAPEUTIC OR IMMUNE ATTACKS, LEADING TO DRUG RESISTANCE. IN ORDER TO PROMPTLY AND ACCURATELY CAPTURE THESE CHANGES AND ADJUST TREATMENT PLANS, REPEATED TUMOR BIOPSIES WOULD BE NEEDED, WHICH IS DIFFICULT TO PERFORM IN ROUTINE CLINICAL PRACTICES GIVEN THE INVASIVE AND BONE-PREDOMINANT NATURE OF MCRPC. THEREFORE, IN ORDER TO IMPROVE MCRPC PROGNOSIS, IT IS HIGHLY IMPORTANT TO DEVELOP NOVEL, NON-INVASIVE LIQUID BIOPSY APPROACHES TO REAL-TIME MONITOR TREATMENT RESPONSE AND GUIDE THE USE OF DIFFERENT TREATMENTS. CIRCULATING TUMOR CELLS (CTCS) ARE SHED FROM TUMORS INTO BLOOD AND HAVE EXTREMELY HIGH MALIGNANT POTENTIAL, AND ARE ARGUABLY THE MOST IMPORTANT SUBSET OF TUMOR CELLS TO MONITOR AND TREAT. CTCS CAN BE NON-INVASIVELY AND REPEATEDLY ENUMERATED IN REAL-TIME, AND HAVE EXHIBITED PROMISING PROGNOSTIC POTENTIALS, AS EVIDENCED BY THE FDA-APPROVAL OF THE CELLSEARCH PLATFORM FOR CTC ENUMERATIONS AS AN INDEPENDENT PROGNOSTIC FACTOR OF SEVERAL METASTATIC CANCERS INCLUDING MCRPC. HOWEVER, NATIONAL GUIDELINES HAVE NOT UNANIMOUSLY ENDORSED THE USE OF CTC ENUMERATION IN ROUTINE CLINICAL PRACTICES, MOSTLY BECAUSE IT REMAINS UNCLEAR WHAT ACTIONS SHOULD BE TAKEN FOR HIGH-RISK PATIENTS WITH ELEVATED CTCS. THESE FACTS HIGHLIGHT THE IMPORTANCE OF MOVING BEYOND CTC ENUMERATION AND TOWARDS IN-DEPTH GENOMIC CHARACTERIZATIONS OF CTCS. LARGE STUDIES ON SINGLE-CELL CTC ANALYSIS HAVE BEEN RARELY REPORTED, PARTLY DUE TO THE SIGNIFICANT CHALLENGES ON SINGLE-CTC DETECTION, ISOLATION, WHOLE GENOME AMPLIFICATION (WGA), AND SEQUENCING BIAS IDENTIFICATION AND CORRECTION. WE HAVE ESTABLISHED A COMPREHENSIVE PIPELINE ON THE ENRICHMENT, ENUMERATION, ISOLATION, WGA, SEQUENCING, AND DATA ANALYSIS OF SINGLE CTCS. BASED ON THREE PCA PATIENT COHORTS AT THE SIDNEY KIMMEL CANCER CENTER, MD ANDERSON CANCER CENTER, AND GEORGE WASHINGTON UNIVERSITY CANCER CENTER, WE WILL CONDUCT GENOMIC PROFILING OF SINGLE CTCS TO IDENTIFY MARKERS OF TREATMENT RESPONSE AND PROGNOSIS. TO OUR BEST KNOWLEDGE, THIS IS THE FIRST LARGE POPULATION-BASED STUDY OF SINGLE-CTC ANALYSIS IN MCRPC. FINDINGS FROM THIS STUDY WILL SIGNIFICANTLY IMPROVE THE POTENTIAL OF THE CLINICAL APPLICATION OF CTCS IN MCRPC MANAGEMENT, BY PRECISELY TAILORING TREATMENT TO THE GENOMIC MAKE-UP OF INDIVIDUAL CTCS FROM INDIVIDUAL PATIENTS.

A NEW LCA MODEL BY POLARITY GENE ABLATION

MGLUR5 HYPOACTIVITY IS INTEGRAL TO GLUTAMATERGIC DYSREGULATION IN SCHIZOPHRENIA

ANALYSIS OF A NOVEL CRIMEAN-CONGO HEMORRHAGIC FEVER VACCINE AND ITS MECHANISM OF PROTECTION IN RODENT MODELS - CRIMEAN-CONGO HEMORRHAGIC FEVER VIRUS (CCHFV) CAUSES SEVERE DISEASE IN HUMANS, WITH FATALITY RATES REACHING 40%. CCHFV IS ENDEMIC TO PARTS OF AFRICA, ASIA, THE MIDDLE EAST, AND EUROPE, SPECIFICALLY TO REGIONS WHERE THE TICK VECTOR, SPECIES OF THE HYALOMMA GENUS, IS PRESENT. CLASSIFIED AS AN NIH/NIAID CATEGORY A AND WHO HIGH- PRIORITY PATHOGEN, CCHFV POSES THE HIGHEST POSSIBLE RISK TO NATIONAL SECURITY AND PUBLIC HEALTH. CCHFV IS A NEGATIVE-SENSE SINGLE-STRANDED RNA VIRUS IN THE ORDER BUNYAVIRALES. CCHFV IS AN EMERGING INFECTIOUS DISEASE, POSING A HIGH RISK OF A WIDESPREAD OUTBREAK. AN INACTIVATED WHOLE VIRUS VACCINE WAS THE ONLY CCHFV VACCINE TO BE TESTED IN HUMANS AND WAS INEFFECTIVE. WE PROPOSE THE USE OF INACTIVATED RABIES VIRUS (RABV)- AND VESICULAR STOMATITIS VIRUS (VSV)-BASED CCHFV VACCINES, AS INACTIVATED RHABDOVIRAL VECTORS HAVE NOT YET BEEN EXPLORED. INACTIVATED RHABDOVIRAL-BASED VACCINES ARE SAFE AND EFFECTIVE AT INDUCING IMMUNITY AND PROTECTION AGAINST MULTIPLE HEMORRHAGIC FEVER VIRUSES, AND A VSV-BASED SURROGATE CHALLENGE VIRUS IS AN EFFECTIVE TOOL IN ANOTHER HEMORRHAGIC FEVER MODEL. THE GOAL OF THIS PROJECT IS TWO-FOLD: FIRST, TO COMPARE RABV- AND VSV-BASED CCHFV/RABV BIVALENT VACCINES IN TERMS OF THEIR PRODUCTION, IMMUNOGENICITY; SECOND, TO ESTABLISH A NON-BSL-4 VSV-BASED SURROGATE MOUSE CHALLENGE SYSTEM FOR CCHFV TO DETERMINE MECHANISM OF PROTECTION. WE HYPOTHESIZE THAT INACTIVATED RHABDOVIRAL-BASED CCHFV VACCINES WILL PROTECT AGAINST THE CCHFV CHALLENGE THROUGH NON-NEUTRALIZING ANTIBODIES DIRECTED AGAINST GP38. TOWARD THIS HYPOTHESIS, WE PROPOSE THREE AIMS: AIM 1: CHARACTERIZATION OF RHABDOVIRAL-BASED CCHFV VACCINE CONSTRUCTS. THIS AIM DOES CHARACTERIZE AND TEST THE IMMUNOGENICITY OF RABV- AND VSV-BASED CCHFV VACCINES AND COMPARE THEM TO THE BULGARIA HUMAN VACCINE AND AN MRNA-BASED VACCINE PROVIDED BY COLLABORATORS. AIM 2: DETERMINE RHABDOVIRAL-BASED CCHFV VACCINE MECHANISM OF PROTECTION BY ESTABLISHING A SURROGATE CHALLENGE VIRUS MODEL. THIS AIM AIMS TO DEVELOP A NON-BSL-4-REQUIRING SURROGATE CHALLENGE MODEL FOR CCHFV AND COMPARE IT TO THE ESTABLISHED BSL-4 WT CCHFV MODEL IN VACCINE EFFICACY STUDIES. AIM 3: EVALUATE THE PROTECTIVE EFFICACY OF RHABDOVIRAL-BASED VACCINE CANDIDATES IN A WILDTYPE CCHFV LETHAL CHALLENGE MODEL USING NEEDLE AND TICK CHALLENGE. THE GOAL OF THIS AIM IS TO DETERMINE THE EFFICACY OF OUR RHABDOVIRAL-BASED VACCINES AND CONTROL VACCINES AGAINST WT CCHFV CHALLENGE IN TWO DIFFERENT LETHAL CHALLENGE MODEL.

THE ROLE OF THE DE-CONDENSED STRUCTURE OF NASCENT CHROMATIN DURING T CELL DIFFERENTIATION

BIASED MUSCARINIC ACETYLCHOLINE RECEPTOR SIGNALING IN OBSTRUCTIVE LUNG DISEASE PATHOLOGY AND THERAPY

NOVEL TUMOR-TARGETED IMMUNOTHERAPIES AGAINST METASTATIC UVEAL MELANOMA

MULTIFUNCTIONAL REGULATION OF PROSTATE CANCER METABOLISM BY SIGMA1 MODULATORS

TARGETING CDK6 EXPRESSION/ACTIVITY IN PH+ AND PH1-LIKE ACUTE LYMPHOBLASTIC LEUKEMIA (ALL) - ABSTRACT PHILADELPHIA-POSITIVE ACUTE LYMHOBLASTIC LEUKEMIA (PH+ ALL) AND PH1-LIKE B-ALL ACCOUNT FOR MOST CASES OF “HIGH-RISK” ADULT B-ALL. CURRENT THERAPIES WITH TYROSINE KINASE INHIBITORS (TKIS) HAVE IMPROVED THE OUTCOME OF PH+ ALL, BUT RESISTANCE TO TKIS DEVELOPS RAPIDLY IN MOST PATIENTS. PH1-LIKE B-ALL IS CURRENTLY TREATED WITH INTENSIVE COMBINATION CHEMOTHERAPY BUT DISEASE RELAPSE IS COMMON WITH A 5-YEAR SURVIVAL IN ONLY ~25% OF PATIENTS. AS A RESULT, THE PROGNOSIS OF PH+ ALL AND PH1-LIKE B-ALL REMAINS DISMAL. IN PREVIOUS STUDIES, WE SHOWED THAT PH+ AND PH1-LIKE ALL CELLS EXHIBIT A SELECTIVE REQUIREMENT FOR CDK6 EXPRESSION WHILE CDK4 EXPRESSION IS DISPENSABLE. CDK6 IS THE CATALYTIC SUBUNIT OF THE CYCLIN D/CDK6 COMPLEX WHICH IS ESSENTIAL FOR THE G1 TO S-PHASE CELL CYCLE TRANSITION AND HAS KINASE-INDEPENDENT GROWTH-PROMOTING EFFECTS IN HEMATOLOGICAL MALIGNANCIES. OUR PRELIMINARY STUDIES INDICATE THAT CDK6 SILENCING IS MORE EFFECTIVE THAN CDK6 ENZYMATIC INHIBITION IN SUPPRESSING PH+ ALL IN MICE. TO BLOCK KINASE-DEPENDENT AND INDEPENDENT EFFECTS OF CDK6, WE HAVE DEVELOPED CDK4/6-TARGETED PROTEOLYSIS-TARGETING CHIMERA (PROTACS) THAT INHIBIT CDK4/6 ENZYMATIC ACTIVITY IN VITRO AND PROMOTE THE PREFERENTIAL DEGRADATION OF CDK6 OVER CDK4 IN PH+ AND PH1-LIKE ALL CELLS, PROVIDING DURABLE SUPPRESSION OF CDK6 FUNCTION. IN THIS PROPOSAL, WE WILL ASSESS THE REQUIREMENT OF CDK6 IN PH+ AND PH1-LIKE ALL BY COMPARING THE EFFECTS OF CDK6 DEGRADATION BY PROTAC YX-2-107 AND PHARMACOLOGICAL INHIBITION USING PALBOCICLIB, AN FDA-APPROVED CDK4/6 INHIBITOR (AIM 1.1). WE WILL ALSO DETERMINE WHETHER THE MORE POTENT LEUKEMIA SUPPRESSION INDUCED BY CDK6 DOWN-REGULATION IN COMPARISON TO CDK6 ENZYMATIC INHIBITION CAN BE EXPLAINED BY CHANGES IN GENE EXPRESSION INDUCED SELECTIVELY BY CDK6 SILENCING. SUCH CHANGES INVOLVE THE HISTONE DEACETYLASE 1(HDAC1) GENE AND SEVERAL OTHERS INVOLVED IN MITOCHONDRIAL METABOLIC PATHWAYS (AIMS 1.2 AND 1.3). ALTHOUGH WE HAVE BEEN ABLE TO ACHIEVE HIGH SPECIFICITY OF CDK6 VERSUS CDK4 TARGETING AND BIOLOGICAL/THERAPEUTIC EFFECTS COMPARABLE/SUPERIOR TO PALBOCICLIB EX VIVO AND IN PDXS OF PH+ ALL, WE WILL CONTINUE TO IMPROVE OUR LEAD COMPOUND PROTAC YX-2-107 BY MEDICINAL CHEMISTRY APPROACHES IN ORDER TO DEVELOP DERIVATIVES WITH ENHANCED IN VIVO EFFICACY. IN AIM 2, WE WILL ASSESS METABOLIC PROPERTIES OF SELECT CDK6-DEGRADING PROTACS AND TEST THEIR BIOLOGICAL/THERAPEUTIC EFFECTS IN PH+ AND PH1-LIKE ALL CELLS EX VIVO AND IN MICE INJECTED WITH DE NOVO OR RELAPSED/TKI-RESISTANT PATIENT-DERIVED PH+/PH1-LIKE ALL CELLS. COLLECTIVELY, OUR PROTAC-BASED APPROACH WHICH LEVERAGES THE EXPERTISE IN CANCER BIOLOGY AND MEDICINAL CHEMISTRY OF THE CALABRETTA AND SALVINO'S LABORATORIES HOLDS PROMISE TO DEVELOP NOVEL AND MORE EFFECTIVE THERAPEUTIC AGENTS FOR THE TREATMENT OF CDK6-DEPENDENT HIGH-RISK B-ALL IN PRE-CLINICAL PDX MODELS AND, POTENTIALLY, IN THE CLINIC.

OCCULT COLORECTAL CANCER ELIMINATION BY GUCY2C-DIRECTED IMMUNOTHERAPY

TRAINING IN TISSUE ENGINEERING AND REGENERATIVE MEDICINE

MOLECULAR COMPOSITION OF THE MITOCHONDRIAL CALCIUM UNIPORTER AND CARDIAC PATHOPHYSIOLOGY

NONINVASIVE, UNIPLEX, MOLECULAR, PATHOMIC URINARY ASSAY FOR DETECTION OF PROSTATE CANCER

PYRIMIDINE METABOLISM IN THE DEVELOPMENT OF AUTOREACTIVE B CELLS AND SLE - PROJECT SUMMARY SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) IS A FEMALE-BIASED DEBILITATING AUTOIMMUNE DISEASE THAT AFFECTS MILLIONS IN AMERICA AND WORLDWIDE. ANTIBODY-FORMING CELLS (AFCS) AND GERMINAL CENTERS (GCS) PROMOTE THE DEVELOPMENT OF AUTOREACTIVE PLASMA CELLS (PCS) AND THE RELEASE OF PATHOGENIC AUTOANTIBODIES (AUTOABS) THAT ARE CRITICAL FOR THE INITIATION AND PROGRESSION OF DISEASE MANIFESTATIONS INCLUDING LUPUS NEPHRITIS. MECHANISMS THAT PROMOTE THE DEVELOPMENT OF AUTOREACTIVE B CELLS WHICH DRIVE SLE PATHOGENESIS ARE INCOMPLETELY UNDERSTOOD. HOWEVER, SUCH KNOWLEDGE IS INSTRUMENTAL FOR DEVELOPING MUCH NEEDED THERAPEUTICS THAT ARE PREFERABLE TO CURRENT SLE TREATMENT OPTIONS THAT GLOBALLY SUPPRESS THE IMMUNE SYSTEM AND INCREASE DISEASE-ASSOCIATED MORBIDITY AND MORTALITY. METABOLIC REPROGRAMMING HAS EMERGED AS A CRITICAL MECHANISM THAT UNDERPINS SLE AUTOIMMUNE RESPONSES AND DISEASE PROGRESSION. VARIOUS METABOLIC PATHWAYS ARE IMPLICATED IN AUTOIMMUNE RESPONSES IN SLE. ALTHOUGH THE IMPORTANCE OF NUCLEOTIDE METABOLISM HAS BEEN DESCRIBED IN CANCER AND PURINES SUCH AS ATP ARE ESSENTIAL FOR CELLULAR METABOLISM, THE ROLE OF PYRIMIDINES IN B CELL METABOLIC REPROGRAMMING AND RESPONSES IN SLE IS UNKNOWN. IMPORTANTLY, DRUGS TARGETING THE PYRIMIDINE PATHWAY ARE FDA APPROVED FOR THE TREATMENT OF CANCER AND RHEUMATOID ARTHRITIS. DRUGS TARGETING THE PYRIMIDINE PATHWAY IS ALSO CONSIDERED FOR MAINTENANCE THERAPY FOR LUPUS NEPHRITIS WITH NO MAJOR TOXICITY. HOWEVER, THE ROLE OF PYRIMIDINE METABOLISM IN AUTOREACTIVE B CELL DEVELOPMENT AND PATHOGENIC AUTOANTIBODY (AUTOAB) PRODUCTION IN AFC AND GC PATHWAYS IN SLE IS NOT KNOWN. THIS GRANT WILL FOCUS ON DELINEATING THE MECHANISMS BY WHICH PYRIMIDINE SYNTHESIS PROMOTES SYSTEMIC AUTOIMMUNITY. THE KNOWLEDGE GAINED WILL BE BENEFICIAL FOR DISSECTING MECHANISMS CRITICAL FOR B CELL DEVELOPMENT AND FUNCTION AND MAY LEAD TO NOVEL THERAPIES FOR SLE AND OTHER B CELL-MEDIATED DISEASES. OUR PRELIMINARY DATA HIGHLIGHTED THE SIGNIFICANCE OF DE NOVO PYRIMIDINE SYNTHESIS IN AUTOREACTIVE B CELL RESPONSES IN A SLE MOUSE MODEL. OUR DATA ALSO INDICATED AN INTERPLAY OF PYRIMIDINE METABOLISM AND B CELL METABOLIC REPROGRAMMING IN SLE-PRONE B CELLS. ADDITIONAL DATA POINTED TO A ROLE FOR MTORC1, A MASTER REGULATOR OF METABOLISM, IN ACTIVATING THE PYRIMIDINE PATHWAY IN SLE-PRONE B CELLS. WE HYPOTHESIZE THAT B CELL-INTRINSIC PYRIMIDINE METABOLISM AND AN INTERPLAY OF DE NOVO PYRIMIDINE SYNTHESIS AND METABOLIC NETWORKS PROMOTE AUTOREACTIVE B CELL DEVELOPMENT IN AFC AND GC PATHWAYS, LEADING TO AUTOAB PRODUCTION AND SLE. THIS HYPOTHESIS WILL BE TESTED BY THREE SPECIFIC AIMS. AIM-1 WILL, FOR THE FIRST TIME, IDENTIFY THE CELL-INTRINSIC MECHANISMS OF PYRIMIDINE METABOLISM IN AUTOREACTIVE B CELL RESPONSES THAT PROMOTE THE FORMATION OF AUTOAB-PRODUCING PLASMA CELLS AND AUTOABS IN SLE. AIM-2 WILL DETERMINE THE INTERPLAY OF PYRIMIDINE SYNTHESIS AND B CELL METABOLIC NETWORKS THAT PROMOTE AUTOREACTIVE B CELL DEVELOPMENT IN SLE. AIM-3 WILL FOCUS ON THE MECHANISM BY WHICH MTORC1 SIGNALING DRIVES THE ACTIVATION OF DE NOVO PYRIMIDINE SYNTHESIS PATHWAY IN SLE-PRONE B CELLS.

TARGETING CELL SENESCENCE IN A NOVEL MODEL OF SPONTANEOUS DISC DEGENERATION - INTERVERTEBRAL DISC DEGENERATION IS THE MAJOR RISK FACTOR ASSOCIATED WITH CHRONIC NECK AND LOW BACK PAIN, UBIQUITOUS HEALTH CONDITIONS THAT AFFECTS MILLIONS OF PEOPLE WORLD-WIDE. THERE IS AN INCOMPLETE UNDERSTANDING OF THE PATHOGENESIS OF DISC DEGENERATION PARTIALLY DUE TO LACK OF AN APPROPRIATE ANIMAL MODEL. CURRENT ANIMAL MODELS PRIMARILY USE TRAUMATIC INSULTS TO PROMOTE DEGENERATION DIFFERING FROM MOST HUMAN CASES OF DISC DEGENERATION. WHILE, A FEW ANIMAL MODELS OF SPONTANEOUS DISC DEGENERATION ARE REPORTED, SEVERAL LIMITATIONS PREVENT THEIR WIDE-SPREAD USE. THUS, THERE IS A GREAT NEED FOR SMALL ANIMAL MODELS THAT ARE MORE REPRESENTATIVE OF HUMAN DISC PATHOLOGY. PREVIOUS STUDIES HAVE INVESTIGATED INBRED STRAINS OF MICE INCLUDING SM/J AND LG/J FOR THEIR REGENERATIVE ABILITY. IN CONTRAST TO LG/J, A “SUPER HEALER” STRAIN, SM/J WAS FOUND TO BE A “POOR HEALER”. HOWEVER, THERE NO STUDIES HAVE INVESTIGATED THE DISC HEALTH STATUS IN THESE STRAINS OF MICE. WE SHOW FOR THE FIRST TIME THAT SM/J MICE EXHIBIT SPONTANEOUS DISC DEGENERATION THAT CAPTURES IMPORTANT FEATURES OF HUMAN DISC PATHOLOGY. BASED ON OUR NOVEL PILOT DATA, WE HYPOTHESIZE THAT THIS DEGENERATIVE PHENOTYPE HAS A STRONG GENETIC BASIS AND DRIVEN BY EARLY CHANGES IN CELL PHENOTYPE, METABOLISM AND SENESCENCE. WE WILL INVESTIGATE THE CELLULAR AND GENETIC MECHANISMS THAT UNDERLIE DISC DEGENERATION IN SM/J MICE IN THREE AIMS. IN AIM 1 WE WILL TEST THE HYPOTHESIS THAT DISC DEGENERATION IN SM/J MICE IS CHARACTERIZED BY COMPROMISED CELL SURVIVAL, ALTERED CELL PHENOTYPE, METABOLISM AND SENESCENCE. WE WILL PERFORM RNA-SEQ AND CHIP-SEQ ANALYSIS OF NP TISSUE OF SM/J AND LG/J MICE. WE WILL ALSO PERFORM METABOLOMICS TO DETERMINE METABOLIC STATUS OF CELLS. THESE STUDIES WILL PROVIDE UNBIASED INSIGHTS INTO TEMPORAL ALTERATIONS IN TRANSCRIPTOME AND METABOLISM TO DELINEATE PATHWAYS AND MECHANISMS CENTRAL TO THE PATHOGENESIS OF DISC DEGENERATION IN SM/J MICE. IN AIM 2 WE WILL TEST THE HYPOTHESIS THAT REMOVAL OF SENESCENT CELLS FROM THE INTERVERTEBRAL DISC SLOWS DOWN THE PROGRESSION OF DISC DEGENERATION IN SM/J MICE. PILOT STUDIES HAVE SHOWN THAT THERE IS AN ACCUMULATION OF SENESCENT CELLS IN SM/J DISCS. WE WILL TREAT EX VIVO ORGAN CULTURED DISCS AND SM/J MICE IN VIVO DURING EARLY AND ESTABLISHED STAGES OF DISC DEGENERATION WITH WELL-CHARACTERIZED SENOLYTIC DRUGS AND ANALYZE METABOLIC, HISTOPATHOLOGICAL AND GENETIC CHANGES. LASTLY IN AIM 3 WE WILL DELINEATE THE GENETIC MECHANISMS THAT CONTRIBUTE TO CELL SENESCENCE AND DISC DEGENERATION IN SM/J MICE. WE WILL PERFORM A GENOME-WIDE ASSOCIATION STUDY (GWAS) USING LG/J X SM/J ADVANCED INTERCROSS LINES (AILS) TO DISCOVER REGIONS OF THE MOUSE GENOME CONTRIBUTING TO DIFFERENCES IN DISC DEGENERATION. FOR THIS PURPOSE, AIL MICE WILL BE GENOTYPED AT 143,000 GIGAMUGA CHIP SINGLE NUCLEOTIDE POLYMORPHISMS (SNPS) AND DISCS FROM SEVERAL MICE WILL BE USED FOR RNA-SEQ ANALYSIS. TOGETHER, THESE DATA WILL BE USED TO IDENTIFY HIGH-RESOLUTION QUANTITATIVE TRAIT LOCI (QTL) FOR DISC TRAITS ~5 GENES PER SUPPORT INTERVAL. THE GENES RESPONSIBLE FOR THESE QTL WILL BE IDENTIFIED. THIS AIM WILL GENERATE CANDIDATE GENES LIKELY CONTRIBUTING TO VARIATION IN THE SENESCENCE AND DISC DEGENERATION BETWEEN LG/J AND SM/J STRAINS.

PROSTATE CANCER DIAGNOSIS BY MULTIPARAMETRIC ULTRASOUND

MONITORING NEOADJUVANT CHEMOTHERAPY OF BREAST CANCER USING 3D SUBHARMONIC AIDED PRESSURE ESTIMATION

PROMOTING REGENERATIVE REPAIR OF AGED CARTILAGE - PROJECT SUMMARY/ABSTRACT AGE IS THE NUMBER ONE RISK FACTOR FOR OSTEOARTHRITIS (OA), HOWEVER, THE MECHANISMS THAT DRIVE AGE-ASSOCIATED JOINT CHANGES AND HOW THESE CONTRIBUTE TO CARTILAGE DAMAGE ARE NOT WELL DEFINED. AS OA JOINT TISSUES AGE, IT IS WELL ESTABLISHED THAT CARTILAGE-FORMING STEM CELLS (CSCS) DISPLAY A SIGNIFICANT DECLINE IN THEIR ABILITY TO DIFFERENTIATE, AND THE JOINT ENVIRONMENT BECOMES MORE INFLAMMATORY. BOTH OF THESE FACTORS TOGETHER UNDERMINE THE ABILITY TO RECOVER FROM INJURIES AND ALSO CONTRIBUTES TO THE FAILURE OF AUTOLOGOUS STEM CELL THERAPIES. THUS, WE PROPOSE AN INNOVATIVE DUAL STRATEGY WHICH WILL ADDRESS BOTH. WE HYPOTHESIZE THAT ACTIVATION OF SIRTUIN 6 (SIRT6) WILL REJUVENATE OLDER CSCS, AND APOPTOSIS SIGNAL REGULATING KINASE 1 (ASK1) INHIBITION WILL REESTABLISH A REGENERATIVE ENVIRONMENT IN THE JOINT. TOGETHER, THIS NOVEL SOLUTION HAS THE POTENTIAL TO GREATLY AUGMENT THE ABILITY FOR CSC DIFFERENTIATION AND CARTILAGE REPAIR IN THE AGED POPULATION. WE WILL TEST THIS HYPOTHESIS AND DETERMINE THE EPIGENETIC MECHANISMS THAT SIRT6 REGULATE IN THE FOLLOWING THREE AIMS. IN AIM 1, WE WILL DETERMINE THE MECHANISMS BY WHICH (A) SIRT6 ACTIVATION AND (B) ASK1 INHIBITION ENHANCE THE CARTILAGE-FORMING DIFFERENTIATION AND PRO-SURVIVAL ACTIVITIES OF AGED CSCS. IN AIM 2, WE WILL DETERMINE THE EFFICACY OF SIRT6 ACTIVATION, IN THE PRESENCE OF ASK1 INHIBITION TO STIMULATE CARTILAGE FORMATION IN AGED, MALE AND FEMALE RODENTS. FINALLY, IN AIM 3, WE WILL DETERMINE THE EFFICACY OF SIRT6 ACTIVATION AND ASK1 INHIBITION TO PROMOTE CARTILAGE REGENERATION AND REPAIR IN RESPONSE TO CHONDRAL INJURY IN A PRE-CLINICAL AGED MINIPIG MODEL. THESE AIMS WILL BE ACHIEVED THROUGH COMPREHENSIVE IN VITRO ANALYSIS OF YOUNG AND OLD (MALE AND FEMALE) CSCS AND MACROPHAGES, IN PROOF-OF-CONCEPT AGING RODENT MODELS (MICE AND RATS), AND A CLINICALLY RELEVANT LARGE ANIMAL CHONDRAL DEFECT MODEL (AGED MINIPIG). IMPORTANTLY, THE FINDINGS FROM THIS PROPOSAL ARE LIKELY TO GIVE UNIQUE AND IMPORTANT INSIGHTS INTO THE ROLE OF EPIGENETIC REGULATION OF AGED CELLS AND THE ROLE OF THE SURROUNDING ENVIRONMENT IN OTHER AGING AND DISEASED TISSUES. IF SUCCESSFUL, THE OVERALL IMPACT OF THIS NOVEL PRO-REGENERATIVE THERAPY TO AMELIORATE THE RAVAGES OF AGING AND JOINT INJURY WOULD REPRESENT A SIGNIFICANT ADVANCEMENT IN THE TREATMENT OF AGE-ASSOCIATED DISEASES, SUCH AS OA. ADDITIONALLY, IT WILL LEAD TO THE DISCOVERY OF NEW DISEASE-MODIFYING TREATMENTS FOR OTHER ORGAN SYSTEMS AND INCREASE THE ABILITY OF THE AGED POPULATION TO LIVE A HEALTHY, MOBILE LIFE.

MECHANISM OF LUNG CANCER RESISTANCE TO TYROSINE KINASE INHIBITOR AND RADIATION TREATMENTS

ANDROGEN EFFECT ON MOTOR/COGNITIVE OUTCOME IN KLINEFELTER SYNDROME

PHILADELPHIA AUTOIMMUNITY CENTER OF EXCELLENCE

MULTISCALE MODEL OF THE VAGAL OUTFLOW TO THE HEART

PHARMACOLOGICAL JAK2 INHIBITION TO OVERCOME ANDROGEN RECEPTOR ABERRATIONS IN PROSTATE CANCER - THERE IS AN UNMET MEDICAL NEED FOR DEVELOPMENT OF MORE EFFICACIOUS THERAPIES FOR CASTRATE-RESISTANT (CR) PROSTATE CANCER (PC). THE CASTRATE RESISTANT STATE OF PC IS INCURABLE AND RESULTS FROM A FAILURE OF ANDROGEN DEPRIVATION THERAPY (ADT), WHICH TARGETS ANDROGEN RECEPTOR (AR) ACTIVITY IN PC CELLS. THIS HAS LED TO DEVELOPMENT OF SECOND-GENERATION AR-TARGETED THERAPIES THAT MORE EFFECTIVELY ANTAGONIZE THE AR LIGAND BINDING DOMAIN (ENZALUTAMIDE) OR REDUCE ANDROGEN SYNTHESIS (ABIRATERONE ACETATE). HOWEVER, AFTER AR-TARGETING THERAPIES, AR AND ITS SPLICE VARIANTS ARE STILL EXPRESSED AT HIGH LEVELS AND REMAIN TRANSCRIPTIONALLY ACTIVE IN CRPC AND DRIVE CASTRATE-RESISTANT GROWTH, ULTIMATELY CAUSING PATIENT DEATH. A MAJOR GAP IN THE FIELD IS THE LACK OF UNDERSTANDING OF TARGETABLE MECHANISMS THAT INDUCE PERSISTENT AR EXPRESSION AND TRANSCRIPTIONAL ACTIVITY IN CRPC. IN THIS MPI PROPOSAL, WE WILL INTERROGATE THE NOVEL CONCEPT THAT JAK2-STAT5 SIGNALING REPRESENTS A CRITICAL DRIVER OF AR GENE EXPRESSION IN PC AND, THEREFORE, PHARMACOLOGICAL TARGETING OF JAK2 SIGNALING BY THE NEW-GENERATION JAK2 INHIBITORS REPRESENTS A NOVEL THERAPEUTIC STRATEGY TO ELIMINATE AR IN CRPC. THIS IS SUPPORTED BY OUR RIGOROUS PRELIMINARY DATA SHOWING THAT ACTIVATION OF JAK2-STAT5 SIGNALING IS A STRONG INDUCER OF THE AR GENE TRANSCRIPTION LEADING TO HIGH AR MRNA AND PROTEIN LEVELS IN PC. LIKEWISE, INHIBITION OF JAK2-STAT5 SIGNALING BLOCKS EXPRESSION OF AR AND AR-VS IN PC CELL LINES, PC XENOGRAFT TUMORS IN VIVO AND IN PATIENT-DERIVED CLINICAL PCS GROWN AS EXPLANT CULTURES EX VIVO. COLLECTIVELY, THESE FINDINGS SUPPORT THE HYPOTHESIS THAT ACTIVATION OF JAK2-STAT5 CRITICALLY PROMOTES CRPC PROGRESSION BY SUSTAINING EXPRESSION OF AR AND CONSTITUTIVELY ACTIVE AR VARIANTS. WE WILL PURSUE TWO AIMS:1) DETERMINE THE MECHANISMS UNDERLYING CONTROL OF AR AND AR VARIANT MRNA EXPRESSION BY JAK2 SIGNALING IN PC, AND THE OVERLAP OF THE JAK2-STAT5 AND AR TRANSCRIPTOMES; 2) DETERMINE THE EFFICACY OF THE NEW-GENERATION JAK2-INHIBITORS FEDRATINIB (FED) AND PACRITINIB (PAC) IN ELIMINATING AR-FL/V7/V9 MRNA AND PROTEIN EXPRESSION IN CRPC AND SUPPRESSING GROWTH OF CRPC IN VITRO AND IN VIVO. THE PROPOSED WORK IS SIGNIFICANT BECAUSE IT WILL DETERMINE THE MECHANISMS BY WHICH JAK2-SIGNALING DRIVES PERSISTENT AR EXPRESSION IN CRPC. MOREOVER, THE PROPOSED WORK WILL ESTABLISH WHETHER JAK2 INHIBITION BY PAC AND FED CAN BLOCK AR EXPRESSION AND GROWTH OF PC RESISTANT TO AR-TARGETED THERAPY. THE NOVELTY OF THE PROPOSED CONCEPT IS THAT JAK2 REGULATION OF AR OPENS AN ENTIRELY NEW AVENUE TO DIRECTLY CONTROL AR LEVELS AND PC GROWTH THROUGH PHARMACOLOGICAL SUPPRESSION WITH NEW-GENERATION JAK2 INHIBITORS CURRENTLY FDA-APPROVED OR IN CLINICAL DEVELOPMENT FOR OTHER PURPOSES. THESE FIRST-IN-THE-FIELD STUDIES WILL HAVE NEAR-TERM IMPACT FOR THERAPY DEVELOPMENT FOR CRPC PATIENTS BECAUSE THEY ARE EXPECTED TO ESTABLISH PAC AND FED AS NOVEL AGENTS TO TARGET AR IN PC, AND TRANSLATE TO A PHASE I/II CLINICAL STUDY IN CRPC.

OPTIMIZING ULTRASOUND ENHANCED DELIVERY OF THERAPEUTICS

INVESTIGATING A DUAL-PATHWAY FRAMEWORK FOR PRAXIS - PROJECT SUMMARY PERFORMING SKILLED ACTIONS RELIES ON TWO FUNDAMENTAL ABILITIES: IMITATING MOVEMENTS TO QUICKLY LEARN NEW BEHAVIORS, AND USING TOOLS TO MORE EFFICIENTLY MANIPULATE OUR ENVIRONMENTS. IMITATION AND TOOL USE ARE CRITICAL FOR MANY ACTIVITIES OF DAILY LIVING; LOSS OF THESE ABILITIES IN THE CLINICAL DISORDER OF LIMB APRAXIA, FREQUENTLY OBSERVED IN MANY NEUROLOGICAL DISORDERS INCLUDING IN ~50% OF INDIVIDUALS WITH LEFT-HEMISPHERE STROKE, IS THE STRONGEST PREDICTOR OF INCREASED CAREGIVER DEPENDENCE AND POOR POST-STROKE RECOVERY. DESPITE THE RECOGNIZED IMPORTANCE OF THESE ABILITIES, THE PROCESSING MECHANISMS UNDERLYING IMITATION AND TOOL USE REMAIN UNCLEAR, IN LARGE PART DUE TO THE CHALLENGES ASSOCIATED WITH STUDYING THEM. IMITATION AND TOOL USE BOTH INVOLVE TRANSLATING CONCEPTUAL COGNITIVE GOALS (I.E., KNOWING HOW TO SERVE A TENNIS BALL) INTO INTRICATE SETS OF MOTOR COMMANDS (I.E., ACCURATELY HITTING A TENNIS BALL WITH A RACKET). HENCE, THE STUDY OF IMITATION AND TOOL USE REQUIRES MERGING MANY CONCEPTS FROM COGNITIVE AND MOTOR NEUROSCIENCE. THIS PROPOSAL AIMS TO BRIDGE THIS COGNITIVE-MOTOR DIVIDE BY DEVELOPING A NEW THEORY OF THE COMMON MECHANISMS SUPPORTING IMITATION AND TOOL USE THAT EXPLAINS HOW CONCEPTUAL GOALS ARE ACTUALLY TURNED INTO MOTOR COMMANDS. THE PROPOSED THEORY BUILDS ON OUR PRIOR RESEARCH BY HYPOTHESIZING TWO INTERACTING PROCESSING ROUTES: (1) A ROUTE THAT SPECIFIES ACTIONS IN TERMS OF THE TRAJECTORY OF THE END-EFFECTOR (E.G., WRITING LETTERS), AFFORDING AN EFFICIENT MEANS OF DEFINING DESIRED TOOL OR HAND MOTION IN AN ABSTRACT (BODY-INDEPENDENT) MANNER, AND (2) A ROUTE THAT SPECIFIES ACTIONS IN TERMS OF BODY CONFIGURATIONS, PROVIDING A MORE DETAILED BUT STILL COMPUTATIONALLY TRACTABLE DESCRIPTION OF THE POSITIONING OF THE ENTIRE LIMB (AND TOOL) THROUGHOUT AN ACTION. WHILE THESE TWO ROUTES TYPICALLY WORK TOGETHER, THEY MAY BE ENGAGED TO DIFFERENT EXTENTS DUE TO TASK DEMANDS (E.G., FOCUSING ON RACKET TRAJECTORIES OR SHOULDER-ELBOW-WRIST POSITIONS DURING A TENNIS SERVE) OR WHEN THE DIFFERENT BRAIN REGIONS SUPPORTING THESE DISTINCT PROCESSING ROUTES BECOME DISRUPTED. THUS, EVIDENCE OF THESE TWO ROUTES WILL BE ESTABLISHED THROUGH NOVEL TASK-SWITCHING PARADIGMS IN HEALTHY INDIVIDUALS, MEASURING BEHAVIORAL IMPAIRMENTS DUE TO TRANSIENT DISRUPTIONS OF RELEVANT BRAIN REGIONS USING TRANSCRANIAL MAGNETIC STIMULATION, OR BY QUANTIFYING CHRONIC DEFICITS IN PATIENTS WITH LEFT-HEMISPHERE STROKES. WE WILL APPLY NOVEL RIGOROUS KINEMATIC ANALYSES TO ASSESS BEHAVIOR, CORRELATE PATIENT IMPAIRMENTS WITH CLINICAL MEASURES OF APRAXIA, AND USE RECENTLY-DEVELOPED MULTIVARIATE LESION-SYMPTOM MAPPING APPROACHES TO CONFIRM THE NEUROANATOMICAL BASES OF THESE ROUTES. TOGETHER, SUCH EFFORTS WILL IDENTIFY AND CHARACTERIZE HOW TRAJECTORY AND BODY-CONFIGURATION PLANNING SUPPORT THE PRODUCTION AND PERCEPTION OF IMITATION AND TOOL USE ACTIONS. RESULTS WILL NOT ONLY ADVANCE OUR UNDERSTANDING OF APRAXIA, HIGHLIGHTING ALTERNATIVE COMPENSATORY PATHWAYS THAT MIGHT BE TARGETED FOR REHABILITATION TREATMENT, BUT SERVES AS A MODEL FOR STUDYING COGNITIVE-MOTOR INTERACTIONS MORE BROADLY.

MECHANISMS OF PROTECTIVE MEMORY CD8 T-CELL INDUCTION BY MRNA-LNP VACCINES - SUMMARY CD8 T-CELLS RECOGNIZE AND KILL VIRUS-INFECTED CELLS DISPLAYING AT THE CELL SURFACE SHORT VIRAL PEPTIDES BOUND TO MAJOR HISTOCOMPATIBILITY (MHC) CLASS I MOLECULES (MHC-I). CD8 T-CELLS CONTRIBUTE TO THE CLEARANCE OF MANY VIRAL INFECTIONS. AFTER AN INFECTION SUBSIDES, AN EXPANDED POPULATION OF “MEMORY” CD8 T-CELLS (M CD8 T-CELLS) MAY CONTRIBUTE TO MORE RAPIDLY CONTROLLING A SECONDARY INFECTION WITH THE VIRUS. VACCINES CAN MIMIC THIS PROCESS. MODIFIED MRNA (MMRNA) ENCAPSULATED IN LIPID NANOPARTICLES (MMRNA-LNP) HAVE EMERGED AS A POWERFUL VACCINE PLATFORM. MMRNA-LNPS HAVE MANY ADVANTAGES AS VACCINES: 1) THEY CAN BE FOCUSED ON THE ANTIGEN OF INTEREST. 2) THEY ARE HIGHLY IMMUNOGENIC. 3) THEY ARE EASY TO MAKE. 4) THEY CAN BE MASS-PRODUCED RAPIDLY. 5) THEY ARE RELATIVELY INEXPENSIVE. THE SWIFT DEVELOPMENT AND APPROVAL OF THE MMRNA-LNP VACCINES TO COMBAT SARS-COV-2 ATTEST TO THEIR POTENTIAL. WHILE IT IS KNOWN THAT MMRNA-LNPS INDUCE CD8 T-CELL RESPONSES, MOST OF THE WORK ON THEIR PROTECTION MECHANISMS HAS FOCUSED ON ABS. THE MCD8 T-CELLS INDUCED BY MMRNA- LNP CAN POTENTIALLY COMPLEMENT AB PROTECTION OR MAY PROVIDE MOST OF THE PROTECTION FOR VIRUSES THAT ARE REFRACTORY TO AB-MEDIATED CONTROL. MMRNA-LNPS COULD ALSO BE USED TO INDUCE CD8 T-CELLS AGAINST CANCER. WE HAVE PUBLISHED THAT THE MCD8 T-CELL RESPONSES INDUCED BY MMRNA-LNPS PROTECT MICE FROM HIGHLY LETHAL MOUSEPOX, A SYSTEMIC VIRAL DISEASE OF THE MOUSE CAUSED BY THE ORTHOPOXVIRUS (OPV) ECTROMELIA VIRUS (ECTV). ECTV IS AN OUTSTANDING MODEL FOR SYSTEMIC VIRAL INFECTIONS IN GENERAL AND FOR OPVS THAT CAN INFECT HUMANS, SUCH AS THE ERADICATED VARIOLA VIRUS (VIRUS OF SMALLPOX) AND FOR MONKEYPOX VIRUS (MPXV), WHICH RECENTLY CAUSED A MAJOR OUTBREAK. IN STILL UNPUBLISHED EXPERIMENTS, WE ALSO FOUND THAT M CD8 T-CELLS INDUCED BY A MINI- MMRNA VACCINE ENCODING FOR ONLY THE MINIMAL, HIGHLY CONSERVED CD8 T-CELL EPITOPE VNFNFNGL OF THE SARS- COV-2 SPIKE PROTEIN PROTECTS WILD-TYPE MICE FROM LETHAL RESPIRATORY INFECTION WITH THE MOUSE-ADAPTED SARS- COV-2 STRAIN MA30, AN OUTSTANDING MODEL FOR SARS-COV-2 AND OTHER GRAVE RESPIRATORY INFECTIONS. HERE WE PROPOSE ELUCIDATING THE MECHANISMS WHEREBY MMRNA-LNPS INDUCE PROTECTIVE MCD8 T-CELLS USING THE ECTV SYSTEMIC AND THE MA30 SARS-COV-2 RESPIRATORY MOUSE MODELS. OUR SPECIFIC AIMS ARE TO: A) SPECIFIC AIM 1. INVESTIGATE THE MECHANISMS OF MHC-I ANTIGEN PRESENTATION AFTER MMRNA-LNP VACCINATION. B) SPECIFIC AIM 2. INVESTIGATE THE ROLES OF TYPE I INTERFERON (IFN-I) AND OTHER PROINFLAMMATORY CYTOKINES IN PROTECTIVE M CD8 T-CELL DEVELOPMENT AFTER MRNA-LNP VACCINATION.

MIR-152/PKM2/SLC7A5 AXIS IN BREAST CANCER DEVELOPMENT, CHEMO- AND RADIATION-TREATMENT RESPONSE - BREAST CANCER (BCA) IS THE LEADING CAUSE OF WOMEN CANCER MORTALITY WORLDWIDE, AND TRIPLE NEGATIVE BREAST CANCER (TNBC) CELLS ACCOUNTS FOR 15-20% BCA. HOWEVER, HIGHER THERAPEUTIC RESISTANCE AND LOWER SURVIVAL OF TNBC PATIENTS REMAIN THE MAJOR HINDER OF BCA TREATMENT. DOXORUBICIN (DOX) AND RADIATION ARE COMMONLY USED FOR TNBC TREATMENT. IN OUR PRELIMINARY STUDY, WE FOUND THAT TNBC SHOWED MUCH LOWER EXPRESSION LEVELS OF MIR-152. HIGHER EXPRESSION LEVELS OF ZESTE HOMOLOGUE 2 (EZH2) AND DNA HYPERMETHYLATION WERE INVOLVED IN MIR-152 SUPPRESSION IN TNBC CELLS. TO UNDERSTAND MECHANISM OF MIR-152 SUPPRESSION, AND ROLE AND MECHANISM OF MIR-152 IN REGULATING CANCER DEVELOPMENT AND DOX- AND RADIATION-INDUCED RESISTANCE, WE PERFORMED A LOT OF PRELIMINARY EXPERIMENTS TO IDENTIFY WHAT MOLECULES THAT WERE POTENTIAL MIR-152 DIRECT TARGETS AS WELL AS THAT WERE UPREGULATED IN DOX-RESISTANT TNBC OR PDX TUMOR TISSUES. WE FOUND THAT PYRUVATE KINASE MUSCLE 2 (PKM2), SOLUTE CARRIER FAMILY 7 MEMBER 5 (SLC7A5), AND POLYPYRIMIDINE TRACT-BINDING PROTEIN (PTBP1) AS DIRECT TARGETS OF MIR-152 WITH POTENTIAL ABILITY TO CONVERSE DOX-MEDIATED RESISTANCE. PKM2, SLC7A5 AND PTBP1 LEVELS WERE INCREASED WHEN MIR-152 EXPRESSION WAS SUPPRESSED IN TNBC CELLS AND PDX MODEL. WE FOUND THAT MIR-152 SUPPRESSION PLAYED AN IMPORTANT ROLE IN MEDIATING TNBC METABOLIC REPROGRAMMING, WARBURG EFFECT SWITCH, TUMOR GROWTH AND DOX RESISTANCE THROUGH PKM2/SLC7A5/PTBP1. WE SHOWED THAT OVEREXPRESSION OF MIR-152 OR PKM2 KNOCKDOWN RENDERED TNBC CELLS MORE SENSITIVE TO RADIATION TREATMENT. THESE NEW FINDINGS CREATE NEW OPPORTUNITY TO INVESTIGATE MECHANISM OF TNBC THERAPEUTIC RESISTANCE. WE HYPOTHESIZE THAT RESISTANCE TO CHEMOTHERAPY AND RADIATION TREATMENT, METABOLIC REPROGRAMMING, AND TNBC DEVELOPMENT ARE INDUCED BY MIR-152 SUPPRESSION AND UPREGULATION OF PKM2, SLC7A5, AND PTBP1. TO TEST THIS CENTRAL HYPOTHESIS, WE WILL PERFORM EXPERIMENTS THROUGH THREE AIMS. IN AIM 1, WE WILL INVESTIGATE ROLE AND MECHANISM OF MIR-152 SUPPRESSION IN TNBC CELLS IN DOXORUBICIN- AND RADIATION-INDUCED RESISTANCE; AND ROLE AND MECHANISM OF PKM2 SWITCH AND INDUCTION VIA ALTERNATIVE SPLICING BY MIR-152 SUPPRESSION TO INDUCE WARBURG EFFECT, THERAPEUTIC RESISTANCE, AND TUMOR GROWTH. IN AIM 2, WE WILL DETERMINE ROLE AND MECHANISM OF MIR- 152/SLC7A5/PTBP1 PATHWAY IN REGULATING METABOLIC REPROGRAMMING AND WARBURG EFFECT SWITCH, AND IN MEDIATING DOXORUBICIN- AND RADIATION-INDUCED THERAPEUTIC RESISTANCE. IN AIM 3, WE WILL DETERMINE WHETHER MIR-152 SUPPRESSION AND PKM2 INDUCTION REGULATE TUMOR ANGIOGENESIS THROUGH CXCL8 EXPRESSION USING A HUMANIZED CHIMERIC TUMOR MODEL; AND WHETHER LEVELS OF MIR-152, PKM2, HIF-1A, HNRNPA3, SLC7A5, AND PTBP1 ARE CORRELATED EACH OTHER, AND ARE CORRELATED WITH THERAPEUTIC RESPONSES AND WITH THE CANCER STAGES AND SURVIVAL. THIS R01 PROJECT WILL IDENTIFY MECHANISMS OF THERAPEUTIC RESISTANCE, AND METABOLIC REPROGRAMMING; AND PROVIDE INFORMATION FOR DEVELOPING NEW TREATMENT OPTION OF TNBC IN THE FUTURE.

PIRNA BIOGENESIS RULED BY PIWI-TUDOR INTERACTION

A MOBILE TXT-BASED INTERVENTION TO IMPROVE ADHERENCE TO ADJUVANT HORMONE THERAPY AND SYMPTOM MANAGEMENT FOR BCA SURVIVORS

TARGETING LATEXIN SIGNALING FOR ENDOTHELIAL BARRIER DYSFUNCTION IN INFLAMMATORY LUNG INJURY - PROJECT SUMMARY THE ACUTE RESPIRATORY DISTRESS SYNDROME (ARDS) IS A COMMON CAUSE OF RESPIRATORY FAILURE IN THE CRITICALLY ILL, ACCOUNTING FOR ~75000 DEATHS/YEAR AND 3.6 MILLION HOSPITAL DAYS. THE EXISTING PARADIGM IS THAT THE ARDS RESULTS FROM WIDESPREAD DYSFUNCTION OF THE PULMONARY ENDOTHELIUM, LEADING TO MICROVASCULAR THROMBOSIS, INTERSTITIAL EDEMA, ALVEOLAR FLOODING AND RESPIRATORY FAILURE, ALTHOUGH THE MECHANISMS LEADING TO ENDOTHELIAL DYSFUNCTION REMAIN UNKNOWN. THIS PROPOSAL ARISES FROM NOVEL OBSERVATIONS LINKING THE UPREGULATION OF LATEXIN TO ENDOTHELIAL DYSFUNCTION IN THE LUNG. WE FOUND THAT HEALTHY LUNG ENDOTHELIUM EXPRESSES LOW LEVELS OF LATEXIN WHEREAS LEVELS MARKEDLY INCREASE IN RESPONSE TO TURBULENT FLOW OR BACTERIAL LPS. FURTHER, WE FOUND THAT ENDOTHELIAL BARRIER FUNCTION WAS ENHANCED BY REDUCING LATEXIN EXPRESSION IN LPS-EXPOSED LUNG ENDOTHELIAL CELLS AND THAT GLOBAL DEFICIENCY OF LATEXIN IN MICE REDUCED LPS-INDUCED PULMONARY EDEMA, LUNG INFLAMMATION AND MORTALITY. MECHANISTICALLY, WE FOUND THAT LATEXIN MEDIATED ITS EFFECTS THROUGH COMPLEX MECHANISMS, INCLUDING BINDING TO THE ENZYME SRC KINASE AND FACILITATING ITS MEMBRANE TRANSLOCATION AND PHOSPHORYLATION OF VE-CADHERIN AT ADHERENS JUNCTIONS (AJ). ADDITIONALLY, MASS SPECTROSCOPY REVEALED THAT LATEXIN PHYSICALLY INTERACTS WITH SEVERAL OTHER PROTEINS, INCLUDING CLATHRIN AND AP2, SUGGESTING A ROLE IN ENDOCYTIC TRAFFICKING, AND SYNTAXIN-3 AND SNAP3, SUGGESTING A ROLE IN EXOCYTOSIS. CONSISTENT WITH THIS LATTER MECHANISM, WE FOUND THAT LATEXIN DEFICIENCY REDUCED VON WILLENBRAND FACTOR SECRETION FROM LUNG ENDOTHELIUM. TAKEN TOGETHER, THESE OBSERVATIONS LEAD US TO PROPOSE THE FOLLOWING CENTRAL HYPOTHESIS REGARDING THE ROLE OF LATEXIN IN LUNG ENDOTHELIAL BIOLOGY AND THE PATHOGENESIS OF ENDOTHELIAL DYSFUNCTION IN ARDS. WE HYPOTHESIZE THAT LATEXIN PLAYS A PATHOGENIC ROLE IN DRIVING AGONIST- INDUCED ENDOTHELIAL DYSFUNCTION IN THE LUNG AND THAT INHIBITING ITS INTERACTIONS WITH OTHER KEY PROTEINS WILL REDUCE THE SEVERITY OF PULMONARY EDEMA, LUNG INFLAMMATION AND MICROVASCULAR THROMBOTIC COMPLICATIONS IN EXPERIMENTALLY-INDUCED ARDS. TO TEST THIS HYPOTHESIS, WE PROPOSE 3 INDEPENDENT BUT MECHANISTICALLY LINKED SPECIFIC AIMS. IN AIM 1, WE WILL ESTABLISH THAT ENDOTHELIAL-SPECIFIC DELETION OF LATEXIN ENHANCES ENDOTHELIAL BARRIER PROTECTION AND REDUCES MICROVASCULAR THROMBOSIS AND MORTALITY TO LPS IN MICE. IN AIM 2, WE WILL DELINEATE THE MOLECULAR MECHANISMS BY WHICH LATEXIN REGULATES VE-CADHERIN MEMBRANE BIOAVAILABILITY AT ENDOTHELIAL AJS BY PERFORMING VARIOUS IN VITRO AND IN VIVO LOSS- AND GAIN-OF-FUNCTION STUDIES FOR GENES LINKED TO SRC-MEDIATED VE-CADHERIN PHOSPHORYLATION AND CLATHRIN-MEDIATED ENDOCYTOSIS IN ECS. IN AIM 3, WE WILL DELINEATE THE MOLECULAR MECHANISMS BY WHICH LATEXIN MEDIATES VWF SECRETION FROM ENDOTHELIAL CELLS, FOCUSING ON ITS ROLE IN SNARE COMPLEX FORMATION. IN TOTO, THIS PROPOSAL WILL ESTABLISH THE MECHANISMS BY WHICH LATEXIN REGULATES KEY PATHOLOGICAL BEHAVIORS IN LUNG ENDOTHELIUM AND PROVIDE DIRECTION FOR DEVELOPING NOVEL THERAPEUTIC APPROACHES FOR INFLAMMATORY VASCULAR DISEASES OF THE LUNG, SUCH AS THE ARDS.

CHROMATIN BIOLOGY OF THE AFRICAN TRYPANOSOME - PROJECT SUMMARY PROTOZOAN PARASITES OF THE GROUP KINETOPLASTIDS ARE RESPONSIBLE FOR MAJOR HUMAN MALADIES SUCH AS FATAL SLEEPING SICKNESS (TRYPANOSOMA BRUCEI, ALSO TERMED THE AFRICAN TRYPANOSOME), CHAGAS DISEASE (T. CRUZI), AND LEISHMANIASIS (LEISHMANIA SPECIES). DUE TO THE LACK OF INEXPENSIVE AND SAFE DRUGS, RISING RESISTANCE AGAINST CURRENT DRUGS, AND LIMITED DRUG DISCOVERY EFFORTS, NOVEL APPROACHES ARE URGENTLY NEEDED TO COMBAT THESE NEGLECTED TROPICAL DISEASES. BECAUSE KINETOPLASTIDS CONSTITUTE ONE OF THE EARLIEST-BRANCHING ORGANISMS IN THE EUKARYOTIC TREE OF LIFE, THEY EXHIBIT NUMEROUS MOLECULAR AND CELLULAR FEATURES THAT ARE DISTINCT FROM METAZOA AND FUNGI, AND THAT CAN BE EXPLOITED FOR PHARMACOLOGICAL INTERVENTION. BY COMBINING STRUCTURAL, BIOCHEMICAL, AND IN VIVO APPROACHES, WE SEEK TO ADDRESS FUNDAMENTAL QUESTIONS IN CHROMATIN BIOLOGY AND GENE REGULATION IN THE MODEL KINETOPLASTID T. BRUCEI. WE ARE PARTICULARLY INTERESTED IN THE STRUCTURE AND MECHANISM OF THE CLOSELY RELATED DOT1A AND DOT1B ENZYMES THAT ARE KEY REGULATORS OF ESSENTIAL FUNCTIONS IN T. BRUCEI AND THAT CATALYZE THE METHYLATION OF HISTONE H3 LYSINE 76 (H3K76) IN THE GLOBULAR NUCLEOSOME CORE REGION. WHILE DOT1A REGULATES CELL-CYCLE PROGRESSION, DOT1B IN ANTIGENIC VARIATION, AN ESSENTIAL MECHANISM FOR THE PARASITE TO EVADE THE HOST’S IMMUNE SYSTEM. DUE TO SIGNIFICANT MECHANISTIC DIFFERENCES OF TRYPANOSOME DOT1A/B TO HUMAN AND YEAST DOT1 ENZYMES, THE MOLECULAR MECHANISMS OF HOW THEY METHYLATE CHROMATIN AND HOW THEY ARE REGULATED REMAIN POORLY UNDERSTOOD. IN AIM 1, WE WILL THEREFORE INVESTIGATE THE MECHANISM OF DOT1A-NUCLEOSOME SUBSTRATE RECOGNITION AND ITS IMPACT ON CELL CYCLE CONTROL. IN AIM 2, WE WILL DECIPHER THE REGULATORY MECHANISM OF DOT1A GOVERNED BY RNASEH2, AN ENZYME THAT IS KNOWN TO CLEAVE RNA IN RNA/DNA HYBRIDS AND THAT HAS BEEN IMPLICATED IN BOTH DNA REPLICATION AND TRANSCRIPTIONAL REGULATION. OUR GOAL IS TO DEFINE THE IMPACT OF RNASEH2 ON DOT1A ACTIVITY, PROVIDE A STRUCTURAL BASIS FOR ITS REGULATORY FUNCTION, AND ELUCIDATE THE MECHANISM OF HOW DOT1A ACTIVITY IS COORDINATED WITH THE CELL CYCLE. THE INTERACTION OF RNASEH2 WITH DOT1A/B IS SPECIFIC TO TRYPANOSOMES, SUGGESTING A NOVEL REGULATORY MECHANISM OF DOT1 ENZYMES. COLLECTIVELY, OUR STUDIES WILL ILLUMINATE THE LONG- STANDING QUESTION OF HOW DOT1A IS RECRUITED TO CHROMATIN AND HOW IT IS REGULATED IN A SPATIOTEMPORAL MANNER. OUR STUDIES WILL YIELD THE FIRST ATOMIC STRUCTURES OF THE FUNDAMENTAL UNIT OF CHROMATIN, THE NUCLEOSOME, OF THE VAST GROUP OF PROTOZOA, WHICH ARE MEDICALLY, ECOLOGICALLY, EVOLUTIONARILY, AND SCIENTIFICALLY IMPORTANT EUKARYOTES. DUE TO THE NOVEL REGULATORY FUNCTION OF RNASEH2, OUR RESULTS WILL BROADEN OUR MECHANISTIC UNDERSTANDING OF DOT1A AND RNASEH2 ENZYMES. BECAUSE T. BRUCEI DOT1-REGULATED PROCESSES ARE ESSENTIAL FOR THE PARASITE, THIS RESEARCH MAY ULTIMATELY HAVE A LARGE IMPACT ON GLOBAL HEALTH BY EXPLOITING THE UNIQUE ATTRIBUTES OF PROTOZOAN DOT1 STRUCTURE AND REGULATION TO INFORM NOVEL THERAPIES FOR SLEEPING SICKNESS AND OTHER DISEASES CAUSED BY KINETOPLASTIDS THAT AFFECT HALF A BILLION OF PEOPLE.

DIACYLGLYCEROL KINASE IN AIRWAY SMOOTH MUSCLE FUNCTIONS

NATIONAL INSTITUTE ON DISABILITY AND REHABILITATION RESEARCH - SPINAL CORD INJURY MODEL DEMONSTRATIONS

REDUCING EMERGENCY DIABETES CARE FOR OLDER AFRICAN AMERICANS

LOCAL?IMMUNE?TOLERANCE?IN?THE?LARGE?INTESTINE?MEDIATED?BY?GPR15

G PROTEIN-COUPLED RECEPTOR KINASES

CAV-3 IN DIABETIC MYOCARDIAL INJURY FOLLOWING ISCHEMIA/REPERFUSION

SUPPORTING WOMEN AND CHILDREN IN SUBSTANCE ABUSE RECOVERY - A FAMILY UNIT APPROACH

MIR-21 REGULATION OF AUTOREACTIVE B CELL AND LUPUS NEPHRITIS DEVELOPMENT - PROJECT SUMMARY MICRORNAS (MIRNAS) ARE CRITICAL POST-TRANSCRIPTIONAL REGULATORS OF GENE EXPRESSION IN IMMUNITY AND AUTOIMMUNITY, ALTHOUGH MANY MIRNAS REMAIN POORLY CHARACTERIZED. ONE SUCH MIRNA, MIRNA-21 (MIR-21), IS UPREGULATED IN SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) PATIENTS AND PROMOTES VARIOUS DISEASES IN MOUSE MODELS. THESE INCLUDE EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE), ASTHMA, CANCER, AND OUR FINDINGS ON ITS ROLE IN TOLL-LIKE RECEPTOR 7 (TLR7)-DRIVEN SLE IN MICE. WE FOUND THAT TLR7 OVEREXPRESSING SLE-PRONE B6.SLE1BYAA MICE DEFICIENT IN MIR-21 (SLE1BYAAMIR-21-/-) HAD REDUCED GERMINAL CENTER (GC) B CELL, T FOLLICULAR HELPER (TFH) CELL AND PLASMA CELL (PC) RESPONSES. THESE RESPONSES IN SLE1BYAAMIR-21-/- MICE WERE STRONGLY ASSOCIATED WITH REDUCED AUTOANTIBODY, AND SPLENIC AND BONE MARROW AUTOANTIBODY-PRODUCING ANTIBODY FORMING CELL (AFC) RESPONSES. THESE DATA SUBSTANTIATE A ROLE FOR MIR-21 IN THE LOSS OF B CELL TOLERANCE AND AUTOIMMUNITY, WHICH NECESSITATES IDENTIFICATION OF MECHANISMS (FOCUS OF THE GRANT) BY WHICH MIR-21 PROMOTES AUTOANTIBODY PRODUCTION AND SLE. THIS WILL FILL GAPS IN KNOWLEDGE AND MAY HELP DISCOVER AN URGENTLY NEEDED TARGETED THERAPY FOR SLE, A DEBILITATING DISEASE THAT AFFECTS MILLIONS OF AMERICANS AND PEOPLE WORLD-WIDE. TLR SIGNALING, ESPECIALLY TLR7, PROMOTES SLE IN HUMANS AND MURINE MODELS, AND AN ATTRACTIVE THERAPEUTIC TARGET. TLR7 SIGNALING ACTIVATES THE NF-KB INFLAMMATORY SIGNALING PATHWAY. TLR7 UPREGULATES MIR-21 EXPRESSION IN B CELLS WHICH CAN TARGET THE NEGATIVE REGULATOR OF NF-KB SIGNALING, PELI1, ALTHOUGH THE ROLE OF MIR-21 IN REGULATING NF- KB SIGNALING IN SLE-PRONE B CELLS BY TARGETING PELI1 IS UNKNOWN. PRELIMINARY DATA SHOWED THAT B CELLS FROM SLE1BYAAMIR-21-/- MICE HAVE INCREASED PELI1 EXPRESSION COMPARED TO CONTROL SLE1BYAA B CELLS. IN ADDITION, PREVIOUS STUDIES IMPLICATED TLR SIGNALING IN METABOLIC REPROGRAMMING, PRIMARILY THROUGH THE ACTIVATION OF THE PI3K/AKT/MTOR PATHWAY IN VARIOUS IMMUNE CONDITIONS, ALTHOUGH THE ROLE OF THIS PATHWAY IN TLR7-MEDIATED METABOLIC REPROGRAMING IN B CELLS IN SLE IS NOT CLEAR. MIR-21 WAS ALSO SHOWN TO REGULATE THE PI3K/AKT/MTOR PATHWAY IN VARIOUS MODELS; HOWEVER, ITS ROLE IN REGULATING B CELL METABOLISM IN TLR7-DRIVEN SLE IS NOT KNOWN. INTERESTINGLY, OUR RNA-SEQ ANALYSIS OF B CELLS FROM A TLR7-INDUCED MODEL SHOWED NEGATIVELY ENRICHED GENES IN THE PI3K/AKT/MTOR PATHWAY IN THE ABSENCE OF MIR-21. ADDITIONAL PRELIMINARY DATA SHOWED INCREASED EXTRACELLULAR ACIDIFICATION (ECAR) AND OXYGEN CONSUMPTION (OCR) RATES IN SLE1BYAA B CELLS OVEREXPRESSING TLR7 COMPARED TO SLE1B B CELLS, INDICATING A MORE ENERGETIC PHENOTYPE IN RESPONSE TO TLR7 SIGNALING. CONVERSELY, SLE1BYAAMIR-21-/- B CELLS EXHIBITED REDUCED ECAR AND OCR COMPARED TO SLE1BYAA B CELLS. WE HYPOTHESIZE THAT MIR-21 PROMOTES TLR7-DRIVEN SYSTEMIC AUTOIMMUNE RESPONSES AND SLE DISEASE BY REGULATING TLR7-NF-KB SIGNALING AND METABOLIC REPROGRAMMING IN B CELLS. PROPOSED HYPOTHESIS AND SPECIFIC AIMS WILL HELP DELINEATE THE MECHANISM BY WHICH MIR-21 DRIVES AUTOIMMUNE RESPONSES AND LUPUS NEPHRITIS AND WILL INFORM US WHETHER MIR-21 OR MIR-21 REGULATED PATHWAY(S) CAN BE A THERAPEUTIC TARGET FOR SLE.

T CELL CONTROL OF MCMV AND TISSUE-LOCALIZED IMMUNE SUPPRESSION

IMPACT OF TBI TUMOR SUPPRESSOR PATHWAY ON DAN CHECK POINTS AND THERAPEUTIC RESPONSE

A POPULATION-BASED APPROACH TO SINGLE CTC ANALYSIS IN METASTATIC BREAST CANCER OUTCOME

FUNCTIONAL ANALYSIS OF NSV-BASED HIV VECTORS

IRF7-MEDIATED DEVELOPMENT OF AUTOREACTIVE B CELLS AND SLE - PROJECT SUMMARY MECHANISMS THAT DRIVE THE DEVELOPMENT OF AUTOREACTIVE B CELLS IN AN AUTOIMMUNE DISORDER SYSTEMIC LUPUS ERYTHEMATOSUS (SLE) IS INCOMPLETELY UNDERSTOOD. OVERALL GOAL OF THE RAHMAN LAB IS TO DELINEATE THE MECHANISMS THAT PROMOTE THE DEVELOPMENT OF AUTOREACTIVE B CELLS, AUTOANTIBODY PRODUCTION AND SLE, A DEBILITATING AUTOIMMUNE DISEASE THAT AFFECTS MILLIONS OF AMERICANS AND PEOPLE WORLDWIDE. DEVELOPING A COMPLETE UNDERSTANDING OF SUCH MECHANISMS WILL HELP DEVELOP TARGETED THERAPIES THAT WOULD BE PREFERABLE TO CURRENTLY AVAILABLE SLE TREATMENT OPTIONS THAT GLOBALLY SUPPRESS THE IMMUNE SYSTEM, AND RESULT IN RECURRENT INFECTIONS AND INCREASE DISEASE-ASSOCIATED MORBIDITY AND MORTALITY. GWAS STUDIES HAVE IDENTIFIED RISK VARIANTS IN THE INTERFERON REGULATORY FACTOR 7 (IRF7) GENE, WHICH INCREASE THE RISK OF DEVELOPING SLE. IRF7 IS THE KEY TRANSCRIPTION FACTOR THAT REGULATES TYPE I INTERFERON (T1-IFN) RESPONSE DOWNSTREAM OF NUCLEIC ACID SENSING TOLL- LIKE RECEPTOR (TLR) THAT PROMOTE SLE IN MICE AND HUMANS. OUR PRELIMINARY DATA HIGHLIGHT THE SIGNIFICANCE OF IRF7 IN AUTOREACTIVE B CELL DEVELOPMENT AND SLE MANIFESTATIONS IN A WELL-CHARACTERIZED FCGRIIB-/- SLE MODEL. PUBLISHED AND PRELIMINARY DATA FROM THE FCGRIIB-/- MODEL INDICATE A CRITICAL ROLE FOR IRF7 AND A MODEST ROLE FOR T1-IFN SIGNALING IN AUTOIMMUNE RESPONSES AND SLE DEVELOPMENT. THESE DATA SUGGEST T1-IFN-DEPENDENT AND -INDEPENDENT ROLES OF IRF7 IN AUTOREACTIVE B CELL AND SLE DEVELOPMENT, AND POINT TO IRF7 AS A POTENTIALLY BETTER THERAPEUTIC TARGET FOR SLE THAN T1-IFN BLOCKING THERAPIES ALONE, ALTHOUGH RECENT CLINICAL TRIALS FOR T1-IFN RECEPTOR BLOCKING THERAPY SHOWED PROMISING OUTCOME. HOWEVER, WE KNOW LITTLE ABOUT THE CELL TYPE-SPECIFIC ROLE OF IRF7 IN SLE DEVELOPMENT, ALTHOUGH ITS ROLE IN T1-IFN PRODUCTION BY PDCS IS WELL ESTABLISHED. IMPORTANTLY, B CELL-INTRINSIC AND -EXTRINSIC ROLES OF IRF7 IN AUTOREACTIVE B CELL DEVELOPMENT VIA EXTRAFOLLICULAR ANTIBODY-FORMING CELL (AFC) AND FOLLICULAR GERMINAL CENTER (GC) PATHWAYS IN SLE ARE NOT KNOWN. ALSO, T1-IFN-INDEPENDENT IRF7 ROLE IN SLE REMAINS UNKNOWN. WE HYPOTHESIZE THAT IRF7 REGULATES B CELL-INTRINSIC AND -EXTRINSIC, AND T1-IFN- DEPENDENT AND -INDEPENDENT MECHANISMS IN AUTOREACTIVE B CELL DEVELOPMENT IN THE AFC AND GC PATHWAYS, LEADING TO PATHOGENIC AUTOANTIBODY PRODUCTION AND SLE. THIS HYPOTHESIS IS SUPPORTED BY OUR PRELIMINARY DATA SHOWING 1) A SIGNIFICANT UPREGULATION OF IRF7 IN AFCS (PLASMABLASTS/PLASMA CELLS), GC B CELLS AND MYELOID CELLS 2) INCREASED IRF7 EXPRESSION IN ACTIVATED B CELLS FROM SLE PATIENTS 3) A DRASTIC REDUCTION IN AUTOIMMUNE AFC AND GC RESPONSES, AUTOANTIBODIES AND IMMUNE COMPLEX DEPOSITION IN SLE-PRONE FCGRIIB-/- MICE DEFICIENT IN IRF7 4) IRF7 INVOLVEMENT IN ALTERED B CELL METABOLIC ACTIVITY IN SLE-PRONE B CELLS, AND PUBLISHED DATA REVEALING 5) A ROLE FOR IRF7 IN MOUSE AND HUMAN SLE. THE CURRENT PROPOSAL WILL HELP DETERMINE B CELL-INTRINSIC AND -EXTRINSIC ROLES AND MECHANISMS BY WHICH IRF7 PROMOTE AUTOREACTIVE B CELL AND SLE DEVELOPMENT, AND WHETHER IRF7 COULD BE A BETTER THERAPEUTIC OPTION FOR SLE THAN T1-IFN OR T1-IFN-RECEPTOR BLOCKING ALONE.

DEVELOPMENT OF AN MRI TEMPLATE & NEUROIMAGING BIOMARKERS OF THE PEDIATRIC SPINAL CORD

REGULATION OF THE CELL DEATH PROGRAM BY DFNA5

A NOVEL STRATEGY FOR TRANSCRIPTIONAL REPROGRAMMING OF LYMPHOID LEUKEMIA CELLS - ABSTRACT LYMPHOID B-ALL AND TCLL LEUKEMIA CONSISTS OF LEUKEMIC BLAST CELLS (LBCS) ARRESTED AT EARLY STAGES OF DIFFERENTIATION WHICH EXHIBIT HIGH PROLIFERATIVE POTENTIAL AND CAPABILITY FOR SELF-RENEWAL. THE IDEA TO INDUCE REPROGRAMMING OF LEUKEMIC AND OTHER CANCER CELLS, LEADING TO CELL MATURATION AND SENESCENCE, GAINED HIGH POPULARITY, BUT ITS CLINICAL APPLICATIONS ARE RARELY SUCCESSFUL AND ARE MAINLY LIMITED TO THE THERAPY OF THE APL LEUKEMIA WITH ATRA AND ARSENIC TRIOXIDE. THIS STRATEGY MAY HAVE BEEN UNSUCCESSFUL DUE TO A GAP IN THE KNOWLEDGE OF THE MECHANISMS THROUGH WHICH TRANSCRIPTIONAL REPROGRAMMING OCCURS. OUR PUBLISHED DATA SUGGEST THAT NORMAL HEMATOPOIETIC PROGENITOR CELLS (HPCS) UNDERGO TRANSIENT DE-CONDENSATION OF CHROMATIN TO ALLOW LINEAGE-SPECIFIC TRANSCRIPTION FACTORS (TFS) TO BIND TO THEIR GENE TARGETS AND TO ACTIVATE NEW TRANSCRIPTIONAL PROGRAMS, LEADING TO CELL DIFFERENTIATION. THIS TRANSIENT DE-CONDENSATION OCCURS THROUGH VERY LOW ACCUMULATION OF H3K27ME3, ON DNA JUST AFTER DNA REPLICATION. H3K27ME3 IS A MARK OF THE MOST CONDENSED ARRAYS OF NUCLEOSOMES IN THE GENOME AND IS FOUND AT REGULATORY REGIONS OF ALL REPRESSED GENES. OUR RESULTS SUGGEST THAT TESTED CULTURED AND PRIMARY LYMPHOID B-ALL AND TCLL CELLS HAVE LOST THIS INHERENT ABILITY TO ‘OPEN’ NASCENT CHROMATIN, THUS CREATING A BARRIER FOR THEIR TRANSCRIPTIONAL REPROGRAMMING. IN THIS PROPOSAL, WE WILL TEST A NEW REPROGRAMMING STRATEGY, WHICH OVERCOMES THESE BARRIERS OF REPROGRAMMING-BASED THERAPIES AND MAY LEAD TO ELIMINATION OF LEUKEMIC CELLS. THE KEY FEATURE OF THIS NEW STRATEGY IS THE FIRST STEP, WHICH INCLUDES PHARMACOLOGICAL INHIBITION OF THE H3K27ME3 HISTONE METHYLTRANSFERASES (HMTS) EZH1/EZH2, THUS CREATING DE- CONDENSED STRUCTURE OF NASCENT CHROMATIN AT REGULATORY REGIONS OF ALL GENES. AT THE SECOND STEP, WE WILL USE SMALL MOLECULES TO ACTIVATE ENDOGENOUS INDUCIBLE TFS, WHICH CAN THEN READILY BIND TO THEIR TARGET GENES DUE TO THE DE-CONDENSED STRUCTURE OF NASCENT CHROMATIN. TUMOR CELLS, INCLUDING LEUKEMIC CELLS, ARE COMMONLY KNOWN TO ACCUMULATE MUTATIONS IN INDUCIBLE TFS AND RECEPTORS; THUS, SCREENS OF SMALL MOLECULE INDUCERS FOR A VARIETY OF TFS/RECEPTORS WILL BE PERFORMED TO DETERMINE THE BEST POSSIBLE INDUCER FOR DISTINCT SUBTYPES OF B-ALL. PRELIMINARY RESULTS SUGGEST THAT INDUCTION BY SMALL MOLECULE INDUCERS LEADS TO TRANSCRIPTIONAL REPROGRAMMING OF CELL LINES AND PRIMARY B-ALL AND TCLL CELLS, CHANGES IN THEIR IMMUNOPHENOTYPE AND APOPTOSIS. MOREOVER, THIS STRATEGY STRONGLY SUPPRESSES LYMPHOID LEUKEMIA BURDEN IN MICE. THE GOAL OF THIS PROJECT IS TO DEVELOP A WIDELY APPLICABLE TREATMENT STRATEGY FOR TRANSCRIPTIONAL REPROGRAMMING AND LOSS OF CELL VIABILITY FOR MANY TYPES OF LYMPHOID LEUKEMIC CELLS. TO THIS END, WE PROPOSE TO: 1. EXTEND AND GENERALIZE THE LYMPHOID LEUKEMIC CELLS REPROGRAMMING APPROACH; 2. EXAMINE THE MECHANISMS AND BIOLOGICAL OUTCOMES OF REPROGRAMMING OF LYMPHOID LEUKEMIC CELLS; 3. EXAMINE THE EFFECTS OF OUR TREATMENT STRATEGY IN VIVO.

CASPASE-1 ACTIVATION BY THE INFLAMMASOMES - INFLAMMASOMES ARE MULTI-PROTEIN COMPLEXES THAT ASSEMBLE TO ACTIVATE CASPASE-1 IN RESPONSE TO TISSUE DAMAGE AND INFECTION BY MICROBIAL OR VIRAL PATHOGENS. ONCE ACTIVATED, CASPASE-1 PROCESSES THE INACTIVE PROFORMS OF INTERLEUKIN-1SS (IL-1SS) AND INTERLEUKIN-18 (IL-18) TO PRODUCE THE ACTIVE PRO-INFLAMMATORY CYTOKINES IL-1SS AND IL- 18, RESPECTIVELY. IN ADDITION, CASPASE-1 PROCESSES THE GASDERMIN PROTEIN GSDMD TO INDUCE PYROPTOSIS OR INFLAMMATORY CELL DEATH. THE NLRP3 INFLAMMASOME IS UNIQUE AMONG THE DIFFERENT INFLAMMASOMES IN THAT IT IS ACTIVATED BY DIVERSE PATHOGEN-ASSOCIATED AND DANGER-ASSOCIATED MOLECULAR PATTERNS (PAMPS AND DAMPS) DERIVED FROM INFECTION WITH MICROBIAL PATHOGENS, OR TISSUE DAMAGE. AS A RESULT, UNCONTROLLED NLRP3 ACTIVATION CAN LEAD TO A NUMBER OF HUMAN INFLAMMATORY DISEASES, INCLUDING GOUT, ARTHRITIS, ATHEROSCLEROSIS, AND TYPE 2 DIABETES. THE MECHANISM OF NLRP3 ACTIVATION BY THESE SEEMINGLY UNRELATED STIMULI IS POORLY UNDERSTOOD BUT IS CURRENTLY BELIEVED TO REQUIRE TWO DISTINCT SIGNALS; A PRIMING SIGNAL OR “SIGNAL 1” PRODUCED BY TOLL-LIKE RECEPTORS (TLRS) AND AN ACTIVATION SIGNAL OR “SIGNAL 2” THAT INDUCES FRAGMENTATION OF TRANS GOLGI NETWORK (TGN) AND BINDING TO NEK7. STUDIES IN THE APPLICANT'S LABORATORY DEMONSTRATED THAT SIGNAL 1 INDUCES POST-TRANSLATIONAL MODIFICATION (PTM) OF NLRP3 AT CRITICAL SITES VIA THE MYD88 AND TRIF SIGNALING PATHWAYS AND PARTIAL OLIGOMERIZATION OF NLRP3. IN THIS APPLICATION, STUDIES ARE PROPOSED TO ELUCIDATE HOW TLR-INDUCED PTM CONTRIBUTES TO POSTTRANSLATIONAL PRIMING OF THE NLRP3 INFLAMMASOME BY EMPLOYING MASS SPECTROMETRY TO IDENTIFY AND CHARACTERIZE ALL CRITICAL CHANGES IN THE PHOSPHORYLATION AND OTHER PTM PROFILE OF NLRP3 INDUCED BY SIGNAL 1, AND INVESTIGATING HOW THESE CHANGES CONTRIBUTE TO ACTIVATION OF NLRP3. ADDITIONAL AIMS WILL INVESTIGATE THE EFFECT OF SIGNAL 1 AND SIGNAL 1-INDUCED PTM ON NLRP3 ASSOCIATION WITH DISPERSED TGN AND NEK7, AND IDENTIFY THE TGN-ASSOCIATED KINASES REQUIRED FOR FINAL ASSEMBLY AND ACTIVATION OF THE INFLAMMASOME. FINALLY, PRELIMINARY EVIDENCE SUGGEST THAT KINASES INVOLVED IN THE REGULATION OF INTRACELLULAR ION HOMEOSTASIS EXERT NEGATIVE CONTROL ON ACTIVATION OF NLRP3 BY SIGNAL 2. THUS, ADDITIONAL EXPERIMENTS WILL INVESTIGATE HOW SIGNALING FROM THESE KINASES IN MACROPHAGES IMPACTS NEK7 PHOSPHORYLATION AND INTERACTION WITH NLRP3, AND HOW GENETIC DEFICIENCY IN THESE KINASES IMPACTS NLRP3-MEDIATED PRO-INFLAMMATORY RESPONSES TO PAMPS AND DAMPS IN VIVO. RESULTS FROM THIS RESEARCH WILL PROVIDE FUNDAMENTAL NEW INSIGHTS INTO THE PATHWAYS THAT REGULATE THE ASSEMBLY AND ACTIVATION OF THE NLRP3 INFLAMMASOME, AND THE CELLULAR MECHANISMS THAT CONTROL ITS ACTIVATION. SUCCESSFUL COMPLETION OF THIS STUDY SHOULD HAVE A HIGH IMPACT ON THE FIELD BY PROVIDING A UNIFYING PARADIGM FOR HOW NLRP3 CAN BE REGULATED BY AN EXCEPTIONALLY DIVERSE GROUP OF ACTIVATING STIMULI. UNDERSTANDING THESE MECHANISMS IS OF GREAT SCIENTIFIC AND HEALTH SIGNIFICANCE AS THIS SHOULD BETTER OUR UNDERSTANDING OF THE MOLECULAR BASIS OF NLRP3-RELATED DISEASES AND SHOULD IN THE LONG TERM HELP IN THE DEVELOPMENT OF THERAPEUTICS TO ALLEVIATE THESE INFLAMMATORY DISEASES.

ER-MITOCHONDRIAL COMMUNICATION IN CALCIUM SIGNALING, ENERGY METABOLISM AND LIVER DISEASE - ABSTRACT THE EFFECTS OF VARIOUS HORMONES ON OXIDATIVE METABOLISM AND OTHER MITOCHONDRIAL FUNCTIONS, IN THE LIVER AND OTHER TISSUES, ARE MEDIATED BY CYTOPLASMIC [CA2+] OSCILLATIONS PROPAGATED TO THE MITOCHONDRIA. CA2+ IS RELEASED TO THE CYTOPLASM FROM THE ENDOPLASMIC RETICULUM (ER) THROUGH THE IP3 RECEPTORS (IP3RS), WHICH, BASED ON FINDINGS FROM US AND OTHERS, EXPOSE MITOCHONDRIA AT ER-MITOCHONDRIA CONTACT SITES (ERMCS) TO HIGH [CA2+] NANODOMAINS TO ATTAIN ACTIVATION OF THE LOW-AFFINITY MITOCHONDRIAL CA2+ UPTAKE SITES. ERMCS WERE ALSO RECOGNIZED IN OTHER PROCESSES INCLUDING LIPID METABOLISM, ORGANELLE DYNAMICS AND AUTOPHAGY. OUR WORK HAS REVEALED THE PHYSICAL SUPPORT OF ERMCS BY TETHERING PROTEINS. WE HAVE CREATED SYNTHETIC MEMBRANE LINKERS TO MEASURE AND PERTURB ERMCS AND LOCAL INTER-ORGANELLE COMMUNICATION IN LIVE CELLS, AND PROVIDED CLUES TO LOCAL CALCIUM AND REACTIVE OXYGEN SPECIES (ROS) SIGNALING. WE HAVE ALSO PROVIDED THESE REAGENTS TO SEVERAL HUNDRED LABORATORIES WORLDWIDE, WHICH TOGETHER HAVE SHOWED THE ROLE OF INTERORGANELLAR CONTACTS IN A RANGE OF PARADIGMS INCLUDING METABOLISM, VESICLE DYNAMICS, NEURONAL AND IMMUNE FUNCTIONS AND LINKED STRUCTURAL OR FUNCTIONAL IMPAIRMENTS OF THE ER-MITOCHONDRIAL COUPLING TO AN ARRAY OF DISORDERS ACROSS ORGANS INCLUDING THE LIVER (E.G. FATTY LIVER). HOWEVER, FUNDAMENTAL QUESTIONS REMAIN UNANSWERED. ERMCS ARE DYNAMICALLY RESTRUCTURED TO MEET THE CONTINUOUSLY CHANGING DEMANDS OF THE CELL, BUT HOW ERMCS ARE FORMED AND DISSOLVED IS YET TO BE DETERMINED. IP3R-MEDIATED FLUCTUATIONS IN [CA2+] MIGHT PROVIDE A MEANS TO CONTROL CONTACT FORMATION, GIVEN THAT ELEVATIONS OF CYTOPLASMIC [CA2+] STOP MITOCHONDRIAL MOVEMENTS CLOSE TO THE ER THROUGH THE CA2+-SENSING MIRO PROTEINS, AND BOTH THE IP3R AND MIRO PROTEINS, HAVE BEEN IMPLICATED AS COMPONENTS OF INTERORGANELLAR COMPLEXES. HOWEVER, THE INTERACTION PARTNERS AND MECHANISMS ARE ELUSIVE. WE HYPOTHESIZE THAT IP3RS AND MIROS MEDIATE ERMC FORMATION IN AN ISOFORM-SPECIFIC AND CA2+-DEPENDENT MANNER TO REGULATE PHYSIOLOGICAL FUNCTIONS OF HEPATOCYTES. AIMS#1&2 WILL TEST THIS HYPOTHESIS USING NOVEL GENETIC AND MICROSCOPIC IMAGING TOOLKITS THAT WILL ENABLE US TO SPECIFICALLY AND SYSTEMATICALLY MEASURE AND PERTURB ERMC FORMING ELEMENTS. THE EFFECT OF GENETIC PERTURBATIONS IN THE LIVER WILL BE TESTED BY NOVEL, IN VIVO IMAGING APPROACHES. THE INTERACTOMES OF IP3R AND MIRO WILL BE EVALUATED PRIMARILY BY UNBIASED PROTEOMICS, BUT BCL-XL WILL ALSO BE SPECIFICALLY TESTED AS A TETHER FORMING PARTNER FOR THE IP3R. FINALLY, A MAJOR LIMITATION IN THE FIELD OF INTER-ORGANELLAR COMMUNICATION RESEARCH IS THAT QUANTITATIVE EVALUATION OF THE GEOMETRY OF NANOMETER SCALE MEMBRANE CONTACTS REMAINS DIFFICULT. IN AIM#3 WE WILL DEVELOP AND CHARACTERIZE METHODS OF MEASURING AND DESCRIBING ORGANELLE INTERFACE GEOMETRY IN 2-AND 3D ELECTRON MICROSCOPY DATA AND UNCOVER THE STRUCTURAL FEATURES MOST RELEVANT TO CALCIUM TRANSFER. THE PROPOSED WORK WILL EXPLORE THE MOLECULAR MECHANISMS OF ERMC DYNAMICS AND THEIR PHYSIOLOGICAL RELEVANCE AND WILL CONTINUE OUR EFFORTS IN CREATING MOLECULAR TOOLS AND METHODS THAT ALLOW MANY INVESTIGATORS TO EXPLORE THE LOCAL COMMUNICATIONS OF ER AND MITOCHONDRIA OR OTHER ORGANELLES.

NEREGULIN 1-ERBB SIGNALING IN SCHIZOPHRENIA

MECHANISMS OF PULSATILE CALCIUM SIGNALING

MEMBRANE TARGETING OF G PROTEINS

VPAC1 RECPETOR-TARGETED PET IMAGING OF PROSTATE CANCER

METTL3 IN CHROMIUM-INDUCED ANGIOGENESIS AND CARCINOGENESIS - EXPOSURE OF HEXAVALENT CHROMIUM [CR(VI)] IS KNOWN TO INDUCE LUNG CANCER. ALTHOUGH THERE IS EMERGING INTEREST IN MECHANISMS OF CR(VI)-INDUCED CARCINOGENESIS, ROLE OF CR(VI) IN INDUCING RNA MODIFICATION IN CARCINOGENESIS IS UNKNOWN. OUR PRELIMINARY STUDIES FOUND THAT HIGHER LEVELS OF METHYLTRANSFERASE LIKE 3 (METTL3) WERE FOUND IN LUNG TISSUES FROM CR(VI)-EXPOSED MICE, SAMPLES FROM CR(VI)-EXPOSED NONSMOKING WORKERS, AND CR(VI)- TRANSFORMED (CR-T) CELLS. TO STUDY UNDERLYING MECHANISM, WE FOUND THAT METTL3 WAS INDUCED BY UPREGULATION OF NRF2 AND SOX4, TWO IMPORTANT TRANSCRIPTION FACTORS. WE FOUND THAT METTL3 WAS IMPORTANT IN REGULATING CR-T CELL PROLIFERATION, TUBE FORMATION AND TUMOR ANGIOGENESIS. METTL3 INDUCED HYPOXIA-INDUCIBLE FACTOR 1 (HIF- 1) EXPRESSION THROUGH SUPPRESSING PHD2, SUGGESTING METTL3/PHD2/HIF-1 SIGNALING WOULD BE IMPORTANT IN CR(VI) CARCINOGENESIS. C-X-C MOTIF CHEMOKINE 5 (CXCL5) AND IL-8 WERE DOWNSTREAM EFFECTORS OF METTL3. OUR WHOLE GENOME EXPRESSION ARRAY ANALYSIS OF BLOOD MONONUCLEAR CELLS (PBMCS) FROM CR(VI)-EXPOSED NONSMOKING WORKERS AND CONTROL SUBJECTS SHOWED THAT METTL3, CXCL5 AND IL-8 WERE AMONG THE MOST UPREGULATED MOLECULES IN CR(VI) EXPOSURE GROUP, WHICH WAS VALIDATED USING RT-QPCR AND ELISA ASSAYS. WE HYPOTHESIZE THAT LONG-TERM CR(VI) EXPOSURE INDUCES METTLE3 OVEREXPRESSION TO REGULATE CELL TRANSFORMATION, TUMOR GROWTH AND ANGIOGENESIS THROUGH METTL3/PHD2/HIF-1 AXIS IN LUNG EPITHELIAL CELLS, AND SOX4 AND NRF-2 ARE TWO KEY UPSTREAM INDUCERS. IN ORDER TO TEST THIS HYPOTHESIS, WE WILL PERFORM THREE SPECIFIC AIMS: AIM 1) TO DETERMINE ROLE AND MECHANISM OF METTL3 UPREGULATION IN CR(VI)-INDUCED CELL TRANSFORMATION, TUMOR GROWTH AND ANGIOGENESIS, AND TO IDENTIFY UPSTREAM REGULATOR(S) OF METTL3 ELEVATION IN CR-T CELLS. AIM 2) TO INVESTIGATE KEY DOWNSTREAM TARGETS AND MOLECULES OF METTL3 IN CR(VI)-INDUCED CELL TRANSFORMATION AND TUMOR GROWTH. AIM 3) TO DETERMINE WHETHER METTL3 UPREGULATION IN CR-T CELLS INDUCES TUMOR ANGIOGENESIS THROUGH CXCR1/2 RECEPTORS AND PARACRINE EFFECT USING HUMANIZED CHIMERIC TUMOR MODEL; TO DETERMINE EXPRESSION LEVELS OF NRF2, SOX4, METTL3, PHD2, HIF-1, CXCL5, AND/OR IL-8 IN PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) AND LUNG TISSUES FROM THE CR(VI)-EXPOSED MICE AND IN PBMCS FROM WORKERS WITH OCCUPATIONAL EXPOSURE TO CR(VI). WE WILL USE A COMBINATION OF MOLECULAR APPROACHES, ANIMAL MODELS, AND BLOOD AND TISSUE SAMPLES FROM HUMAN SUBJECTS AND MICE TO DEFINE THE ROLE AND MECHANISMS OF NEW METTL3/PDH2/HIF-1 AXIS INDUCED BY NRF2 AND SOX4 IN MEDIATING CELL TRANSFORMATION, TUMOR GROWTH AND ANGIOGENESIS, AND DETERMINE THE POSSIBLE CORRELATIONS WITH CR(VI) INTERNAL EXPOSURE DOSES IN WORKERS AND IN MICE VIA LEVELS OF THESE MOLECULES. WE WILL ALSO INVESTIGATE THE EFFECTS OF DOWNSTREAM EFFECTORS OF METTL3/PDH2/HIF-1 AXIS, AND THEIR RECEPTORS IN CR-T CELL-INDUCING ANGIOGENESIS. THESE STUDIES WILL HELP US UNDERSTAND UNDERLYING MECHANISMS OF CR(VI) IN INDUCING TUMOR GROWTH AND ANGIOGENESIS, AND IDENTIFY NEW BIOMARKERS FOR EARLY DETECTION OF CR(VI) EXPOSURE AND CANCER PREVENTION.

ALTERNATIVE DIVERSION TO BEHAVIORAL HEALTH CRISES IN NORTHEASTERN PENNSYLVANIA. - MONROE COUNTY IS A HIGH-NEED SERVICE AREA IN RURAL PENNSYLVANIA. POLICE ARE OFTEN THE PRIMARY RESPONDERS TO BEHAVIORAL HEALTH EMERGENCIES. THE PUBLIC SAFETY ANSWERING POINT (PSAP) REPORTED THAT TO DATE IN 2022 THERE WERE 49 SUICIDE ATTEMPTS AND 650 CALLS THAT CAME THROUGH FOR OTHER BEHAVIORAL HEALTH CONCERNS. TO BETTER MEET THE NEEDS OF INDIVIDUALS IN MONROE COUNTY AND DIVERT UNNEEDED RESPONSE FROM LAW ENFORCEMENT, THOMAS JEFFERSON UNIVERSITY AND MONROE COUNTY’S MOBILE CRISIS PROVIDER, RESOURCES FOR HUMAN DEVELOPMENT (RHD), ARE PROPOSING TO PARTNER WITH THE LOCAL PSAP AND POLICE DEPARTMENTS TO ESTABLISH AN ALTERNATIVE DISPATCH PROGRAM AND ENHANCE RHD’S INFRASTRUCTURE TO RESPOND TO MOBILE CRISIS NEEDS USING BEST PRACTICES OUTLINED IN NATIONAL GUIDELINES FOR BEHAVIORAL HEALTH CRISIS CARE: BEST PRACTICE. THE TOTAL POPULATION OF MONROE COUNTY IN 2020 WAS 169,273 INDIVIDUALS, WITH A RACIAL COMPOSITION OF 76% WHITE, 17% BLACK, 2% ASIAN, 3% TWO OR MORE RACES, WITH 18% IDENTIFYING AS HISPANIC AND 2% AS TWO OR MORE ETHNICITIES. INDIVIDUALS PRIMARILY SPEAK ENGLISH (85%), FOLLOWED BY SPANISH (~10%), INDO-EUROPEAN (~5%), AND ASIAN/ISLANDER (1.5%). INDIVIDUALS IDENTIFY EQUALLY AS MALE AND FEMALE. THE MEDIAN AGE IS 43.3, WITH 63% OF THE POPULATION BETWEEN 18-64 YEARS OF AGE. THE MEDIAN HOUSEHOLD INCOME IS $68,734, WITH 10.7% LIVING BELOW POVERTY LINE (U.S. CENSUS BUREAU, 2020). WHILE RHD’S GOALS FOR CRISIS EXPANSION EXTEND BEYOND SUPPORTS FOR MOBILE CRISIS RESPONSE, THE FOLLOWING GOALS AND OBJECTIVES WILL DRIVE THE FOCUS OF THIS GRANT PROGRAM: 1. ENHANCE MOBILE CRISIS RESPONSE TEAM TO MEET NATIONAL GUIDELINES FOR BEHAVIORAL HEALTH CRISIS CARE: BEST PRACTICE FOR MONROE COUNTY, 2. MINIMIZE LAW ENFORCEMENT INVOLVEMENT AND INVOLUNTARY TRANSPORT BY DEVELOPING AN ALTERNATIVE DISPATCH PROGRAM WHERE MOBILE CRISIS WILL RESPOND TO BEHAVIORAL HEALTH CRISES WITHOUT LAW ENFORCEMENT, UNLESS SPECIAL CIRCUMSTANCES WARRANT INCLUSION, 3. MAINTAIN COUNTYWIDE METRICS FOR MOBILE CRISIS, PSAP, 988, AND LAW ENFORCEMENT BEHAVIORAL HEALTH CALLS, 4. HELP IDENTIFY SERVICE NEEDS, SERVICE GAPS, AND OPPORTUNITIES TO IMPROVE EQUITY IN MONROE COUNTY BY APPLYING A COMMUNITY CRISIS MAPPING MODEL (E.G., CRISIS INTERCEPT MAPPING), AND 5. INCREASE QUALIFIED WORKFORCE CAPACITY BY DEVELOPING AND IMPLEMENTING EVIDENCE- BASED TRAINING OPPORTUNITIES FOR MOBILE CRISIS RESPONSE STAFF, PSAP, AND FIRST RESPONDERS. THE PROPOSED PROJECT TEAM, SPECIFICALLY THOMAS JEFFERSON UNIVERSITY AND RHD, HAVE COLLABORATED ON NUMEROUS FEDERAL GRANTS TO IMPROVE CRISIS INTERVENTION SERVICES IN MONROE COUNTY. THE PENNSYLVANIA OFFICE OF MENTAL HEALTH AND SUBSTANCE ABUSE SERVICES, LOCAL PSAP, AND 3 BOROUGH POLICE DEPARTMENTS HAVE AGREED TO SUPPORT THE PROPOSED PROJECT ACTIVITIES AND CONTRIBUTE TO THE COLLECTION OF ALL RELEVANT KEY PERFORMANCE INDICATORS, INCLUDING BOTH INDIVIDUAL LEVEL DATA AS WELL AS AGGREGATE FIGURES (I.E., AVERAGE NUMBER SERVED PER 8-HOUR SHIFT). WHILE IT IS DIFFICULT TO DETERMINE EXACTLY HOW MANY RESIDENTS OF MONROE COUNTY WILL BE IMPACTED BY THESE INITIATIVES, WE BASED OUR PROJECTIONS ON RHD’S ANNUAL NUMBER OF UNIQUE MOBILE CRISIS INTERVENTIONS AND ACCOUNTED FOR GROWTH IN YEARS 2-4 DUE TO GRANT PROJECT ACTIVITIES. WE ANTICIPATE THAT WE WILL SERVE 2,785 UNDUPLICATED PEOPLE IN THE 4 YEARS OF THE PROJECT.

ELUCIDATING THE MECHANISM IN THE REGULATION OF RNA-BINDING PROTEIN PHASE SEPARATION

PREVENTING COGNITIVE DECLINE IN AFRICAN AMERICANS WITH MILD COGNITIVE IMPAIRMENT.

PREVENTING SPINAL INFECTION: ULTRASOUND TRIGGERED PROPHYLAXIS

BIO-PSYCHO-SOCIAL DRIVERS OF DISPARITIES IN LIVER DISEASE PROGRESSION AMONG KOREAN AMERICANS WITH HEPATITIS B INFECTION - HEPATITIS B VIRUS (HBV) INFECTION IS GLOBALLY THE MOST COMMON AND SERIOUS VIRAL INFECTION OF THE LIVER, CAUSING PREMATURE DEATHS FROM LIVER DISEASES OR LIVER CANCER. CHRONIC HEPATITIS B (CHB) INFECTION AND LIVER CANCER CAUSED BY HBV AMONG ASIAN AMERICANS IS ONE OF THE MOST SERIOUS—BUT FREQUENTLY NEGLECTED—HEALTH DISPARITY ISSUES IN THE U.S. HEPATOCARCINOGENESIS FROM HBV INFECTION TO HEPATOCELLULAR CARCINOMA (HCC) DEVELOPMENT HAS BEEN INCREASINGLY RECOGNIZED AS A MULTIFACTORIAL, MULTI-LEVEL AND MULTI-STEP PROCESS. LESS IS KNOWN ABOUT THE MULTIPLE FACTORS AND THE UNDERLYING MECHANISMS RESPONSIBLE FOR CHRONIC LIVER DISEASE PROGRESSION IN HIGH RISK ASIAN AMERICANS. EVOLVING MODELS OF LIVER DISEASE POSIT A STRONG ROLE FOR VIRUS, CHRONIC INFLAMMATION AND OXIDATIVE DNA DAMAGE. CO-FACTORS THAT MAY INFLUENCE THESE DISEASE PROCESSES INCLUDE EXOGENOUS AND ENDOGENOUS FACTORS. THE APPLICATION OF A MULTI-LEVEL FRAMEWORK, HOWEVER, IS RELATIVELY NEW IN THE STUDY OF CHRONIC LIVER DISEASE PROGRESSION (E.G., CHB PHENOTYPE, LIVER DISEASE SEVERITY) AMONG CHB PATIENTS. MOREOVER, THERE IS GREAT VARIANCE IN THE HEPATIC DISEASE PROGRESS AMONG CHB PATIENTS, AND THE PSYCHOSOCIAL FACTORS CONTRIBUTING TO OR RESPONSIBLE FOR BIOLOGICAL SYSTEM IMPAIRMENT HAVE NOT BEEN IDENTIFIED. THE GOAL OF THIS STUDY IS TO LONGITUDINALLY EXAMINE THE MULTIPLE PATHWAYS TO DISPARITIES IN CHRONIC LIVER DISEASE PROGRESSION USING TWO UNIQUE CLINIC-BASED PATIENT COHORTS OF CHB KOREAN AMERICANS RECEIVING CARE FROM KOREAN-AMERICAN HEPATOLOGISTS. WE WILL RECRUIT AND ENROLL 600 KOREAN AMERICAN CHB PATIENTS. MEDICAL CHART REVIEW, STRUCTURED PATIENT INTERVIEWS, BIOMARKERS, QUALITATIVE, AND GEO-SPATIALLY REFERENCED DATA WILL BE INTEGRATED TO EXPLORE MULTI-LEVEL MODELS OF LIVER DISEASE PROGRESSION OVER A 10- YEAR CLINICAL WINDOW, WITHIN A GROUP AT HIGH RISK FOR ADVERSE OUTCOMES. THE SPECIFIC AIMS ARE: (1) TO ESTIMATE THE PREVALENCE OF CHB PHENOTYPES AND LIVER DISEASE SEVERITY, AND ASSOCIATED COVARIATES AT THE INITIAL VISIT; (2) IDENTIFY HOW MULTIPLE FACTORS (E.G., SOCIAL ENVIRONMENTAL, PSYCHOSOCIAL, BEHAVIORAL, CLINICAL AND BIOLOGICAL ATTRIBUTES) ARE ASSOCIATED WITH VARIATION IN LIVER DISEASE PROGRESSION; (3) TO EXAMINE THE MODERATING AND MEDIATING EFFECTS OF THESE FACTORS ON THE RELATIONSHIP BETWEEN CHB PHENOTYPES AND ADVERSE LIVER DISEASE OUTCOMES (E.G., HCC); AND (4) TO UNDERSTAND CARE-SEEKING BEHAVIORS AND DYNAMICS OF CARE WITHIN AN ETHNICALLY CONCORDANT LIVER DISEASE CARE MODEL USING AN EXPLANATORY MIXED METHODS APPROACH. FINALLY, COMPARISON TO CLINICAL DATA FROM LARGE CHB COHORTS IN US AND KOREA WILL STRENGTHEN GENERALIZABILITY. INSIGHTS GAINED FROM THIS PROPOSED STUDY WILL PROVIDE THE GROUNDWORK TO UNDERSTAND THE MULTI-LEVEL AND MULTI-STEP PROCESS OF CHRONIC LIVER DISEASE AND LIVER CANCER AMONG HIGH RISK POPULATION WITH CHB PATIENTS, AND DEVELOP POTENTIAL INTERVENTIONS AT MULTIPLE LEVELS (POLICY, SOCIAL-BEHAVIORAL AND CLINICAL) TO REDUCE DISPARITIES IN LIVER DISEASE PROGRESSION.

THE PATHOGENIC RELATIONSHIP BETWEEN NEURONAL ACTIVITY AND C9ORF72-LINKED NEURODEGENERATION

ROLE OF IMMUNE RECEPTOR CLUSTERING IN CONTROLLING EFFICACY OF ANTIBODY-DEPENDENT FC?RIIIA-MEDIATED CYTOTOXICITY BY NK CELLS

MULTISUBUNIT VIRAL ATPASES THAT COUPLE ATP-HYDROLYSIS TO GENOME TRANSLOCATION

REGULATION OF G PROTEIN-COUPLED RECEPTOR SIGNALING AND TRAFFICKING

EARLY LIFE ENVIRONMENT MODIFIES BEHAVIORAL, EPIGENETIC, AND TRANSCRIPTIONAL OUTCOMES FROM DEVELOPMENTAL LEAD EXPOSURE

DYNAMIN-RELATED PROTEIN DRP1 REGULATES CARDIAC EXCITATION-CONTRACTION-BIOENERGETICS COUPLING

TYROSINE PHOSPHORYLATION IN LENS CELL DIFFERENTIATION

CHARACTERIZATION OF BETA-ARRESTIN-BIASED BETA 2-ADRENERGIC RECEPTOR SIGNALING IN CARDIOVASCULAR FUNCTION

TUMOR MICROENVIRONMENT METABOLISM IN INVASIVE DUCTAL CARCINOMA OF THE BREAST

SYNOVIAL FLUID AND JOINT SEPSIS

MECHANISMS OF HIV-1 MEMBRANE FUSION

MODELING MULTISCALE CONTROL OF LIVER REGENERATION

STRUCTURAL-FUNCTIONAL ZONING OF THE MITOCHONDRION IN CARDIAC CA2+, ROS, AND ENERGETICS REGULATION

STRESS-ADAPTATION IN OBESITY-ASSOCIATED PANCREATIC CANCER

MITOCHONDRIAL CALCIUM AND NEURONAL HEALTH - ABSTRACT PROPAGATION OF CALCIUM (CA2+) SIGNALS TO THE MITOCHONDRIA THROUGH THE CA2+ UNIPORTER HAS LONG BEEN CONSIDERED CENTRAL TO NEURONAL FUNCTION. THE DISCOVERY OF THE MOLECULAR COMPONENTS OF THE UNIPORTER INCLUDING THE CA2+ SENSING REGULATOR MICU1, WAS FOLLOWED BY THE IDENTIFICATION OF MANY MICU1 LOSS-OF-FUNCTION PATIENTS WHO DISPLAY PROGRESSIVE NEUROLOGICAL DISORDER, DOMINATED BY MOTOR AND LEARNING IMPAIRMENTS, WHICH WERE RECAPITULATED IN MOUSE MODELS CREATED BY US AND OTHERS. EMERGING EVIDENCE INDICATES THAT ACUTE MICU1 PROTEOLYTIC DEGRADATION BY YME1L ALSO OCCURS DURING HYPOXIA, SUGGESTING THAT DYSREGULATION OF MITOCHONDRIAL CA2+ HOMEOSTASIS MAY ALSO PLAY A ROLE IN ISCHEMIA/STROKE. HOWEVER, THE NEURONAL PATHOGENESIS ASSOCIATED WITH MICU1 DEFICIENCY AND, MORE BROADLY, THE ROLE OF UNIPORTER REGULATION IN HEALTHY NEURONS, REMAIN POORLY UNDERSTOOD. MICU1 FORMS DIMERS WITH ITS PARALOGS, MICU2 AND MICU3, AND WITH ITSELF. MICU2 HAS ALSO BEEN LINKED TO HUMAN NEUROLOGICAL IMPAIRMENTS BUT IS SCARCE IN ADULT NEURONS, WHEREAS MICU3 IS ABUNDANT IN ADULT BRAIN. OUR CENTRAL HYPOTHESIS IS THAT CONTROL OF MITOCHONDRIAL CA2+ UPTAKE BY MICU1/2/3 IS ESSENTIAL FOR COORDINATING MITOCHONDRIAL FUNCTION WITH SYNAPTIC ACTIVITY. THIS CONTROL ALSO PREVENTS MITOCHONDRIAL CA2+ OVERLOAD AND OXIDANT DYSREGULATION THAT LEAD TO NEURONAL STRESS. FURTHER, WE POSTULATE DYNAMIC TUNING OF NEURONAL MICU-DEPENDENT GATEKEEPING: DURING DEVELOPMENT—BY A SWITCH FROM PRIMARILY MICU1-MICU2 TO MICU1-MICU3 DIMERS DURING EARLY LIFE—AND UNDER HYPOXIC STRESS, BY SPECIFIC PROTEOLYTIC DEGRADATION OF MICU1, WITH POTENTIALLY PATHOGENIC CONSEQUENCES. TO TEST THESE IDEAS, WE WILL USE A NEURON-SPECIFIC MICU1 KNOCKOUT MOUSE WHICH WE FOUND TO DISPLAY NEURODEGENERATION CHARACTERIZED BY MITOCHONDRIAL CA2+ OVERLOAD, ALTERED MITOCHONDRIAL AND NEURONAL ULTRASTRUCTURE, AND MOTOR AND COGNITIVE IMPAIRMENTS. WE HAVE ALSO GENERATED MICE WITH MICU2 AND MICU3 LOSS AND HAVE OBTAINED MICU1 AND MICU2 PATIENT-DERIVED CELLS. WE HAVE SET UP ADVANCED FUNCTIONAL IMAGING AND LARGE VOLUME 3D ULTRASTRUCTURE CAPACITIES. THUS, WE ARE WELL POSITIONED TO STUDY THE NEURONAL PATHOGENESIS ASSOCIATED WITH THE LOSS OF EACH OR MULTIPLE MICU ISOFORMS. IN AIM#1 WE WILL TEST THE HYPOTHESIS THAT NEURONS DERIVED FROM MICU1, MICU2 OR MICU3-DEFICIENT MICE AND MICU1 AND MICU2-DEFICIENT PATIENT FIBROBLAST-DERIVED NEURONS HAVE DISTINCT IMPAIRMENTS IN CA2+ SIGNALING, MITOCHONDRIAL DYNAMICS AND SYNAPTIC VESICLE RELEASE. IN AIM#2 WE WILL TEST IF NEURONAL MICU1/2/3 LOSS PROMOTES MITOCHONDRIAL OXIDANT PRODUCTION AND OXIDANT-MEDIATED CELL INJURY. THE RESULTS WILL HELP TO DECIDE IF OXIDANTS ARE RELEVANT TO MICU-LINKED NEURONAL/BRAIN INJURY. IN AIM#3 WE WILL TEST THE HYPOTHESIS THAT THE RELATIVE ABUNDANCE OF THE DIFFERENT MICU ISOFORMS IS DYNAMIC IN NEURONS DURING DEVELOPMENT, AND THE LOSS OF EACH ISOFORM SPECIFICALLY AFFECTS NEUROANATOMY AND BRAIN FUNCTION PRIMARILY IN THE MOTOR AREAS AND THE HIPPOCAMPUS. IN AIM#4 WE WILL TEST IF ISCHEMIA-REPERFUSION INDUCES YME1L-MEDIATED TURNOVER OF MICU1 TO PROMOTE MITOCHONDRIAL CA2+ OVERLOAD INJURY AND TO SENSITIZE THE UNIPORTER TO INHIBITORS. THESE STUDIES ARE EXPECTED TO DECIDE IF MICU1 LOSS IS A PATHOGENIC COMPONENT AND THERAPY-MODIFYING FACTOR IN BRAIN HYPOXIA.

NON-CANONICAL MAPK SIGNALING IN YEAST

INTEGRATION OF EPIGENETIC AND NON-CODING RNA MECHANISMS IN LEUKEMIA

EXPLOITING EMERGING IDEAS IN G PROTEIN-COUPLED RECEPTOR BIOLOGY AND PHARMACOLOGY TO TREAT ASTHMA - OVERALL PROGRAM ABSTRACT FOR THIS NHLBI PPG WE HAVE ASSEMBLED A HIGHLY COLLABORATIVE TEAM OF EXPERTS IN G PROTEIN-COUPLED RECEPTOR (GPCR) BIOLOGY, AIRWAY SMOOTH MUSCLE (ASM) BIOLOGY, AND ASTHMA BIOLOGY TO PURSUE THE THEME OF “EXPLOITING EMERGING IDEAS IN G PROTEIN-COUPLED RECEPTOR BIOLOGY AND PHARMACOLOGY TO TREAT ASTHMA.” THE PROGRAM EXPECTS TO ACHIEVE THE FOLLOWING GOALS: 1) IDENTIFY NOVEL TARGETS AND TARGETING MOLECULES INVOLVING GPCR REGULATION OF ASM CONTRACTION AND PROLIFERATION; 2) DELINEATE THE MECHANISMS BY WHICH THESE NOVEL TARGETS/TARGETING MOLECULES REGULATE THESE ASM FUNCTIONS; AND 3) ESTABLISH THE THERAPEUTIC UTILITY OF NOVEL TARGETING MOLECULES IN IN VIVO MODELS OF ASTHMA. THESE GOALS WILL BE ACCOMPLISHED ACROSS 4 SCIENTIFIC PROJECTS, TWO SCIENTIFIC CORES, AND AN ADMINISTRATIVE CORE, TO TEST THE CENTRAL HYPOTHESIS THAT NOVEL MODES OF GPCR REGULATION CAN BE TARGETED TO EFFECT SUPERIOR MANAGEMENT OF ASM FUNCTION AND ASTHMA. EACH PROJECT TAKES ADVANTAGE OF A ROBUST EXPERIMENTAL STRATEGY THAT INCORPORATES MULTIPLE CELL, TISSUE, AND IN VIVO MODELS OF ASM FUNCTION, COMPLEMENTED BY CUTTING-EDGE BIOCHEMICAL ANALYSES TO DETAIL MECHANISMS OF ACTION. PROJECT 1 WILL TEST THE ABILITY OF, AND MECHANISMS EFFECTED BY, RECENTLY-DEVELOPED BIASED Β2AR LIGANDS TO MANAGE ASM CONTRACTILITY AND THE ASTHMA PHENOTYPE, WHILE (WITH CORE A) LEVERAGING RECENT INSIGHT IN Β2AR STRUCTURE-ACTIVITY RELATIONSHIP TO DEVELOP NEW LIGANDS SUPERIOR TO IN-HAND LEADS. PROJECT 2 PIVOTS AWAY FROM THE RECEPTOR LOCUS AS A MEANS TO REGULATE GPCR CONTROL OF ASM, TO EXPLORE THE ROLE OF THE CAMP TRANSPORT INHIBITOR ABBC1, POSITING THAT ABBC1- DEPENDENT PATHWAYS INHIBIT Β-AGONIST EFFECTS ON HASM E-C COUPLING, AND ARE UPREGULATED IN ASTHMA PATIENTS. PROJECT 3 SIMILARLY EXPLORES REGULATION OF DOWNSTREAM GQ-COUPLED RECEPTOR SIGNALING BY DIACYLGLYCEROL KINASE (DGK), WHOSE TARGETING EFFECTIVELY INHIBITS PRO-CONTRACTILE AND PRO-MITOGENIC GPCR SIGNALING IN ASM TO CONTROL THE ASTHMA PHENOTYPE. PROJECT 4 BREAKS NEW GROUND IN THE EXPLORATION OF CLASS C GPCRS, CLARIFYING THE ROLES OF GPRC5A AND GPRC5B IN ASM AND ASTHMA BIOLOGY, AND IN COLLABORATION WITH CORE A, DEVELOPING NEW LIGANDS TO SKEW THE SIGNALING COMPETITION BETWEEN THESE 2 RECEPTORS TOWARDS THERAPEUTICALLY BENEFICIAL GPRC5A SIGNALING AND AWAY FROM PATHOGENIC GPRC5B SIGNALING. THE FOUR PROJECTS WILL BE SUPPORTED BY CORE A, WHICH WILL EMPLOY HIGH-THROUGHPUT SCREENING OF DIVERSE LIBRARIES OF ARRAYED AND EXPRESSED SMALL MOLECULES, AND FURTHER EMPLOY STRUCTURAL BIOLOGY, COMPUTATIONAL MODELING AND VIRTUAL SCREENING TO LEVERAGE SCREENING RESULTS, PREDICT AND TEST NOVEL DRUG-LIKE MOLECULES. CORE B WILL PROVIDE ALL DE-IDENTIFIED HUMAN CELL AND TISSUE MODELS TO STUDY NOVEL MECHANISMS REGULATING EC COUPLING IN HASM. CORE C WILL PROVIDE ADMINISTRATIVE SUPPORT FOR THE PROGRAM. EACH PROJECT ALSO COLLABORATES WITH OTHER PROJECTS TO TEST COMBINATION STRATEGIES THAT TARGET RECEPTORS (Β2AR, GPR5A, GPR5B) IN CONJUNCTION WITH DOWNSTREAM REGULATORS (ABCC1, DGK). THE STRENGTHS OF THIS PROGRAM ARE THE COMMON FOCUS ON A SINGLE THEME AND THE PRODUCTIVE WORKING RELATIONSHIP AMONG INVESTIGATORS WITH THE ABILITY TO APPLY CUTTING-EDGE GPCR BIOLOGY TO KEY QUESTIONS IN ASTHMA BIOLOGY AND PHARMACOLOGY.

CLINICALLY TRANSLATABLE ULTRASOUND-SENSITIVE MICROBUBBLE APPROACHES FOR OVERCOMING TUMOR HYPOXIA

DISRUPTION OF OSMOREGULATION PROMOTES DEGENERATIVE DISC DISEASE

IMPROVING MEDICATION ADHERENCE IN OLDER AFRICAN AMERICANS WITH DIABETES

MULTI-MODALITY DETECTION OF RCC RECURRENCE POST ABLATION - PROJECT SUMMARY: RENAL CELL CARCINOMA (RCC) ACCOUNTS FOR 3-5% OF ALL CANCERS. LOCALIZED ABLATION IS A RECOMMENDED TREATMENT FOR T1A RENAL CANCERS (APPROXIMATELY 25% OF ALL RCC) AND PARTICULARLY IMPORTANT FOR THE TREATMENT OF PATIENTS WHO ARE POOR SURGICAL CANDIDATES DUE TO DIMINISHED RENAL FUNCTION OR WITH A SOLITARY KIDNEY. POST-ABLATION IMAGING PROTOCOLS FOR DETECTION OF RECURRENCE VARY, BUT MAINLY RELY ON CONTRAST-ENHANCED COMPUTED TOMOGRAPHY (CECT) OR CONTRAST-ENHANCED MAGNETIC RESONANCE IMAGING (CEMRI) AT 3-12 MONTHS INTERVALS. HOWEVER, DETECTION OF RELAPSE BASED ON DETECTION OF CONTRAST ENHANCEMENT WITHIN THE ABLATION CAVITY CAN BE CHALLENGING ON BOTH MODALITIES – DUE TO MOST IMAGING FINDINGS OF RECURRENCE BEING NONSPECIFIC BECAUSE OF CHANGES IN TUMOR SIZE, ENHANCEMENT FROM FAT NECROSIS AND SCARRING, AND THE DEVELOPMENT OF CALCIFICATIONS FOLLOWING THERAPY. ADDITIONALLY, THESE TECHNIQUES CAN BE PROBLEMATIC DUE TO MODALITY AND CONTRAST AGENT CONTRAINDICATIONS. AS AN ALTERNATIVE TO CECT AND CEMRI, CONTRAST-ENHANCED ULTRASOUND (CEUS) HAS GAINED ACCEPTANCE FOR A VARIETY OF CLINICAL APPLICATIONS. IN SMALL PILOT STUDIES, OUR GROUP HAS DEMONSTRATED THE ABILITY OF CEUS TO MONITOR FOR BOTH RESIDUAL AND RECURRENT RCC FOLLOWING PERCUTANEOUS ABLATION WITH 100% SENSITIVITY AND NO ADVERSE EVENTS. WHILE ENCOURAGING, OUR FINDINGS WERE BASED ON A RELATIVELY SMALL SAMPLE SIZE AND, WITH AN OVERALL SPECIFICITY OF THE TECHNIQUE THAT RANGED FROM 75-96%, PRIMARILY DUE TO AN INABILITY TO PROPERLY IDENTIFY THE BOUNDARIES OF THE ABLATION CAVITY FROM THE SURROUNDING RENAL CORTEX. TO ASSESS THE CLINICAL PROMISE OF CEUS, WE PROPOSE A LARGER, FULLY POWERED STUDY, RECRUITING PATIENTS ACROSS MULTIPLE INSTITUTIONS AND USING MULTI-MODALITY IMAGING TECHNIQUES TO OVERCOME THESE LIMITATIONS AND ENABLE CLINICAL ADOPTION. IN OUR FIRST SPECIFIC AIM, WE WILL CHARACTERIZE THE SENSITIVITY, SPECIFICITY, POSITIVE AND NEGATIVE PREDICTIVE VALUE, AND INTER-READER AGREEMENT OF BOTH 2D CEUS AND CONTRAST-ENHANCED CROSS-SECTIONAL IMAGING FOR DETECTING RECURRENT RCC FOLLOWING ABLATION; WE WILL USE A COMBINATION OF LONG-TERM FOLLOW-UP IMAGING AND TISSUE PATHOLOGY AS A REFERENCE STANDARD, INSTEAD OF RELYING ON CECT OR CEMRI. AT EACH SITE, BLINDED RADIOLOGISTS WITH VARYING EXPERIENCE LEVELS IN CEUS WILL INTERPRET EXAMS, ENABLING US TO DEFINE THESE PARAMETERS WHILE ALSO QUANTIFYING READER VARIABILITY. IN THE SECOND AIM, WE WILL EVALUATE THE POTENTIAL IMPROVEMENT TO THE QUALITATIVE ASSESSMENTS OF 2D CEUS WHEN FUSED WITH THE PATIENT'S PRE-TREATMENT CROSS-SECTIONAL IMAGING, WHICH WE HYPOTHESIZE WILL PROVIDE BETTER DELINEATION OF THE INITIAL TUMOR BOUNDARY. IN THE THIRD AIM, WE WILL ASSESS THE POTENTIAL ADDED VALUE OF USING MULTI-MODALITY VOLUMETRIC CEUS FOR DETECTING RCC RECURRENCE POST ABLATION AND ALSO AN ADVANCED DOPPLER NON-CONTRAST TECHNIQUE AS AN EXPLORATORY SUB-AIM. FINALLY, WE WILL EXPLORE USE OF QUANTITATIVE PARAMETERS EXTRACTED FROM 2D/3D CEUS EXAMS TO DETERMINE IF THIS IMPROVES THE OVERALL PERFORMANCE OF CEUS. AT THE STUDY'S CONCLUSION, WE HOPE TO ADDRESS THE CURRENT LIMITATIONS IN RCC RECURRENCE SURVEILLANCE IMAGING WITH FULLY POWERED CEUS STUDY TO SUPPORT CLINICAL ADOPTION, WHILE ALSO ADDRESSING PRIOR LIMITATIONS THROUGH THE USE OF MULTI-MODALITY AND QUANTITATIVE IMAGING.

REGULATION OF CLATHRIN AND ADAPTOR FUNCTION

DISCOVERY OF GRAM-NEGATIVE PERMEABLE CHEMICAL PROBES FOR TRNA METHYLATION

A COMPARISON OF BUPRENORPHINE VERSUS MORPHINE IN THE TREATMENT OF THE NEONATAL AB

MODELING CENTRAL AUTONOMIC REGULATORY NETWORK ADAPTATION TO HYPERTENSION

PRIMARY CARE TRAINING AND ENHANCEMENT - LANGUAGE AND DISABILITY ACCESS - ADDRESS: 1015 WALNUT ST., SUITE 401, PHILADELPHIA, PA 19107 PROJECT DIRECTOR NAME: MARY M. STEPHENS, MD, MPH CONTACT NUMBERS: 215-503-7136 (PHONE), 215-955-0640 (FAX) EMAIL ADDRESS: MARY.STEPHENS@JEFFERSON.EDU FUNDS REQUESTED: $2,763,252; $555,269 (YEAR 1), $550,169 (YEAR 2), $555,202 (YEAR 3), $550,169 (YEAR 4), $552,443 (YEAR 5) THE DEPARTMENT OF FAMILY AND COMMUNITY MEDICINE OF SIDNEY KIMMEL MEDICAL COLLEGE AT THOMAS JEFFERSON UNIVERSITY IS PROPOSING THE CREATION OF THE PROGRAM: JEFFERSON EDUCATION AND TRAINING: PRIMARY HEALTHCARE IN IDD AND LANGUAGE ACCESS (JET: PHILA). THIS POGRAM WOULD ADDRESS BOTH FOCUS AREAS (LANGUAGE ASSISTANCE FOR INDIVIDUALS WITH LIMITED ENGLISH PROFICIENCY (LEP) AND CARE FOR INDIVIDUALS WITH PHYSICAL DISABILITIES AND/OR INTELLECTUAL AND DEVELOPMENTAL DISABILITIES (IDD)) AND WOULD PROVIDE TRAINING OPPORTUNITIES FOR BOTH MEDICAL STUDENTS AND FAMILY MEDICINE RESIDENTS. THE GOAL OF JET: PHILA IS TO INCREASE PRIMARY CARE ACCESSIBILITY BY HELPING OUR TRAINEES GAIN KNOWLEDGE AND DEVELOP SKILLS TO BECOME COMPETENT AND RESPECTFUL PROVIDERS WHO WILL DELIVER QUALITY CARE TO THEIR PATIENTS WITH LEP AND IDD. INDIVIDUALS WITH LEP OR IDD FACE SIGNIFICANT HEALTH DISPARITIES. MANY OF THESE DISPARITIES ARE A DIRECT RESULT OF THE VARYING LEVELS OF KNOWLEDGE AND THE BIASES THAT EXIST AMONG THEIR PROVIDERS. AT THIS TIME, EDUCATION AND SKILLS RELATED TO WORKING WITH EITHER OF THESE GROUPS IS NOT A REQUIREMENT OR CORE COMPETENCY OF EITHER THE MEDICAL SCHOOL OR RESIDENCY CURRICULUM. TO ADDRESS THIS, JET: PHILA WOULD OFFER ELECTIVES FOR MEDICAL STUDENTS, AREAS OF CONCENTRATION FOR RESIDENTS, LANGUAGE IMMERSION COURSES, SERVICE-LEARNING ACTIVITIES, AND YEARLY CONFERENCES. JET: PHILA'S SPECIFIC GOALS INCLUDE: -DEVELOP AND IMPLEMENT AN ON-GOING ACADEMIC AND CLINICAL CURRICULUM FOR GRADUATE MEDICAL EDUCATION (GME) AND UNDERGRADUATE MEDICAL EDUCATION (UME) TRAINEES FOCUSED ON THE CARE OF INDIVIDUALS WITH LEP AND IDD. -FACILITATE TRAINEES P ARTICIPATION IN STAKEHOLDER INTERVIEWS, INTERPROFESSIONAL ACTIVITIES, HEALTH EDUCATION AND LITERACY CLASSES, AND PATIENT JOURNEY ACTIVITIES. -EXPAND ON AND FOSTER EXISTING RELATIONSHIPS WITH WYSS WELLNESS CENTER, PUENTES DE SALUD, ESPERANZA HEALTH CENTER, THE ARC OF PHILADELPHIA, SPECIAL OLYMPICS OF PA, CAROUSEL CONNECTIONS, VISION FOR EQUALITY, OFFICE OF DEVELOPMENTAL PROGRAMS, AND HMS SCHOOL FOR CEREBRAL PALSY IN PHILADELPHIA, PA. WE HAVE ASSEMBLED A TEAM OF FACULTY MEMBERS WITH YEARS OF EXPERIENCE AND A WEALTH OF KNOWLEDGE ON CARING FOR PATIENTS WITH LEP AND IDD. IN ADDITION, THE TEAM INCLUDES A NUMBER OF RESEARCH PROFESSIONALS WITH PROJECT MANAGEMENT, PROGRAM EVALUATION, AND DATA ANALYSIS SKILLS. OVER THE COURSE OF THIS FIVE YEAR GRANT, OUR GOAL IS TO TRAIN ABOUT 48 RESIDENTS AND 80 MEDICAL STUDENTS ACROSS THE TWO FOCUSES WITH THE EDUCATION AND SKILLS THAT ARE CURRENTLY MISSING FROM THE CURRICULA, SO THAT THEY ARE PREPARED TO CARE FOR THEIR FUTURE PATIENTS. ADDITIONALLY, WE HOPE THAT THE REACH OF THIS GRANT WILL EXTEND BEYOND THESE TRAINEES AS THEY SHARE WHAT THEY LEARN WITH THEIR CLASSMATES BY PRESENTING ON THEIR EXPERIENCES AND CREATING DELIVERABLES THAT CAN BE ACCESSED BY ANYONE WHO IS INTERESTED. WE ALSO PLAN TO SUPPORT TRAINEE AND FACULTY PRESENTATION AT LOCAL AND NATIONAL CONFERENCES. FURTHER, OUR GOAL IS TO FOSTER OUR ALREADY EXISTING RELATIONSHIPS WITH OUR COMMUNITY PARTNERS AND ENGAGE THEM IN THE CONFERENCES AND WORKSHOPS THAT WE HAVE PLANNED.

MECHANISMS OF UVEAL MELANOMA DORMANCY AND TARGETED THERAPY TOLERANCE

USP22 FUNCTION IN ADVANCED PROSTATE CANCER

MONODISPERSE MICROBUBBLES FOR NONINVASIVE PRESSURE ESTIMATION - PROJECT SUMMARY THE CURRENT CLINICAL STANDARD FOR QUANTIFYING FLUID PRESSURES RELIES ON THE INVASIVE PLACEMENT OF PRESSURE CATHETERS OR NEEDLES. THESE MEASURES ARE COSTLY AND NOT WITHOUT RISK, THEREBY REDUCING HOW OFTEN DATA IS COLLECTED. ULTRASOUND CONTRAST AGENTS (UCA) ARE GAS-FILLED MICROBUBBLES THAT, WHEN INSONTATED AT A FUNDAMENTAL FREQUENCY (F0), ACT AS NONLINEAR OSCILLATORS, GENERATING SIGNAL COMPONENTS RANGING FROM THE SUBHARMONIC (F0/2) THROUGH HIGHER HARMONICS. THE SUBHARMONIC AMPLITUDE OF UCA EXHIBITS A LINEAR RELATIONSHIP WITH HYDROSTATIC PRESSURE, LEADING TO THE TECHNIQUE OF SUBHARMONIC-AIDED PRESSURE ESTIMATION (SHAPE). SHAPE OPTIMIZATIONS TO DATE HAVE RELIED PRIMARILY ON EMPIRICAL EVIDENCE TO IDENTIFY OPTIMAL ACOUSTIC PARAMETERS AND SELECT A COMMERCIALLY AVAILABLE UCA. CURRENTLY, SHAPE PROVIDES UP TO 14 DB REDUCTION IN THE SUBHARMONIC AMPLITUDE OVER A PRESSURE INCREASE OF 180 MMHG (0.6 DB/KPA). CLINICAL TRIALS USING SHAPE FOR THE DIAGNOSIS OF PORTAL PRESSURES, CARDIAC PRESSURES, AND INTERSTITIAL TUMORAL PRESSURES DURING THERAPY HAVE ALL SHOWN SUCCESS. HOWEVER, LARGE VARIATIONS IN SHAPE HAVE BEEN OBSERVED AT LOWER FLUID PRESSURES, INDICATING A NEED TO IMPROVE THE TECHNIQUE'S OVERALL SENSITIVITY. USING A VARIATION OF THE RAYLEIGH–PLESSET EQUATION, OUR GROUP AND OTHERS HAVE MODELED THE SHAPE RESPONSE OF INDIVIDUAL COMMERCIAL BUBBLES AND IDENTIFIED POTENTIAL SENSITIVITIES > 2 DB/KPA USING OPTIMIZED ACOUSTIC PARAMETERS. THUS, THE POTENTIAL EXISTS TO MORE THAN TRIPLE THE CURRENT SENSITIVITY OF SHAPE, THEREBY GREATLY REDUCING THE OVERALL ERRORS ASSOCIATED WITH LOWER PRESSURE MEASUREMENTS. MONODISPERSE MICROBUBBLES CAN BE CREATED USING EITHER BUOYANCY SEPARATION OF EXISTING UCAS OR MICROFLUIDIC TECHNIQUES. WE HYPOTHESIZE THESE AGENTS WILL ALLOW US TO BETTER REFINE PREVIOUS MODELING EFFORTS, WHILE ALSO GREATLY IMPROVING THE OVERALL SENSITIVITY OF SHAPE BY TAILORING THE UCA TO ITS APPLICATION. TO SUPPORT THIS HYPOTHESIS, WE RECENTLY SHOWED THAT MONODISPERSE UCA NEARLY DOUBLED THE SENSITIVITY OF SHAPE (EVEN WITHOUT FULL ACOUSTIC OPTIMIZATION). THIS PROPOSAL WILL BE A FIRST STEP TOWARDS THE LONG-TERM GOAL OF TRANSLATING SHAPE-SPECIFIC UCA INTO CLINICAL TRIALS FOR IMPROVING THE OVERALL SENSITIVITY OF SHAPE AS A NONINVASIVE PRESSURE ESTIMATION TECHNIQUE. AS PART OF THIS APPLICATION, WE PROPOSE TO TEST THE IN VITRO SENSITIVITY OF SHAPE USING MONODISPERSE UCA USING TWO FABRICATION APPROACHES, TO REFINE AND VALIDATE OUR PRIOR MODELS OF SHAPE WITH EMPIRICAL EVIDENCE FROM MONODISPERSE UCA, AND FINALLY, TO DETERMINE THE ABILITY OF A CUSTOMIZED, MONODISPERSE UCA TO IMPROVE THE SENSITIVITY OF SHAPE IN IN VIVO MODELS OF CARDIAC PRESSURES AND PORTAL HYPERTENSION. AT THE CONCLUSION OF THIS PROJECT, WE WILL HAVE DEVELOPED AND VALIDATED A SHAPE-SPECIFIC UCA, CAPABLE OF IMPROVING THE SENSITIVITY OF SHAPE. THESE FINDINGS ARE EXPECTED TO REDUCE THE VARIABILITY OF SHAPE AS A NONINVASIVE CLINICAL MEASURE OF FLUID PRESSURES, ENABLING SAFER AND MORE AVAILABLE CLINICAL CARE.

DEVELOPMENT OF A NOVEL LUNG FUNCTION IMAGING MODALITY FOR COMPREHENSIVE MANAGEMENT OF LUNG CANCER

TRNA-DERIVED NON-CODING RNAS IN ASM FUNCTION AND IN ASTHMA

NRSA FOR PRIMARY MEDICAL CARE

CONTRAST-ENHANCED ULTRASOUND EVALUATION OF FOCAL LIVER LESIONS IN PATIENTS WITH CIRRHOSIS OR OTHER RISK FACTORS FOR DEVELOPING HCC

MECHANISMS OF CELL DEATH IN CUTANEOUS MELANOMA - PROJECT SUMMARY THE INCIDENCE OF CUTANEOUS MELANOMA IS RISING. WHILE SMALL MOLECULE TARGETED INHIBITORS AND IMMUNE CHECKPOINT ANTIBODIES HAVE INCREASED LONG-TERM SURVIVAL IN ADVANCED-STAGE CUTANEOUS MELANOMA, MANY PATIENTS STILL DO NOT BENEFIT AND REGIMENS ARE ASSOCIATED WITH HIGH TOXICITY. WE ARE STUDYING THE DETERMINANTS OF TREATMENT RESPONSE AND MECHANISMS OF RESISTANCE IN MELANOMA. FROM OUR STUDIES, WE AIM TO GENERATE PRE- CLINICAL DATA FOR NEW COMBINATIONS THAT DELAY/PREVENT THE ONSET OF ACQUIRED RESISTANCE WHILE MINIMIZING PATIENT TOXICITIES IN ORDER TO IMPROVE PATIENT SURVIVAL AND QUALITY OF LIFE. MULTIPLE CLINICAL TRIALS HAVE EMANATED FROM OUR WORK (NCT03580382, NCT02012231, NCT02683395). TUMOR IMMUNOGENICITY, DEFINED AS THE ABILITY OF THE TUMOR ITSELF TO TRIGGER AN ANTI-TUMOR ADAPTIVE IMMUNE RESPONSE, IS ONE OF THE MOST IMPORTANT DETERMINANTS OF SUCCESSFUL ANTI-CANCER THERAPY. THE IMMUNOGENICITY OF A TUMOR DEPENDS ON ITS ANTIGENICITY, CONFERRED BY NEO- ANTIGENS, AND ALSO BY ADJUVANT EFFECTS TRIGGERED BY DAMAGE-ASSOCIATED MOLECULAR PATTERNS (DAMPS) RELEASED FROM STRESSED OR DYING TUMOR CELLS DURING A PROCESS CALLED IMMUNOGENIC CELL DEATH (ICD). WE RECENTLY DISCOVERED A SIGNALING PATHWAY THAT ALLOWS EFFICIENT RELEASE OF DAMPS FROM DYING CELLS BY SWITCHING APOPTOSIS INTO A POTENTIALLY IMMUNOGENIC FORM OF CELL DEATH CALLED PYROPTOSIS. MECHANISTICALLY, ACTIVATION OF CASPASE-3 DURING APOPTOSIS LEADS TO CLEAVAGE OF GASDERMIN E (GSDME), GENERATING A PORE-FORMING GSDME-N FRAGMENT. GSDME-N PORES ALLOW RELEASE OF INTRACELLULAR DAMPS SUCH AS HMGB1, DNA, AND ATP. THE ABILITY OF THIS NOVEL PATHWAY TO SWITCH APOPTOSIS INTO PYROPTOSIS SUGGESTS THAT GSDME-INDUCED PYROPTOSIS IS LIKELY A KEY EFFECTOR OF CANCER CELL IMMUNOGENICITY AND MAY DETERMINE THEIR SUCCESSFUL RESPONSE TO VARIOUS ANTI-CANCER THERAPIES. SUPPORTING THIS HYPOTHESIS, OUR PRELIMINARY DATA REVEALED THAT EFFICIENT BRAFI + MEKI-INDUCED ANTI- TUMOR IMMUNE RESPONSES IN MELANOMA CELLS ARE DEPENDENT, AT LEAST IN PART, ON THE PYROPTOTIC ACTIVITY OF GSDME. THE GOALS OF THIS APPLICATION ARE TO DEFINE MECHANISMS UNDERLYING BRAFI + MEKI REGULATION OF GSDME AND PYROPTOSIS IN MELANOMA AND TO INVESTIGATE HOW GSDME-INDUCED PYROPTOSIS ALTERS THE EFFECTS OF IMMUNE CHECKPOINT INHIBITORS. THE STANDARD OF CARE FOR MELANOMAS IS IMMUNE CHECKPOINT INHIBITION, SPECIFICALLY ANTI-PD-1 (PEMBROLIZUMAB AND NIVOLUMAB) AND ANTI-CTLA-4 (IPILIMUMAB). IMMUNE CHECKPOINT INHIBITORS ARE EFFICACIOUS IN SOME MELANOMA PATIENTS; HOWEVER, MANY DO NOT RESPOND. OTHER PATIENTS WHO INITIALLY RESPOND, ULTIMATELY PROGRESS. THIS PROPOSAL IS DESIGNED TO UTILIZE TARGETED THERAPIES TO OPTIMIZE UP-FRONT IMMUNE CHECKPOINT INHIBITORS AS WELL AS INVIGORATING THE IMMUNE SYSTEM IN RESISTANT TUMORS. THUS, WE AIM TO DEVELOP NEW THERAPEUTIC STRATEGIES THAT WILL ADDRESS CLINICAL UNMET NEEDS IN THE MELANOMA FIELD.

SMARCAL1 FUNCTION IN REPLICATION STRESS AND DISEASE

SPINAL CORD INJURY MODEL SYSTEMS PROGRAM: REGIONAL SPINAL CORD INJURY CENTER OF THE DELAWARE VALLEY

ATTENTION AND EXECUTIVE FUNCTIONING IN AGING AND PARKISONISM

PET IMAGING OF KRAS2 ACTIVATION TO GUIDE EGFR TARGETING IN CANCER THERAPY

INFLAMMATORY MONOCYTES IN THE DRAINING LYMPH NODE OF ECTV INFECTED MICE

CHARACTERIZATION OF COCAINE INDUCED SIGNALING PATHWAYS THAT ENHANCES HIV TRANSCRIPTION

LAY HEALTH WORKER MODEL TO REDUCE LIVER CANCER DISPARITIES IN ASIAN AMERICANS

MOLECULAR NEUROGENETICS OF THE BRAINSTEM NEURONAL SOURCE OF CARDIOPROTECTIVE VAGAL OUTFLOW - WE AIM TO IDENTIFY, PREDICT AND CONTROL THE CENTRAL BRAINSTEM NEURONS THAT INTEGRATE INTEROCEPTIVE INPUTS TO PRODUCE AND DETERMINE THE ACTIVITY OF THE VAGUS NERVE IN REGULATING THE HEART. THE LEVEL OF “VAGAL OUTFLOW” IS STRONGLY ASSOCIATED WITH THE HEALTH OF THE HEART. INSUFFICIENT VAGAL OUTFLOW CONTRIBUTES TO MANY FORMS OF HEART DISEASE, WHICH APPEAR PREVENTABLE AND, POTENTIALLY, REVERSIBLE BY INCREASES IN CARDIOPROTECTIVE VAGAL OUTFLOW. WE AIM TO STUDY THE CENTRAL NEURONS WITHIN THE DORSAL MOTOR NUCLEUS OF THE VAGUS (DMV) THAT ARE AN IMPORTANT SOURCE OF CARDIOPROTECTIVE VAGAL OUTFLOW TO THE HEART, AS THESE ARE ASSOCIATED WITH SOME OF THE MOST DEVASTATING DISEASES OF THE HEART. WE WILL DETERMINE THE CARDIOPROTECTIVE MOLECULAR MECHANISMS IN DMV NEURONS IN ORDER TO USE THIS INFORMATION TO INTERVENE AND AFFECT THOSE MOLECULAR MECHANISMS IN WAYS THAT ALLOW CONTROL OF THE VAGAL OUTFLOW. WE WILL ACCOMPLISH THIS THROUGH AN INTEGRATED EXPERIMENTAL AND COMPUTATIONAL ANALYSIS OF THE RESPONSES OF MICRORNAS AND MRNAS IN DMV NEURONS (E.G., GORKY ET AL. 2021). WE HAVE SUCCESSFULLY FOLLOWED THIS APPROACH USING MICRORNA REGULATION IN A PARALLEL PROJECT ON HYPERTENSION (DECICCO ET AL. 2015; GORKY, DEDICCO ET AL. IN REVIEW), AND SEEK TO EMULATE THAT APPROACH HERE. WE HAVE PRELIMINARY DMV DATA INTEGRATING INTEROCEPTIVE INPUTS IN CARDIAC ISCHEMIA AND REMOTE ISCHEMIC PRECONDITIONING (RIPC) SUGGESTING THIS APPROACH WILL BE APPLICABLE FOR VAGAL CARDIOPROTECTION IN MALE AND FEMALE RATS. WE HYPOTHESIZE THAT THE VAGAL OUTFLOW THAT DRIVES CARDIOPROTECTION IN RIPC DERIVES FROM AN ALTERED MOLECULAR ACTIVITY WITHIN THE DORSAL MOTOR NUCLEUS OF THE VAGUS (DMV), MEDIATED BY DIFFERENTIAL MICRORNA REGULATION IN DMV NEURONS. AIM 1 WILL SEEK TO RENORMALIZE THE MOLECULAR STATE OF DMV NEURONS FOLLOWING LEFT ANTERIOR DESCENDING CORONARY ARTERY (LAD) LIGATION BY MODULATING THE MICRORNA REGULATORY NETWORKS WITHIN THE DMV. AIM 2 WILL IDENTIFY THE DYNAMIC TRAJECTORY OF RIPC-INDUCED MICRORNA AND GENE REGULATORY NETWORKS IN DMV TO PREDICT RIPC-INDUCED MICRORNA CONTROL POINTS IN DMV THAT CAN POTENTIALLY EXTEND THE MOLECULAR CARDIOPROTECTIVE EFFECT BEYOND THE CURRENTLY DESCRIBED 24-HOUR EFFICACY WINDOW POST RIPC. AIM 3 WILL TEST THE HYPOTHESIS THAT MICRORNAS REGULATED IN RESPONSE TO RIPC, AND THEIR TARGETS PUTATIVELY CONTRIBUTING TO CARDIOPROTECTION, ARE CO-REGULATED IN ONE OR MORE SPECIFIC SUBSETS OF DMV NEURONS. WE HAVE ASSEMBLED AN INTERDISCIPLINARY TEAM FOR THIS PROJECT. DR. VADIGEPALLI IS A SYSTEMS BIOLOGIST WITH SKILLS IN THE ANALYSIS OF HIGH-DIMENSIONAL DATASETS TO DERIVE PREDICTIONS OF TRANSCRIPTIONAL NETWORK MODULES AND OF MICRORNA NETWORK REGULATORS. DR. SCHWABER HAS EXTENSIVE EXPERIENCE WITH THE INTEGRATIVE CIRCUIT NEUROANATOMY AND NEUROPHYSIOLOGY OF THE CENTRAL MECHANISMS OF VAGAL CARDIAC ACTIVITY. DR. BRAILOIU IS AN EXPERT ON NEURONAL PROCESSES IN THE BRAINSTEM AUTONOMIC NUCLEI AND BRINGS TECHNICAL EXPERTISE ON SINGLE NEURON ISOLATION AND ANALYSIS. THESE STUDIES WILL TAKE STATE-OF-THE-ART MOLECULAR NEUROGENETICS OF SYSTEMS BIOLOGY AND CONTROL SYSTEMS APPROACHES, TO ENABLE TEST OF OUR NEW CENTRAL NEURONAL CARDIOPROTECTION HYPOTHESIS, REVEALING BRAINSTEM NEURONAL DRUGGABLE TARGETS FOR TREATMENT OF HEART DISEASE.

CROSSTALK CA2+ SIGNALING BETWEEN RYANODINE RECEPTORS TYPE 1 AND 2 IN THE PATHOGENESIS OF CARDIAC HYPERTROPHY AND HEART FAILURE - THE UNDERLYING MOLECULAR MECHANISMS FOR HEART FAILURE (HF) ARE MULTIFACTORIAL, WITH ENERGY DYSREGULATION AND OXIDATIVE STRESS APPEARING TO BE THE KEY CAUSES. THE WELL-BALANCED ENERGY CONSUMPTION/GENERATION OF THE WORKING HEART IS THOUGHT TO BE ACHIEVED BY CA2+ ENTRY VIA MITOCHONDRIAL CA2+ UNIPORTER (MCU) WHICH STIMULATES ENZYMES IN THE TRICARBOXYLIC ACID (TCA) CYCLE FOR ATP GENERATION, REFERRED TO AS EXCITATION-BIOENERGETICS (EB) COUPLING. SURPRISINGLY, KNOCKOUT OF MCU IN THE HEART RESULTS IN MINIMAL DEFECTS IN BIOENERGETICS, SUGGESTING THAT OTHER CA2+ TRANSPORTERS ALSO MAY PARTICIPATE IN EB COUPLING. WE HAVE PREVIOUSLY SHOWN THAT RYANODINE RECEPTOR TYPE 1 (RYR1) IS EXPRESSED IN CARDIAC MITOCHONDRIA (MRYR1), PLAYING A KEY ROLE IN EB COUPLING. SEVERAL GROUPS HAVE CONFIRMED OUR FINDINGS, INCLUDING RECENT REPORTS SHOWING THAT CA2+ RELEASE FROM THE SARCOPLASMIC RETICULUM (SR) IS TUNNELED TO MRYR TO STIMULATE ATP PRODUCTION. WE HAVE ALSO OBTAINED NEW DATA SHOWING THAT RYR1 EXPRESSION IS INCREASED IN MOUSE AND HUMAN HYPERTROPHIED HEART. THE MITOCHONDRIA ISOLATED FROM RYR1 OVER EXPRESSION (OE) MOUSE HEARTS HAVE HIGHER CA2+ CONCENTRATIONS ([CA2+]M) AND INCREASED ROS GENERATION. THIS RYR1 OE MOUSE ALSO SHOWS CARDIAC HYPERTROPHY AND HIGHER FREQUENCY OF CA2+ SPARKS, SUGGESTING LEAKIER RYR2 FOR CA2+. TAKEN TOGETHER, WE PROPOSE A NOVEL CENTRAL HYPOTHESIS: HYPERTROPHIC STIMULI LEAD TO MRYR1 OVEREXPRESSION IN HEART TO COMPENSATE FOR INCREASED ENERGY DEMANDS BY PROMOTING MITOCHONDRIAL CA2+ UPTAKE FOR EB COUPLING (AIM 1). HOWEVER, CHRONIC INCREASES IN MITOCHONDRIAL CA2+ UPTAKE CAUSES A SUSTAINED HIGH LEVEL OF ROS GENERATION VIA RIESKE IRON-SULFUR PROTEIN (RISP) AT COMPLEX III (AIM 2). THE INCREASED ROS WOULD OXIDIZE AND THUS ACTIVATE NEARBY SR RYR2 FURTHER INCREASE MITOCHONDRIAL CA2+ LOADING DUE TO CONSTANT CA2+ LEAK FROM SR. THIS VICIOUS CYCLE OF MRYR1[CA2+]MROSSR RYR2 CA2+ LEAK[CA2+]M (: LEADS TO, : INCREASES), WHICH CAUSES MITOCHONDRIAL DYSFUNCTION INCLUDING OPENING OF MITOCHONDRIAL PERMEABILITY TRANSITION PORE. CONSEQUENTLY, THE HEART IS FAILING BECAUSE OF THE INADEQUATE ENERGY FOR PUMPING AND HIGHER MYOCYTE DEATH AND INJURY (AIM 3). THE SUCCESSFUL RESEARCH OUTCOME FROM THIS PROPOSAL WILL PROVIDE A NEW PARADIGM FOR HF: THAT RYR1 OE IN THE STRESSED HEART IS AN INITIAL ADAPTIVE MECHANISM THAT SEQUENTIALLY BECOME MAL-ADAPTIVE, WHICH SUBSEQUENTLY LEADS TO HF. THIS NEW INFORMATION WILL HIGHLIGHT A POTENTIALLY INNOVATIVE THERAPEUTIC STRATEGY FOR DEVELOPMENT OF NOVEL INHIBITORS THAT ARE MORE SELECTIVE FOR RYR1 THAN FOR RYR2 – SUCH AS DANTROLENE WHICH HAS ALREADY BEEN FREQUENTLY USED FOR TREATING MALIGNANT HYPERTHERMIA PATIENTS – AS EFFECTIVE TREATMENTS OF CARDIAC HYPERTROPHY AND HF.

SOD1/BC1-2 COMPLEX: A ROLE IN REGULATING MOTOR NEURON CELL DEATH

PATHOGENESIS OF INFLAMMATION-DRIVEN INTERVERTEBRAL DISC HERNIATION: THE ROLE OF SYNDECAN 4

MODEL DEVELOPMENT FOR PREDICTION OF SURGICAL OUTCOME IN TEMPORAL LOBE EPILEPSY PATIENTS: INCORPORATION OF THE CORRELATION BETWEEN POST-SURGICAL REORGANIZATION PHENOTYPES AND PRE-SURGICAL DATA

THE AR N/C INTERACTION IN SBMA - MECHANISTIC ROLE AND THERAPEUTIC POTENTIAL

THE ROLE OF M6A-RNA METHYLATION IN MEMORY FORMATION AND RECALL AND ITS MODULATION AND INFLUENCE ON LONG-TERM OUTCOMES AS A CONSEQUENCE OF EARLY LIFE LEAD EXPOSURE - REGULATION OF TRANSCRIPTION IS CENTRAL TO PROPER NERVOUS SYSTEM DEVELOPMENT AND FUNCTIONING. DYSREGULATION OF TRANSCRIPTIONAL AND POST-TRANSCRIPTIONAL REGULATORY PATHWAYS ARE ASSOCIATED WITH VARIOUS NEURODEVELOPMENTAL DISEASES AND DISORDERS INCLUDING DEVELOPMENTAL LEAD (PB) EXPOSURE, WHICH OFTEN RESULTS IN SIGNIFICANT AND PERSISTENT COGNITIVE AND BEHAVIORAL DEFICITS. WE HAVE PREVIOUSLY REPORTED PB-INDUCED METHYLATION CHANGES AT GENE PROMOTER REGIONS AND EFFECTS OF PB ON VARIOUS POST-TRANSLATIONAL HISTONE MODIFICATIONS (PTHMS) IN HIPPO- CAMPUS (HIPP) AND MEDIAL PREFRONTAL CORTEX (MPFC) IN ANIMALS WITH PB-INDUCED COGNITIVE DYSFUNCTION, WITH SEX, AMOUNT OF PB EXPOSURE, AND DEVELOPMENTAL WINDOW OF EXPOSURE AS EFFECT MODIFIERS. WHILE PRIOR WORK HAS FOCUSED ON EFFECTS OF PB ON DNA-RELATED MECHANISMS, AN ADDITIONAL MOLECULAR TRANSCRIPTIONAL CONTROL MECHANISM THAT HAS NOT BEEN STUDIED IN THIS REGARD IS RNA MODIFICATION. PRELIMINARY DATA FROM OUR LAB SUPPORT THE HYPOTHESIS THAT DIRECT RNA METHYLATION VIA N6-METHYLADENOSINE (M6A/M), MAY BE AN IMPORTANT MECHANISM CONTRIBUTING TO THE FUNCTIONALLY ALTERED TRANSCRIPTOME AFTER DEVELOPMENTAL PB EXPOSURE. DIRECT MODIFICATIONS OF RNA APPEAR TO PROVIDE AN ADDITIONAL LAYER OF CONTROL OVER RNA FUNCTION FOR THE FINE-TUNING OF TRANSCRIPTOMIC RESPONSES TO THE ENVIRONMENT, CRITICAL FOR NORMAL PLASTICITY AND MEMORY FUNCTION. CONSIDERING THE NEGATIVE IMPACT THAT PB EXPOSURE HAS ON NEURAL PLASTICITY AND MEMORY, WE SUGGEST THAT THERE IS A POTENTIALLY IMPORTANT ROLE OF M6A/M IN POST-TRANSCRIPTIONAL REGULATION OF GENE-SPECIFIC RESPONSES ASSOCIATED WITH PB-INDUCED PLASTICITY/ MEMORY IMPAIRMENTS. OUR HYPOTHESIS IS THAT THIS ABUNDANT EPITRANSCRIPTOMIC MARK, M6A/M, NOT ONLY PLAYS AN IMPORTANT ROLE IN MODULATING MEMORY PROCESSES UNDER NORMAL CIRCUMSTANCES BUT IS ALTERED BY EARLY PB EXPOSURE WITH AN ADVERSE INFLUENCE ON TRANSCRIPTIONAL AND BEHAVIORAL RESPONSES TO PB. WE WILL TEST THIS HYPOTHESIS IN THE FOLLOWING AIMS: AIM1: EXAMINE THE EXTENT TO WHICH TRANSCRIPTOME-WIDE M6A/M PROFILES IN MPFC AND HIPP CA1 AND ARE ALTERED BY DEVELOPMENTAL PB EXPOSURE IN MALES AND FEMALES AND ASSOCIATED WITH EXPRESSION OF MEMORY DEFICITS. HYPOTHESIS: THERE WILL BE BRAIN REGION-DISTINCT TRANSCRIPTOME-WIDE ENRICHMENT PROFILES OF M6A/M (DETECTED BY M6A-ECLIP (ENHANCED UV-CROSSLINKING AND IMMUNOPRECIPITATION), PARTICULARLY ASSOCIATED WITH MEMORY/PLASTICITY-RELATED GENES AND PATHWAYS, THAT ARE ABERRANTLY MODULATED IN PB-EXPOSED, MEMORY-IMPAIRED ANIMALS; AIM2: EXAMINE THE FUNCTIONAL SIGNIFICANCE OF M6A/M MODIFICATION IN MPFC AND CA1 IN PB-EXPOSED ANIMALS THROUGH MANIPULATION OF M6A/M LEVELS. AS PRELIMINARY DATA INDICATE DECREASED M6A LEVELS AND INCREASED FTO EXPRESSION IN PB-EXPOSED RATS WITH MEMORY DEFICITS, WE WILL EXPLORE THE POTENTIAL EFFICACY OF VIRAL-MEDIATED TARGETED KNOCKDOWN OF FTO ON PB-INDUCED MEMORY IMPAIRMENTS AND M6A/M LEVELS AND MEMORY/PLASTICITY-RELATED GENE EXPRESSION IN MPFC AND CA1. WE HYPOTHESIZE THAT BY INCREASING M6A/M LEVELS WE WILL AT LEAST PARTIALLY AMELIORATE THE PB-INDUCED MEMORY IMPAIRMENT.

WNT PATHWAY-DRIVEN ANTI-ESTROGEN THERAPY RESISTANCE IN BREAST CANCER - PROJECT SUMMARY THE MOST AGGRESSIVE CASES OF ESTROGEN RECEPTOR-POSITIVE (ER+) BREAST CANCERS (BC) ARE DIAGNOSED IN YOUNG WOMEN (<50 YEARS-OF-AGE). THIS PROJECT IS FOCUSED ON AN UNDERSTUDIED, AGGRESSIVE, AND THERAPY-RESISTANT FORM OF ER+ BC THAT IS CHARACTERIZED BY PREDOMINANTLY ER+ LUMINAL CANCER CELLS INTERSPERSED WITH RARE CLUSTERS OF ER-NEGATIVE STEM/PROGENITOR CELLS THAT EXPRESS THE BASAL CYTOKERATIN-5 (CK5). PATIENTS WITH THIS MIXED ‘LUMINO- BASAL BREAST CANCER’ (LBBC) PHENOTYPE REPRESENT A PREVIOUSLY UNRECOGNIZED POPULATION WITH UNFAVORABLE PROGNOSIS. THESE TUMORS ARE AGGRESSIVE DUE TO THEIR INHERENT ER-INDEPENDENT ENDOCRINE ESCAPE PATHWAY, WHICH INVOLVES RAPID EXPANSION OF THE ER-/CK5+ CANCER CELL POPULATION IN RESPONSE TO ANTIESTROGEN THERAPIES. OUR DATA REVEAL THAT LBBC SHARES THE YOUNGER AGE DISTRIBUTION AND AGGRESSIVE FEATURES OF BASAL-LIKE BC, A BC SUBTYPE THAT IS ASSOCIATED WITH PREGNANCY AT EARLY AGES WITHOUT BREASTFEEDING – FACTORS THAT ARE MORE PREVALENT AMONG BLACK AND POORER WOMEN AND THUS MAY HELP EXPLAIN DISPARITIES IN BC. INTRIGUINGLY, THE BASAL-LIKE FEATURES OF LBBC ARE PROMOTED BY THE PREGNANCY HORMONE PROGESTERONE AND SUPPRESSED BY THE LACTOGENIC HORMONE PROLACTIN. COMPELLING PILOT DATA INDICATES THAT PROGESTINS OR ANTI-ESTROGENS EXPAND THE THERAPY- RESISTANT CK5+ STEM/PROGENITOR CELL POPULATION IN LBBC THROUGH A WNT11-PLZF-BCL6-DEPENDENT SIGNALING NETWORK AND IS REVERSED BY PROLACTIN. THE PROPOSED STUDIES WILL MECHANISTICALLY TEST THE GENERAL REQUIREMENT OF THIS WNT11-PLZF-BCL6 SIGNALING NETWORK REQUIRED FOR ANTIESTROGEN AND PROGESTERONE-DRIVEN EXPANSION OF THE CK5+ CELL POPULATION AND LBBC PROGRESSION (AIM 1). BECAUSE THE CK5+ CELL POPULATION IS OFTEN RARE IN UNTREATED TUMORS, MORE ROBUST DIAGNOSTIC CLASSIFIERS FOR LBBC THAN CK5 ALONE WILL BE DEVELOPED (AIM 2). FINALLY, EFFORTS WILL EXPLORE THE EFFICACY OF TARGETED AGENTS TO DISRUPT THIS SIGNALING NETWORK IN VIVO IN PRECLINICAL MODELS TO PROVIDE RATIONALE FOR NEW THERAPEUTIC STRATEGIES FOR LBBC (AIM 3). IN SUM, WE WILL MECHANISTICALLY VALIDATE A NEWLY DISCOVERED PATHWAY THAT PROMOTES THE THERAPY-RESISTANT LBBC PHENOTYPE, DEVELOP BETTER DIAGNOSTIC METHODS, AND EXPLORE THERAPEUTIC STRATEGIES TO OVERCOME ENDOCRINE RESISTANCE OF THESE TUMORS. THE PROJECT HOLDS POTENTIAL FOR HIGH IMPACT PROGRESS THAT CAN HELP REDUCE BREAST CANCER MORTALITY AND MORBIDITY.

DETERMINING THE ROLE OF AR TRANSCRIPTIONAL FUNCTION IN SBMA

OPTIMIZING EP RECEPTOR SUBTYPE TARGETING FOR ASTHMA THERAPY

FROM MTDNA STRESS TO CELLULAR IMMUNITY: TRIGGERS, MECHANISMS AND EFFECTORS - PROJECT SUMMARY: MITOCHONDRIA ARE THE EVOLUTIONARY PRODUCT OF THE ENDOSYMBIOSIS BETWEEN THE ANCESTRAL EUKARYOTIC CELL AND AN OBLIGATE AEROBE BACTERIUM, WHICH BROUGHT NEW FUNCTIONALITIES TO EUKARYOTIC CELLS. DURING THEIR EVOLUTION, MITOCHONDRIA TRANSFERRED MORE THAN 99% OF THEIR KNOWN GENETIC MATERIAL TO THE NUCLEUS – WITH THE EXCEPTION OF A SMALL, MULTI-COPY GENOME REFERRED TO AS MTDNA. IN HOMO SAPIENS, THE 16.6 KBP OF MTDNA IS CIRCULAR AND ENCODES FOR 13 MEMBERS OF THE OXIDATIVE PHOSPHORYLATION CHAIN (OXPHOS) AND OTHER STRUCTURAL RNAS. BEYOND THEIR CANONICAL ROLE IN THE GENERATION OF ATP THROUGH OXPHOS, THE MITOCHONDRIA ARE ALSO CRITICAL STAKEHOLDERS IN SEVERAL CELLULAR PROCESSES FROM METABOLITE FLUXES AND CALCIUM SIGNALING TO CELL DEATH AND AGING. RECENTLY, MITOCHONDRIAL STRESS HAS BEEN IMPLICATED IN THE ABERRANT ACTIVATION OF INNATE IMMUNITY, MEDIATED BY THE RELEASE OF ORGANELLAR COMPONENTS, SUCH AS MITOCHONDRIAL NUCLEIC ACIDS, RECOGNIZED BY CYTOSOLIC SENSORS AS FOREIGN AND POTENTIALLY DANGEROUS. ABERRANT IMMUNITY HAS DEEP IMPLICATIONS IN HUMAN HEALTH AND IS A DRIVER OF HUMAN DISEASES, SUCH AS NEUROINFLAMMATION AND AUTOIMMUNITY. MITOCHONDRIAL IMMUNITY HAS BEEN STUDIED BOTH IN VITRO AND IN VIVO, BUT THE COMPLETE REPERTOIRE OF ITS TRIGGERS AND EFFECTORS HAS YET TO BE CHARACTERIZED. I RECENTLY UNCOVERED A PREVIOUSLY UNKNOWN SOURCE OF STRESS CONDUCIVE TO ABERRANT IMMUNITY: STRESS TO THE MTDNA IN THE FORM OF DOUBLE-STRANDED BREAKS (MTDSBS). THE PRESENCE OF THIS STRESSOR IS RELAYED TO THE CYTOSOLIC COMPARTMENT VIA MITOCHONDRIAL HERNIATION, A RECENTLY DESCRIBED FORM OF BAX/BAK MEDIATED ORGANELLE PERMEABILIZATION, THAT EXPOSES MITOCHONDRIAL CONTENTS TO THE CYTOSOL. AFTER MTDSBS, MITOCHONDRIAL RNA – RATHER THAN THE RECIPIENT OF THE STRESS, MTDNA – INITIATED THE INNATE IMMUNITY CASCADE BY ACTIVATING THE SENSOR RIG-I. OUR PROPOSED RESEARCH PLAN BUILDS ON THIS PAST WORK TO ASK ESSENTIAL QUESTIONS: (1) WHICH SOURCES OF MTDNA STRESS ARE CONDUCIVE TO ABERRANT IMMUNITY AND WHAT IS THE IMPACT OF DYSFUNCTIONAL MITOCHONDRIAL TRANSCRIPTION OR TRANSLATION; (2) HOW IS MITOCHONDRIAL HERNIATION REGULATED AND HOW DOES IT DIFFER FROM OTHER FORMS OF MITOCHONDRIAL PERMEABILIZATION; AND (3) WHAT ARE THE DISTINCTIVE FEATURES OF MITOCHONDRIAL RNA ACTIVATION OF RIG-I AND WHERE DO THEY ORIGINATE? OUR GOAL IS TO UNDERSTAND HOW MITOCHONDRIA INTEGRATE AND TRANSLATE STRESS SIGNALS, PARTICULARLY IN THE CONTEXT OF INNATE IMMUNITY. MY LAB WILL PROBE DIFFERENT SOURCES OF MTDNA STRESS AND INTERROGATE THE MECHANISMS, EFFECTORS, AND MITOCHONDRIAL MOIETIES ENGAGING THE CYTOSOLIC SENSORS OF IMMUNITY. RIG-I IS A KEY PROTEIN FOR THE DEFENSE AGAINST VIRUSES, BUT ITS ABERRANT ACTIVATION IS INVOLVED IN BOTH AUTOIMMUNE CONDITIONS AND THE BENEFICIAL ANTI-TUMOR RESPONSES ELICITED BY CANCER TREATMENTS. BY BUILDING UPON OUR RECENT FINDINGS AND PRIOR EXPERIENCE, AS WELL AS BY ESTABLISHING COLLABORATIONS AND SEEKING THE ASSISTANCE OF SENIOR INVESTIGATORS WITH ADVANCED EXPERTISE IN BOTH MITOCHONDRIAL BIOLOGY AND THE KEY TECHNOLOGIES PROPOSED, OUR RESEARCH PROGRAM AIMS TO HAVE LONG-LASTING IMPACTS ON THE FOUNDATIONAL AND TRANSLATIONAL KNOWLEDGE OF CELLULAR STRESS RESPONSES GOING BEYOND MITOCHONDRIAL BIOLOGY INTO A PATHWAY WITH FURTHER IMPLICATIONS IN HUMAN HEALTH AND DISEASE: INNATE IMMUNITY.

IMPAIRMENT OF THE GLIAL GLUTAMATE TRANSPORTER GLT1 IN ALS

SPECIALIZED TOOLS AND AUTO-UPDATABLE SCALABLE INTERACTIVE DATABASES TO STUDY ISOMIRS, TRFS AND RRFS IN HUMAN AND MOUSE - THIS PROJECT FOCUSES ON THREE CATEGORIES OF SMALL RNAS: THE ISOFORMS OF MICRORNAS (MIRNAS) THAT ARE KNOWN AS ISOMIRS; THE FRAGMENTS THAT ARE DERIVED FROM TRANSFER RNAS (TRNAS) AND ARE KNOWN AS TRFS; AND, THE FRAG- MENTS THAT ARE DERIVED FROM RIBOSOMAL RNAS (RRNAS) AND ARE KNOWN AS RRFS. ISOMIR, TRFS, AND RRFS HAVE SEVERAL IMPORTANT PROPERTIES THAT WARRANT THEIR DETAILED STUDY: (1) THEY ACCOUNT FOR ~80% OF ALL SMALL RNAS IN A CELL. (2) THEY REGULATE THE ABUNDANCE OF MESSENGER RNAS (MRNAS) AND PRO- TEINS. (3) THEIR EXPRESSION PATTERNS DEPEND ON CELLULAR “CONTEXT” (E.G., TISSUE TYPE, DISEASE TYPE). (4) IN HU- MANS, THEIR EXPRESSION PATTERNS ADDITIONALLY DEPEND ON “PERSONAL ATTRIBUTES” (E.G., SEX, GENETIC ANCESTRY, AGE). TO CORRECTLY MINE ISOMIRS, TRFS, AND RRFS FROM RNA-SEQ DATA THE IDEAL TOOLS MUST ADDRESS SEVERAL COMPLI- CATING FACTORS. FIRST, THE SAME SHORT SEQUENCE (E.G., TRF) CAN ARISE FROM DIFFERENT PARENTAL RNAS. THESE PARENTAL RNAS CAN BELONG TO THE SAME SUB-TYPE (E.G., DIFFERENT TRNA ISODECODERS OF THE SAME TRNA ISOACCEPTOR) OR DIFFERENT SUB-TYPES (E.G., ISODECODERS FROM DIFFERENT TRNA ISOACCEPTORS). SECOND, THE SEQUENCES OF MANY ISO- MIRS, TRFS, AND RRFS CAN ALSO BE FOUND IN UNRELATED REGIONS OF THE GENOME. THIRD, PARALOGUES AND/OR INCOMPLETE COPIES OF MIRNAS, TRNAS, AND RRNAS RIDDLE THE NUCLEAR GENOMES OF MANY ORGANISMS INCLUDING HUMAN AND MOUSE. THE DETAILS OF THESE COMPLICATING FACTORS ARE SPECIFIC TO THE RNA TYPE AND TO THE GENOME. CONSEQUENTLY, THE IDEAL TOOLS MUST BE TARGET-GENOME-SPECIFIC. THE COMPLICATING FACTORS AND THE NEED FOR GENOME SPECIFICITY APPEARED IN THE LITERATURE ONLY RECENTLY. AS A RESULT, MOST AVAILABLE TOOLS TO DATE HAVE BEEN GENERAL-PURPOSE AND DO NOT ACCOUNT FOR THESE COMPLICATIONS. NOT SURPRISINGLY, MOST AVAILABLE DATABASES WERE BUILT USING GENERAL-PURPOSE TOOLS. WITHOUT REALIZING THE UNDERLYING SHORTCOMINGS, MANY RESEARCHERS RELIED ON THE INFORMATION PROVIDED BY THESE TOOLS AND DATABASES TO DESIGN EXPERIMENTS AND ANALYZE THEIR DATA. IN TURN, THIS HAS LED TO MANY PUBLISHED ARTICLES THAT UNINTENTIONALLY DESCRIBE FINDINGS OF UNCLEAR VALUE ABOUT MOLECULES THAT ARE NOT ALWAYS ISOMIRS, TRFS, OR RRFS. WE WILL ADDRESS THESE GAPS AS FOLLOWS. IN AIM 1, WE WILL BUILD SPECIALIZED TOOLS THAT ADDRESS THE PECULIARITIES OF EACH RNA TYPE AND ACCURATELY MINE ISOMIRS, TRFS, AND RRFS FROM HUMAN AND MOUSE RNA-SEQ DATA. THE TOOLS WILL BE ROBUST, SELF-CONTAINED, AND USER-FRIENDLY. IN AIM 2, WE WILL BUILD SPECIALIZED DATABASES TO ORGANIZE AND PROVIDE EASY ACCESS TO INFORMATION ABOUT ISOMIRS, TRFS, AND RRFS THAT WE HAVE ALREADY COMPILED BY MINING 50,000 PUBLIC DATASETS. IN AIM 3, WE WILL BUILD A SYSTEM THAT AUTO-IDENTIFIES NEWLY-ADDED DATASETS TO NIH’S SRA, PROFILES AND ANNOTATES EACH DATASET’S ISOMIRS, TRFS, AND RRFS, AND UPDATES THE DATABASES WITH THE NEW INFOR- MATION EACH MONTH. IN AIM 4, WE WILL CREATE EDUCATIONAL MATERIAL DESCRIBING BEST PRACTICES TO HELP RESEARCHERS BENEFIT MAXIMALLY FROM THIS FRAMEWORK, AND BUILD A SYSTEM TO ALLOW THEM TO INTERACT WITH ONE ANOTHER AND SUBMIT THEIR FEEDBACK. LASTLY, WE WILL VALIDATE EXPERIMENTALLY SELECT SMALL RNAS IMPLICATED IN BREAST CANCER METASTASIS.

CHEMOPREVENTION OF UPPER AERODIGESTIVE TRACT CANCER BY DIETARY ZINC

MITOCHONDRIAL CALCIUM SIGNALING IN CELL DEATH.

ASSISTIVE TECHNOLOGY TO SUPPORT DEVELOPMENT AND LEARNING IN YOUNG CHILDREN BIRTH - 3

S100A1: A NOVEL MODULATOR OF MYOCARDIAL INFARCT INFLAMMATION AND REGENERATION

DEFINING THE ROLE OF WEST NILE VIRUS-HOST PROTEIN INTERACTIONS IN EVADING ANTIVIRAL IMMUNITY

RB FUNCTION IN PROSTATE CANCER PROGRESSION

INVESTIGATING THE ROLE AND REGULATION OF THE MAP7 FAMILY PROTEINS IN AXONAL MORPHOGENESIS AND FUNCTION - PROJECT ABSTRACT MICROTUBULES ARE PROTEIN POLYMERS THAT ARE ESSENTIAL FOR NEURONAL CELL MORPHOGENESIS DURING DEVELOPMENT. MICROTUBULE ASSEMBLY AND FUNCTION RELY ON THE REGULATION OF LATTICE-BOUND MICROTUBULE-ASSOCIATED PROTEINS (MAPS). MANY KNOWN MAPS INFLUENCE MICROTUBULE STABILITY AND ORGANELLE TRANSPORT, AND SOME OF THEM ARE HIGHLY ASSOCIATED WITH NEURODEGENERATION. THE PROPOSED STUDY WILL INVESTIGATE A UNIQUE FAMILY OF MAPS THAT INTERACT WITH BOTH MICROTUBULES AND THE MOTOR PROTEIN KINESIN-1. OUR RECENT STUDY OF ITS FOUNDING MEMBER MAP7 PROVIDES THE FIRST DEMONSTRATION OF THE NEURONAL FUNCTION OF THIS LESS-WELL UNDERSTOOD MAP. MAP7 IS DEVELOPMENTALLY REGULATED TO PROMOTE AXONAL BRANCH FORMATION OF SENSORY NEURONS IN THE DORSAL ROOT GANGLION (DRG). USING PRIMARY NEURONAL CELL CULTURE, WE HAVE FOUND THAT MAP7 REGULATES BRANCH FORMATION AND GROWTH. FURTHER CELL BIOLOGICAL ANALYSIS HAS REVEALED THAT MAP7 REGULATES MICROTUBULE STABILITY AND KINESIN-1-MEDIATED ORGANELLE TRANSPORT, TWO PROCESSES THAT ARE CRITICAL TO AXONAL MORPHOGENESIS AND FUNCTION. RECENTLY, WE EXPAND OUR STUDIES IN SEVERAL DIRECTIONS. FIRST, WE ANALYZED ANOTHER MAP7D1, A CLOSEST MAP7 HOMOLOG AND FOUND THAT IT HAS SIMILAR PROPERTIES AND REDUNDANT FUNCTION AS MAP7 IN CULTURE. SECOND, WE DEVELOPED A NEW ASSAY TO SHOW THE POTENTIAL ROLE OF MAP7 IN REGULATING TRANSPORT DURING BRANCH GROWTH. THIRD, IN THE STUDY OF MAP7 MOUSE MUTANTS, WE FOUND A POTENTIAL NEW ROLE OF MAP7 IN NOCICEPTION. FOLLOWING THESE FINDINGS, WE HYPOTHESIZE THAT THE MAP7 FAMILY PROTEINS PROVIDE A NOVEL MECHANISM AND PLAY MULTIPLE ROLES IN AXONAL BRANCH DEVELOPMENT AND FUNCTION BY REGULATING ORGANELLE TRANSPORT. TO TEST THIS HYPOTHESIS, WE ASK THREE QUESTIONS: 1) ARE BOTH MAP7/MAP7D1 REQUIRED FOR BRANCH DEVELOPMENT OF DRG NEURONS? 2) HOW DOES MAP7 REGULATE ORGANELLE TRANSPORT FOR BRANCH GROWTH AND NOCICEPTION? 3) IS THE NOCICEPTION FUNCTION SPECIFIC FOR MAP7 EXPRESSED IN DRG NEURONS AND ITS INTERACTION WITH KINESIN-1? ANSWERING THESE QUESTIONS WILL NOT ONLY EXPAND OUR KNOWLEDGE OF THIS NOVEL FAMILY OF MAPS BUT ALSO ADDRESS FUNDAMENTAL QUESTIONS REGARDING MICROTUBULE- BASED INTRACELLULAR TRANSPORT IN NEURONAL DEVELOPMENT AND FUNCTION. MORE IMPORTANTLY, OUR PROPOSED STUDIES WITH A FOCUS ON DRG SENSORY NEURONS WILL ALLOW US TO EXPLORE THE ROLE OF TRANSPORT REGULATION IN NOCICEPTION AND THUS BRING US CLOSER TO MEDICALLY RELEVANT PROBLEMS, SUCH AS PAIN. GIVEN THE IMPORTANCE OF NOCICEPTION AND PAIN IN HUMAN HEALTH, OUR PROPOSED STUDIES WITH A FOCUS ON A FUNDAMENTAL CELL BIOLOGICAL PROBLEM ARE THUS HIGHLY RELEVANT TO THE NIH MISSION OF INVESTIGATING BRAIN FUNCTIONS AND DISORDERS.

ROLE OF GATA2 SIGNALING NETWORK IN LETHAL PROSTATE CANCER

FUNCTIONAL DIVERSITY OF COMPARTMENTALIZED CALCIUM SIGNALING IN AIRWAY SMOOTH MUSCLE

STRUCTURAL AND DYNAMIC ANALYSIS OF GRK INTERACTION WITH G PROTEIN-COUPLED RECEPTORS

MITOCHONDRIAL DYNAMICS IN ALCOHOL-INDUCED TISSUE INJURY

REPLICATION STRESS IN HEMATOPOIESIS AND LYMPHOMAGENESIS

NOVEL MECHANISMS AND REGULATION OF MAMMALIAN DOUBLE-STRAND BREAK REPAIR - PROJECT SUMMARY. CELL PROLIFERATION UNDER NORMAL AND ADVERSE ENVIRONMENTAL CONDITIONS DEPENDS ON MULTIPLE DNA DAMAGE RESPONSE (DDR) PATHWAYS. REPLICATIVE STRESS AND CELLULAR EXPOSURE TO GENOTOXIC AGENTS CAN LEAD TO DOUBLE- STRAND BREAKS (DSBS) WHICH ARE THE MOST LETHAL DNA LESIONS. MAJOR DSB REPAIR (DSBR) PATHWAYS INCLUDE ERROR- PRONE NON-HOMOLOGOUS END-JOINING (NHEJ) AND ACCURATE HOMOLOGY-DIRECTED REPAIR (HDR) WHICH PROMOTES GENOME INTEGRITY AND TUMOR SUPPRESSION. MICROHOMOLOGY-MEDIATED END-JOINING (MMEJ), ALSO REFERRED TO AS ALTERNATIVE END-JOINING (ALT-EJ) AND POLYMERASE THETA-MEDIATED END-JOINING (TMEJ), IS THE MOST RECENTLY DISCOVERED DSBR PATHWAY THAT CONTRIBUTES TO GENOME INSTABILITY AND CANCER CELL SURVIVAL. OVER THE LAST 15-20 YEARS, BASIC RESEARCH OF DSBR HAS ARGUABLY LED TO SOME OF THE MOST IMPORTANT ADVANCES IN THE HISTORY OF BIOMEDICAL SCIENCE AND BIOTECHNOLOGY, SUCH AS THE DEVELOPMENT OF POLY ADP RIBOSE 1 (PARP1) INHIBITORS AS PRECISION MEDICINE FOR HDR-DEFICIENT CANCERS, AND GENOME ENGINEERING INVOLVING CRISPR-CAS9 WHICH WAS THE SUBJECT OF THE 2020 NOBEL PRIZE IN CHEMISTRY. DR. POMERANTZ’S LAB HAS STRONGLY CONTRIBUTED TO ADVANCING OUR KNOWLEDGE OF THE MOLECULAR MECHANISMS OF MMEJ BASED DSBR OVER THE PAST 10 YEARS, AND HAS SIGNIFICANTLY CONTRIBUTED TO ADVANCING OUR UNDERSTANDING OF RNA-TEMPLATED DSBR (RNA-DNA REPAIR) OVER THE PAST 5 YEARS. FOR EXAMPLE, DR. POMERANTZ’S LAB RECENTLY IDENTIFIED A NEW MMEJ REPAIR PATHWAY INVOLVING POLL, AND DISCOVERED THAT HUMAN POLQ EXHIBITS REVERSE TRANSCRIPTASE ACTIVITY AND PROMOTES RNA-DNA REPAIR IN HUMAN CELLS. TO SIGNIFICANTLY ADVANCE OUR BASIC KNOWLEDGE OF SOME OF THE MOST INTRIGUING AND IMPORTANT AREAS OF DSBR, THE POMERANTZ LAB PROPOSES TO CONTINUE TO ADDRESS THE FOLLOWING THREE QUESTIONS: 1. DOES RNA DIRECTLY CONTRIBUTE TO DSBR? 2. HOW ARE DSBR PROTEINS STRUCTURALLY AND CHEMICALLY REGULATED? 3. DO UNDISCOVERED DSBR PATHWAYS EXIST AND CONTRIBUTE TO GENOME INSTABILITY/INTEGRITY?

THE ROLE OF CHROMATIN STRUCTURE IN DIFFERENTIATION OF HEMATOPOIETIC STEM CELLS

PREVENTION OF ANTHRACYCLINE-INDUCED CARDIOTOXICITY BY NUR77 ACTIVATION - PROJECT SUMMARY DOXORUBICIN (DOX), AN ANTHRACYCLINE CHEMOTHERAPEUTIC, IS A WELL-ESTABLISHED AND HIGHLY EFFECTIVE CHEMOTHERAPY DRUG COMMONLY USED TO TREAT MULTIPLE CANCER TYPES, BUT ITS USE IS LIMITED BY CARDIOTOXICITY. PATIENTS WITH DOX THERAPY OFTEN DEVELOP DELAYED CARDIOMYOPATHY FOR WHICH NO EFFECTIVE THERAPIES EXIST. THE MOLECULAR MECHANISMS OF THIS DRUG TOXICITY REMAIN POORLY UNDERSTOOD AND ARE LIKELY TO BE MULTIFACTORIAL. DNA DAMAGE, MITOCHONDRIAL DYSFUNCTION, P53 ACTIVATION, AND EXCESSIVE CARDIOMYOCYTE APOPTOSIS ARE MAJOR CONTRIBUTORS TO THE PATHOGENESIS OF DOX-INDUCED CARDIAC TOXICITY, BUT WHETHER A PHARMACOLOGICAL STRATEGY CAN SIMULTANEOUSLY WORK TO REVERSE ALL OF THESE EVENTS IN PATIENTS THAT RECEIVE DOX TREATMENT HAS YET TO BE UNCOVERED. OUR LABORATORIES HAVE A LONG TRACK RECORD IN STUDYING THE ROLE OF NUCLEAR RECEPTOR 4A (NR4A) BIOLOGY IN CARDIOVASCULAR HOMEOSTASIS AND DISEASE. IN RECENT WORK, WE HAVE GENERATED A SIGNIFICANT BODY OF EVIDENCE TO SUPPORT THE NOVEL CONCEPT THAT REDUCED CARDIAC EXPRESSION OF NUR77, A FAMILY MEMBER OF NR4A RECEPTORS, PLAYS A CAUSAL ROLE IN THE PATHOGENESIS OF DOX CARDIOMYOPATHY. WE HAVE FOUND THAT 1) CARDIAC EXPRESSION OF NUR77 IS SIGNIFICANTLY REDUCED IN DOX TREATED CARDIOMYOCYTES AND HEARTS AND THAT 2) CARDIAC SPECIFIC OVEREXPRESSION OF NUR77 EFFECTIVELY PREVENTS DOX INDUCED CARDIAC DYSFUNCTION, CARDIOMYOCYTE APOPTOSIS, AND FIBROSIS. IN ADDITION, WE HAVE IDENTIFIED DNA TOPOISOMERASE IISS-BINDING PROTEIN 1 (TOPBP1) AS A KEY DOWNSTREAM EFFECTOR OF NUR77 IN THE HEART, AND HAVE FOUND THAT ITS EXPRESSION IS SUBSTANTIALLY REDUCED IN DOX-TREATED HEARTS. MORE IMPORTANTLY, OUR PRELIMINARY DATA INDICATE THAT NUR77 PREVENTS CARDIAC TOXICITY BY INDUCING TOPBP1-TOPOISOMERASE IISS (TOP2SS) INTERACTIONS, WHICH IN TURN EFFECTIVELY SHIELDS TOP2SS/GENOMIC DNA FROM DOX-INDUCED DAMAGE IN THE HEART. THESE DATA LED US TO HYPOTHESIZE THAT NUR77 SERVES AS AN IMPORTANT CARDIOPROTECTIVE AGENT AND THAT TARGETED ACTIVATION OF NUR77 CAN EFFECTIVELY PREVENT DOX-INDUCED CARDIOMYOPATHY WHILE HAVING MINIMAL TO NO EFFECTS ON ITS ANTI-CANCER BEHAVIORS. WE WILL TEST THIS HYPOTHESIS IN THE FOLLOWING THREE AIMS. AIM 1 WILL PERFORM LOSS-OF-FUNCTION STUDIES BY EMPLOYING NUR77 CARDIAC SPECIFIC KNOCKOUT MICE (CNUR77-KO) TO DETERMINE WHETHER CARDIAC SPECIFIC DELETION OF NUR77 EXACERBATES DOX-INDUCED CARDIOTOXICITY THROUGH ENHANCED APOPTOSIS, FIBROSIS, AND INFLAMMATION IN THE HEART. AIM 2 WILL DETERMINE THE UNDERLYING MECHANISMS BY WHICH NUR77 EXHIBITS ITS CARDIOPROTECTIVE EFFECTS AGAINST DOX-INDUCED CARDIAC INJURY. CARDIAC SPECIFIC TOPBP1 TRANSGENIC AND KNOCKOUT MICE WILL BE USED TO ASSESS THE CONTRIBUTION OF TOPBP1 TO THE NUR77-INITIATED CARDIAC PROTECTION IN MOUSE MODELS OF DOX CARDIOTOXICITY. AIM 3 WILL STUDY WHETHER PHARMACOLOGICAL ACTIVATION OF NUR77 HOLDS THE CAPACITY TO BLOCK DOX CARDIOMYOPATHY WITHOUT COMPROMISING ANTI-CANCER ACTIVITY OF THE DRUG. SUCCESSFUL COMPLETION OF THESE AIMS WILL GUIDE US TO DEVELOP NEW THERAPEUTIC AGENTS TO PREVENT SELECTIVE TOXICITY OF ANTICANCER DRUGS TO THE HEART. THE PREDICTED UTILITY AND FEASIBILITY OF SUCH TARGETING IN HUMANS FAVORS THE TRANSLATIONAL POTENTIAL AND ULTIMATE IMPACT OF THE PROPOSED STUDIES.

REGULATION OF PLATELET REACTIVITY BY S1P SIGNALING

INVESTIGATING A NEW VULNERABILITY IN ORAL SQUAMOUS CELL CARCINOMA - SUMMARY ORAL SQUAMOUS CELL CARCINOMA (OSCC) IS THE MOST COMMON ORAL MALIGNANCY AND SUBTYPE OF HEAD AND NECK CANCER AND FREQUENTLY IDENTIFIED BY DENTISTS DURING ROUTINE CARE. OSCC IS THE SIXTH MOST COMMON CANCER GLOBALLY WITH INCIDENCE CONTINUING TO RISE. CURRENT THERAPIES FOR OSCC ARE INADEQUATE AND OSCC ARE RESISTANT TO IMMUNE CHECKPOINT INHIBITION (ICI), RESULTING IN FREQUENT REOCCURRENCE AND POOR RATES OF PATIENT SURVIVAL. THE DEVELOPMENT OF TARGETED THERAPIES FOR OSCC IS HAMPERED BY THE LACK OF COMMON DRIVING ONCOGENE MUTATIONS, BUT INACTIVATION OF THE P53 TUMOR SUPPRESSOR IS FREQUENT IN HUMAN PAPILLOMAVIRUS (HPV) NEGATIVE OSCC, WHICH CONTRIBUTES TO TREATMENT RESISTANCE, IMMUNE CELL SUPPRESSION, AND REDUCED PATIENT SURVIVAL. THEREFORE, NEW AND INNOVATIVE TREATMENT APPROACHES ARE DESPERATELY NEEDED FOR OSCC AND PARTICULARLY THOSE WITH INACTIVATED P53. THUS, WE SOUGHT TO IDENTIFY NEW VULNERABILITIES IN HPV-NEGATIVE OSCC. IT WAS BELIEVED THAT IF P53 WAS INACTIVATED, CANCER CELLS DID NOT REQUIRE MDM2, WHICH BINDS AND REGULATES P53. HOWEVER, CHALLENGING THIS LONG-HELD BELIEF, OUR RESULTS WITH CANCERS ARISING IN P53-NULL MICE SHOWED THAT THE MDM2 ONCOGENE WAS REQUIRED FOR THE SURVIVAL OF P53-NULL MURINE CANCERS. CAPITALIZING ON OUR PARADIGM-SHIFTING DISCOVERY IN MICE, OUR PRELIMINARY DATA REVEAL MDM2 IS A PREVIOUSLY UNKNOWN AND UNEXPECTED VULNERABILITY IN P53-INACTIVATED HUMAN OSCC. WE PROPOSE THAT TARGETED LOSS OF MDM2 WILL EFFECTIVELY ELIMINATE OSCC CELLS BY ACTIVATING P53- INDEPENDENT MECHANISMS THAT SIGNAL FOR APOPTOSIS AND PATHWAYS THAT CONTRIBUTE TO INCREASING IMMUNOGENICITY. PRELIMINARY RESULTS WITH A NOVEL MDM2-TARGETED DEGRADER WE DESIGNED AND SYNTHESIZED SUPPORT THIS HYPOTHESIS. TO FURTHER TEST OUR HYPOTHESIS, WE PROPOSE TO 1) INVESTIGATE THE CYTOTOXICITY MECHANISM(S), THE REQUIRED PATHWAYS, AND IN VIVO EFFICACY WITH TARGETED MDM2 LOSS WITH OUR MDM2 PROTAC IN OSCC CELLS WITH VARIOUS P53 ALTERATIONS; AND 2) INVESTIGATE IMPROVING THE IMMUNOGENICITY OF OSCC TUMORS AND OVERCOMING ICI RESISTANCE WITH SEVERAL APPROACHES THAT INCLUDE OUR MDM2 PROTAC AND IMMUNE COMPETENT SYNGENEIC MOUSE MODELS. COMPLETION OF THIS STUDY WILL UNCOVER THE SPECIFIC REQUIREMENTS OF MDM2 IN OSCC, SIGNIFICANTLY INCREASING THE MECHANISTIC UNDERSTANDING OF THE P53-INDEPENDENT FUNCTIONS OF MDM2, AND PROVIDE CRITICAL PRE-CLINICAL EVALUATION OF POTENTIAL NOVEL THERAPY APPROACHES FOR OSCC THAT INCREASE IMMUNOGENICITY AND OVERCOME ICI RESISTANCE.

DEVELOPMENT OF BEST-IN-CLASS POL-THETA INHIBITORS - PROJECT SUMMARY. BRCA1 AND BRCA2 FACILITATE DNA DOUBLE-STRAND BREAK (DSB) REPAIR VIA HOMOLOGOUS RECOMBINATION (HR). YET BRCA1/2 ARE OFTEN MUTATED IN CANCERS, SUCH AS BREAST/OVARIAN/PROSTATE/PANCREATIC CARCINOMAS. BRCA-DEFICIENT CANCER CELLS ARE THEREFORE HR-DEFECTIVE, AND SUSCEPTIBLE TO AGENTS THAT INDUCE DNA DAMAGE AND/OR PREVENT DSB REPAIR. FOR INSTANCE, POLY-ADP RIBOSE POLYMERASE 1 (PARP1) INHIBITORS (PARPI) PREFERENTIALLY KILL BRCA-DEFICIENT CANCER CELLS AND ARE APPROVED TO TREAT HR-DEFICIENT CANCERS. HOWEVER, ONLY ~60% PATIENTS RESPOND TO PARPI AND DRUG RESISTANCE IS A MAJOR PROBLEM. THUS, DEVELOPING SECOND-GENERATION PRECISION MEDICINES THAT TARGET HR-DEFICIENT CANCERS AND/OR OVERCOME PARPI RESISTANCE IS URGENTLY NEEDED. DNA POLYMERASE THETA (POLQ), A DNA REPAIR FACTOR IMPORTANT FOR MICROHOMOLOGY-MEDIATED END-JOINING (MMEJ) OF DSBS, HAS BEEN RECENTLY VALIDATED AS A DRUGGABLE TARGET IN BRCA-DEFICIENT CANCERS. POLQ IS ESSENTIAL FOR BRCA-DEFICIENT CANCER CELLS, BUT IS DISPENSABLE FOR BRCA-PROFICIENT CELLS, AND POLQ NULL MICE EXHIBIT NO MAJOR PHENOTYPES. RECENTLY REPORTED POLQ INHIBITORS (POLQI) SHOWED PREFERENTIAL KILLING OF BRCA-DEFICIENT CELLS. YET, THESE EARLY STAGE INHIBITORS HAVE SIGNIFICANT DEFICIENCIES (I.E. POOR ADME AND PK PARAMETERS, OR LOW POTENCY) WHICH MAY LIMIT THEIR EFFICACY IN CLINICAL STUDIES. WE HAVE DEVELOPED A NOVEL, ORAL BIOAVAILABLE POLQI THAT EXHIBITS 5.1 NM IC50 AGAINST POLQ POLYMERASE (POLQ-POL), SELECTIVELY KILLS BRCA-DEFICIENT CELLS, AND EXHIBITS BETTER PHARMACOKINETICS THAN RECENTLY PUBLISHED POLQI. OUR COLLABORATIVE TEAM (POMERANTZ, JOHNSON, AND CHILDERS LABS) PLANS TO FURTHER IMPROVE AND DEMONSTRATE PROOF-OF-PRINCIPLE OF OUR NOVEL POLQI AS THE POTENTIALLY BEST-IN-CLASS POLQI BY DEVELOPING THE FOLLOWING AIMS: 1. OPTIMIZATION OF POLQI VIA A PRODRUG STRATEGY; 2. BIOCHEMICAL AND CELLULAR CHARACTERIZATION OF POLQI; 3. IN VIVO EFFICACY OF OPTIMIZED POLQI IN BRCA-DEFICIENT SOLID TUMORS. WE EXPECT THAT SUCCESSFUL COMPLETION OF THESE AIMS WILL ENABLE THE DEVELOPMENT OF A POTENTIALLY BEST-IN-CLASS POLQI PRODRUG CANDIDATE.

BCLW IN LYMPHOMA SURVIVAL AND RESISTANCE TO TARGETED BCL2 FAMILY THERAPIES

ROLE OF PATHOGENIC PARKINSONIAN MUTATIONS IN THE SEEDING AND PROPAGATION OF ALPHA-SYNUCLEIN IN THE CNS

MOLECULAR AND CELLULAR MECHANISMS THAT UNDERLIE SYNAPTIC MATURATION

MEMBRANE TRAFFICKING IN INNATE IMMUNITY TO BACTERIAL PATHOGENS

TARGETING VISCERAL PAIN THROUGH INTESTINAL NEUROPOD CELL GUCY2C SIGNALING - VISCERAL HYPERSENSITIVITY (VH) AND PAIN IN DISORDERS OF GUT-BRAIN INTERACTION (DGBI), LIKE CONSTIPATION-TYPE IRRITABLE BOWEL SYNDROME (IBS-C) OR CHRONIC IDIOPATHIC CONSTIPATION (CIC), AFFLICTS >10% OF THE POPULATION WITH REDUCING QUALITY OF LIFE AT A COST OF ~$30B/Y IN THE U.S. THE ETIOLOGY OF VH IN DBGI IS NOT KNOWN. VH REFLECTS RECRUITMENT OF SILENT DORSAL ROOT GANGLIA (DRG) VISCERAL AFFERENTS, REDUCING THEIR THRESHOLD TO FIRE, AND INCREASING THEIR RATE OF FIRING, ACTION POTENTIALS. CURRENT ANALGESICS, INCLUDING NON-STEROIDAL ANTI-INFLAMMATORY DRUGS AND OPIATES, ARE INADEQUATE, WITH POOR EFFICACY AND SIDE EFFECTS, HIGHLIGHTING THE CLINICAL NEED FOR NOVEL THERAPEUTICS. THE INTESTINAL RECEPTOR GUANYLYL CYCLASE C (GUCY2C) BINDS COGNATE PEPTIDES AT THE EXTRACELLULAR DOMAIN THAT ACTIVATE AN INTRACELLULAR CATALYTIC DOMAIN, CONVERTING GTP TO CYCLIC GMP, THE DOWNSTREAM EFFECTOR. GUCY2C IS THE RECEPTOR FOR DIARRHEAGENIC BACTERIAL HEAT-STABLE ENTEROTOXINS (STS), AND THE HOMOLOGOUS HORMONE UROGUANYLIN IN SMALL INTESTINE AND THEIR RECEPTOR ACTIVATION INDUCES SECRETION (DIARRHEA FOR STS). THIS SECRETION IS THE BASIS FOR TREATING DBGI CONSTIPATION SYNDROMES WITH LINACLOTIDE (LINZESS™), AN ST ANALOG, OR PLECANATIDE (TRULANCE™), A UROGUANYLIN ANALOG. BEYOND SECRETION, GUCY2C AGONISTS RELIEVE PAIN IN DBGI PATIENTS AND RODENTS, REDUCING VISCERAL NOCICEPTION PRODUCED BY COLORECTAL DISTENSION (CRD). WHILE VISCERAL PAIN RELIEF BY AGONISTS IS MEDIATED BY GUCY2C, MECHANISMS, AND THEIR ROLE IN THE PATHOPHYSIOLOGY OF VH, ARE UNKNOWN. INTERESTINGLY, GUCY2C AGONISTS ARE FORMULATED FOR ACTIVITY CONFINED TO SMALL INTESTINE. OUR RECENT STUDIES REVEALED THAT DBGI PATIENTS WITH VH LOSE UROGUANYLIN, SILENCING GUCY2C IN SMALL INTESTINE. ADDITIONALLY, WE DISCOVERED THAT GUCY2C IS OVER-EXPRESSED BY SMALL INTESTINE NEUROPOD CELLS, WHICH SYNAPSE WITH DRG NEURONS CONTROLLING GUT-SPINAL CORD SIGNALING. SILENCING GUCY2C PRODUCED SPONTANEOUS VH IDENTICAL TO THAT PRODUCED BY INFLAMMATION. IN CLOSE AGREEMENT, NEUROPOD CELLS PRODUCE DRG NEURON HYPEREXCITABILITY WHICH IS ELIMINATED BY GUCY2C SIGNALING. THESE DATA SUGGEST A NOVEL ANATOMICAL HYPOTHESIS IN WHICH NEUROPOD CELLS IN SMALL INTESTINE CONTROL THE EXCITABILITY OF DRG NEURONS AFFERENT TO THE SPINAL CORD WHICH, IN TURN, INHIBIT NOCICEPTIVE SIGNALING FROM CRD. THE PATHOPHYSIOLOGICAL HYPOTHESIS SUGGESTS THAT VH IN DBGI REFLECTS UROGUANYLIN LOSS, SILENCING GUCY2C IN NEUROPOD CELLS IN SMALL INTESTINE, DISRUPTING THE CONTROL OF DRG NEURON EXCITABILITY WHICH AMPLIFIES COLORECTAL NOCICEPTION. THE THERAPEUTIC HYPOTHESIS SUGGESTS THAT GUCY2C-CGMP SIGNALING CAN BE SELECTIVELY AMPLIFIED ONLY IN NEUROPOD, BUT NOT OTHER INTESTINAL, CELLS TO SUPPRESS NEURON EXCITABILITY CONTROLLING VISCERAL PAIN WITHOUT PRODUCING SECRETION AND DIARRHEA, THE MAJOR THERAPEUTIC LIMITATION TO ORAL GUCY2C AGONISTS. PROPOSED STUDIES WILL DEFINE KEY ELEMENTS OF A NEW GUT-SPINAL CORD AXIS IN WHICH GUCY2C DRIVES NEUROTRANSMISSION BY NEUROPOD CELLS IN SMALL INTESTINE TO REGULATE NEURON EXCITABILITY CONTROLLING VISCERAL PAIN IN THE COLORECTUM. THE POTENTIAL TO TRANSLATE THESE INSIGHTS INTO NEW THERAPEUTIC PARADIGMS FOR VH TARGETING NEUROPOD CELLS TO MAXIMIZE ANALGESIA, BUT MINIMIZE DIARRHEA, IS HIGHLIGHTED BY THE AVAILABILITY OF ORAL GUCY2C AGONISTS TO TREAT CONSTIPATION.

GERIATRIC EDUCATION CENTERS

ROLE OF STX11 AND STXBP2 IN LYTIC GRANULE EXOCYTOSIS IN HEALTH AND DISEASE

THE ROLE OF THE AR INTERACTOME IN SBMA

MOLECULAR MECHANISMS OF MITOCHONDRIAL CA2+ TRANSPORT

TRAINING GRANT IN CELLULAR, BIOCHEMICAL AND MOLECULAR SCIENCES - PROJECT SUMMARY/ABSTRACT THIS APPLICATION REQUESTS SUPPORT FOR A PREDOCTORAL TRAINING PROGRAM IN CELLULAR, BIOCHEMICAL AND MOLECULAR SCIENCES (CBMS) AT THOMAS JEFFERSON UNIVERSITY. WE REQUEST 5 YEARS OF FUNDING TO SUPPORT TRAINING IN THE USE OF BIOCHEMICAL, CELLULAR AND MOLECULAR STRATEGIES TO ADDRESS IMPORTANT BIOLOGICAL QUESTIONS. THE PROGRAM COMBINES DIDACTIC COURSEWORK TO LAY CONCEPTUAL FOUNDATIONS, SEMINAR AND SMALL GROUP COURSES AND MEETINGS TO ENHANCE SCIENTIFIC COMMUNICATION, RIGOROUS, EVIDENCE-BASED, RESEARCH TRAINING WITH A DIVERSE ARRAY OF MENTORS TO ENHANCE CRITICAL THINKING SKILLS, AND CAREER DEVELOPMENT ACTIVITIES TO PROVIDE INFORMATION AND ACCESS TO A BROAD SET OF BIOMEDICAL RESEARCH CAREERS. ALL MENTORS IN THIS PROGRAM HAVE ESTABLISHED RESEARCH PROGRAMS AND SIGNIFICANT EXPERIENCE IN MENTORING. THE PROGRAM WILL BE EVALUATED THROUGH BOTH INTERNAL AND EXTERNAL REVIEW MECHANISMS TO ENSURE THAT THE TRAINEES RECEIVE THE BEST POSSIBLE TRAINING EXPERIENCE. THE CBMS TRAINING PROGRAM INCLUDES FACULTY RESEARCH MENTORS FROM 10 BASIC AND CLINICAL DEPARTMENTS (BIOCHEMISTRY & MOLECULAR BIOLOGY, CANCER BIOLOGY, MEDICAL ONCOLOGY, MEDICINE, DERMATOLOGY & CUTANEOUS BIOLOGY, MICROBIOLOGY & IMMUNOLOGY, NEUROSCIENCE, ORTHOPEDIC SURGERY, PATHOLOGY, ANATOMY & CELL BIOLOGY, AND PHARMACOLOGY & EXPERIMENTAL THERAPEUTICS) AND IS ADMINISTERED BY THE PRINCIPAL INVESTIGATORS AND ADMINISTRATIVE COMMITTEES. THE CBMS TRAINING PROGRAM IS FOCUSED ON DELIVERING BROAD, CROSS-DISCIPLINARY TRAINING IN THE MOLECULAR REGULATION OF CELLULAR FUNCTION AND DYSFUNCTION IN A SAFE, DIVERSE, AND INCLUSIVE ENVIRONMENT TO PRODUCE SCIENTISTS WITH STRONG EXPERTISE IN CRITICAL SCIENTIFIC REASONING, RIGOROUS AND REPRODUCIBLE EXPERIMENTAL DESIGN, QUANTITATIVE APPROACHES, AND EXCELLENCE IN DATA ANALYSIS AND INTERPRETATION. OVERALL, THE CBMS TRAINING PROGRAM WILL PROVIDE OUTSTANDING CROSS-DISCIPLINARY PREDOCTORAL TRAINING IN THE USE OF MECHANISTIC APPROACHES TO ADDRESS IMPORTANT SCIENTIFIC QUESTIONS IN HEALTH AND DISEASE. CAREER DEVELOPMENT PROGRAMS AND PLANS TO ENHANCE DIVERSITY WITHIN THE BIOMEDICAL RESEARCH WORKFORCE WILL NOT ONLY ENHANCE THE CBMS TRAINING PROGRAM BUT WILL ENRICH AND ELEVATE THE OVERALL PREDOCTORAL TRAINING ENVIRONMENT AT THOMAS JEFFERSON UNIVERSITY.

INTEGRIN-MEDIATED MECHANISMS OF PROSTATE CANCER PROGRESSION

PROTEOGLYCAN REGULATION OF TUMOR ANGIOGENESIS AND ENDOTHELIAL CELL AUTOPHAGY

REGENERATION OF NUCLEUS PULPOSUS BY TISSUE ENGINEERING

NONINVASIVE PORTAL PRESSURE MEASUREMENTS IN CHILDREN