Rosalind Franklin University Of Medicine & Science

MODULATION OF THE BNST ACTIVITY BY OXYTOCIN - ROLE IN STRESS, FEAR AND ANXIETY

SUSTAINED V1A RECEPTOR ACTIVATION FOR PROLONGED HEMODYNAMIC SUPPORT AND NEUROLOGICAL PROTECTION AFTER NONCOMPRESSIBLE HEMORRHAGE AND TRAUMATIC BRAIN

STRUCTURE AND FUNCTION OF THE YEAST ATPASE

URINARY PROTEOMIC BIOMARKERS OF ADAPTIVE BONE FORMATION AND STRESS INJURIES IN MILITARY RECRUITS.

REPROGRAMMING CELL FATE FOR REPAIR

MOLECULAR CLONING OF EPITHELIAL K CHANNELS

PARALLEL MATURATION OF SOCIAL BEHAVIORS AND AMYGDALA CIRCUITS

HIV INFECTION AND LATENCY IN ASTROCYTES

EFFECTS OF CHRONIC STRESS ON AMYGDALA PHYSIOLOGY

MOLECULAR BASIS OF MULTIDRUG BINDING AND TRANSPORT BY THE MATE TRANSPORTERS

MIR-132, A NEW PLAYER IN HIV/NEUROAIDS

NEURAL SUBSTRATE OF COGNITIVE AND EMOTIONAL DEFICITS IN PSYCHOPATHY

MOLECULAR MODULATOR OF RPA AND RAD51 IN MAINTAINING GENOME STABILITY

A LONGITUDINAL AND EXPERIENCE SAMPLING INVESTIGATION OF REJECTION SENSITIVITY AND ITS ROLE IN SEXUAL MINORITY ADOLESCENTS' MENTAL HEALTH - PROJECT SUMMARY SEXUAL MINORITY ADOLESCENTS (SMA) ARE DISPROPORTIONATELY AFFECTED BY MENTAL HEALTH PROBLEMS SUCH AS DEPRESSION AND SUICIDALITY RELATIVE TO THEIR HETEROSEXUAL PEERS. THESE DISPARITIES ARE GREATEST DURING ADOLESCENCE AND PERSIST INTO ADULTHOOD, AND ARE GENERALLY ATTRIBUTED TO THE UNIQUE INTERPERSONAL STRESSORS THAT THEY FACE (E.G., SEXUAL ORIENTATION-RELATED REJECTION). GIVEN THAT REJECTION IS A ROBUST RISK FACTOR FOR MENTAL HEALTH PROBLEMS, ESPECIALLY DURING ADOLESCENCE, IT HAS HISTORICALLY BEEN THE FOCUS OF RESEARCH ON SM INDIVIDUALS' MENTAL HEALTH. HOWEVER, IN ADDITION TO EXPERIENCES OF REJECTION, EXPECTATIONS OF REJECTION AND THE EMOTIONS THAT ACCOMPANY THEM ALSO PLAY IMPORTANT ROLES IN MENTAL HEALTH. THE REJECTION SENSITIVITY (RS) THEORY PROPOSES THAT EARLY EXPERIENCES OF REJECTION CAN LEAD TO A DISPOSITION TO ANXIOUSLY EXPECT, READILY PERCEIVE, AND INTENSELY REACT TO REJECTION, WHICH CAN THEN COMPROMISE ONE'S MENTAL HEALTH. DESPITE EVIDENCE THAT RS IS ASSOCIATED WITH A RANGE OF MENTAL HEALTH PROBLEMS AMONG SM ADULTS, OUR UNDERSTANDING OF ITS DEVELOPMENT AND ROLE IN SMA'S MENTAL HEALTH IS LIMITED IN SEVERAL IMPORTANT WAYS (E.G., NEARLY ALL PRIOR STUDIES HAVE BEEN CROSS- SECTIONAL STUDIES OF ADULTS, AND THERE HAS BEEN A LACK OF ATTENTION TO DIFFERENT EMOTIONS THAT CAN ACCOMPANY EXPECTATIONS OF REJECTION AS WELL AS UNDERLYING MECHANISMS AND PROTECTIVE FACTORS). GIVEN THESE GAPS, THE GOALS OF THE PROPOSED R01 ARE TO DETERMINE THE ROLE OF SEXUAL ORIENTATION-RELATED REJECTION IN THE DEVELOPMENT OF DIFFERENT TYPES OF RS AND THEIR RESPECTIVE MENTAL HEALTH CONSEQUENCES AMONG SMA, TO IDENTIFY THE MECHANISMS THROUGH WHICH RS INFLUENCES MENTAL HEALTH IN THIS POPULATION, AND TO IDENTIFY PROTECTIVE FACTORS IN THE DEVELOPMENT OF RS. WE WILL RECRUIT A COHORT OF 500 SMA (AGES 14-17) TO PARTICIPATE IN A LONGITUDINAL AND EXPERIENCE-SAMPLING STUDY USING METHODS ESTABLISHED IN OUR TEAM'S PRIOR WORK. DATA WILL BE COLLECTED AT FOUR BIANNUAL ASSESSMENTS (BASELINE, 6-, 12-, 18-MONTHS) TO DETERMINE THE LONGITUDINAL RELATIONS AMONG SEXUAL ORIENTATION-RELATED REJECTION, RS (ANXIOUS AND ANGRY), AND MENTAL HEALTH (DEPRESSION SYMPTOMS AND CLINICAL ELEVATIONS, SUICIDAL IDEATION, AND REACTIVE AGGRESSION). IN ADDITION, PARTICIPANTS WILL COMPLETE A 6- WEEK EXPERIENCE-SAMPLING STUDY (4 ASSESSMENTS PER DAY, ADMINISTERED IN TWO 3-WEEK BURSTS) TO EXAMINE ANTECEDENTS AND CONSEQUENCES OF RS AS THEY OCCUR IN DAILY LIFE. THESE DATA WILL BE USED TO ACCOMPLISH THREE SPECIFIC AIMS: (1) DETERMINE THE LONGITUDINAL AND DAILY RELATIONS AMONG SEXUAL ORIENTATION-RELATED REJECTION, RS (ANXIOUS AND ANGRY), AND MENTAL HEALTH (DEPRESSION, SUICIDAL IDEATION, AND REACTIVE AGGRESSION); (2) IDENTIFY MECHANISMS (SOCIAL WITHDRAWAL, SELF- AND OTHER-BLAME IN RESPONSE TO REJECTION, AND SEXUAL ORIENTATION CONCEALMENT) UNDERLYING THE LONGITUDINAL AND DAILY RELATIONS BETWEEN RS AND MENTAL HEALTH PROBLEMS; AND (3) EXAMINE SEXUAL ORIENTATION-RELATED SOCIAL SUPPORT AND ACCEPTANCE AS PROTECTIVE FACTORS IN THE DEVELOPMENT OF RS. THE PROPOSED R01 WILL ADVANCE OUR UNDERSTANDING OF THE DEVELOPMENT OF RS, ITS CONSEQUENCES, THE UNDERLYING MECHANISMS, AND PROTECTIVE FACTORS, WHICH WILL INFORM INTERVENTIONS TO IMPROVE SMA'S MENTAL HEALTH.

STRUCTURE AND MECHANISM OF THE MITOCHONDRIAL ATP SYNTHASE AND BATTEN DISEASE GENE PRODUCT, CLN3P

POSITIVE ALLOSTERIC MODULATOR PNU-120596 TO TREAT HIV-ASSOCIATED NEUROCOGNITIVE DISORDERS - ABSTRACT HIV INFECTION OF THE CENTRAL NERVOUS SYSTEM (CNS) OCCURS IN A MAJORITY OF HIV-INFECTED INDIVIDUALS. IT CAUSES HIV- ASSOCIATED NEUROCOGNITIVE DISORDERS (HAND) EVEN IN THE ERA OF COMBINATION ANTIRETROVIRAL THERAPY (CART). THERE ARE CURRENTLY NO THERAPIES FOR HAND. WE HAVE FOCUSED ON HIV INFECTION AND PATHOGENESIS IN THE CNS OVER THE PAST 25 YEARS. OUR STUDIES HAVE HELPED ESTABLISH CELL-FREE AND CELL-CELL CONTACT-MEDIATED HIV INFECTION OF ASTROCYTES AND A CRITICAL ROLE OF HIV TAT PROTEIN IN HIV NEUROPATHOGENESIS. THE LATTER INCLUDES THE CREATION OF DOXYCYCLINE-INDUCIBLE BRAIN-SPECIFIC HIV TAT TRANSGENIC MICE (ITAT) AS AN IMPORTANT SURROGATE HAND MODEL. WE HAVE RECENTLY UNCOVERED THAT ACTIVATION OF Α7 NICOTINIC ACETYLCHOLINE RECEPTOR (NACHR) BY ITS POSITIVE ALLOSTERIC MODULATOR (PAM) PNU-120596 AMELIORATES HAND NEUROLOGY AND NEUROPATHOLOGY USING THE ITAT MODEL, SUGGESTING THE THERAPEUTIC POTENTIAL OF Α7 NACHR PAM AGAINST HAND. THESE STUDIES ARE SIGNIFICANT BECAUSE TAT PROTEIN CONTINUES TO BE DETECTED IN THE BRAIN OF HIV-INFECTED INDIVIDUALS TREATED WITH CART. IN ADDITION, WE HAVE SHOWN THAT Α7 NACHR PAM INHIBITS HIV REPLICATION AND REACTIVATION. THESE FINDINGS UNDOUBTEDLY CALL FOR FURTHER CHARACTERIZATION OF THE MECHANISMS OF Α7 NACHR PAM-MEDIATED NEUROPROTECTION AGAINST TAT NEUROTOXICITY AND THE MECHANISMS BY WHICH Α7 NACHR PAM ACTIVATION LEADS TO INHIBITION OF HIV REPLICATION AND REACTIVATION. THE CENTRAL HYPOTHESIS FOR THIS PROPOSAL IS THAT Α7 NACHR PAM HOLD GREAT PROMISE AS CART ADJUNCTIVE THERAPEUTICS TO TREAT HIV NEUROPATHOGENESIS AND HAND. TO TEST THIS HYPOTHESIS, WE PROPOSE THE FOLLOWING THREE INTERRELATED SPECIFIC AIMS: (1) TO FURTHER CHARACTERIZE THE MECHANISMS OF Α7 NACHR/PAM-MEDIATED NEUROPROTECTION AGAINST TAT NEUROTOXICITY; (2) TO DEFINE THE MECHANISMS OF Α7 NACHR/PAM-MEDIATED INHIBITION OF HIV REPLICATION AND LATENT HIV REACTIVATION; AND (3) TO DETERMINE THE IN VIVO EFFECTS OF Α7 NACHR/PAM ACTIVATION ON HIV NEUROPATHOGENESIS AND HIV REPLICATION AND LATENCY IN THE CNS AND PERIPHERY. WE WILL USE A COMBINED MOLECULAR, CELLULAR, BIOCHEMICAL, AND GENETIC APPROACH, INCLUDING THE USE OF PRIMARY MOUSE AND HUMAN CULTURES, ITAT MICE AND Α7 NACHR KNOCKOUT MICE, THE INFECTIOUS AND REPLICATION-COMPETENT ECOHIV MODEL, AND SINGLE NUCLEUS ADVANCED GENOMICS TECHNOLOGIES. THE ANSWERS SOUGHT HAVE FUNDAMENTAL SIGNIFICANCE FOR OUR UNDERSTANDING OF NOT ONLY THIS CRITICAL AND PERVASIVE PROTEIN HIV TAT AND ITS ROLE IN HIV NEUROPATHOGENESIS BUT ALSO THE MOLECULAR MECHANISMS OF HIV TRANSCRIPTION INHIBITION BY Α7 NACHR/PAM ACTIVATION. THE FINDINGS FROM THE PROPOSED STUDIES ARE EXPECTED TO PROVIDE MECHANISTIC AND PRE-CLINICAL EVIDENCE TO SUPPORT THE USE OF Α7 NACHR PAM AS HAND THERAPEUTICS AND INFORM A NEW INTERVENTION STRATEGY FOR HIV NEUROPATHOGENESIS. THE ENORMOUS AMOUNT OF INFORMATION AVAILABLE ON HIV TAT, HIV INFECTION AND PATHOGENESIS OF THE CNS, AND THE ROLES OF Α7 NACHR/PAM IN OTHER NEURODEGENERATIVE DISEASES, AND THE RESULTS OBTAINED FROM OUR RECENT STUDIES MAKE THE ACCOMPLISHMENT OF THESE AIMS PRACTICAL.

ROLE OF NON-MYELOID CELLS' PYROPTOSIS IN MELIOIDOSIS - ACTIVATION OF CASPASE-1 AND CASPASE-11 IN THE CONTEXT OF INFLAMMASOMES LEADS TO PROCESSING AND SECRETION OF IL-1SS AND IL-18 AND TRIGGERS A LYTIC CELL DEATH TERMED PYROPTOSIS. INFLAMMASOME ACTIVATION AND PYROPTOSIS HAVE BEEN SHOWN TO PROTECT THE HOST FROM INFECTION WITH A RANGE OF PATHOGENS INCLUDING BURKHOLDERIA PSEUDOMALLEI, A GRAM-NEGATIVE BACTERIUM USED AS A MOUSE MODEL OF MELIOIDOSIS. HOWEVER, IT IS ALSO BECOMING CLEAR THAT INFLAMMASOME ACTIVATION AND PYROPTOSIS CAN BE DELETERIOUS TO THE HOST BY TRIGGERING LETHAL EICOSANOID STORM, MEDIATING LETHAL ENDOTOXIC SHOCK, AND CAUSING DISSEMINATED INTRAVASCULAR COAGULATION (DIC). WE HAVE RECENTLY SHOWN THAT THE ONLY FUNCTIONAL INFLAMMASOME EXPRESSED BY EPITHELIAL AND ENDOTHELIAL CELLS IS CASPASE-11, AND THAT IN A MODEL OF MELIOIDOSIS CASPASE-11-DEPENDENT PYROPTOSIS OF LUNG EPITHELIAL CELLS IS PROTECTIVE. BASED ON OUR PRELIMINARY RESULTS AND THE PUBLISHED LITERATURE WE HYPOTHESIZE THAT ACTIVATION OF CASPASE-11 IN EPITHELIAL CELLS WILL PROTECT THE HOST FROM PNEUMONIC MELIOIDOSIS THROUGH INDUCTION OF PYROPTOSIS AND PREFERENTIAL PRODUCTION OF ANTI-INFLAMMATORY EICOSANOIDS LIKE PROSTAGLANDIN E2. IN CONTRAST, CASPASE-11 ACTIVATION IN ENDOTHELIAL CELLS MAY PROVE DELETERIOUS BECAUSE IT MAY DAMAGE BARRIER INTEGRITY, INDUCE DIC AND EXCESSIVE INFLAMMATION THROUGH RELEASE OF TISSUE FACTOR AND LTB4. THUS, STUDYING THE FUNCTION OF CASPASE-11 SELECTIVELY IN EPITHELIAL OR ENDOTHELIAL CELLS WILL ALLOW US TO IDENTIFY THE DELETERIOUS AND THE BENEFICIAL RESPONSES AND THEREFORE POTENTIALLY INDICATE THERAPEUTIC AVENUES TO INHIBIT THE FORMER AND ENCOURAGE THE LATTER. IN AIM 1 WE WILL FOCUS ON THE ROLE OF CASPASE-11 IN EPITHELIAL CELLS. WE WILL TEST THE ROLE OF CASPASE-11 AND GASDERMIN D IN THE PRODUCTION OF EICOSANOIDS BY LUNG EPITHELIAL CELLS. WE WILL ALSO TEST THE SUSCEPTIBILITY TO MELIOIDOSIS OF MICE THAT LACK EXPRESSION OF CASPASE-11 IN LUNG EPITHELIAL CELLS AND DETERMINE THE ROLE OF EPITHELIAL-DERIVED EICOSANOIDS IN MELIOIDOSIS. IN AIM 2 WE WILL STUDY THE ROLE OF CASPASE-11 IN ENDOTHELIAL CELLS. WE WILL EXAMINE HOW CASPASE-11 CONTROLS PRODUCTION OF EICOSANOIDS AND RELEASE OF TISSUE FACTOR IN ENDOTHELIAL CELLS. WE WILL TEST THE SUSCEPTIBILITY TO MELIOIDOSIS OF MICE THAT LACK EXPRESSION OF CASPASE-11 IN ENDOTHELIAL CELLS AND ESTABLISH THE ROLE OF TISSUE FACTOR, PLATELET ACTIVATING FACTOR, AND DIC IN THE PATHOGENESIS OF MELIOIDOSIS.

ROLE OF NEUTROPHILS AND ELASTASE IN MELIOIDOSIS

MOLECULAR REGULATION OF TLR2-MEDIATED AUTOIMMUNE INFLAMMATION

NOVEL DRUG DISCOVERY FOR AD TARGETING RYANODINE CALCIUM CHANNELS

AMYGDALA AND PAIN-ASSOCIATED MOOD DISORDERS

SEX DIFFERENCES IN AMYGDALA FUNCTION

MECHANISMS OF STIMULUS-INDUCED NETWORK FOCUSING - PROJECT SUMMARY LARGE-SCALE RECORDINGS ARE DISCOVERING THAT INDIVIDUAL NEURONS IN SENSORY, COGNITIVE, AND MOTOR NETWORKS OFTEN PARTICIPATE VARIABLY, EVEN WHEN PRESENTED WITH IDENTICAL INPUTS. THE REASON FOR SUCH VARIABILITY IS UNCLEAR, AND AN ACTIVE TOPIC OF DEBATE IN THE FIELD. DOES IT REFLECT RANDOMNESS IN NEURONAL PARTICIPATION, OR IS IT AN ADAPTIVE FEATURE THAT PLAYS AN ESSENTIAL ROLE IN HEALTHY BRAIN FUNCTION? THE SCIENTIFIC PREMISE OF THIS APPLICATION IS THE LATTER—THAT VARIABLY PARTICIPATING NEURONS REFLECT THE OPERATION OF A “FOCUSING” MECHANISM INNATE TO MANY NETWORKS THAT ALLOWS THEM TO RAPIDLY AND FLEXIBLY REARRANGE WHICH NEURONS ARE CALLED UPON TO PROCESS SPECIFIC INFORMATION IN THE CONTEXT OF THE MOMENT. THIS HYPOTHESIS EMERGED UNEXPECTEDLY FROM OUR LARGE-SCALE RECORDINGS OF THE RHYTHMIC ESCAPE SWIM NETWORK OF THE MARINE MOLLUSK TRITONIA DIOMEDEA. WE WERE SURPRISED TO DISCOVER THAT DURING THE INITIAL SECONDS OF RESPONDING TO AN UNEXPECTED AVERSIVE SENSORY INPUT, TRITONIA'S SWIM MOTOR PROGRAM RAPIDLY TUNES ITSELF, PULLING MANY INITIALLY-SILENT NEURONS INTO THE BURSTING POPULATION AND DRIVING OTHERS OUT, APPARENTLY OPTIMIZING ITSELF FOR ESCAPE. IN THIS TRITONIA CASE, THE “FOCUSED” STATE IS THEN MAINTAINED FOR SEVERAL MINUTES, ENABLING A STRONGER, FASTER-ONSET MOTOR PROGRAM SHOULD THE SAME STIMULUS RECUR. MANY STUDIES IN VERTEBRATES HAVE REPORTED RAPID GROWTH IN THE SIZE OF RESPONDING NETWORKS WITH REPEATED STIMULATION, BUT THE MECHANISMS AND PURPOSE OF SUCH PHENOMENA ARE POORLY UNDERSTOOD. THIS PROJECT'S GOAL IS TO UNCOVER THE MECHANISMS UNDERLYING WHAT MAY BE AN IMPORTANT VERSATILITY PROCESS FOR HEALTHY FUNCTION IN MANY BRAIN NETWORKS—ONE THAT ALLOWS THEM TO RAPIDLY RE-ALLOCATE NEURONS TO SUIT A SPECIFIC CONTEXT, AND THEN HOLD THAT FOCUSED STATE FOR A SUSTAINED PERIOD OF TIME. THE PROJECT HAS 2 SPECIFIC AIMS: AIM 1 WILL MAP THE RE-ALLOCATING NEURONS AND ADDRESS SEVERAL ISSUES REGARDING THE PHENOMENOLOGY OF THIS POORLY UNDERSTOOD NETWORK FOCUSING PROCESS. AIM 2 WILL DETERMINE THE CELLULAR MECHANISMS DRIVING THE RAPID RE- ALLOCATION OF NEURONS INTO AND OUT OF THE BURSTING ESCAPE SWIM NETWORK AS IT FOCUSES. THE PRINCIPLES OF RAPID NETWORK FOCUSING TO BE INVESTIGATED HERE MAY PROMOTE NOVEL APPROACHES FOR TREATING OR PREVENTING DECLINES IN COGNITIVE FUNCTION IN AGING AND DISEASE.

THE CONTRIBUTION OF PERIVASCULAR ADIPOSE TISSUE MACROPHAGES TO MICROVASCULAR DYSFUNCTION IN OBESITY

¿¿INTRAOSSEOUS ERYTHROPOIETIN FOR ACUTE TISSUE PROTECTION IN BATTLEFIELD CASUALTIES SUFFERING HYPOVOLEMIC SHOCK"

LIPOXINS AND ASPIRIN TRIGGERED LIPOXINS IN KSHV LATENCY AND PATHOGENESIS

MOLECULAR BASIS OF COATS PLUS DISEASE - THE COATS PLUS SYNDROME IS A RARE AND LIFE-THREATENING GENETIC DISORDER CHARACTERIZED BY MULTI-SYSTEM DEVELOPMENTAL DEFECTS THAT LEAD TO BILATERAL EXUDATIVE RETINOPATHY, RETINAL TELANGIECTASIAS, GROWTH RETARDATION, INTRACRANIAL CALCIFICATIONS, BONE ABNORMALITIES, GASTROINTESTINAL VASCULAR ECTASIAS, AND COMMON EARLY-AGING PATHOLOGICAL FEATURES. LIKE MANY OTHER DEVELOPMENTAL DISORDERS, COATS PLUS IS CAUSED BY DEFECTS IN GENES INVOLVED IN MAINTAINING GLOBAL GENOME INTEGRITY. SPECIFICALLY, IT IS CAUSED BY LOSS-OF-FUNCTION MUTATIONS IN THE HUMAN CTC1/STN1/TEN1 (CST) COMPLEX, WHICH IS A TRIMERIC COMPLEX THAT PREFERENTIALLY BINDS TO G-RICH SSDNA OR SS-DS DNA JUNCTIONS AND IS CRITICAL FOR PREVENTING GENOME INSTABILITIES ARISING FROM REPLICATION PERTURBATION. WE HOPE TO AID IN BETTER UNDERSTANDING OF DISEASE DEVELOPMENT AND DESIGNING OF EFFECTIVE THERAPEUTIC STRATEGIES BY INVESTIGATING THE MECHANISMS GOVERNING GENOME STABILITY UNDER REPLICATION STRESS. IN RESPONSE TO FORK STALLING, SIGNALING CASCADES ACTIVATE MULTIPLE PATHWAYS INCLUDING FORK REVERSAL, TRANSLESION SYNTHESIS, REPRIMING DOWNSTREAM OF STALLED SITES, AND DORMANT ORIGIN FIRING TO RESCUE STALLED REPLICATION. ACTIVITIES OF THESE PATHWAYS NEED TO BE TIGHTLY REGULATED TO ENSURE REPLICATION FIDELITY. THE OBJECTIVES OF THIS PROPOSAL IS TO DELINEATE A NOVEL SIGNALING PATHWAY IN RESPONSE TO REPLICATION STRESS, ELUCIDATE HOW IT REGULATES PROTEIN INTERPLAYS AND RECRUITMENT AT STALLED FORKS, AND UNDERSTAND THE MECHANISM REGULATING THE REPRIMING PATHWAY. IN AIM 1, WE HYPOTHESIZE THAT A CALCIUM-DEPENDENT SIGNALING PATHWAY PHOSPHORYLATES STN1 TO ACTIVATE CST AT STALLED FORKS TO PROTECT THE STABILITY OF STALLED FORKS. WE WILL ELUCIDATE THIS NEW SIGNALING PATHWAY AND DETERMINE HOW THIS PATHWAY ANTAGONIZES UNSCHEDULED NASCENT STRAND DNA DEGRADATION AND REGULATES FORK PROTECTION. IN AIM 2, WE WILL INVESTIGATE HOW THIS SIGNALING PATHWAY REGULATES THE INTERPLAY OF SINGLE-STRAND DNA BINDING PROTEINS AT FORKS AND OTHER FORK BINDING PROTEINS. IN AIM 3, WE WILL INVESTIGATE THE MECHANISM FOR RESTRICTING EXCESSIVE REPRIMING TO PREVENT SSDNA GAP FORMATION AND GENOME INSTABILITY. WE WILL COMBINE HIGHLY SENSITIVE CELL-BASED ANALYSES, SINGLE-MOLECULE AND POWERFUL BIOCHEMICAL ASSAYS TO ACCOMPLISH THE GOALS OF THE PROPOSED RESEARCH. WE EXPECT THAT OUR EFFORTS WILL IDENTIFY NEW FACTORS AND PATHWAYS REGULATING THE RESCUE OF STALLED REPLICATION AND THE PRESERVATION OF GENOME STABILITY.

MECHANISMS UNDERLYING STRESS RESILIENCE

STRUCTURAL AND FUNCTIONAL ANALYSIS OF GLUCOSE TRANSPORTERS

INTRACELLULAR ORGANELLE DEFICITS DRIVING ALZHEIMER'S DISEASE

THE ROLE OF INHIBITORY MICROCIRCUITS IN THE NEURAL CONTROL OF BREATHING

SEROTONIN RECEPTORS THAT POTENTIATE ADDICTION-RELATED BEHAVIORAL AND MOLECULAR EFFECTS INDUCED BY METHYLPHENIDATE PLUS SSRI EXPOSURE

STRUCTURAL STUDIES ON PROKARYOTIC POTASSIUM CHANNELS

ROLE OF NLRP3-INFLAMMASOME IN ALUM'S ADJUVANTICITY

INTERCELLULAR COMMUNICATION IN THE LENS

MYOGENIC CELL LINEAGES AND MUSCLE FIBER TYPE FORMATION

TARGETING SMN2 ALTERNATIVE SPLICING FOR THE TREATMENT OF SPINAL MUSCULAR ATROPHY

ROLE OF NEUREXIN IN SOCIAL ADAPTATION AND AMYGDALA PLASTICITY - PROJECT SUMMARY SOCIAL RELATIONSHIPS BENEFIT MENTAL AND PHYSICAL HEALTH, WHILE POOR QUALITY SOCIAL INTERACTIONS ARE ASSOCIATED WITH WORSE HEALTH AND HIGHER MORTALITY. THE ABILITIES THAT UNDERLIE SOCIAL FUNCTION ARE IMPAIRED IN AUTISM SPECTRUM DISORDER (ASD). THERE HAS BEEN SUBSTANTIAL PROGRESS IN IDENTIFYING GENES THAT ARE ASSOCIATED ASD. BUT THE LINK BETWEEN ASD GENETIC FACTORS AND CORE SOCIAL DISRUPTIONS IS UNCLEAR. EVEN IN NON-ASD INDIVIDUALS IT IS NOT CLEAR HOW THESE GENES INFLUENCE SOCIAL BEHAVIOR. TO UNDERSTAND HOW THESE GENES INFLUENCE SOCIAL BEHAVIOR, WE MUST UNCOVER HOW THEY IMPACT THE NEURAL CIRCUITS THAT UNDERLIE SOCIAL FUNCTION. MANY KEY SOCIAL FUNCTIONS ARE TRACED TO A NETWORK OF CONNECTED BRAIN REGIONS THAT IS SIMILAR IN RODENTS AND PRIMATES. THIS PRESENTS AN OPPORTUNITY TO DETERMINE THE IMPACT OF HIGH-RISK ASD GENES ON THE FUNCTION OF KEY CONNECTIONS WITHIN THIS SOCIAL NETWORK. PRIOR RESEARCH DEMONSTRATES THAT PREFRONTAL CORTICAL (PFC) CONNECTIONS TO AMYGDALA, PARTICULARLY ANTERIOR CINGULATE (ACC) EXCITATORY PROJECTIONS TO BASOLATERAL AMYGDALA (BLA), PLAY A FUNDAMENTAL ROLE IN ENCODING SOCIAL INFORMATION AND MOTIVATING SOCIAL BEHAVIORS. SOCIAL BEHAVIOR MUST BE FLEXIBLE IN RESPONSE TO SHIFTS OF EXTRINSIC CONSPECIFIC BEHAVIOR AND INTRINSIC SOCIAL DRIVE. ACC-BLA GUIDES BEHAVIORAL ADAPTATIONS DURING THESE SHIFTS. THIS BEHAVIORAL FLEXIBILITY LIKELY REQUIRES SYNAPTIC PLASTICITY. NRXN1 (NRXN1 IN RODENTS) IS A HIGH CONFIDENCE ASD GENE, AND NRXN1 DISRUPTION REPLICABLY IMPAIRS SOCIAL BEHAVIORS IN RODENTS AND WEAKENS PFC-BLA SYNAPSES. THERE IS MUCH KNOWN ABOUT THE IMPORTANCE OF NEUREXIN 1 PROTEIN IN SYNAPTIC FUNCTIONS, BUT IT IS NOT UNDERSTOOD WHY NRXN1 DISRUPTION LEADS TO SOCIAL IMPAIRMENTS. IN THIS PROJECT WE PROPOSE THAT NEUREXIN 1 AT PFC-BLA SYNAPSES IS POSITIONED TO INFLUENCE SYNAPTIC PLASTICITY DURING SOCIAL EXPERIENCE. A GOAL OF THIS RESEARCH IS TO DETERMINE IF NEUREXIN 1 IMPACTS RODENT SOCIAL BEHAVIORS BY PREFERENTIAL MODULATION OF ACC-BLA SYNAPSES. WE HYPOTHESIZE THAT NRXN1 HAS CIRCUIT-SPECIFIC ACTIONS ON ACC-BLA AND, THROUGH THIS, NRXN1 CONTRIBUTES TO FLEXIBILITY OF SOCIAL BEHAVIOR AND ACC-BLA PLASTICITY. THIS WILL BE TESTED USING A COMBINATION OF ACUTE INTERFERENCE WITH NEUREXIN 1, GENE KNOCKOUTS AND CIRCUIT-SPECIFIC KNOCKDOWN IN RATS, WHILE MEASURING SOCIAL BEHAVIORS THAT RELY ON PFC-BLA INTEGRITY, BLA RESPONSES TO SOCIAL STIMULI, AND PFC-BLA SYNAPTIC FUNCTION. THESE STUDIES WILL PROVIDE US WITH A NEW UNDERSTANDING OF THE ROLE OF NEUREXIN 1 IN MODULATION OF A PFC-BLA CIRCUIT THAT IS REQUIRED FOR SOCIAL FLEXIBILITY. THIS NOVEL INFORMATION WOULD HELP BRIDGE A KNOWN GENETIC SOCIAL INFLUENCER WITH THE NEUROBIOLOGY OF SOCIAL FUNCTION.

EARLY EVENTS OF IN VITRO KSHV INFECTION

REGULATION OF VOLUME IN THE LENS

CARES ACT FUNDING OVERSIGHT-FINANCE DEPARTMENT

DOPAMINE AND GLUTAMATE RECEPTOR INTERACTIONS

ANTIGENS OF KAPOSI'S SARCOMA ASSOCIATED HERPESVIRUS

ROLE OF LMTK2 IN CFTR TRAFFICKING

BAX/BAK-MEDIATED MITOCHONDRIAL OUTER MEMBRANE PERMEABILIZATION IN APOPTOSIS

MECHANISMS OF NUCLEAR BODY BIOGENESIS

A GENETIC STUDY OF BK CHANNEL TRAFFICKING AND REGULATION IN C. ELEGANS

NEURONAL CALCIUM DYSREGULATION IN AGING AND ALZHEIMER'S DISEASE

MECHANISM OF OXYGEN SENSING BY CHEMORECEPTOR CELLS

CARES ACT FUNDING OVERSIGHT - FINANCE DEPARTMENT

IDENTIFICATION OF SPLICE VARIANTS AS MOLECULAR MARKERS IN PARKINSON'S DISEASE

MOLECULAR BASIS OF SUBSTRATE TRANSLOCATION IN THE DRUG/H+ ANTIPORTER 1 FAMILY - MOLECULAR BASIS OF SUBSTRATE TRANSLOCATION IN THE DRUG/H+ ANTIPORTER 1 FAMILY SUMMARY INTEGRAL MEMBRANE PROTEINS KNOWN AS MULTIDRUG TRANSPORTERS EXTRUDE THERAPEUTIC DRUGS OF DIVERSE CHEMICAL STRUCTURES ACROSS CELL MEMBRANES, IMPEDING THE TREATMENT OF HUMAN CANCERS, INFECTIOUS DISEASES, AND NEUROLOGICAL DISORDERS. CURRENTLY WE LACK A DEEP AND MECHANISTIC UNDERSTANDING OF HOW THESE PROTEINS EXPORT DRUGS OR HOW THEY CAN BE THWARTED. WE WILL STUDY THE STRUCTURE AND MECHANISM OF A MODEL MULTIDRUG TRANSPORTER, MDFA FROM ESCHERICHIA COLI, WHICH COUPLES THE INFLUX OF H+ TO THE EFFLUX OF VARIOUS ANTIMICROBIALS AND BELONGS TO THE UBIQUITOUS DRUG/H+ ANTIPORTER 1 (DHA1) FAMILY. MDFA ORTHOLOGUES ARE PRESENT IN MANY PATHOGENIC MICROORGANISMS, AND THE OVEREXPRESSION OF E. COLI MDFA CAN LEAD TO ANTIMICROBIAL RESISTANCE IN CLINICAL PATIENTS. THUS, MDFA REPRESENTS AN IMPORTANT TARGET FOR THERAPEUTIC EXPLOITATION TO OVERCOME MULTIDRUG RESISTANCE. FURTHERMORE, THE SLC18 ANTIPORTERS, WHICH ARE THE HUMAN COUNTERPARTS OF MDFA IN THE DHA1 FAMILY, CONDUCT THE H+-DEPENDENT VESICULAR TRANSPORT OF MONOAMINE NEUROTRANSMITTERS, POLYAMINE NEUROMODULATORS, AND NEUROTOXINS. THE HUMAN SLC18 ANTIPORTERS ARE ESSENTIAL FOR BRAIN FUNCTION AND PROMISING THERAPEUTIC TARGETS FOR BATTLING ALCOHOLISM, AUTISM SPECTRUM DISORDERS, BIPOLAR DISORDER, HUNTINGTON DISEASE, MAJOR DEPRESSIVE DISORDER, PARKINSON’S DISEASE, SCHIZOPHRENIA, AND TOURETTE SYNDROME. OUR LONG-TERM OBJECTIVE IS TO UNDERSTAND HOW THE DHA1 MULTIDRUG TRANSPORTERS AND HUMAN SLC18 ANTIPORTERS TRANSLOCATE THEIR SUBSTRATES AND HOW THEIR FUNCTION CAN BE MODULATED FOR POTENTIAL THERAPEUTIC BENEFIT. NOTABLY, PRIOR BIOCHEMICAL STUDIES HAVE SUGGESTED THAT MDFA TRANSLOCATES CERTAIN SUBSTRATES VIA A NON-CANONICAL MECHANISM. DRAWING UPON THESE DATA AND OUR EXPERIENCE IN MEMBRANE PROTEIN STRUCTURAL BIOLOGY, WE WILL ACCOMPLISH TWO AIMS: (1) TO ELUCIDATE THE MOLECULAR BASIS FOR SIMULTANEOUS TRANSLOCATION OF TWO MONO-CATIONIC SUBSTRATES BY A DHA1; (2) TO REVEAL THE STRUCTURAL MECHANISM FOR NON-CANONICAL, DHA1- MEDIATED EXTRUSION OF DI-CATIONIC THERAPEUTICS. BY COMBINING CRYSTALLOGRAPHIC AND BIOCHEMICAL STUDIES, WE WILL ACQUIRE NEW INSIGHTS INTO HOW A DHA1 TRANSLOCATES TWO SUBSTRATES CONCURRENTLY, HOW A DHA1 INHIBITOR DIFFERS FROM THE SUBSTRATE, AND HOW A DHA1 EXPORTS A THERAPEUTIC DRUG IN TWO CONSECUTIVE AND YET DIFFERENT DEPROTONATION/PROTONATION CYCLES. THE NEW CONCEPTUAL FRAMEWORK AND DHA1 STRUCTURES OBTAINED FROM THIS STUDY WILL SERVE AS A STEPPING-STONE TOWARD DEVISING NOVEL STRATEGIES TO EVADE OR INHIBIT THE CLINICALLY RELEVANT MULTIDRUG TRANSPORTERS, WHICH MAY RESCUE THERAPEUTIC EFFICACY AGAINST MULTIDRUG-RESISTANT CELLS AND HALT THE SPREAD OF UNTREATABLE INFECTIONS. FURTHERMORE, OUR WORK WILL OFFER A SPRINGBOARD FOR THE MECHANISTIC STUDIES OF HUMAN SLC18 ANTIPORTERS, WHICH WILL SHED NEW LIGHT ON HOW THEY UTILIZE THE ELECTROCHEMICAL H+ GRADIENT TO TRANSLOCATE MONOAMINE NEUROTRANSMITTERS, NEUROTOXINS, OR POLYAMINE NEUROMODULATORS, ACROSS VESICULAR MEMBRANES.

ADOLESCENT COCAINE ABUSE: ELECTROPHYSIOLOGY & BEHAVIOR

CORRECTION OF HEARING AND VESTIBULAR DEFECTS IN A MOUSE MODEL FOR DEAFNESS

STEM CELLS FOR BRAIN REPAIR

DEVELOPMENT OF SLOW SKELETAL MUSCLE FIBERS

HEPATITUS C VIRUS-INDUCED INFLAMMASOME AND LIPID METABOLISM

IMAGING CIRCUIT STRUCTURE AND MEMORIES IN A MULTIFUNCTIONAL NETWORK

A TARGETED PREEMPTIVE APPROACH TO ADDRESSING MITOCHONDRIAL TOXICITY OF NUCLEOSIDE

EFFECTS OF REPEATED STRESS ON AMYGDALA FUNCTION DURING ADOLESCENCE

ROLE OF TAM RECEPTORS IN MODULATING HUMORAL IMMUNITY AGAINST PARASITIC INFECTIONS - ABSTRACT UNLIKE MANY INFECTIOUS DISEASES, NATURALLY ACQUIRED IMMUNITY AGAINST PLASMODIUM DO NOT REPRESENT DURABLE STERILIZING IMMUNITY. WHILE THE GRADUAL ACQUISITION OF PROTECTIVE OR CLINICAL IMMUNITY FOLLOWING REPEAT INFECTIONS IS A SOLACE FOR PEOPLE LIVING IN ENDEMIC REGIONS, THESE SYMPTOM-LESS CARRIERS BECOME CRITICAL RESERVOIRS OF THE PATHOGEN, FACILITATING INADVERTENT TRANSMISSION. THE OVERARCHING GOAL OF OUR RESEARCH PROGRAM IS TO IDENTIFY MOLECULAR AND CELLULAR IMMUNE MECHANISMS AND TARGETS THAT ARE IN PLAY DURING THE PATHOGENESIS OF MALARIA RESULTING IN SUBOPTIMAL IMMUNE RESPONSE THAT FAILS TO INDUCE STERILIZING IMMUNITY AND TO DEVISE IMMUNOMODULATORY HOST-DIRECTED INTERVENTIONS AGAINST THIS LIFE-THREATENING DISEASE. OUR RECENT OBSERVATIONS POINT TOWARDS SOME KEY PATHOPHYSIOLOGICAL FEATURES OF THE DISEASE CONTRIBUTING TO THIS DYSREGULATED AND SUBOPTIMAL INDUCTION OF HUMORAL IMMUNE RESPONSE. THIS PROJECT REVOLVES AROUND THE CENTRAL HYPOTHESIS THAT SYSTEMIC HYPOXIA SUBSEQUENT TO MALARIA-INDUCED HEMOLYTIC ANEMIA INDUCES THE EXPRESSION OF TAM RECEPTORS ON B CELLS TO DRIVE THE ACCUMULATION OF EXTRAFOLLICULAR PLASMABLASTS, THAT IN TURN EXERCISE AN IMMUNOSUPPRESSIVE FUNCTION BY ACTING AS A NUTRIENT SINK. THIS PROPOSAL EMPLOYS A COMBINATION OF GENETIC, BIOCHEMICAL AND BONE MARROW CHIMERIC APPROACHES TO INVESTIGATE THE POTENTIAL OF INTERRUPTING THE HYPOXIA-TAM RECEPTORS(S)- PLASMABLAST-NUTRIENT SINK AXIS TO IMPROVE THE OVERALL QUALITY AND MAGNITUDE OF ANTI-PLASMODIUM HUMORAL IMMUNE RESPONSE. IN THIS PROPOSAL WE ALSO PLAN TO REPURPOSE AN IMMUNOSTIMULATORY DRUG, IN STAGE 2 CLINICAL TRIALS THAT BLOCK AXL, ONE OF THE TAM RECEPTORS AND TO INVESTIGATE HOW AXL DEFICIENCY/BLOCKADE MAY REPROGRAM PLASMODIUM SPECIFIC GERMINAL CENTER B CELLS BY INDUCING TRANSCRIPTIONAL AND EPIGENETIC CHANGES. BY SUCCESSFULLY COMPLETING THE EXPERIMENTS PROPOSED IN THIS PROJECT, WE HOPE TO DISCERN (I) THE PRECISE MECHANISM BY WHICH TAM RECEPTOR GOVERN THE DIFFERENTIATION OF PLASMABLASTS (II) HOW THE PATHOPHYSIOLOGY OF THE DISEASE (SUCH AS ANEMIA) ITSELF CAN ALTER THE IMMUNE LANDSCAPE AND (III) THE DIFFERENT INTERVENTIONS TO OPTIMIZE THE IMMUNE RESPONSE TO HARNESS ITS FULL POTENTIAL.

SPLICE VARIANT BIOMARKERS FOR PARKINSON'S DISEASE

THE SEX SPECIFIC IMPACT OF ANXIETY ON ALZHEIMER'S DISEASE PROGRESSION - PROJECT SUMMARY / ABSTRACT FOR THIS K99/R00, I WILL SPECIFICALLY ADDRESS THE ROLE OF ANXIETY AND ANATOMICAL SEX ON ALZHEIMER’S DISEASE (AD) PROGRESSION, SPECIFICALLY MEMORY LOSS. AD, A DEBILITATING NEURODEGENERATIVE AND MENTAL DISORDER, STANDS ALONE AS ONE OF THE TEN LEADING CAUSES OF DEATH IN THE UNITED STATES THAT CANNOT BE PREVENTED, SLOWED, OR CURED. FURTHERMORE, NEUROPSYCHIATRIC DISTURBANCES, SUCH AS DEPRESSION AND ANXIETY, ARE OBSERVED IN 90% OF AD PATIENTS AND ARE FREQUENT IN THOSE AT RISK FOR AD. ALTHOUGH MOST AD STUDIES HAVE BEEN PERFORMED USING MALE MICE, RECENT EVIDENCE SUGGESTS THAT FEMALES ARE MORE SUSCEPTIBLE TO DEPRESSION, ANXIETY, AND AD WHEN COMPARED TO MALES. IN FACT, TWO-THIRDS OF AD PATIENTS ARE WOMEN. HERE, WE AIM TO IDENTIFY THE NEURAL ENSEMBLES LINKING ANXIETY AND MEMORY LOSS FOLLOWING AD PROGRESSION BY UTILIZING BEHAVIORAL STUDIES, OPTOGENETICS, WHOLE-BRAIN MICROSCOPY, AND IN VIVO CA2+ IMAGING IN FEMALE AND MALE MICE. THESE STUDIES REPRESENT A NUMBER OF FIRSTS IN THE AD FIELD: 1) THE FIRST TO TEST THE CONTROVERSIAL HYPOTHESIS THAT ANXIETY CAN BE A PREDICTOR OF AD IN FEMALES; 2) THE FIRST TO INVESTIGATE SEX DIFFERENCES ACROSS THE WHOLE BRAIN AS THE DISEASE PROGRESSES; 3) THE FIRST TO USE IN VIVO CA2+ IMAGING TO TAG AN INDIVIDUAL MEMORY AND ASSES NEURONAL ACTIVITY DURING BEHAVIOR IN AD MICE; AND 4) THE FIRST TO TEST THE THERAPEUTIC POTENTIAL OF TARGETING NEURAL CORRELATES OF MEMORY AND ANXIETY IN AD MICE. IN AIM 1, BEHAVIORAL DIFFERENCES WILL BE CORRELATE ANXIETY-LIKE BEHAVIOR WITH MEMORY LOSS ACROSS NUMEROUS AGES AS AD PROGRESSES. IN AIM 2, THE INDIVIDUAL NEURONS CORRESPONDING TO A MEMORY WILL BE INVESTIGATED BY UTILIZING A TRANSGENIC LINE, THE ARCCREERT2 MICE BRED WITH AN AD LINE (APP/PS1). THIS MOUSE LINE ALLOWS FOR THE INDELIBLE LABELING OF CELLS EXPRESSING THE IMMEDIATE EARLY GENE (IEG) ARC/ARG3.1 AND ALLOWS FOR A COMPARISON BETWEEN THE CELLS THAT ARE ACTIVATED DURING THE ENCODING OF AN EXPERIENCE AND THOSE THAT ARE ACTIVATED DURING THE RETRIEVAL OF THE CORRESPONDING MEMORY. WE WILL USE WHOLE BRAIN IMAGING TO FIND NOVEL BRAIN REGIONS OF INTEREST THAT ARE ALTERED DURING AD. IN AIM 3, I WILL RESCUE IMPAIRED NEURAL NETWORKS IN AD X ARCCREERT2 MICE USING OPTOGENETIC STIMULATION IN COMBINATION WITH IN VIVO CA2+ IMAGING. THE OUTCOME OF THIS TARGETED RESCUE WILL PROVIDE DIRECT EVIDENCE THAT DISRUPTED NEURAL ENSEMBLES RESULTS IN THE COGNITIVE DECLINE AND ANXIETY-LIKE BEHAVIOR OBSERVED IN FEMALE AD MICE. MY OVERALL CAREER GOAL IS TO LEAD AN INDEPENDENT RESEARCH GROUP EXAMINING THE UNDERLYING MECHANISMS OF NEURODEGENERATION AND TO DETERMINE HOW SEX IMPACTS DISEASE PROGRESSION WITH THE LONG-TERM GOAL OF CREATING PERSONALIZED THERAPEUTICS. THIS K99/R00 WILL THUS PROVIDE ME PROTECTED TIME AND MENTORSHIP TO ACQUIRE THE TECHNICAL AND CONCEPTUAL SKILLS TO SUCCESSFULLY ACHIEVE MY CAREER GOALS AND ESTABLISH AN INDEPENDENT RESEARCH PROGRAM GEARED TOWARDS ANSWERING CLINICALLY DRIVEN RESEARCH QUESTIONS IN THE AREA OF NEURODEGENERATIVE DISEASES.

STRUCTURE AND FUNCTION OF THE U3 RNA-PROTEIN COMPLEX

MECHANISMS UNDERLYING REWARD-RELATED SYNAPTOGENESIS

SUMMER RESEARCH FELLOWSHIP PROGRAM FOR PODIATRIC MEDICAL STUDENTS

IMPROVING DIABETIC FOOT ULCER OFFLOADING: A PILOT STUDY ON THE IMPACT OF REMOVABLE CAST WALKER DESIGN FACTORS ON USABILITY - ABSTRACT WITHIN THEIR LIFETIME, OVER 30% OF PEOPLE LIVING WITH DIABETES WILL DEVELOP A DIABETIC FOOT ULCER (DFU), MANY OF WHICH WILL NEVER HEAL AND MAY REQUIRE AMPUTATION. REMOVABLE CAST WALKERS (RCWS) ARE COMMONLY PRESCRIBED TO OFFLOAD (TREAT) DFUS. HOWEVER, ADHERENCE WITH RCWS IS LOW, WHICH IS A SERIOUS CONCERN GIVEN THAT LOW ADHERENCE PREDICTS POOR DFU HEALING. THERE IS AN URGENT NEED TO ADDRESS FACTORS THAT HINDER ADHERENT USE OF RCWS TO PROMOTE DFU HEALING. OUR LONG-TERM GOAL IS TO OPTIMIZE OFFLOADING ADHERENCE (OA), AND SUBSEQUENT DFU HEALING OUTCOMES, BY CONSIDERING NOT ONLY HOW MUCH THE DEVICE OFFLOADS THE DFU, BUT ALSO HOW MUCH THE DEVICE’S DESIGN IMPACTS THE PATIENTS’ EXPERIENCES WITH THE DEVICE. THE OBJECTIVE OF THIS PROPOSAL IS TO PROVIDE PILOT/FEASIBILITY DATA TO INFORM A DEFINITIVE RANDOMIZED CONTROLLED TRIAL (RCT) THAT WILL EVALUATE THE IMPACT OF EXISTING RCW DESIGNS ON OA AND DFU HEALING AND, SECONDARILY, WILL FACILITATE DEVELOPMENT OF A PREDICTIVE MODEL TO GUIDE PATIENT-SPECIFIC PRESCRIPTION OF OFFLOADING DEVICES. OUR WORKING HYPOTHESIS IS THAT HEALING OUTCOMES WITH A GIVEN RCW WILL BE PREDICTED BY OBJECTIVE, BIOMECHANICAL MARKERS OF RCW USABILITY (E.G., MEASURES OF STABILITY) AND BY RCW-INDUCED CHANGES IN PATIENT-REPORTED FACTORS, WITH THE PREDICTIVE RELATIONSHIP MEDIATED BY THE EFFECT OF THESE MEASURES ON OA. WE WILL CONSIDER THREE SPECIFIC AIMS TO ACHIEVE THE OBJECTIVE AND PROVIDE EVIDENCE TO SUPPORT OUR HYPOTHESIS AND OUR ABILITY TO CONDUCT A LARGER FOLLOW-UP RCT: AIM 1: TO EXAMINE THE FEASIBILITY OF CONDUCTING AN RCT TO EVALUATE THE EFFECTS OF RCW FORM ON OA AND DFU HEALING; AIM 2: TO COLLECT PRELIMINARY DATA TO QUANTIFY THE EFFECT OF RCW FORM ON BIOMECHANICAL MARKERS OF USABILITY AND PATIENT-REPORTED ADHERENCE FACTORS IN INDIVIDUALS WITH ACTIVE DFU; AND AIM 3: TO COLLECT PRELIMINARY DATA TO EVALUATE THE STRENGTH OF ASSOCIATIONS BETWEEN OA AND BOTH BIOMECHANICAL MARKERS OF USABILITY AND PATIENT-REPORTED ADHERENCE FACTORS. ACROSS TWO STUDY CENTERS, PARTICIPANTS WILL BE RANDOMIZED TO USE ONE-OF-TWO RCW DESIGNS FOR FOUR WEEKS IN CONJUNCTION WITH WEEKLY STANDARD-OF-CARE TREATMENT. THROUGHOUT TREATMENT, OA WILL BE MONITORED USING A NOVEL DUAL ACCELEROMETER METHOD AND DFU HEALING WILL BE QUANTIFIED VIA PLANIMETRIC WOUND AREA MEASUREMENTS. AT THE START OF TREATMENT PARTICIPANTS WILL COMPLETE A SERIES OF SURVEYS TO ASSESS PATIENT-REPORTED FACTORS (E.G., DEPRESSION). PATIENTS WILL REPEAT SURVEYS AT THE END OF TREATMENT AND ALSO PERFORM SEVERAL TESTS WITHIN A MOTION CAPTURE LAB TO ASSESS ASPECTS OF RCW USABILITY (E.G., STABILITY & ENERGETIC COSTS OF WALKING). THE CONTRIBUTION OF THIS WORK WILL BE SIGNIFICANT AS IT REPRESENTS THE NEXT STEP IN A CONTINUUM OF RESEARCH EXPECTED TO IMPROVE DFU HEALING RATES AND REDUCE DFU COMPLICATIONS INCLUDING AMPUTATION, THEIR ASSOCIATED MEDICAL COSTS AND NEGATIVE IMPACT ON WELL-BEING. THE PROPOSED STUDY IS INNOVATIVE AS IT CHALLENGES CURRENT PRACTICE AND GUIDELINES BY TAKING A PATIENT-CENTERED APPROACH TO DFU HEALING, WHICH CONSIDERS AN OFFLOADING DEVICES’ IMPACT ON THE PATIENT EXPERIENCE, RATHER THAN SOLELY CONSIDERING THE DEVICES’ FUNCTIONAL OFFLOADING CAPACITY.

UNDERSTANDING AND REDUCING HIV RISK BEHAVIOR AND SUBSTANCE USE AMONG SELF-IDENTIFIED BISEXUAL ADOLESCENT MEN

IDENTIFICATION OF NOVEL COMPONENTS OF THE DYSTROPHIN COMPLEX USING C. ELEGANS

NEURAL CIRCUITS FOR DECISION MAKING - PROJECT SUMMARY DIFFERENT PEOPLE FACING SIMILAR SITUATIONS MAKE DIFFERENT CHOICES, AND DIFFERENT CHOICES RESULT IN DIVERGENT LIFE TRAJECTORIES. DESPITE THE PERVASIVE IMPACT ON AN INDIVIDUAL’S LIFE, THE NEURAL BASIS OF IDIOSYNCRATIC DECISION-MAKING REMAINS LARGELY UNKNOWN. BEGINNING TO ADDRESS THIS IMPORTANT TOPIC, WE RECENTLY CHARACTERIZED IDIOSYNCRATIC CHOICE BEHAVIOR QUANTITATIVELY AND IDENTIFIED A POSTERIOR NETWORK COMPRISED OF THE CINGULATE CORTEX, POSTERIOR PARIETAL CORTEX, AND STRIATUM IN WHICH THE IDIOSYNCRATIC CHOICE BIASES ARE LIKELY PROCESSED, TRANSMITTED, AND INTEGRATED WITH OTHER DECISION VARIABLES. THE POSTERIOR NETWORK IMBUING IDIOSYNCRASY MIGHT OPERATE IN PARALLEL TO THE WELL-KNOWN FRONTAL NETWORK FOR RULE- AND VALUE-BASED DECISION-MAKING AND THE SENSORIMOTOR NETWORK FOR STIMULUS-RESPONSE ASSOCIATION, ALL OF WHICH DYNAMICALLY CONTRIBUTE TO THE FINAL CHOICE. HOWEVER, THE DETAILED CIRCUIT MECHANISMS IN THE POSTERIOR NETWORK HAVE YET TO BE DISCOVERED. WE PROPOSE A SERIES OF EXPERIMENTS IN MICE TO ELABORATE INFORMATION FLOWS AND PROCESSING IN THE POSTERIOR NETWORK, UTILIZING WELL-ESTABLISHED IN-LAB TOOLS INCLUDING BEHAVIOR MODELING, ANTEROGRADE- AND RETROGRADE-TRANSSYNAPTIC LABELING, OPTOGENETICS, AND TWO-PHOTON IMAGING. UNDERSTANDING THE NEURAL CIRCUITS GOVERNING THE MOMENT-BY-MOMENT IDIOSYNCRATIC HISTORY BIAS WILL SHED LIGHT ON THE NEURAL ORIGINS OF IDIOSYNCRATIC DECISIONS. FURTHERMORE, LIFE-INTERFERING, MALADAPTIVE CHOICE BEHAVIOR MIGHT BE IN PART A MANIFESTATION OF EXTREMELY DEVIATED IDIOSYNCRATIC BIAS, THUS ELABORATING THE LESS KNOWN POSTERIOR NETWORK MAY BRING NEW INSIGHTS INTO PATHOLOGICAL DECISION-MAKING IN NEUROLOGICAL CONDITIONS SUCH AS AGING, DEMENTIA, AND ADDICTION.

IDENTIFYINGCOMMONUNDERLYINGMECHANISMSDRIVINGSYNAPTIC DEFICITS BTW TBI AD

STIMULUS-INDUCED NEURAL NETWORKS

CRF-OXYTOCIN INTERACTION IN THE REGULATION OF STRESS AND AFFECT

EFFECT OF MENTHOL TO NON-MENTHOL CIGARETTE SWITCHING ON SUBCLINICAL INFLAMMATORY BIOMARKERS OF CARDIOVASCULAR HEALTH: SIMULATING A MENTHOL CIGARETTE BAN - PROJECT SUMMARY/ABSTRACT THE PROPOSED K01 APPLICATION IS DESIGNED TO PROVIDE NANCY JAO, PH.D., WITH THE MENTORED RESEARCH AND TRAINING NECESSARY TO TRANSITION INTO AN INDEPENDENT CLINICAL SCIENTIST WITH A TOBACCO REGULATORY RELEVANT PROGRAM OF RESEARCH. THE FDA HAS LONG INDICATED INTEREST IN BANNING MENTHOL AS A CHARACTERIZING FLAVOR IN COMBUSTIBLE CIGARETTES DUE TO ITS ROLE IN FACILITATING THE INITIATION AND USE OF TOBACCO PRODUCTS AND INCREASING THE NUMBER OF SMOKING-RELATED DEATHS. SMOKING-INDUCED INFLAMMATION IS A LEADING PATHWAY BY WHICH CIGARETTE SMOKING CONTRIBUTES TO INCREASED CARDIOVASCULAR DISEASE (CVD) MORBIDITY AND MORTALITY IN SMOKERS. ELEVATIONS IN BIOMARKERS OF INFLAMMATION CAN BE DETECTED EARLY, EVEN IN ASYMPTOMATIC INDIVIDUALS, AS A SUBCLINICAL INDICATOR OF CVD RISK. BASIC SCIENCE STUDIES HAVE SHOWN THAT MENTHOL FLAVORING CAN CAUSE INCREASES IN INFLAMMATORY RESPONSE AND DYSFUNCTION BEYOND THE EFFECTS OF SMOKING. HOWEVER, IT IS UNKNOWN WHETHER MENTHOL CIGARETTE (MC) USE MAY ELEVATE BIOMARKERS OF INFLAMMATION IN SMOKERS AND INCREASE CVD RISK COMPARED TO NON-MENTHOL CIGARETTE (NMC) USE. ADDITIONALLY, WHILE IT IS EXPECTED THAT A MC BAN WILL IMPROVE HEALTH OUTCOMES FOR SMOKERS WHO QUIT OR SWITCH TO NON-COMBUSTIBLE PRODUCTS, IT IS UNKNOWN WHETHER THERE WILL BE REDUCTIONS IN INFLAMMATION FOR THOSE WHO SWITCH FROM MC TO NMC SMOKING. IN STUDY 1, WE AIM TO EVALUATE DIFFERENCES IN BIOMARKERS OF SYSTEMIC INFLAMMATION AND CVD RISK BETWEEN MC AND NMC SMOKERS IN NATIONALLY-REPRESENTATIVE, LONGITUDINAL POPULATION ASSESSMENT OF TOBACCO AND HEALTH (PATH) STUDY. IN STUDY 2, WE AIM TO EXAMINE HOW SWITCHING FROM MC TO NMC SMOKING MAY IMPACT BIOMARKERS OF SYSTEMIC INFLAMMATION, SMOKING BEHAVIOR, AND SUBJECTIVE RESPONSES RELATED TO SMOKING. MC SMOKERS (N=68) WILL BE RECRUITED FOR A FIVE-WEEK STUDY, WITH ONE-WEEK OF BASELINE OF MC SMOKING (PHASE 1), FOLLOWED BY FOUR WEEKS OF SWITCHING TO STUDY-PROVIDED, BRAND-MATCHED NMCS (PHASE 2). BIOMARKERS OF SYSTEMIC INFLAMMATION (E.G., HSCRP, INTERLEUKIN CYTOKINES) AND TOBACCO EXPOSURE (E.G., COTININE, CARBON MONOXIDE) WILL BE ANALYZED FROM BLOOD SAMPLES BEFORE, DURING, AND AFTER SWITCHING. ECOLOGICAL MOMENTARY ASSESSMENT (EMA) METHODS WILL ALSO BE GATHERED TO MEASURE PATTERNS OF SMOKING AND SMOKING-RELATED SUBJECTIVE RESPONSES. THE PROPOSED RESEARCH IS SIGNIFICANT AND DIRECTLY TARGETS FDA'S INTEREST IN UNDERSTANDING THE IMPACT OF FLAVORINGS ON THE HEALTH EFFECTS OF TOBACCO USE. THROUGHOUT THE FIVE-YEAR AWARD, DR. JAO WILL BE MENTORED BY AN IMPRESSIVE MENTORSHIP TEAM IN (1) TRANSLATIONAL RESEARCH IN BIOBEHAVIORAL AND HEALTH EFFECTS OF TOBACCO USE, PARTICULARLY RELATING TO BIOMARKERS OF INFLAMMATION AND CVD RISK; (2) LONGITUDINAL RESEARCH METHODS AND ADVANCED STATISTICAL ANALYSES, INCLUDING UTILIZATION OF THE PATH STUDY DATASET AND EMA METHODOLOGY; AND (3) DESIGN AND IMPLEMENTATION OF HUMAN CLINICAL TRIAL STUDIES WITH TOBACCO REGULATORY IMPLICATIONS. UNDERSTANDING THE IMPACT OF MC USE ON THESE SENSITIVE SUBCLINICAL BIOMARKERS PRIOR TO CVD DIAGNOSIS CAN HELP THE FDA IDENTIFY AND QUANTIFY CARDIOVASCULAR HEALTH HAZARDS OF MC USE, PARTICULARLY FOR THOSE WHO MAY CONTINUE TO USE COMBUSTIBLE CIGARETTES IF A MC BAN IS IMPLEMENTED BY THE FDA.

IDENTIFICATION OF SPLICE VARIANTS AS A BIOSIGNATURE FOR PARKINSON'S DISEASE

SOCS: ITS REGULATION AND ROLE IN ONCOGENESIS

NEURAL EP24.15- A MODEL FOR NEUROPEPTIDASE FUNCTION

ENTRY OF HHV-8 INTO THE TARGET CELLS

NETWORK ANALYSIS FOR IDENTIFYING DISEASE MECHANISMS AND THERAPEUTIC TARGETS FOR DEMENTIA

BLOOD RNA BIOMARKERS OF PARKINSON'S DISEASE AND PROGRESSIVE SUPRANUCLEAR PALSY

PSYCHOSTIMULANTS AND PLASTICITY

SSARP FUNDING OVERSIGHT - FINANCE DEPARTMENT

LTQ ORBITRAP VELOS MASS SPECTROMETER WITH ETD

STRUCTURE/FUNCTION OF NERVE GROWTH FACTOR/NEUROTROPHINS

TARGETING BALANCE CONFIDENCE AS A STRATEGY TO INCREASE INTEGRATION AND IMPROVE OUTCOMES IN USERS OF LOWER-LIMB PROSTHESES

BASAL GANGLIA OUTPUT AND PSYCHOSTIMULANT ABUSE

DEVELOPING AND IMPROVING INSTITUTIONAL ANIMAL RESOURCES AT ROSALIND FRANKLIN UN.

ENHANCING COGNITION IN SCHIZOPHRENIA

CHARACTERIZATION OF STRIATAL NITRIC OXIDE SIGNALING

NEP-LIKE ENDOPEPTIDASES IN TRAUMATIC BRAIN INJURY AND THE ASSOCIATED DEMENTIAS

GENE THERAPY TARGETING STRIATAL CGMP SIGNALING IN EXPERIMENTAL PARKINSONISM

HOMEOSTATIC REGULATION OF CAV2 VOLTAGE-GATED CALCIUM CHANNELS AT SYNAPSES - THE HOMEOSTATIC PLASTICITY ALLOWS NEURAL CIRCUITS TO RESTORE BASELINE FUNCTION BY REBALANCING PRE- OR POST- SYNAPTIC FUNCTION. IMPAIRED HOMEOSTATIC RESPONSES UNDERLIE THE PATHOPHYSIOLOGY OF A VARIETY OF NEUROLOGICAL AND PSYCHIATRIC DISEASES. THE CORE PRINCIPLE OF HOMEOSTATIC PLASTICITY IS THAT SYNAPTIC STRENGTH IS SCALED UP OR DOWN PROPORTIONALLY TO THE DEGREE OF NEURAL DISTURBANCE. WITHOUT THIS SYNAPTIC SCALING, INFORMATION FLOW CAN BE LOST OR DISTORTED. ONE OF THE KEY FACTORS THAT REGULATE NEUROTRANSMITTER RELEASE AND THUS SYNAPTIC STRENGTH IS THE ABUNDANCE OF VOLTAGE-GATED CAV2 CALCIUM CHANNELS AT THE PRESYNAPTIC TERMINAL. TO ELUCIDATE THE RELATIONSHIP BETWEEN CAV2/UNC-2 CHANNEL ABUNDANCE AND NEUROTRANSMITTER RELEASE, WE USED THE NEMATODE C. ELEGANS, A SIMPLE GENETIC MODEL ORGANISM THAT POSSESSES HIGHLY CONSERVED CAV2 CHANNELS AND ACTIVE ZONE PROTEINS. USING CRISPR/CAS9, WE ENDOGENOUSLY TAGGED CAV2/UNC-2 CHANNELS WITH GFP AND INTRODUCED EITHER A GAIN- OR LOSS-OF-FUNCTION MUTATION. WHEN WE COMPARED WILD-TYPE AND UNC-2 MUTANT FORMS, WE FOUND THAT CHANNEL FUNCTION INVERSELY CORRELATES WITH THEIR ABUNDANCE. HOWEVER, OUR PRELIMINARY DATA SHOWED THAT SYNAPTIC VESICLE (SV) EXOCYTOSIS, BUT NOT CHANNEL FUNCTION ITSELF, DICTATES UNC-2 ABUNDANCE. LOSS OF UNC-13 FUNCTION, WHICH CAUSES AN SV EXOCYTOSIS DEFECT, INCREASES CHANNEL ABUNDANCE. IN CONTRAST, AN OPEN SYNTAXIN MUTATION, WHICH ENHANCES SV EXOCYTOSIS BY BYPASSING SV PRIMING, DECREASES CHANNEL ABUNDANCE. BASED ON THESE FINDINGS, WE HYPOTHESIZE THAT A HIGH RATE OF NEUROTRANSMITTER RELEASE NEGATIVELY REGULATES CV2/UNC-2 CHANNEL LEVELS. THIS NEGATIVE FEEDBACK REGULATORY MECHANISM PROVIDES PRESYNAPTIC NEURONS WITH THE ABILITY TO SCALE UP OR DOWN NEUROTRANSMITTER RELEASE BY ADJUSTING CAV2 CHANNEL LEVELS INDEPENDENTLY OF POSTSYNAPTIC INPUTS. TO FURTHER VALIDATE OUR HYPOTHESIS, WE WILL EMPLOY A WIDE ARRAY OF SV EXOCYTOSIS MUTATIONS ALONGSIDE OPTOGENETIC AND CHEMICAL GENETIC TOOLS TO DELVE DEEPER INTO THE MECHANISMS CONTROLLING PRESYNAPTIC UNC-2 LEVELS. CONCURRENTLY, WE AIM TO IDENTIFY THE CORE ACTIVE ZONE PROTEINS CRUCIAL FOR ACTIVITY-DEPENDENT UNC-2 ABUNDANCE MODULATION. COMPLETION OF THE PROJECT WILL VALIDATE A PREVIOUSLY UNIDENTIFIED FORM OF PRESYNAPTIC HOMEOSTATIC PLASTICITY THAT OPERATES INDEPENDENTLY OF POST-SYNAPTIC INPUTS.

SELECTIVE TARGETING OF CNS CELLS IN VIVO FOR MULTIPLE TRANSGENE DELIVERY AND NEURONAL REPROGRAMMING - PROJECT SUMMARY/ABSTRACT VIRAL VECTORS ENJOY WIDESPREAD USE FOR GENE TRANSFER TO MULTIPLE CELL TYPES AND ORGAN SYSTEMS IN BOTH RESEARCH STUDIES AND, INCREASINGLY, IN THERAPEUTIC STRATEGIES. FOR STUDIES OF THE CNS, THE ABILITY OF THE VIRAL VECTOR TO EFFECTIVELY AND SELECTIVELY INFECT THE DESIRED CELL TYPES WITHIN THE CNS PLAYS A CRITICAL ROLE IN THE SUBSEQUENT INTERPRETATION OF STUDY OUTCOMES. HOWEVER, MOST VIRAL VECTORS ARE SOMEWHAT PROMISCUOUS IN THEIR CELL TYPE SPECIFICITY. ALTHOUGH A PARTICULAR VECTOR PSEUDOTYPE MAY PRIMARILY INFECT ONE TYPE OF CELL, THE ADDITIONAL “OFF- TARGET” INFECTION OF OTHER CELL TYPES MAY INTRODUCE UNWANTED VARIANCE TO THE STUDY OUTCOMES. IN SOME RECENT STUDIES, THE INTENDED INFECTION OF ASTROCYTES ALSO RESULTED IN THE UNWANTED INFECTION OF NEURONS, UNDERMINING THE STUDIES’ CONCLUSIONS. THUS, THERE IS A NEED TO IMPROVE THE CURRENT STATE OF CELL-TYPE TROPISM TO ONE OF CELL- TYPE SELECTIVITY TO SPECIFICALLY AND EXCLUSIVELY TARGET THE DESIRED CELL TYPE AND ELIMINATE OFF-TARGET EFFECTS. A SIMILAR UNMET NEED IS THE LACK OF A TOOL TO SELECTIVELY TARGET A PREVIOUSLY INFECTED CELL FOR A SUBSEQUENT “FOLLOW- UP” GENE DELIVERY. IN THIS EXPLORATORY/DEVELOPMENTAL R21 MECHANISM PROJECT, WE PROPOSE TO BUILD UPON THE POPULAR EXPRESSION OF THE AVIAN TVA RECEPTOR USED TO ENABLE ENVA-PSEUDOTYPED RABIES VIRUS DELIVERY OF A FLUORESCENT REPORTER GENE FOR TRANSSYNAPTIC LABELING IN THE CNS. OUR AIMS ARE IN THE CONTEXT OF OUR INTEREST TO PERFORM DIRECT IN VIVO CELLULAR REPROGRAMMING OF GLIAL PROGENITOR CELLS INTO NEURONS. IN AIM 1, WE WILL USE OUR CURRENT RETROVIRAL APPROACH TO INFECT PROLIFERATING GLIAL PROGENITOR CELLS IN THE ADULT BRAIN TO EXPRESS THE AVIAN TVA RECEPTOR. WE WILL THEN BE ABLE TO SELECTIVELY TARGET THE TVA-EXPRESSING GLIAL PROGENITOR CELLS FOR GENE TRANSFER USING ENVA-PSEUDOTYPED LENTIVIRUS. IN AIM 2, WE WILL DIRECTLY TARGET THE GLIAL PROGENITOR CELLS BY PSEUDOTYPING LENTIVIRUS TO EXCLUSIVELY BIND TO A CELL-SPECIFIC MEMBRANE RECEPTOR. THIS WILL FACILITATE THE DEMONSTRATION OF TARGETED INFECTION OF GLIAL PROGENITOR CELLS THROUGH SELECTIVE AND EXCLUSIVE GENE TRANSFER. INCLUDING TVA IN THE CELL SPECIFIC RECEPTOR-PSEUDOTYPED LENTIVIRUS CONSTRUCT WILL PERMIT SUBSEQUENT SELECTIVE TARGETING BY ENVA-PSEUDOTYPED LENTIVIRUS FOR DELIVERY OF SUBSEQUENT REPROGRAMMING FACTORS OR OTHER GENE TRANSFER.

PHARMACOLOGICAL INHIBITION OF NEP-LIKE ENZYMES IN TRAUMATIC BRAIN INJURY AND DEMENTIA - ABSTRACT RECENTLY, MY LABORATORY HAS FOUND THAT THE NEP-LIKE ENDOPEPTIDASES, NEPRILYSIN (NEP) AND NEP2, HAVE A ROLE IN ACUTE RECOVERY AFTER TRAUMATIC BRAIN INJURY (TBI). THIS HAS INTERESTING AND EXCITING IMPLICATIONS FOR THE TBI FIELD AS WELL AS FOR UNDERSTANDING RISK FACTORS FOR DEVELOPING DEMENTIA. THE CAUSAL LINKS BETWEEN TBI AND DEMENTIA ARE BECOMING BETTER UNDERSTOOD. MANY OF THE PATHOLOGICAL HALLMARKS OF ALZHEIMER’S DISEASE (AD) ARE SHARED WITH EARLY CHANGES IN TBI. OF NOTE ARE THE RAPID ELEVATIONS OF THE AMYLOID-BETA (AΒ) PEPTIDE AFTER TBI. NATURAL MECHANISMS OF CLEARANCE OF AΒ ARE IMPORTANT IN SLOWING THE PROGRESSION OF AD AND SO THESE CLEARANCE MECHANISMS MAY ALSO BE IMPORTANT IN TBI. BOTH NEP AND ITS HOMOLOG NEP2 DEGRADE AΒ AND ARE IMPORTANT FOR CONTROLLING CEREBRAL LEVELS OF THE PEPTIDE. WE PROPOSE A ROLE FOR THESE ENZYMES IN THE PROCESSES FOLLOWING TBI. TO FURTHER INVESTIGATE THE ROLE(S) OF NEP/NEP2 IN TBI WE WILL USE OUR APP HUMANIZED TRANSGENIC MICE AND ALTER THE ACTIVITY OF THESE ENZYMES IN A TBI CONTEXT IN BOTH SHORT (AIM 1) AND LONG-TERM (AIM 2) ASSESSMENTS. THIS PROJECT WILL DETERMINE THE IMPORTANCE OF NEP-LIKE ENZYMES IN THE ACUTE PATHOLOGY POST TBI AS WELL AS IN THE DEVELOPMENT DEMENTIA.

NEUROPEPTIDE CIRCUITS AND STRESS REGULATION IN THE BASOLATERAL AMYGDALA

GENERATION OF PARKINSON-DERIVED HUMAN GLIAL PROGENITOR CELLS FOR DOPAMINERGIC NEURONAL CONVERSION - PROJECT SUMMARY/ABSTRACT PARKINSON’S DISEASE (PD) IS CLASSICALLY CHARACTERIZED BY THE PROGRESSIVE LOSS OF DOPAMINERGIC NEURONS LEADING TO THE EMERGENCE OF DEBILITATING MOTOR, AND NON-MOTOR SYMPTOMS. AS THE CNS DOES NOT REPLACE NEURONS LOST TO INJURY OR DISEASE, A PRIMARY FOCUS IN DEVELOPING THERAPEUTIC STRATEGIES FOR PD HAS CENTERED AROUND THE IDEA OF GRAFTING REPLACEMENT DOPAMINERGIC NEURONS. EARLY NEUROSURGICAL APPROACHES USING FETAL VENTRAL MIDBRAIN ALLOGRAFTS PRODUCED SOME MIXED, BUT ENCOURAGING RESULTS. AS FETAL DONOR TISSUE PRESENTS CHALLENGES IN REGARD TO SOURCING AND ETHICS, VARIABILITY, IMMUNOGENICITY, AND COST, RECENT EFFORTS HAVE FOCUSED ON THE PROSPECT OF USING INDUCED PLURIPOTENT CELLS DRIVEN TO A DOPAMINERGIC NEURONAL FATE. THE ADVANTAGES OF THIS PRECISION MEDICINE APPROACH, WHERE THE PATIENT’S OWN CELLS ARE ENGINEERED INTO THE REPLACEMENT NEURONS, IS OFFSET BY THE EFFORT AND COST TO PRODUCE AND DIFFERENTIATE CELLS UNDER CGMP CONDITIONS, WITH THE ULTIMATE DELIVERY REQUIRING INVASIVE NEUROSURGERY. WE PROPOSE TO BYPASS THE NEED TO GENERATE DOPAMINERGIC NEURONS EXOGENOUSLY, BY USING AN ALTERNATIVE APPROACH WHERE GLIAL PROGENITOR CELLS ALREADY PRESENT IN THE PARKINSON’S PATIENT’S BRAIN COULD BE DIRECTLY REPROGRAMMED TO BECOME DOPAMINERGIC NEURONS. IN EXPLORING THIS APPROACH, ACCESS TO ADULT HUMAN GLIAL PROGENITOR CELLS FROM NON-AFFECTED OR PD-PATIENTS IS A LIMITATION AND REPROGRAMMING OF ADULT HUMAN GLIAL PROGENITOR CELLS INTO DOPAMINERGIC NEURONS HAS NOT YET BEEN REPORTED. WE HAVE SUCCESSFULLY TARGETED RAT OLIGODENDROCYTE PROGENITOR CELLS (OPCS) IN VIVO TO REPROGRAM THEM INTO A NEURONAL LINEAGE. WITH THIS EXPLORATORY/DEVELOPMENTAL R21 MECHANISM AWARD, WE PROPOSE TO LINEAGE REPROGRAM HUMAN OPCS INTO DOPAMINERGIC NEURONS TO ULTIMATELY REPLACE NEURONS LOST IN PD. IN AIM 1, WE WILL TEST THE HYPOTHESIS THAT PD- DERIVED FIBROBLASTS DIRECTLY REPROGRAMMED INTO HUMAN OPCS WITHOUT PASSING THROUGH A PLURIPOTENT STATE RETAIN THEIR EPIGENETIC SIGNATURE AND CAN MODEL THE HUMAN OPCS WE EVENTUALLY INTEND TO TARGET FOR NEURONAL REPLACEMENT IN THE PD BRAIN. AIM 2 WILL TEST THE HYPOTHESIS THAT TEMPORAL SEQUENTIAL REPROGRAMMING CAN DIRECT ADULT-DERIVED HUMAN OPCS FROM NON-AFFECTED AND PD-PATIENTS TO BECOME DOPAMINE NEURONS. THESE IN VITRO STUDIES, DETERMINING THE TIMING AND SEQUENCE OF LINEAGE SPECIFICATION INSTRUCTIONS TO GENERATE DOPAMINERGIC NEURONS, WILL LAY THE NECESSARY FOUNDATION FOR THE FUTURE STEP USING DIRECT IN VIVO DELIVERY TO MODEL A REPROGRAMMING NEURONAL REPLACEMENT THERAPY FOR PD. THIS PROJECT ALSO HAS IMPLICATIONS FOR TESTING THE CAPACITY TO DRIVE THE GENERATED HUMAN OPCS TO COMPLETE DIFFERENTIATION AS OLIGODENDROCYTES AND TO THEN ADDRESS MYELINATION DEFICIENCIES IN PARKINSON’S DISEASE.

IDENTIFICATION OF A NOVEL TUMOR SUPPRESSOROF MELANOMA AND UV-INDUCED GENOME INSTABILITY - SKIN CANCER IS ONE OF THE MOST COMMON CANCERS IN THE US AND IMPOSES A HIGH ECONOMIC BURDEN. MOST SKIN CANCERS, INCLUDING MALIGNANT MELANOMA, ARE CAUSED BY ULTRAVIOLET (UV) LIGHT-INDUCED DNA DAMAGE AND GENOME INSTABILITY. IT IS WELL KNOWN THAT UV RADIATION (UVR)-INDUCED BULKY DNA ADDUCTS ARE BARRIERS FOR NORMAL REPLICATION PROGRESSION, AND THEIR FORMATION CAUSES REPLICATION FORK STALLING THAT IS A MAJOR DRIVING FORCE OF GENOME INSTABILITY. FAILURE TO STABILIZE STALLED FORKS AND RESUME STALLED REPLICATION OFTEN CAUSES FORK COLLAPSE, GENERATING DNA BREAKS AND GENOME INSTABILITIES THAT LEAD TO TUMORIGENESIS. HOWEVER, THE MECHANISM UNDERLYING HOW GENOME STABILITY IS MAINTAINED AND HOW STALLED REPLICATION IS RESCUED AFTER UV EXPOSURE IS POORLY UNDERSTOOD. UNDERSTANDING SUCH MECHANISM IS THUS IMPORTANT FOR UNDERSTANDING EARLY EVENTS IN MELANOMAGENESIS. MOREOVER, ENHANCING REPLICATION STRESS LEVELS IN TUMOR CELLS MAY OFFER A PROMISING CANCER THERAPEUTIC APPROACH, IN PARTICULAR FOR TREATING CANCERS HARBORING MUTATIONS IN REPLICATION STRESS RESPONSE GENES. THUS, OBTAINING AN IN-DEPTH UNDERSTANDING ON REPLICATION STRESS SUPPRESSION AND FORK REPAIR MAY ASSIST IN DEVELOPING NOVEL APPROACHES TO FACILITATE TARGETED THERAPY OF MELANOMA. THE LONG-TERM GOAL OF OUR RESEARCH PROGRAM IS TO DELINEATE THE MECHANISMS FOR MAINTAINING GENOME STABILITY IN RESPONSE TO EXPOSURE TO ENVIRONMENTAL GENOTOXINS. PI’S LAB HAS PIONEERED IN IDENTIFYING THE CST COMPLEX - A TRIMERIC PROTEIN COMPLEX CONSISTING OF CTC1, STN1, TEN1 THAT BINDS TO SSDNA WITH HIGH AFFINITY – AS AN IMPORTANT PLAYER IN MAINTAINING GLOBAL GENOME INTEGRITY UPON REPLICATION PERTURBATION. OUR RECENT DATA SUGGEST THE POTENTIAL INVOLVEMENT OF CST IN SUPPRESSING UVR-INDUCED GENOME INSTABILITY. THE GOAL OF THIS PROPOSAL IS TO TEST THE HYPOTHESIS THAT CST PLAYS AN IMPORTANT ROLE IN REGULATING REPLICATION REINITIATION WHEN FORKS ARE BLOCKED BY UV-INDUCED BULKY DNA ADDUCTS. CST DYSFUNCTION MAY ELEVATE UVR-INDUCED GENOME INSTABILITY AND INCREASE MELANOMA FORMATION. IN AIM 1, WE WILL DETERMINE HOW CST FACILITATES DNA SYNTHESIS WHEN UV-INDUCED BULKY LESIONS BLOCK REPLICATION PROGRESSION. IN AIM 2, WE WILL USE A NEW MOUSE MODEL TO DETERMINE WHETHER SPECIFIC DISRUPTION OF STN1 IN MATURE MELANOCYTES PROMOTES UVR-INDUCED MELANOMA PRODUCTION IN VIVO. IT IS EXPECTED THAT RESULTS FROM THE PROPOSED RESEARCH WILL OFFER NOVEL INSIGHTS INTO OUR UNDERSTANDING OF GENOME PROTECTION AFTER UV DAMAGE AND POTENTIALLY IDENTIFY A NOVEL TUMOR SUPPRESSOR OF MELANOMA, THUS FACILITATING THE DEVELOPMENT OF NEW APPROACHES FOR MELANOMA THERAPY.

STUDIES ON PROBENECID PRODRUGS THAT PROTECT AGAINST MITOCHONDRIAL TOXICITY OF TENOFOVIR - ABSTRACT: ALMOST 38 MILLION INDIVIDUALS WORLDWIDE ARE INFECTED WITH THE HUMAN IMMUNODEFICIENCY VIRUS (HIV) CAUSATIVE AGENT OF ACQUIRED IMMUNODEFICIENCY DISEASE (AIDS), 1.2 MILLION OF THEM BEING AMERICANS. WITH NO VACCINE AND NO CURE AVAILABLE, HIV/AIDS PATIENTS MUST TAKE ANTI-HIV DRUGS THROUGHOUT THE REST OF THEIR LIVES. THUS, THE LONG-TERM USE OF HIGHLY ACTIVE ANTIRETROVIRAL THERAPY (HAART)DRUGS THAT REVOLUTIONIZED HIV/AIDS AIDS PATIENTS CAN ALSO LIVE AS LONG AS NON-INFECTED PEOPLE, HAS BEEN ACCOMPANIED BY SERIOUS MITOCHONDRIAL TOXICITIES, WITH FATAL CONSEQUENCES THAT LED TO DISCONTINUATION OF MANY OF THE EARLY EFFECTIVE NUCLEOSIDE REVERSE TRANSCRIPTASE (RT) INHIBITOR (NRTIS) BACKBONE COMPONENTS, LIKE DDC AND D4T. THE DEVELOPMENT OF THE NUCLEOTIDE RT INHIBITORS (NTRTIS) PARTICULARLY TENOFOVIR (TFV), HAS BEEN INNOVATIVE IN THE HIV RT INHIBITORS FIELD, TO THE EXTENT THAT IT HAS GAINED WIDESPREAD USE WORLDWIDE AND IS INCORPORATED INTO ALMOST ALL FIRST-LINE HIV/AIDS THERAPEUTIC COMBINATION REGIMENS, AS WELL AS USED FOR PRE-EXPOSURE PROPHYLACTICS (PREP). TFV IS MARKETED AS TWO PRODRUGS TENOFOVIR DISOPROXIL FUMARATE (TDF) AND TENOFOVIR ALAFENAMIDE (TAF). HOWEVER, LONG-TERM USE OF THE TDF PRODRUG CAUSES KIDNEY TOXICITY STEMMING FROM THE ACTIVE PARENT TFV. TFV-ASSOCIATED KIDNEY TOXICITY IS SAID TO HAVE CONSTITUTED THE SINGLE REFERRALS FOR SPECIALIST RENAL SERVICES WHEN TFD WAS FIRST INTRODUCED. MOREOVER, THIS RENAL DECLINE WAS IRREVERSIBLE IN ABOUT 30 % OF PATIENTS EVEN AFTER CESSATION OF TFV THERAPY. ALTHOUGH, TAF, WHICH WAS INTRODUCED MORE RECENTLY IS MORE POTENT AND LESS TOXIC THAN TDF, CASE STUDIES SHOW IT HAS KIDNEY TOXICITY POTENTIAL WHICH MAY MANIFEST UPON LONG-TERM USE. MOREOVER, IT CAUSES TROUBLING WEIGHT GAIN THAT COULD EXACERBATE METABOLIC SYNDROME. THESE PROBLEMS MEAN THAT TAF CANNOT ALWAYS REPLACE TDF. THE MECHANISMS OF TFV TOXICITY ARE NOT WELL UNDERSTOOD, BUT DAMAGE TO RENAL PROXIMAL TUBULAR MITOCHONDRIAL HAS BEEN STRONGLY IMPLICATED. THE GOAL OF THIS RESEARCH IS TO GAIN BETTER UNDERSTANDING AND DEVELOP EFFECTIVE APPROACHES TO ADDRESSING TFV KIDNEY TOXICITY. THE SPECIFIC AIMS ARE: 1) TO CONFIRM THAT HUMAN MITOCHONDRIAL MEMBRANES EXPRESS FUNCTIONAL ORGANIC ANION TRANSPORTERS 1 AND 3 (HOAT1 AND HOAT3), AND SHOW THEIR ABILITY TO IMPORT TFV INTO MITOCHONDRIA, AND, 2) TO DEVELOP AND INVESTIGATE THE PROTECTIVE ABILITY OF MITOCHONDRIALLY TARGETED HOAT1 AND 3 INHIBITOR PRODRUGS AGAINST TFV TOXICITY. WE HAVE DEMONSTRATED THE EXPRESSION OF THE TFV TRANSPORTERS HOAT1 AND HOAT3 IN THE MITOCHONDRIAL MEMBRANES OF HUMAN RENAL PROXIMAL TUBULE EPITHELIAL CELLS, SUPPORTING OUR HYPOTHESIS THAT TFV ENTERS MITOCHONDRIA THROUGH HOAT1 AND 3 TO CAUSE MITOCHONDRIAL TOXICITY. FOR AIM 2, WE HAVE SYNTHESIZED AN INITIAL HOAT1, AND 3 INHIBITOR PRODRUG AND SHOWN THAT IT IS PROTECTIVE AGAINST TFV MITOCHONDRIAL TOXICITY. A COMPREHENSIVE MULTIDISCIPLINARY APPROACH INVOLVING MEDICINAL CHEMISTRY, BIOCHEMISTRY, FUNCTIONAL GENOMICS, AND PHARMACOLOGY WILL BE APPLIED, USING HUMAN KIDNEY PROXIMAL TUBULE CELLS. OUR INITIALLY SYNTHESIZED TARGETED PRODRUG REVERSES TFV-INDUCED MITOCHONDRIAL TOXICITY. THE SUCCESS OF THIS PROJECT WILL GIVE NEW INSIGHTS INTO TFV-INDUCED TOXICITY AND PROVIDE BIOLOGICAL PROBES AND/OR POTENTIAL DRUG LEADS.

INFLAMMATORY CYTOKINES AND PYROPTOSIS IN CORONAVIRUS INFECTION

VALIDATING NOVEL RYANODINE RECEPTOR-TARGETED COMPOUNDS FOR AD THERAPEUTICS

IMPROVING VACCINATION AGAINST TYPE-2 T-INDEPENDENT ANTIGENS - SUMMARY INFECTIONS WITH ENCAPSULATED BACTERIA ARE RESPONSIBLE FOR THE DEATH OF MILLIONS OF CHILDREN AND THE ELDERLY EACH YEAR. CAPSULAR POLYSACCHARIDES BEHAVE AS T-INDEPENDENT ANTIGENS (TI) AND VACCINATION BASED ON THESE CARBOHYDRATES PROVIDES PROTECTION AGAINST THESE INFECTIONS. THE EFFICACY OF VACCINES AGAINST TI ANTIGENS IS SEVERELY LIMITED BY THEIR INABILITY TO INDUCE RECALL RESPONSES UPON REVACCINATION OR INFECTION. THIS PHENOMENON DEPENDS ON INHIBITION OF MEMORY B CELLS BY ANTIGEN-SPECIFIC IGG GENERATED IN THE PRIMARY RESPONSE. THE MECHANISM BEHIND THIS INHIBITORY ACTIVITY HOWEVER, REMAINS UNCLEAR. THE EFFICACY OF VACCINES CRITICALLY DEPENDS ON ADJUVANTS THAT CAN PROMOTE STRONG MEMORY AND INFLUENCE IG CLASS SWITCH RECOMBINATION (CSR). HOWEVER, EFFECTIVE ADJUVANTS FOR TI ANTIGENS VACCINATION ARE NOT AVAILABLE. THE GOAL OF OUR PROPOSAL IS TO ANSWER TWO OUTSTANDING QUESTIONS REGARDING THE RESPONSE TO TI ANTIGENS: WHAT IS THE MECHANISM OF THE INHIBITION OF RECALL RESPONSES? AND HOW CAN CSR AND GENERATION OF MEMORY B CELLS CAN BE MANIPULATED TO IMPROVE VACCINATION? FOR THESE STUDIES WE WILL USE A MOUSE MODEL OF TULAREMIA AND WILL USE FRANCISELLA TULARENSIS LPS, A MODEL TI ANTIGEN THAT WE SHOWED ACTIVATES B1 CELLS. IN AIM 1 WE WILL TEST THE HYPOTHESIS THAT IGG-LPS IMMUNE COMPLEXES INHIBIT MEMORY B1 CELLS REACTIVATION AND IGM PRODUCTION THROUGH COMPLEMENT ACTIVATION. WE WILL TEST WHETHER COMPLEMENT DEPLETION OR ABSENCE COULD RESTORE THE RECALL RESPONSE TO VACCINATION WITH TI ANTIGENS AND RESULT IN A MORE EFFECTIVE VACCINE FOR TULAREMIA. IN AIM 2 WE WILL TEST THE HYPOTHESIS THAT NOVEL ADJUVANTS CAN IMPROVE RECALL RESPONSES TO TI ANTIGEN BY MANIPULATING CSR IN B1 CELLS AND/OR IMPROVING GENERATION OF MEMORY B1. BASED ON OUR PRELIMINARY RESULTS WE WILL TEST A NUMBER OF POTENTIAL ADJUVANT TREATMENTS WITH THE GOAL OF ACHIEVING SUSTAINED IGM AND INCREASED LPS-SPECIFIC IGA PRODUCTION BUT REDUCED IGG (RESPONSIBLE FOR INHIBITION OF RECALL RESPONSE). THESE ADJUVANTS ARE ALSO EXPECTED TO INCREASED NUMBERS OF MEMORY B1 CELLS. IF SUCCESSFUL, OUR STUDIES WILL CREATE PROOF OF CONCEPTS THAT CAN INFORM FUTURE DEVELOPMENT OF MORE EFFECTIVE VACCINATION STRATEGIES FOR TI ANTIGENS.

THE DEVELOPMENT OF GENE THERAPEUTIC APPROACHES TO SUPPRESS CEREBRAL INFLAMMATION IN DEMENTIA - THERE IS AN URGENT NEED FOR THE DEVELOPMENT OF NEW AND EFFECTIVE THERAPEUTIC APPROACHES TO ALZHEIMER’S DISEASE (AD). THE FIELD OF GENE THERAPY HAS PROGRESSED SIGNIFICANTLY IN THE LAST 10 YEARS AND IS BEGINNING TO ENTER THE CLINIC FOR DISEASE TREATMENT. GENE TRANSFER APPROACHES HAVE THE ADVANTAGE OF BEING MORE TARGETED TO SPECIFIC PATHWAYS AND REQUIRE FAR FEWER INTERVENTIONS COMPARED TO MORE TRADITIONAL APPROACHES. IN THE AD FIELD, IT HAS BECOME CLEAR THAT INFLAMMATION IS A KEY COMPONENT CONTRIBUTING TO AD PATHOLOGY. ONE MAJOR ROUTE FOR INFLAMMATION IS THROUGH ACTIVATION OF INFLAMMASOMES, SENSORS OF CELLULAR INSULTS. THEREFORE, WE PROPOSE THE DEVELOPMENT OF GENE TRANSFER APPROACHES TO INHIBIT INFLAMMASOME FUNCTION. THIS WILL BE DONE THROUGH THE USE OF A BRAIN ADMINISTERED DOMINANT-NEGATIVE (DN) INHIBITOR OF THE INFLAMMASOME COMPLEX (AIM 1) OR THROUGH THE USE OF A PERIPHERALLY DELIVERED BRAIN-TARGETED DN INHIBITOR OF THE INFLAMMASOME COMPLEX (AIM 2). ADENO- ASSOCIATED VIRAL (AAV) VECTORS WILL BE USED TO FACILITATE DN INHIBITOR GENE EXPRESSION AS THEY PROVIDE AN EFFICIENT AND SAFE VECTOR SYSTEM. WE WILL USE A RAT TRANSGENIC MODEL OF AD-LIKE AMYLOIDOSIS TO TEST THESE APPROACHES AND ASSESS LEARNING/MEMORY IN ADDITION TO NEUROCHEMICAL AND IMMUNOHISTOLOGICAL MEASURE ASSOCIATED WITH AD. THIS PROPOSAL WILL EXPLORE NOVEL THERAPEUTIC APPROACHES TO AD THROUGH TARGETING A KEY INFLAMMATORY DISEASE PATHWAY IN A RELEVANT ANIMAL MODEL.

REGULATION OF INTESTINAL MICROBIOTA BY B1 B CELLS DURING LUNG INFECTION

EPIGENETIC MECHANISMS OF DIABETIC METABOLIC MEMORY AS STUDIES IN ADULT ZEBRAFISH

INTERFERON GAMMA INDUCIBLE PROTEIN 16 AND KSHV GENE EXPRESSION

NMDA RECEPTOR PLASTICITY IN NUCLEUS ACCUMBENS SPINES DURING COCAINE WITHDRAWAL

FUNCTIONAL EXPRESSION OF CFTR AND OTHER MULTIDOMAIN PROTEINS

TOWARD SUICIDAL AUTOMATION OF PORPHYRIC LEISHMANIA FOR PHOTODYNAMIC VACCINATION

HEPATITIS C VIRUS AND LIVER FIBROGENESIS

UTILIZING GENE-LEVEL BIOMARKERS OF AD TO IDENTIFY PATHOPHYSIOLOGICAL MECHANISMS IN HUMAN NEURONS - ABSTRACT SUMMARY RECENT ADVANCES AD THERAPEUTICS INCREASES THE URGENCY TO IDENTIFY INDIVIDUALS AT RISK FOR DEVELOPING ALZHEIMER’S DISEASE (AD). THIS EFFORT CAN LIKELY BE ACCELERATED USING STEM CELL BIOLOGY APPROACHES WHICH HAVE LED TO THE ABILITY TO GENERATE HUMAN INDUCED NEURONS (HIN) DIRECTLY FROM EASILY OBTAINABLE PATIENT CELLS SUCH AS FIBROBLASTS. THIS SERVES AS A POWERFUL TOOL FOR STUDYING AGING AND DISEASE-RELATED PROCESSES IN CLINICALLY RELEVANT CELL TYPES. META-ANALYSIS STUDIES OF EXISTING TRANSCRIPTOMIC DATABANKS FROM AD PATIENTS AND HEALTHY INDIVIDUALS WITHOUT DEMENTIA (NONAD) HAVE ALSO PROVIDED NEW INSIGHT INTO UNDERLYING PATHOPHYSIOLOGICAL DRIVERS CONTRIBUTING TO A DIAGNOSIS OF AD. WHILE THESE GENE EXPRESSION PROFILES HAVE BEEN IDENTIFIED FOR AD RISK WHICH CARRYOVER FROM THE INDIVIDUAL TO THEIR DIRECTLY TRANSFORMED NEURONS (HUMAN INDUCED NEURON, HIN). THIS IS A CRUCIAL MISSING LINK NEEDED TO CAPTURE THE PHYSIOLOGICAL OUTCOMES OF THESE GENE NETWORK PROFILES IN ORDER TO IDENTIFY ASSOCIATED MECHANISMS OF METABOLIC STRESS, PATHOLOGICAL PROTEIN AGGREGATION AND SYNAPTIC PATHOPHYSIOLOGY – ALL KEY FEATURES OF AD. THEREFORE, WE WILL APPLY NEUROPHYSIOLOGICAL AND CELLULAR FUNCTIONAL ANALYSES TO HINS DERIVED FROM REPRESENTATIVE INDIVIDUALS DIAGNOSED WITH SPORADIC OR FAMILIAL AD, AS WELL AS AGE/SEX-MATCHED NONAD CONTROLS, TO IDENTIFY FUNCTIONAL CONSEQUENCES OF THESE GENOMIC VARIATIONS IN THE HUMAN POPULATION. OUR OVERALL OBJECTIVE IS TO IDENTIFY GENE-PATHWAY BASED BIOMARKERS OR RISK FACTORS FOR AD AND DEFINE THE NEURONAL PATHOPHYSIOLOGICAL PHENOTYPES ASSOCIATED WITH THESE GENE PATHWAYS. WE WILL TEST THE HYPOTHESIS THAT SYNAPTIC SIGNALING, METABOLISM AND PROTEIN HANDLING PROCESSES IN NEURONS DERIVED FROM THE AD POPULATION WILL REFLECT THE ALTERATIONS IN GENE PATHWAYS THAT DISTINGUISH HEALTHY AGING PROCESSES FROM AD PATHOGENESIS, AND THUS CAN SERVE AS IMPORTANT BIOMARKERS. IN AIM 1 WE WILL IDENTIFY MITOCHONDRIAL FUNCTIONAL DEFICITS ASSOCIATED WITH ALTERATIONS IN GLUCOSE METABOLISM GENE PATHWAYS IN AD. IN AIM 2 WE WILL IDENTIFY PATHOLOGICAL MANIFESTATIONS OF ALTERED GENE PATHWAYS REGULATING AUTOPHAGY AND PROTEIN MISHANDLING IN AD PATIENTS. IN AIM 3 WE WILL IDENTIFY NEUROPHYSIOLOGICAL AND SYNAPTIC SIGNALING EFFECTS CORRESPONDING WITH ALTERED GENE NETWORKS IN AD THAT UNDERLIE COGNITIVE DECLINE. THE COMBINATION OF OUR EXPERTISE IN INDUCED NEURON PRODUCTION AS WELL AS BIOINFORMATICS AND NEUROPHYSIOLOGY ASSAYS MAKES THIS A POWERFUL AND INNOVATIVE PROPOSAL THAT WILL PROVIDE IMPORTANT INSIGHTS INTO RISK FACTORS AND BIOMARKERS OF AD. THIS COMES AT A CRUCIAL TIME IN THE AD FIELD WHERE IT IS IMPERATIVE TO IDENTIFY AT RISK INDIVIDUALS EARLY IN ORDER TO MAXIMIZE BENEFITS OF NEW THERAPEUTICS BECOMING AVAILABLE.

STRUCTURE/FUNCTION OF MITOCHONDRIAL CITRATE CARRIER

IDENTIFICATION OF GENES RESPONSIBLE FOR SARCOLEMMAL INTEGRITY IN C. ELEGANS

PHOTO-INACTIVATION OF LEISHMANIA FOR SAFE AND EFFECTIVE DELIVERY OF SURROGATE VAC

ROLE OF ROS AND NRF2 IN HHV-8 INFECTION AND PATHOGENESIS

THE ROLE OF MITOCHONDRIAL STRESS RESPONSE IN ALCOHOL-MEDIATED NEUROTOXICITY - CHRONIC, EXCESSIVE ALCOHOL USE CAUSES REGIONAL STRUCTURAL BRAIN DAMAGE AND COGNITIVE DISORDERS. CHRONIC ALCOHOL MISUSE IS ALSO ASSOCIATED WITH A WIDE ARRAY OF MOVEMENT IMPAIRMENTS. CHRONIC ALCOHOL CONSUMPTION ALONE, OR TOGETHER WITH ALCOHOLIC HEPATIC ENCEPHALOPATHY, CAN CAUSE VARIOUS TYPES OF TREMOR, ASTERIXIS, AND CEREBELLAR DYSFUNCTION. WHILE ALCOHOL-INDUCED BRAIN DAMAGE HAS BEEN EXPLAINED BY ALCOHOL’S EFFECTS ON NEURAL EXCITABILITY OR NUTRITIONAL DEFICIENCY, WE DO NOT FULLY UNDERSTAND THE PATHOGENIC MECHANISM AT THE MOLECULAR AND CELLULAR LEVELS BY WHICH ALCOHOL EXERTS ITS TOXICITY AND DAMAGES THE NERVOUS SYSTEM. THE NEMATODE C. ELEGANS IS AN AMENABLE MODEL ORGANISM THAT CAN BE USED FOR DISSECTING THE PATHOLOGICAL MECHANISM OF ALCOHOLIC NEUROTOXICITY. OUR STUDY IN C. ELEGANS SHOWS THAT WHILE ALCOHOL STRONGLY INDUCES MANY CONSERVED CELLULAR STRESS RESPONSES, ITS MAIN TOXIC EFFECTS ARE CENTERED ON MITOCHONDRIAL FUNCTION. REMARKABLY, WE FIND THAT PERPETUAL MITOCHONDRIAL UNFOLDED PROTEIN RESPONSE (UPRMT) SPARES FROM A MOTOR FUNCTION DEFICIT CAUSED BY CHRONIC ALCOHOL EXPOSURE. MOREOVER, A GENETIC MANIPULATION THAT SPECIFICALLY INDUCES NEURON-SPECIFIC UPRMT ACTIVATION IS SUFFICIENT TO PROTECT AGAINST ALCOHOL-MEDIATED MOVEMENT IMPAIRMENT. WE HYPOTHESIZE THAT ACTIVATION OF A BRANCH OF UPRMT IN A SELECT NEURAL POPULATION PROTECTS AGAINST ALCOHOLIC MOVEMENT DISORDERS. TO TEST THIS HYPOTHESIS, WE PROPOSE TWO SPECIFIC AIMS: AIM 1 WILL INVESTIGATE HOW PERPETUAL UPRMT PROTECTS AGAINST ALCOHOL-MEDIATED MOVEMENT IMPAIRMENT; AND AIM 2 WILL IDENTIFY THE MECHANISM BY WHICH NEURONAL UPRMT LEADS TO THE PROTECTION AGAINST ALCOHOL-MEDIATED MOVEMENT IMPAIRMENT. A DETAILED ANALYSIS OF ALCOHOL-MEDIATED UPRMT AND ITS INTRA- AND INTERCELLULAR SIGNALING IN THE CONTEXT OF ALCOHOL-MEDIATED MOVEMENT IMPAIRMENT WILL LEAD TO EFFECTIVE DRUGGABLE TARGETS THAT PROTECT AGAINST, OR AMELIORATE, ALCOHOLIC MOVEMENT IMPAIRMENT.

ALCOHOL-INDUCED MUSCLE DAMAGE IN C. ELEGANS

INTRACELLULAR ORGANELLE DEFICITS DRIVING ALZHEIMER'S DISEASE - ALZHEIMER’S DISEASE (AD) IS A DEVASTATING NEURODEGENERATIVE DISORDER CHARACTERIZED BY AGGREGATION OF Β-AMYLOID (AΒ) PEPTIDES, NEUROFIBRILLARY TANGLES COMPOSED OF HYPERPHOSPHORYLATED TAU, AND A PROGRESSIVE LOSS OF COGNITIVE FUNCTION. WHILE MUCH IS KNOWN REGARDING THE BIOCHEMICAL COMPOSITION AND STRUCTURE OF AMYLOID AND TAU IN AD, RELATIVELY LESS FOCUS HAS BEEN PLACED ON THE INTRACELLULAR HANDLING OF DETRIMENTAL PROTEIN PRODUCTS, AND MORE SPECIFICALLY, HOW UPSTREAM DEFICITS IN ORGANELLE FUNCTION CAN ACCELERATE THE DISEASE PROCESS AND DIRECTLY CONTRIBUTE TO MEMORY IMPAIRMENTS. WHILE NEURONS RELY ON DEDICATED ORGANELLES TO EXECUTE SPECIFIC FUNCTIONS AND SUSTAIN HEALTH, SEVERAL IN PARTICULAR HAVE BEEN LINKED TO AD PATHOPHYSIOLOGY, INCLUDING THE ER, WHICH IS IMPORTANT FOR PROTEIN ASSEMBLY AND INTRACELLULAR CALCIUM SIGNALING; LYSOSOMES, WHICH ARE CRITICAL FOR BREAKING DOWN AND REMOVING THE CELLULAR DEBRIS AND MISFOLDED PROTEINS COLLECTED BY AUTHOPHAGOSOMES; AND MITOCHONDRIA, WHICH ARE RESPONSIBLE FOR THE BIOENERGETICS OF THE CELL (MUSTALY ET AL., 2018). IN THE GLOBAL OPERATIONS OF MAINTAINING NEURONAL VIABILITY, THE FUNCTIONS OF THESE ORGANELLES ARE HIGHLY INTER-DEPENDENT, AND THEY ARE OFTEN PHYSICALLY COUPLED TO ONE ANOTHER. DESPITE THE CLOSE COUPLING, THEIR RESPECTIVE ROLES IN CONTRIBUTING TO AD HAVE TYPICALLY BEEN STUDIED IN ISOLATION. FOR EXAMPLE, THERE ARE COMPELLING STUDIES DETAILING ASPECTS OF ER, LYSOSOME, OR MITOCHONDRIAL DYSFUNCTION IN AD, YET SUBSTANTIALLY LESS IS UNDERSTOOD ABOUT HOW ALTERED INTERACTIONS AMONG THESE ORGANELLES CAN LEAD TO PATHOGENIC CASCADES. THIS ISOLATIONIST APPROACH MAY LEAD TO CRITICAL OVERSIGHTS IN UNDERSTANDING KEY PROCESSES IN AD AND MISSING POTENTIAL THERAPEUTIC OPPORTUNITIES. THUS, THE OVERALL GOAL OF THIS STUDY IS TO IDENTIFY MECHANISMS UNDERLYING DEFICIENCIES IN SPECIFIC ORGANELLE FUNCTIONS AND EXAMINE HOW THIS AFFECTS THEIR INTERACTIONS, CHARACTERIZE HOW THIS POTENTIATES AD PATHOLOGY, AND ESTABLISH THE UPSTREAM DRIVERS OF THIS CASCADE FOR CONSIDERATION AS A THERAPEUTIC TARGET. THIS WILL BE ACCOMPLISHED THROUGH THE FOLLOWING AIMS: AIM I: DETERMINE IF THE AD-ASSOCIATED DISRUPTION IN ER FUNCTION DISRUPTS LYSOSOMAL DYNAMICS AND CLEARANCE OF AGGREGATED PROTEINS. AIM II: DETERMINE THE MECHANISM BY WHICH EXCESS ER-CA2+ RELEASE DISRUPTS MITOCHONDRIAL FUNCTION AND DEGRADATION. AIM III: ESTABLISH UPSTREAM DRIVERS OF INTRACELLULAR PATHOGENIC CASCADES AND DETERMINE IF TARGETING ER-HOMEOSTASIS WILL RESOLVE LYSOSOMAL AND MITOCHONDRIAL DEFECTS. THE PROPOSED STUDY WILL HAVE A SIGNIFICANT IMPACT ON THE FIELD AS IT WILL PROVIDE NEW MECHANISTIC INFORMATION ABOUT HOW MISAGGREGATED PROTEINS ACCUMULATE IN AD AND ARE ASSOCIATED WITH ALTERED ER SIGNALING. MOREOVER, TARGETING SPECIFIC INTRACELLULAR ORGANELLES MAY REVEAL EFFECTIVE NEW TREATMENT STRATEGIES FOR AD.

ANTI-NUCLEOLIN APTAMER AS1411: APPLICATIONS IN KAPOSI'S SARCOMA ASSOCIATED HERPES VIRUS (KSHV) BIOLOGY - KAPOSI’S SARCOMA-ASSOCIATED HERPESVIRUS (KSHV), A MEMBER OF THE HUMAN -HERPESVIRUS FAMILY IS ALSO TERMED AS HUMAN HERPESVIRUS TYPE 8 (HHV-8). KSHV IS AN ETIOLOGICAL AGENT OF AN ENDOTHELIAL TUMOR CALLED KAPOSI’S SARCOMA (KS) AND A HIGHLY AGGRESSIVE LYMPHOPROLIFERATIVE B CELL LYMPHOMA CALLED PRIMARY EFFUSION LYMPHOMA (PEL). KS IS THE MOST COMMON VASCULAR MALIGNANCY CAUSING HIGH MORBIDITY AND MORTALITY IN HIV-INFECTED PATIENTS. PEL IS A RARE TUMOR OF HEMATOPOIETIC AND LYMPHOID TISSUES AND IS ASSOCIATED WITH A POOR PROGNOSIS. PEL CASES HAVE BEEN DRAMATICALLY REDUCED IN THE US SINCE THE WIDESPREAD USE OF COMBINED ANTIRETROVIRAL THERAPY (CART) BUT KSHV STILL CAUSES SIGNIFICANT MORTALITY IN THE DEVELOPING WORLD. THERE IS A CRITICAL NEED IN KSHV TARGETING SPECIFIC ANTIVIRAL DRUGS, WHICH ARE VERY WELL TOLERATED WITH NO SEVERE ADVERSE EVENTS FOR IMMUNOCOMPROMISED PATIENTS WITH KSHV ASSOCIATED MALIGNANCIES. KSHV EXHIBITS TWO DISTINCT PHASES IN ITS LIFE CYCLE. DURING LATENCY, A MINIMAL NUMBER OF LATENCY GENES SUCH AS VFLIP (ORF71), VCYCLIN (ORF72), LATENCY-ASSOCIATED NUCLEAR ANTIGEN-1 (LANA-1; ORF73), AND KAPOSIN ARE EXPRESSED. BY CONTRAST, DURING THE LYTIC PHASE, KSHV EXPRESSES A WIDE ARRAY OF IMMEDIATE-EARLY (IE), DELAYED-EARLY (DE), AND LATE GENES; UNDERGO ACTIVE REPLICATION AND PRODUCE VIRION PROGENY. KSHV HAS BEEN SHOWN TO UTILIZE MULTIPLE HOST GROWTH FACTORS, CYTOKINES, ANGIOGENIC FACTORS, AND CELL SIGNALING AND METABOLISM-RELATED PROTEINS FOR CREATING A BENEFICIAL ENVIRONMENT FOR ITS REPLICATION, SURVIVAL, AND LATENCY. OUR EXCITING STUDIES DISCOVERED THAT: 1) KS SKIN LESIONS EXPRESS REMARKABLY ROBUST EXPRESSION OF NUCLEOLIN COMPARED TO HEALTHY SKIN TISSUE; 2) KSHV DE NOVO INFECTION IN PRIMARY ENDOTHELIAL CELLS INDUCED A HIGH LEVEL OF NUCLEOLIN AND PHOSPHO-NUCLEOLIN, AND MORE IMPORTANTLY, 3) INCUBATION OF KSHV INFECTED CELLS WITH G- QUADRUPLEX FORMING ANTI-NUCLEOLIN APTAMER AS1411 REDUCED KSHV LATENT (ORF73) AND INCREASED LYTIC (ORF50) GENE EXPRESSION AND WE OBTAINED SIMILAR RESULTS IN NUCLEOLIN SILENCED KSHV INFECTED PEL CELLS. 4) AS1411 TREATMENT WAS EFFICACIOUS IN INDUCING CELL DEATH IN KSHV INFECTED PEL CELL LINES. BASED ON OUR PRELIMINARY RESULTS, WE HYPOTHESIZE THAT KSHV INDUCES HOST FACTOR NUCLEOLIN TO SUPPORT ITS LATENCY AND LIFE CYCLE AND TARGETING NUCLEOLIN WITH AS1411 WOULD HAVE THERAPEUTIC POTENTIAL IN KSHV ASSOCIATED MALIGNANCIES. TO TEST THIS HYPOTHESIS, WE HAVE FORMULATED TWO SPECIFIC AIMS IN WHICH WE WILL 1) TO DETERMINE THE REGULATORY MECHANISMS OF NUCLEOLIN EXPRESSION UPON KSHV INFECTION AND ITS ROLE IN KSHV LATENCY, AND 2) TO EVALUATE THE CHEMOTHERAPEUTIC POTENTIAL OF USING AS1411 TO TREAT KS AND PEL. OUR STUDIES ARE SIGNIFICANT AND WILL HAVE A POSITIVE IMPACT BY ADVANCING THE UNEXPLORED AND NOVEL TARGETED THERANOSTIC FIELD OF APTAMERS IN KSHV BIOLOGY AND UNDERSTANDING THEIR ANTIVIRAL AND ANTICANCER POTENTIAL CAN ALSO BE APPLIED TO OTHER VIRAL MALIGNANCIES.

WALK-RELATED PERFORMANCE FATIGABILITY AS A NOVEL MARKER OF FALL RISK IN LOWER LIMB PROSTHESIS USERS WITH DIABETES

MACHINE LEARNING DISCOVERY OF NOVEL BRAINSTEM NUCLEI CONTROLLING OROFACIAL BEHAVIORS - PROJECT SUMMARY/ABSTRACT THE BRAINSTEM MEDULLARY RETICULAR FORMATION GOVERNS VITAL MOTOR BEHAVIORS, SUCH AS BREATHING, VOCALIZATION, SWALLOWING, AND CHEWING, AS WELL AS EXPRESSIVE MOVEMENTS OF THE FACE AND MOUTH. CRITICAL FOR UNDERSTANDING THESE NEURAL CIRCUITS IS IDENTIFICATION AND LOCALIZATION OF RETICULAR SUBPOPULATIONS CONTROLLING THESE MYRIAD FUNCTIONS. A KEY OBSTACLE TO IDENTIFYING THESE NUCLEI IS THE LACK OF CLEAR CYTOARCHITECTONIC BOUNDARIES AND MOLECULAR MARKERS WITHIN THE RETICULAR FORMATION DELINEATING FUNCTIONAL NUCLEI. OUR CENTRAL HYPOTHESIS IS THAT FUNCTIONAL SUBPOPULATIONS WITHIN THE MEDULLARY RETICULAR FORMATION THAT PLAY DISTINCT ROLES IN OROFACIAL MOTOR BEHAVIORS CAN BE IDENTIFIED BASED ON THEIR GENE EXPRESSION PROFILE AND OROFACIAL MOTOR RESPONSES USING MACHINE LEARNING CLUSTERING AND CELL TYPE-SPECIFIC, SPATIALLY-SELECTIVE OPTOGENETIC MANIPULATION. TESTING THIS IDEA IS A CRITICAL STEP TOWARDS UNDERSTANDING CONTROL AND COORDINATION OF OROFACIAL MOTOR BEHAVIORS AND THEIR DISRUPTION IN NEURODEVELOPMENTAL AND NEURODEGENERATIVE CONDITIONS, SUCH AS RETT SYNDROME, AUTISM SPECTRUM DISORDERS, AND PARKINSON’S DISEASE, THAT CAUSE ATYPICAL FACIAL MOVEMENTS AND SPEECH AND SOMETIMES FATAL DISORDERS IN SWALLOWING AND BREATHING. THE APPROACH COMPRISES TWO AIMS: (AIM 1) CLUSTER MEDULLARY RETICULAR FORMATION INTO MACHINE LEARNING-DEFINED NUCLEI (ML NUCLEI) THAT REPRESENT FUNCTIONALLY-DISTINCT OROFACIAL CONTROL MODULES USING A LARGE SCALE, PUBLICLY AVAILABLE SINGLE CELL SPATIAL TRANSCRIPTOMIC ATLAS FROM THE ALLEN INSTITUTE; AND (AIM 2) MAP OROFACIAL RESPONSES TO SYSTEMATIC, CELL TYPE-SPECIFIC OPTOGENETIC MANIPULATION OF MEDULLARY RETICULAR FORMATION FOR DELINEATION OF FUNCTIONALLY DISTINCT OROFACIAL CONTROL MODULES. IN PRELIMINARY DATA, WE EXTRACT, PROCESS, CLUSTER, AND ANALYZE A SINGLE CELL SPATIAL TRANSCRIPTOMIC ATLAS FROM AN ADULT MOUSE TO SHOW THE FEASIBILITY AND VALIDITY OF OUR APPROACH. WE FURTHER DEMONSTRATE OUR TECHNICAL CAPACITY TO CARRY OUT AND ANALYZE OROFACIAL KINEMATICS IN RESPONSE TO RETICULAR FORMATION INJECTIONS IN AWAKE, HEAD-FIXED ADULT MICE. THE EXPECTED OUTCOME OF THE EXPLORATORY PROJECT IS DETERMINATION AND FUNCTIONAL VALIDATION OF PRECISE BOUNDARIES FOR PREVIOUSLY IDENTIFIED AND NOVEL SUBPOPULATIONS UNDERLYING CONTROL OF BEHAVIORS CRITICAL FOR SURVIVAL. THE OVERALL IMPACT OF THIS PROPOSAL WILL BE A HIGHLY INNOVATIVE APPLICATION OF MACHINE LEARNING TO NEUROSCIENCE DISCOVERY, AKIN TO AN UNBIASED NEUROANATOMICAL “SCREEN,” THAT MAY BE APPLIED TO TISSUES THROUGHOUT THE BODY AND THE ELUCIDATION OF FUNDAMENTAL PRINCIPLES GOVERNING THE ORGANIZATION OF THE BRAINSTEM THAT WILL ENABLE DEVELOPMENT OF THERAPIES FOR TREATING BRAINSTEM DISORDERS AND IMPROVE SURGICAL INTERVENTIONS REQUIRING A HIGHER RESOLUTION UNDERSTANDING OF BRAINSTEM STRUCTURE AND FUNCTION.

MOLECULAR AND CELLULAR REGULATION OF CAV2 VOLTAGE-GATED CALCIUM CHANNELS - CAV2 VOLTAGE-GATED CALCIUM CHANNELS MEDIATE CALCIUM INFLUX AT PRESYNAPTIC AXON TERMINALS, TRIGGERING NEUROTRANSMITTER RELEASE. GENETIC VARIANTS IN CAV2 CHANNELS ARE ASSOCIATED WITH MULTIPLE NEUROLOGICAL AND NEURODEVELOPMENTAL CONDITIONS, INCLUDING ATAXIA, SEIZURES, AND AUTISM. CAV2 CHANNEL LEVELS ARE UPREGULATED OR DOWNREGULATED IN VARIOUS NEUROLOGICAL DISEASES, INJURIES, AND PATHOLOGICAL CONDITIONS; HOWEVER, THE MOLECULAR MECHANISMS GOVERNING THEIR TRAFFICKING AND DEGRADATION REMAIN LARGELY UNKNOWN. UNLIKE MAMMALS, WHERE MULTIPLE GENES ENCODE SUBUNITS, LEADING TO REDUNDANCY AND COMPENSATORY MECHANISMS, THE NEMATODE C. ELEGANS HAS A SIMPLIFIED CAV2 CHANNEL ORGANIZATION, WITH EACH SUBUNIT ENCODED BY A SINGLE GENE. MOREOVER, IMPAIRMENT IN C. ELEGANS CAV2 CHANNELS LEADS TO EASILY OBSERVABLE DISRUPTIONS IN RHYTHMIC BODY MOVEMENTS, PROVIDING MEASURABLE INDICATORS OF CHANNEL FUNCTION AND ABUNDANCE. THUS, WE PROPOSE THAT C. ELEGANS IS A RAPID AND EFFICIENT MODEL TO IDENTIFY GENES THAT REGULATE CAV2 CHANNELS. THROUGH AN UNBIASED FORWARD GENETIC SCREEN IN A SENSITIZED BACKGROUND, WE IDENTIFIED AT LEAST FIVE NOVEL GENES THAT REGULATE CAV2 CHANNEL LEVELS. IN THIS PROPOSAL, WE AIM TO MOLECULARLY CHARACTERIZE THESE NEWLY IDENTIFIED GENES AND ELUCIDATE THEIR ROLES IN CAV2 TRAFFICKING AND DEGRADATION. THE EXPECTED OUTCOMES OF THIS STUDY ARE TO ADVANCE OUR UNDERSTANDING OF CAV2 CHANNELOPATHIES AND TO UNCOVER POTENTIAL NEW THERAPEUTIC TARGETS FOR MODULATING CAV2 CHANNEL LEVELS.

CELLULAR AND SYNAPTIC SIGNALING MECHANISMS DRIVING EARLY AD PATHOLOGY

HHV-8, ANGIOGENESIS AND INFLAMMATION

ROLE OF THE INFLAMMASOME IN HEPATITIS C VIRUS PATHOGENESIS

AFFERENTS MODULATING VTA ACTIVITY AND THEIR PLASTICITY AFTER SELF-ADMINISTRATION

DEVELOPING SELECTIVE INHIBITORS AND PROBES FOR CONCENTRATIVE NUCLEOSIDE TRANSPORTERS - ABSTRACT: NUCLEOSIDE TRANSPORTERS (NTS) MEDIATE THE CELLULAR TRANSPORT OF PHYSIOLOGICAL NUCLEOSIDES AND MANY SYNTHETIC DERIVATIVES LIKE THE REVERSE TRANSCRIPTASE INHIBITORS (NRTIS) USED IN HIV/AIDS THERAPY. THE EMERGING TOXICITY AND ADVERSE EFFECTS LIKE POTENTIALLY IRREVERSIBLE KIDNEY TOXICITY, WEIGHT GAIN/METABOLIC SYNDROME AND POTENTIAL FETAL ABNORMALITIES PLAGUING THE MOST POPULAR HIV/AIDS DRUGS TENOFOVIR DISOPROXIL FUMARATE (TDF), TENOFOVIR ALAFENAMIDE (TAF) AND INTEGRASE INHIBITOR DOLUTEGRAVIR, MAKES RELYING ON NRTIS, WHICH CONSTITUTE THE BACKBONES OF MANY HIV/AID COMBINATION THERAPIES, ATTRACTIVE. AS SUCH IT IS IMPERATIVE TO SEEK TO OPTIMIZE THEIR THERAPEUTIC OUTCOMES. HUMAN (H) CONCENTRATIVE NUCLEOSIDE TRANSPORTERS (HCNTS) FAMILY COMPRISES THREE MEMBERS, HCNT1, 2 AND 3, THAT MEDIATE SODIUM-DEPENDENT NUCLEOSIDE TRANSPORT. IN CONTRAST TO THEIR UBIQUITOUSLY EXPRESSED HUMAN EQUILIBRATIVE NUCLEOSIDE TRANSPORTERS (HENTS) COUNTERPARTS, HENT1, HENT2, HENT3 AND HENT4, HCNTS ARE RESTRICTED IN TISSUE DISTRIBUTION, BEING PREVALENT IN ABSORPTIVE TISSUES. UNLIKE HENTS WITH BROAD SUBSTRATE SPECIFICITY, CNTS HAVE LIMITED SUBSTRATE SPECIFICITIES, WITH HCNT1 AND HCNT2 PREFERRING PYRIMIDINE AND PURINE NUCLEOSIDE SUBSTRATES, RESPECTIVELY, WHILE HCNT3 TRANSPORTS BOTH NUCLEOSIDE CLASSES. THEY OCCUR ON THE LUMINAL SIDE OF KIDNEYS AND ARE PRINCIPAL DRIVERS OF REABSORPTION OF NUCLEOSIDE DRUGS LIKE THE NRTIS, WHICH COULD RESULT IN TOXICITIES. THE TIME IS NOW TO OPTIMIZE NRTI THERAPIES, AND ONE WAY IS TO ADDRESS THEIR POTENTIAL MITOCHONDRIAL TOXICITY (MITOTOXICITY), WHICH IS VERY TROUBLING IN EARLY DRUGS LIKE ZALCITABINE (DDC) AND DIDANOSINE (DDI), LIMITING USE. THE CURRENT FREQUENTLY USED NRTIS, ZIDOVUDINE (AZT), LAMIVUDINE (3TC) AND EMTRICITABINE (FTC) ALSO HARBOR MITOTOXICITY, PARTICULARLY AZT, THAT HAS NOT BEEN ADDRESSED. THE GOALS OF THIS RESEARCH ARE TO DEVELOP HCNT SUBTYPE SELECTIVE INHIBITORS FOR WHICH THERE IS A WOEFUL LACK, HAMPERING THE STUDY OF HCNTS’ BIOLOGY AND PHARMACOLOGY, AND FOR THERAPEUTIC APPLICATIONS LIKE BLOCKING DRUG REABSORPTION TO MITIGATE AGAINST MITOTOXICITY. UNLIKE THEIR HENT COUNTERPARTS, FOR WHICH THERE ARE SPECIFIC INHIBITORS WITH IC50’S DOWN TO NM LEVELS, HCNTS LACK THEM. OF NOTE, THE STANDARD HCNT INHIBITOR, PHLORIDZIN (PHZN), HAS HCNT1 INHIBITORY IC50 AS HIGH AS 250 ΜM, WITH LOW SUBTYPE SELECTIVITY, AND ALSO INHIBITS SODIUM-GLUCOSE TRANSPORTERS (SGLTS) EVEN MORE POTENTLY. WE ARE APPLYING A MULTIDISCIPLINARY APPROACH COMPRISING STRUCTURE- AND LIGAND-BASED DESIGN, SYNTHESIS, CELL-BASED BIOASSAYS AND ADMET TO DISCOVER AND OPTIMIZE POTENT HCNT SUBTYPE SELECTIVE/SPECIFIC INHIBITORS. WE HAVE ALREADY IDENTIFIED LEAD COMPOUNDS WITH IC50 VALUES DOWN TO 2 ΜM, AND UP TO 25-FOLD IMPROVEMENT OVER PHLORIDZIN AGAINST HCNT1. OUR SPECIFIC AIMS ARE: 1) TO DISCOVER AND OPTIMIZE POTENT SUBTYPE SELECTIVE HCNT INHIBITORS, TO BE USED AS BIOLOGICAL TOOLS, AND 2) TO USE HCNT3-SPECIFIC/SELECTIVE INHIBITOR FOR PROOF-OF-CONCEPT THAT TARGETING HCNT3 CAN DECREASE NRTI REUPTAKE AND MITIGATE AGAINST MITOCHONDRIAL TOXICITY. A MULTIDISCIPLINARY APPROACH COMBINING COMPUTATIONAL AND SYNTHETIC CHEMISTRY AND CELL-BASED BIOLOGICAL TESTING WILL BE APPLIED. THE SUCCESS OF THE PROJECT WILL MAKE AVAILABLE HIGHLY ANTICIPATED PROBE COMPOUNDS FOR STUDYING THE BIOLOGY OF AND PHARMACOLOGY OF THIS IMPORTANT TRANSPORTERS THAT ARE CRITICAL TO THE SUCCESS OF NUCLEOSIDE DRUG THERAPIES.

MATERNAL EXPERIENCE AND STRESS RESILIENCE IN FEMALES

ROLE OF CYCLOOXYGENASE-2 AND PGE2 IN KSHV PATHOGENESIS

ASSESSING THE ROLE OF THE SOCKET TRIM LINE ON REACTIVE BALANCE IN PEOPLE WITH TRANSFEMORAL AMPUTATION

DURABILITY TESTING OF 3D-PRINTED PROSTHETIC SOCKETS.

PHYSIOLOGY OF INSECT AMINO ACID TRANSPORT

PHYSIOLOGY OF INSECT AMINO ACID TRANSPORT

CRITICAL ROLES OF ENDOGENOUS TLR SIGNALING IN DRIVING TH17 EFFECTOR RESPONSES

DO COCAINE AND CHRONIC STRESS CONVERGE IN THE BASOLATERAL AMYGDALA

ELUCIDATING THE ROLE OF HEPATIC KETOGENESIS IN PANCREATIC CANCER CACHEXIA AND RECOVERY - PROJECT SUMMARY/ABSTRACT 80% OF PATIENTS WITH PANCREATIC DUCTAL ADENOCARCINOMA (PDAC) DEVELOP CACHEXIA, WHICH RESULTS IN DISPROPOR- TIONATE SKELETAL MUSCLE MASS LOSS, RELATIVE TO CALORIC DEFICITS. CACHEXIA CAUSES FUNCTIONAL IMPAIRMENT AND INELI- GIBILITY FOR ANTI-TUMOR THERAPEUTIC INTERVENTIONS, LEADING TO DRASTICALLY LOWER PATIENT QUALITY OF LIFE AND INCREASED MORTALITY. SUPPLEMENTAL NUTRITION IS NOT SUFFICIENT TO PREVENT TISSUE LOSS, AND THERE IS AN ABJECT LACK OF TARGETED ANTI-CACHECTIC THERAPEUTICS. THIS IS LARGELY DUE TO AN INCOMPLETE UNDERSTANDING OF HOW EFFECTORS, SUCH AS INTER- LEUKIN 6 (IL-6), DRIVE CACHEXIA, AND MECHANISTIC STUDIES THAT POORLY REFLECT THE CLINICAL SCENARIOS OF PDAC SURVI- VORSHIP. IMPROVING THE QUALITY OF LIFE AND SURVIVORSHIP OF PATIENTS WITH PDAC REQUIRES THAT WE ADDRESS A FUNDA- MENTAL KNOWLEDGE GAP IN THE MECHANISMS OF CACHEXIA PERSISTENCE AFTER CANCER RECOVERY. OUR RECENT WORK SHOWS THAT SYSTEMIC INFLAMMATION IN PDAC PREVENTS THE LIVER FROM ADAPTING TO KETOGENIC METABOLISM DURING NUTRITIONAL SCARCITY, WHICH LEADS TO MUSCLE LOSS. LOSS OF HEPATIC SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION 3 (STAT3) IS SUFFICIENT TO RESTORE KETOGENIC POTENTIAL AND PREVENT MUSCLE LOSS IN PDAC MICE. THIS WORK IS THE FIRST TO SHOW THAT IMPROVING HEPATIC FATTY ACID OXIDATION PREVENTS CACHEXIA PROGRESSION. TO IMPROVE THE TRANSLATION OF THESE FINDINGS WE MUST ADDRESS TWO UNMET NEEDS: 1) THE MECHANISM OF STAT3-DRIVEN DOWN REGULATION OF LIPID OXIDATION AND 2) STUDIES THAT ADDRESS CACHEXIA RECOVERY IN CANCER SURVIVORS. BASED ON OUR PRIOR WORK, WE HYPOTHESIZE THAT STAT3 SIGNALING ACTS ON THE LIVER TO INDUCE METABOLIC CHANGES THAT PERSIST AFTER CANCER RECOVERY, AT THE DETRIMENT OF THE MUSCLE. THIS PROJECT IS INNOVATIVE BECAUSE IT USES ADVANCED EPIGENETIC TECHNIQUES COMBINED WITH A NOVEL MOUSE MODEL OF PDAC SURVIVORSHIP TO CLEARLY DEFINE THE EPIGENETIC REPROGRAMMING EVENTS THAT PERSIST AFTER TUMOR CLEARANCE AND INCREASE VULNERABILITY OF MUSCLE TO NUTRITIONAL STRESS. THE SIGNIFICANCE OF THIS PROJECT LIES IN PRE-CLINICAL TESTING OF METABOLIC INTERVENTIONS TO PROTECT MUSCLE DURING PDAC RECOVERY, DEFINING A PREVIOUSLY UNDESCRIBED ACTION OF STAT3 IN HEPATIC METABOLISM CONTROL, AND LINKING LONG-TERM METABOLIC DYSFUNCTION TO EPI- GENETIC REPROGRAMMING DURING PDAC CACHEXIA. THE LONG-TERM GOAL OF OUR RESEARCH IS TO IMPROVE THE QUALITY OF LIFE, FROM DIAGNOSIS THROUGH SURVIVORSHIP, OF PATIENTS WITH CANCER CACHEXIA THROUGH A MECHANISTIC UNDERSTANDING OF INTER-ORGAN METABOLIC EVENTS.

ANGER EXPRESSION, ENDOGENOUS OPIOIDS AND ACUTE PAIN

A MODIFIABLE BIOMECHANICAL EXPLANATION FOR OBESITY-RELATED FALLS BY OLDER ADULTS

MOLECULAR CLONING OF EPITHELIAL K CHANNELS

GLUTAMATE AND PSYCHOSTIMULANT-INDUCED NEUROADAPTATIONS

IMPACT OF SOCIAL DRIVE ON SOCIAL FEAR LEARNING AND AMYGDALA FUNCTION DURING DEVELOPMENT

VIOLENCE EXPOSURE AND HIV RISK IN ADOLESCENT WOMEN OF COLOR

PROTEIN TRANSLATION AND THE INCUBATION OF COCAINE CRAVING

NURSE ANESTHETIST TRAINEESHIPS

PLASTICITY IN NUCLEUS ACCUMBENS SPINES DURING INCUBATION OF COCAINE CRAVING

PAPILLOMAVIRUS INFECTION/ROLE/MINOR CAPSID PROTEIN L2

COCAINE-INDUCED AMPAR PLASTICITY: MODULATION BY METABOTROPIC GLUTAMATE RECEPTORS?

TARGETING CGMP SIGNALING IN EXPERIMENTAL PARKINSONISM

THE ROLE OF ACTIVITY-REGULATED CYTOSKELETAL-ASSOCIATED PROTEIN IN AMPHETAMINE PLA

THE ROLE OF SIGNAL TRANSDUCTION CASCADES IN MEDIATING PAPILLOMAVIRUS INFECTION.

NSL - GRADUATE NURSING - NEW

MICRO RNAS OF EXOSOMES FOR EARLY DETECTION OF MAMMARY NEOPLASMS

GRADUATE RESEARCH FELLOWSHIP PROGRAM (GRFP) -THE NATIONAL SCIENCE FOUNDATION (NSF) GRADUATE RESEARCH FELLOWSHIP PROGRAM (GRFP) IS A HIGHLY COMPETITIVE, FEDERAL FELLOWSHIP PROGRAM. GRFP HELPS ENSURE THE VITALITY AND DIVERSITY OF THE SCIENTIFIC AND ENGINEERING WORKFORCE OF THE UNITED STATES. THE PROGRAM RECOGNIZES AND SUPPORTS OUTSTANDING GRADUATE STUDENTS WHO ARE PURSUING RESEARCH-BASED MASTER'S AND DOCTORAL DEGREES IN SCIENCE, TECHNOLOGY, ENGINEERING, AND MATHEMATICS (STEM) AND IN STEM EDUCATION. THE GRFP PROVIDES THREE YEARS OF FINANCIAL SUPPORT FOR THE GRADUATE EDUCATION OF INDIVIDUALS WHO HAVE DEMONSTRATED THEIR POTENTIAL FOR SIGNIFICANT RESEARCH ACHIEVEMENTS IN STEM AND STEM EDUCATION. THIS AWARD SUPPORTS THE NSF GRADUATE FELLOWS PURSUING GRADUATE EDUCATION AT THIS GRFP INSTITUTION. THIS AWARD REFLECTS NSF'S STATUTORY MISSION AND HAS BEEN DEEMED WORTHY OF SUPPORT THROUGH EVALUATION USING THE FOUNDATION'S INTELLECTUAL MERIT AND BROADER IMPACTS REVIEW CRITERIA.- SUBAWARDS ARE NOT PLANNED FOR THIS AWARD.

RAPID NETWORK REORGANIZATION WITH LEARNING

SYNAPTIC ADAPTATIONS UNDERLYING THE INCUBATION OF COCAINE CRAVING

THE IDENTIFICATION AND MANIPULATION OF RNA SPLICING FACTOR REGULATORS IN SPINAL M

ROLE OF TARPS IN ADDICTION-RELATED PLASTICITY

LONGITUDINAL WHITE MATTER CHANGES IN INDIVIDUALS AT RISK FOR ALZHEIMER'S DISEASE

SYNTHESIS AND DEGRADATION OF SYNAPTIC PROTEINS DURING COCAINE WITHDRAWAL

A LONGITUDINAL INVESTIGATION OF INTERSECTIONAL DISCRIMINATION, ALCOHOL USE OUTCOMES, AND UNDERLYING MECHANISMS AMONG BISEXUAL PEOPLE OF COLOR - PROJECT SUMMARY BISEXUAL AND OTHER MULTI-GENDER ATTRACTED PEOPLE (E.G., PANSEXUAL, QUEER; “BI+”) ARE DISPROPORTIONATELY AFFECTED BY ADVERSE ALCOHOL USE OUTCOMES (E.G., HEAVY EPISODIC DRINKING, DRINKING CONSEQUENCES, ALCOHOL USE DISORDER [AUD] SYMPTOMS) COMPARED TO BOTH HETEROSEXUAL AND GAY/LESBIAN PEOPLE, AND DISCRIMINATION HAS BEEN IMPLICATED AS A KEY DRIVER OF THESE DISPARITIES. PEOPLE OF COLOR (POC) ARE ESPECIALLY LIKELY TO IDENTIFY AS BI+, AND BI+ POC ARE SUBJECTED TO UNIQUE FORMS OF DISCRIMINATION AT THE INTERSECTION OF SEXUAL ORIENTATION AND RACE/ETHNICITY. STILL, VERY FEW STUDIES HAVE EXAMINED RISK FOR ALCOHOL USE OUTCOMES AMONG BI+ POC, LEAVING CRITICAL GAPS IN OUR UNDERSTANDING OF RISK IN THIS POPULATION. IN PARTICULAR, THERE HAS BEEN A LACK OF ATTENTION TO THE RANGE OF DISCRIMINATORY EXPERIENCES AFFECTING BI+ POC (I.E., BI+, RACIAL/ETHNIC, AND INTERSECTIONAL DISCRIMINATION, SUCH AS HETEROSEXISM WITHIN ONE’S RACIAL/ETHNIC COMMUNITY AND RACISM WITHIN THE LGBTQ+ COMMUNITY) AND THEIR ASSOCIATIONS WITH ALCOHOL USE OUTCOMES. EVIDENCE INDICATES THAT BI+ DISCRIMINATION IS ASSOCIATED WITH HEAVY EPISODIC DRINKING, DRINKING CONSEQUENCES, AND AUD SYMPTOMS, BUT THESE STUDIES HAVE NOT ATTENDED TO OTHER FORMS OF DISCRIMINATION THAT AFFECT BI+ POC AND THEIR EFFECTS ON ALCOHOL USE OUTCOMES OVER TIME. THE PROPOSED STUDY WILL ADDRESS THESE GAPS BY EXAMINING PROSPECTIVE ASSOCIATIONS BETWEEN DISCRIMINATION (BI+, RACIAL/ETHNIC, AND INTERSECTIONAL) AND ALCOHOL USE OUTCOMES AMONG BI+ POC, TESTING THEORY- DRIVEN MEDIATORS AND MODERATORS OF THESE ASSOCIATIONS, AND IDENTIFYING SUBGROUPS WITH THE GREATEST RISK BASED ON SIMILAR DISCRIMINATION PROFILES. TO DO SO, THE PROPOSED STUDY WILL USE DATA FROM A LONGITUDINAL STUDY OF 500 BI+ ADULTS, WHICH INCLUDES 300 BI+ POC. PARTICIPANTS WILL COMPLETE 4 SURVEYS AT 6-MONTH INTERVALS, INCLUDING MEASURES OF: HEAVY EPISODIC DRINKING, DRINKING CONSEQUENCES, AND AUD SYMPTOMS; BI+, RACIAL/ETHNIC, AND INTERSECTIONAL DISCRIMINATION; COPING MOTIVES AND IDENTITY CONFLICT; AND BI+ AND RACIAL/ETHNIC IDENTITY AFFIRMATION. THE PRIMARY GOALS OF THE PROPOSED STUDY ARE TO: 1) EXAMINE PROSPECTIVE ASSOCIATIONS BETWEEN DISCRIMINATION, UNDERLYING MECHANISMS, AND ALCOHOL USE OUTCOMES AMONG BI+ POC; 2) TEST IDENTITY-RELATED PROTECTIVE FACTORS AS MODERATORS OF ASSOCIATIONS BETWEEN DISCRIMINATION AND ALCOHOL USE OUTCOMES; AND 3) IDENTIFY LATENT PROFILES BASED ON SHARED DISCRIMINATION EXPERIENCES TO IDENTIFY SUBGROUPS OF BI+ POC AT GREATEST RISK. FINDINGS FROM THIS RESEARCH WILL PROVIDE ESSENTIAL INSIGHTS INTO RISK FOR ADVERSE ALCOHOL USE OUTCOMES, UNDERLYING MECHANISMS, PROTECTIVE FACTORS, AND WHO IS AT GREATEST RISK AMONG BI+ POC AND ESTABLISH FOUNDATIONAL LONGITUDINAL EVIDENCE TO INFORM INTERVENTIONS TO REDUCE DISPARITIES IN THIS POPULATION. THIS AWARD WILL ALSO PROVIDE THE APPLICANT WITH TRAINING IN INTERSECTIONAL BI+ HEALTH RESEARCH, ALCOHOL USE RESEARCH, ADVANCED LONGITUDINAL STATISTICAL ANALYSIS, AND GRANT WRITING NECESSARY TO LAUNCH HIS CAREER AS AN INDEPENDENT SCIENTIST.

NURSE ANESTHETIST TRAINEESHIPS

NURSE ANESTHETIST TRAINEESHIPS

NURSE ANESTHETIST TRAINEESHIPS

NURSE ANESTHETIST TRAINEESHIPS

NURSE ANESTHETIST TRAINEESHIPS

NURSE ANESTHETIST TRAINEESHIPS

COCAINE-INDUCED PLASTICITY IN THE NUCLEUS ACCUMBENS

HEADACHE RESEARCH SUMMIT

CONFERENCE SUPPORT FOR 16TH INTERNATIONAL WORKSHOP ON KSHV AND RELATED AGENTS, PU

LDS - ALLOPATHIC MEDICINE - OTHER ADMIN CHANGES

HPSL - PODIATRY - OTHER ADMIN CHANGES

HPSL - PODIATRY - OTHER ADMIN CHANGES

NURSE ANESTHETIST TRAINEESHIPS