New York Medical College

HORMONAL REGULATION OF BLOOD PRESSURE

ENDOTHELIUM AND VASCULAR FUNCTION

INTERDISCIPLINARY LEADERSHIP EDUCATION (LEND)

SYNAPTIC MODULATION OF MESOPONTINE CHOLINERGIC NEURONS

MODULATION AND REGULATION OF ROMK CHANNELS IN KIDNEY

INFLAMMATION AND INFECTION IN ATHEROSCLEROSIS

ROLE OF LITAF IN INFLAMMATORY PROCESSES

CANNABINOID MODULATION OF EV COMPOSITION AND FUNCTION IN HIV/SIV INFECTION

MODIFICATION OF DNA POLYMERASE DELTA BY A NOVEL MECHANISM DURING REPLICATION STRE

PH2 OF T-CELL DEPL FAMILIAL HAPLOIDENTICAL SCT FOR TX HI-RISK SICKLE CELL ANEMIA

REGULATION OF CORONARY COLLATERAL GROWTH IN THE METABOLIC SYNDROME

REGULATION OF VASCULAR SMOOTH MUSCLE CELL PHENOTYPE BY A NOVEL ISOFORM OF GLUCOSE-6-PHOSPHATE DEHYDROGENASE - VASCULAR DISEASES CONTINUE TO BE A MAJOR CAUSE OF DEATH IN THE US AND WORLDWIDE. IT HAS BEEN PROPOSED THAT METABOLIC REPROGRAMMING AND INCREASED GLUCOSE-6-PHOSPHATE DEHYDROGENASE (G6PD) ACTIVITY AND EXPRESSION CONTRIBUTE TO THE PATHOGENESIS OF FATAL ANGIOPROLIFERATIVE VASCULOPATHIES. MOREOVER, SOME STUDIES SUGGEST INDIVIDUALS WITH A LOSS-OF- FUNCTION G6PD (MEDITERRANEAN OR AFRICAN) VARIANT – S188F (G6PDS188F; TYPE A-; SEVERE DEFICIENCY) OR N126D (G6PDN126D; TYPE A; MILD DEFICIENCY) NONSYNONYMOUS SINGLE NUCLEOTIDE POLYMORPHISM – HAVE LOWER FREQUENCIES OF CORONARY ARTERY DISEASE. HOWEVER, G6PD-DRIVEN PATHOGENIC AND G6PD VARIANT-ASSOCIATED PROTECTIVE MECHANISMS AFFECTING VASCULAR DISEASES REMAIN ELUSIVE. WE THEREFORE PROPOSE TO DETERMINE POTENTIAL MECHANISMS, DRIVEN BY A NEWLY DISCOVERED G6PD ISOFORM IN THE NUCLEUS OF VASCULAR SMOOTH MUSCLE CELLS (VSMCS), THAT CONTRIBUTE TO PATHOGENIC LARGE ARTERY STIFFNESS AND REMODELING. BASED ON STRONGLY SUPPORTING PRELIMINARY RESULTS, WE HYPOTHESIZED THAT NUCLEAR G6PD IS A MODULATOR OF EPIGENETIC MODIFIERS AND IS A TRANSCRIPTION REGULATOR IN VSMCS. CONSEQUENTLY, THE LOSS-OF-FUNCTION G6PD (S188F, N126D) VARIANTS BLOCK MALADAPTIVE MODIFICATIONS OF THE EPIGENOME, REDUCING LARGE ARTERY ELASTANCE AND REMODELING ELICITED BY OBESITY/METABOLIC SYNDROME AND BALLOON-INJURY. WE WILL TEST THIS HYPOTHESIS IN THREE SPECIFIC AIMS. IN AIM 1, WE WILL TEST THE HYPOTHESIS THAT G6PD AND/OR G6PD-COORDINATED REDOX IN THE NUCLEUS CONTROLS THE EXPRESSION AND ACTIVITY OF EPIGENETIC MODIFIERS (DNA METHYLTRANSFERASES (DNMT) AND DNA (TET) AND HISTONE (JARID) DEMETHYLASES) AND TRANSCRIPTION OF GENES THAT ENCODE PROTEINS INVOLVED IN REGULATING THE DIFFERENTIATION (CONTRACTILE) AND DEDIFFERENTIATION (PRO-INFLAMMATORY, -THROMBOTIC, AND -PROLIFERATIVE) PHENOTYPES IN VSMCS. IN AIM 2, WE WILL DETERMINE WHETHER LOSS-OF-FUNCTION G6PD VARIANTS DETACH FROM EPIGENETIC MODIFIERS TO INCREASE DNA METHYLATION, SUPPRESS HISTONE3-LYSINE4 TRIMETHYLATION, AND REDUCE TRANSCRIPTION OF GENES THAT CONFER MALADAPTIVE (PRO- INFLAMMATORY, -THROMBOTIC, AND -PROLIFERATIVE) PROPERTIES TO VSMCS. IN AIM 3, WE WILL DETERMINE WHETHER G6PD VARIANT RATS EXPRESS FEWER MALADAPTIVE EPIGENETIC (HISTONE3-LYSINE4 TRIMETHYLATION) CHANGES AND DEVELOP LESS LARGE ARTERY ELASTANCE (STIFFNESS) AND VASCULAR REMODELING THAN WILD-TYPE RATS FED A HIGH-FAT DIET (A MODEL OF OBESITY/METABOLIC SYNDROME) OR SUBJECTED TO CAROTID ARTERY BALLOON-INJURY. THE RESULTS FROM GAIN-OF-FUNCTION AND LOSS-OF-FUNCTION STUDIES OF THIS PROJECT WILL REVEAL THE DIRECT EFFECT OF G6PD ON GENE EXPRESSION ASSOCIATED WITH PATHOGENIC VASCULAR REMODELING AND LARGE ARTERY STIFFNESS, WHICH LEAD TO HEART FAILURE AND DEATH. WE FORESEE TWO SIGNIFICANT IMPACTS ON VASCULAR BIOLOGY: [1] LINKAGE OF HERETOFORE UNKNOWN G6PD-DEPENDENT SUBCELLULAR REDOX IN THE NUCLEUS DIRECTLY TO THE FUNDAMENTAL TRANSCRIPTIONAL MECHANICS AND GENE TRANSCRIPTION IN VASCULAR PATHOBIOLOGY AND [2] DEVELOPMENT OF NEW TREATMENTS TARGETING REDOX SIGNALING TO REDUCE LARGE ARTERY STIFFNESS AND REMODELING.

BIOCHEMICAL STUDIES OF HUMAN DNA POLYMERASE DELTA

ESTROGEN AND EDHF IN NO DEFICIENCY

GENOMIC AND IMMUNOMIC MECHANISMS OF RESISTANCE DURING DARATUMUMAB POST STEM CELL TRANSPLANTATION IN PATIENTS WITH POOR-RISK T CELL LYMPHOBLASTIC LYMPHOMA LEUKEMIA

MOLECULAR MECHANISMS OF 20-HETE-MEDIATED VASCULAR DYSFUNCTION AND HYPERTENSION

ROLE OF KIR4.1 IN REGULATING NCC AND ROMK IN DCT - PROJECT SUMMARY/ABSTRACT KIR4.1/KIR5.1 IN DISTAL CONVOLUTED TUBULE (DCT) PLAYS A KEY ROLE IN MEDIATING THE EFFECT OF LOW K+ (LK) OR HIGH K+ (HK) AND LOW SODIUM (LS) OR HIGH SODIUM (HS) INTAKE ON NCC THROUGH CL--SENSITIVE WITH-NO-LYSINE KINASE (WNK) WHICH REGULATES STE20-PROLINE-ALANINE RICH KINASE (SPAK), A KEY KINASE FOR CONTROLLING NCC ACTIVITY. ALTHOUGH MANY DETAILS OF THIS SIGNALING MECHANISM EXPLAINING THE EFFECTS OF CHRONIC DIETARY K+ OR NA+ INTAKE HAVE BEEN ELUCIDATED, WE HAVE DEMONSTRATED THAT CHRONIC PERTURBATIONS REMODEL THE DISTAL NEPHRON. THUS, THE OBSERVED CHANGES MAY NOT REFLECT THE INITIAL DYNAMIC RESPONSE OF KIR4.1/KIR5.1 AND NCC IN THE DCT. IMPORTANTLY, DIETARY INTAKE IS TYPICALLY EPISODIC, LEADING TO HIGHLY DYNAMIC ELECTROLYTE EXCRETORY PATTERNS. THE DCT COMPRISES AN EARLY PART (DCT1) AND A LATE PART (DCT2). WHILE BASOLATERAL KIR4.1/KIR5.1 AND APICAL NCC ARE EXPRESSED ALONG BOTH DCT1 AND DCT2, THE EPITHELIAL SODIUM CHANNEL (ENAC) AND ROMK ARE ONLY DETECTED IN THE DCT2. THIS RAISES THE POSSIBILITY THAT THE RESPONSE OF KIR4.1/KIR5.1 AND NCC TO SHORT-TERM DIETARY K+ OR NA+ MAY DIFFER BETWEEN DCT1 AND DCT2, CONTRIBUTING TO RAPID HOMEOSTATIC EFFECTS. WE NOW HYPOTHESIZE THAT DIETARY K+-INTAKE INDUCED REGULATION OF KIR4.1/KIR5.1, WNK AND NCC IS INITIATED PREDOMINANTLY IN THE DCT1, WHEREAS DIETARY NA+-INDUCED OR ANGII-APPLICATION-INDUCED REGULATION OF KIR4.1/KIR5.1, WNK, NCC IS STARTED MAINLY IN THE DCT2 VIA TYPE 1A ANGIOTENSIN II RECEPTOR (AT1AR). THIS HYPOTHESIS IS SUPPORTED BY SEVERAL LINES OF PRELIMINARY DATA:1) OVERNIGHT LK INTAKE STIMULATES KIR4.1/KIR5 ACTIVITY IN THE DCT1 BUT NOT IN THE DCT2; 2) OVERNIGHT LS INTAKE INCREASES KIR4.1/KIR5.1 CURRENTS PREDOMINANTLY IN THE DCT2 BUT TO A LESSER DEGREE IN THE DCT1; 3) DELETION OF AT1AR IN THE KIDNEY ABOLISHES THE OVERNIGHT- LS-INDUCED STIMULATION OF KIR4.1/KIR5.1 OF THE DCT AND NCC; 4) ACUTE APPLICATION OF ANGII STIMULATES KIR4.1/KIR5.1 ONLY IN DCT2 AND ENHANCES NCC FUNCTION; 5) OVERNIGHT ANGII INFUSION ROBUSTLY STIMULATES KIR4.1/KIR5.1 ACTIVITY IN DCT2 BUT TO A LESS DEGREE IN THE DCT1; 6) OVERNIGHT ANGII INFUSION STIMULATES WNK4 AND NCC EXPRESSION/ACTIVITY IN KCNJ10-/- MICE BUT NOT IN KIR4.1 KNOCKOUT MICE. THE THREE SPECIFIC AIMS OF THE PROPOSAL ARE: 1) TEST WHETHER DIETARY K+ AND NA+ INDUCED REGULATION OF KIR4.1/KIR5.1, WNK AND NCC IS DIFFERENT BETWEEN DCT1 AND DCT2; 2) TEST WHETHER ACUTE OR OVERNIGHT ANGII TREATMENT ACTIVATES KIR4.1/KIR5.1 AND NCC PREDOMINATELY IN THE DCT2 BUT NOT IN THE DCT1 AND THAT THE ACTIVATION OF KIR4.1/KIR5.1 IS ESSENTIAL FOR ACUTE BUT NOT CHRONIC ANGII-INDUCED STIMULATION OF WNK4 AND NCC 3) TEST WHETHER DELETION OF AT1AR IMPAIRS THE STIMULATORY EFFECT OF OVERNIGHT LS BUT NOT OVERNIGHT LK ON KIR4.1/KIR5.1, WNK4 , PNCC AND K+ HOMEOSTASIS. THE PHYSIOLOGICAL SIGNIFICANCE OF THE PROPOSAL IS TO ELUCIDATE THE MECHANISMS UNDERLYING DUAL REGULATION OF THE DCT, SUCH THAT DCT1 BEHAVES LIKE A “K+-SENSOR” AND DCT2 BEHAVES LIKE A “NA+-SENSOR”. THIS DIFFERENTIAL RESPONSE IS ESSENTIAL FOR NORMAL PHYSIOLOGICAL HOMEOSTASIS DURING SHORT-TERM ALTERATIONS IN K+ OR NA+ INTAKE.

PROBES FOR CYTOMETRY

B. BURGDORFERI HEMATOGENOUS DISSEMINATION

ALDOSTERONE SYNTHASE & HYPERTENSION

REGULATION OF VASCULAR SMOOTH MUSCLE CELL PHENOTYPE BY A NOVEL ISOFORM OF GLUCOSE-6-PHOSPHATE DEHYDROGENASE

CARDIOVAGAL BAROREFLEX DEFICITS IMPAIR NEUROVASCULAR COUPLING AND COGNITION IN POSTURAL TACHYCARDIA SYNDROME

ENDOTHELIAL DYSFUNCTION, NITRIC OXIDE AND RENAL FAILURE

NON-CODING VARIANTS OF ANGIOTENSINOGEN GENE AND HYPERTENSION

ESTROGENIC SIGNALING UPSTREAM AND DOWNSTREAM OF MTOR

IMPORTANCE OF PANNEXIN TO ASTROCYTE AND NEURONAL ATP

GPR75 IN OBESITY-DRIVEN CARDIOVASCULAR AND METABOLIC COMPLICATIONS - ABSTRACT SUMMARY OBESITY AND THE ASSOCIATED METABOLIC COMPLICATIONS INCLUDING METABOLIC SYNDROME AND DIABETES SIGNIFICANTLY CONTRIBUTE TO CARDIOVASCULAR MORBIDITIES. THE OVERALL GOAL OF THIS PROPOSAL IS AIMED AT IDENTIFYING CELLULAR MECHANISMS THAT LINK ACTIVATION OF THE G-PROTEIN COUPLED RECEPTOR 75 (GPR75) TO OBESITY-DRIVEN METABOLIC AND CARDIOVASCULAR COMPLICATIONS. IN THE LAST FUNDING PERIOD, WE PROVIDED STRONG EVIDENCE THAT GPR75 IS THE RECEPTOR TO WHICH 20-HYDROXYEICOSATETRAENOIC ACID (20-HETE), A VASOACTIVE AND PRO-INFLAMMATORY LIPID MEDIATOR, BINDS AND THROUGH WHICH IT TRIGGERS ITS ACTIONS. 20-HETE HAS BEEN IMPLICATED IN THE DEVELOPMENT AND PROGRESSION OF CARDIOVASCULAR AND METABOLIC DISEASES INCLUDING HYPERTENSION, STROKE, MYOCARDIAL INFARCTION, DIABETES AND METABOLIC SYNDROME. OUR RECENT STUDIES CLEARLY DEMONSTRATED THAT 20-HETE- MEDIATES VASCULAR DYSFUNCTION IN HYPERTENSION AND DIET-DRIVEN METABOLIC (E.G., INSULIN RESISTANCE) AND CARDIOVASCULAR (E.G., HYPERTENSION) COMPLICATIONS THROUGH ITS PAIRING AND ACTIVATION OF GPR75. THE ADIPOSE TISSUE AND THE VASCULAR ENDOTHELIUM ARE KEY TO THE ENSUING CARDIOMETABOLIC DISORDERS IN OBESITY. THEY BOTH EXPRESS GPR75 AND ARE PRIME TARGETS FOR 20-HETE BIOACTIONS. IN THE ENDOTHELIUM, 20-HETE, THROUGH ITS BINDING TO GPR75, UNCOUPLES ENOS, INDUCES ACE AND ACTIVATES THE NF-B INFLAMMATORY PROGRAM. 20-HETE STIMULATES ADIPOCYTE HYPERTROPHY AND INTERFERES WITH ADIPOCYTE INSULIN SIGNALING, GLUCOSE HOMEOSTASIS AND MITOCHONDRIA FUNCTION. ENDOTHELIAL DYSFUNCTION IN HYPERTENSIVE SUBJECTS AND OBESITY IN HUMANS AND ANIMAL MODELS ARE ACCOMPANIED BY SEVERAL FOLD INCREASES IN CIRCULATING 20-HETE LEVELS. ACCORDINGLY, IT IS REASONABLE TO ASSUME THAT 20-HETE THROUGH ITS ACTIONS ON THE VASCULAR ENDOTHELIUM AND ADIPOSE TISSUE ACTS AS A SIGNIFICANT DETERMINANT OF OBESITY-DRIVEN CARDIOVASCULAR AND METABOLIC COMPLICATIONS. IMPORTANTLY, IN A LARGE-SCALE EXOME SEQUENCING OF ~600,000 INDIVIDUALS, WE REVEALED THAT LOSS OF FUNCTION GPR75 VARIANTS ARE ASSOCIATED WITH LEANNESS AND PROTECTION FROM OBESITY (SCIENCE, 2021). THIS FINDING, WHICH HAS BEEN VALIDATED IN GPR75 NULL MICE (SCIENCE, 2021 AND PRELIMINARY RESULTS), COMPELLINGLY IMPLICATES THE 20-HETE-GPR75 PAIRING IN THE PATHOGENESIS OF OBESITY AND ITS CARDIOMETABOLIC COMPLICATIONS AND CALLS FOR IDENTIFICATION OF THE UNDERLYING MECHANISMS. CONSEQUENTLY, WE HYPOTHESIZED THAT 20-HETE-GPR75 PAIRING PROMOTES DIET-DRIVEN CARDIOMETABOLIC COMPLICATIONS (E.G., HYPERGLYCEMIA, INSULIN RESISTANCE, HYPERTENSION) BY A MECHANISM THAT IMPAIRS ENDOTHELIAL AND ADIPOCYTE FUNCTION. WE FURTHER ARGUE THAT 20-HETE-GPR75 PAIRING GOVERNS A CROSSTALK BETWEEN ADIPOCYTES AND ENDOTHELIAL CELLS WHICH CONTRIBUTES TO CARDIOVASCULAR AND METABOLIC COMPLICATIONS IN OBESITY. THIS HYPOTHESIS WILL BE TESTED IN THREE SPECIFIC AIMS THAT ASSESS THE CONTRIBUTION OF ENDOTHELIAL- AND ADIPOCYTE-SPECIFIC GPR75 TO DIET-DRIVEN OBESITY AND EXAMINE POTENTIAL MECHANISMS GOVERNING ENDOTHELIAL-ADIPOCYTE CROSSTALK THAT ARE DEPENDENT ON 20-HETE-GPR75 PAIRING.

IPSC DERIVED HUMAN CORTICAL INTERNEURONS AS DEVELOPMENTAL MODEL OF SCHIZOPHRENIA

GLYCOCALYX REPAIR IN SEPSIS USING LIPOSOMAL CARRIERS OF PREASSEMBLED GLYCOCALYX

ROLES OF HEAT SHOCK TRANSCRIPTIONAL FACTOR 1 IN CELL PROLIFERATION INDEPENDENT OF THE HEAT SHOCK RESPONSE

MECHANISMS OF VASOVAGAL SYNCOPE

MYOCYTE REPOLARIZATION AND CARDIAC DYSFUNCTION WITH AGE

HIGH RESOLUTION FUNCTIONAL MAPS OF ENHANCER RNA AND SUBCELLULAR RNA GRANULES IN HUMAN IMMUNE CELLS - PROJECT ABSTRACT/SUMMARY MY RESEARCH FOCUSES ON THE PRECISE UNDERSTANDING OF THE REGULATION OF RNA IN LIVING CELLS AND HUMAN DISEASES. MAPPING WHERE THE REGULATIONS TAKE PLACE IS CRITICAL IN IDENTIFYING FACTORS AND ELEMENTS THAT ALTER RNA EXPRESSION. FURTHER ENHANCING THE GENOMIC RESOLUTION AND THE SPATIOTEMPORAL RESOLUTION OF RNA EXPRESSION HAVE BEEN A STRONG DRIVER OF LEADING DISCOVERIES IN MOLECULAR AND DISEASE MECHANISMS. MY FIRST FOCUS IS TO DEFINE THE HIGH-RESOLUTION MAPS OF THE TRANSCRIPTIONAL ENHANCERS IN PERIPHERAL BLOOD IMMUNE CELLS, USING ENHANCER RNA (ERNA) AS THE GUIDE. ERNA TRANSCRIPTION IS A NOVEL MECHANISM OF WHICH THE RNA IS SYNTHESIZED FROM ENHANCER SEQUENCES THEMSELVES OTHER THAN THEIR TARGET GENES. ERNA HAS EXPANDED THE PREVIOUS UNDERSTANDING OF HOW ENHANCERS ARE CONSTRUCTED AND ALLOWED MAPPING IMPORTANT SUBREGIONS WITHIN THE ENHANCERS IN HIGHER RESOLUTION. ERNA BASED MAPS OF ENHANCERS WILL GUIDE US TO MORE EFFICIENTLY DISCOVER GENOMIC ELEMENTS LINKED TO THE DYSREGULATION OF DISEASE GENES, AND SPECIFICALLY DISSECT CAUSAL GENETIC VARIATIONS OR MUTATIONS. THIS WILL HAVE A WIDESPREAD IMPACT ON MAJOR HUMAN DISEASES, AND MY SPECIFIC FOCUS IS ON IMMUNOMETABOLIC DISEASES AND THE IMPACT OF ENHANCER VARIATIONS AT THE ERNA START SITES. THE SECOND FOCUS IS ON THE SUBCELLULAR COMPARTMENTS THAT LEADS TO DIFFERENTIAL RNA TRAFFICKING, PROCESSING, AND DECAY. MEMBRANELESS ORGANELLES, SUCH AS STRESS GRANULES, ARE SUGGESTED TO BE THE NOVEL MECHANISM OF GENE EXPRESSION CONTROL THROUGH PHASE SEPARATION AND SEQUESTRATION. I WILL EXPLORE THE IDEA THAT SPATIAL COMPARTMENTALIZATION CONFERS SPECIFICITY OF TEMPORAL GENE EXPRESSION. IN PARTICULAR, I WILL DISSECT THE SPECIFICITY OF RNA PROCESSING IN STRESS GRANULES IN HIGH TEMPORAL RESOLUTION IN IMMUNE CELL ACTIVATION AND EXHAUSTION AND EXPLORE THEIR LINKAGE TO THE DYSREGULATION OF DISEASE ASSOCIATED GENES. THIS WILL LEAD TO THE DISCOVERY OF NOVEL THERAPEUTIC TARGETS ON THE SPECIFICITY FACTORS DURING THE PHASE SEPARATION PROCESS. TO ADDRESS THESE, MY GROUP HARNESSES THE POWER OF HIGH THROUGHPUT RNA SEQUENCING METHODS, AND FURTHER TAILOR THEM. WE DEVELOPED NOVEL RNA SEQUENCING METHODS SUCH AS PRECISION RUN-ON SEQUENCING, CHROMATIN RUN-ON SEQUENCING, TAIL END DISPLACEMENT SEQUENCING, AND RNA GRANULE SEQUENCING. THESE METHODS ELUCIDATE MULTIPLE ASPECTS OF RNA MECHANISM IN HIGH THROUGHPUT: NASCENT TRANSCRIPTION, POLY(A) TAIL MODIFICATION, AND RNA SEQUESTRATION. WE WILL CONTINUE TO IMPROVE AND DEVELOP NOVEL RNA ANALYSIS STRATEGIES TO DISSECT ERNA TRANSCRIPTION, RNA MODIFICATIONS, AND COMBINING THESE METHODS WITH SUBCELLULAR COMPARTMENTALIZATION IN HIGH SPATIAL AND TEMPORAL RESOLUTION IN COST EFFICIENT MANNERS. THESE INNOVATIONS WILL LEAD TO ENHANCEMENTS IN REFINING GENETIC MAPS AND HIGHER SPATIOTEMPORAL RESOLUTION IN ANALYZING RNA EXPRESSION. OUR MISSION IS TO DISCOVER CRITICAL MECHANISMS AND CHECKPOINTS OF RNA LIFESPAN, FOCUSING ON ERNA AND RNA GRANULES. THIS WILL PROVIDE FOUNDATIONS TO APPLY THE IDEAS AND METHODS OF HIGH-RESOLUTION RNA MAPPING IN FURTHER COLLABORATIVE EFFORTS FOR ADVANCING THE PRECISION MEDICINE OF HUMAN DISEASE.

GOLGI BLOCKADE IN PULMONARY HYPERTENSION

SENSORY NEUROMODULATION OF PANCREATIC BETA CELLS

POSTDOCTORAL TRAINING IN GENERAL, PEDIATRIC AND PUBLIC HEALTH DENTISTRY AND DENTAL HYGIENE

LOCAL VASOCONSTRICTION IN POSTURAL TACHYCARDIA SYNDROME

COMMUNITY PROJECT FUNDING/CONGRESSIONALLY DIRECTED SPENDING - CONSTRUCTION

REGULATION OF NKCC2 ISOFORMS AND BLOOD PRESSURE BY TUMOR NECROSIS FACTOR-ALPHA - WE PREVIOUSLY SHOWED THAT TUMOR NECROSIS FACTOR-ALPHA (TNF) PRODUCED WITHIN THE KIDNEY IS PART OF A MECHANISM THAT REGULATES RENAL FUNCTION AND THE BLOOD PRESSURE (BP) RESPONSE TO INCREASES IN DIETARY SALT INTAKE. OUR RECENT STUDIES SUGGEST THAT TNF EFFECTS IN THE KIDNEY ARE EVIDENT IN THE MEDULLARY (M) THICK ASCENDING LIMB (TAL), PROXIMAL TUBULE (PT), AND CORTICAL (C) TAL/MACULA DENSA (MD) REGIONS OF THE NEPHRON. HOWEVER, THE CELLULAR SOURCES WITHIN THE KIDNEY THAT PRODUCE THE TNF THAT ACCOUNTS FOR THESE EFFECTS HAVE NOT BEEN DETERMINED, NOR HAVE THE MOLECULAR MECHANISMS BEEN IDENTIFIED. THUS, WE DEVELOPED TWO MOUSE MODELS IN WHICH TNF HAS BEEN GENETICALLY DELETED IN THE: 1) TAL, AND 2) DISTAL NEPHRON DOWNSTREAM OF THE PT, WHICH WILL BE USED TO UNDERSTAND THE ROLE OF TNF PRODUCED BY RENAL EPITHELIAL CELLS AS PART OF AN EMERGING INTRATUBULAR TNF SYSTEM THAT ATTENUATES INCREASES IN BP INDUCED BY HIGH SALT (HS) INTAKE. WE ALSO HAVE TAILORED A COMPLEMENTARY APPROACH, USING PT- AND TAL-SPECIFIC TNF SILENCING LENTIVIRUS CONSTRUCTS, TO SPECIFICALLY INHIBIT TNF PRODUCTION BY THESE CELL TYPES. THE GENETIC AND LENTIVIRUS APPROACHES WILL BE USED IN TANDEM TO DETERMINE THE MECHANISM BY WHICH TNF REGULATES NA+-K+-2CL- (NKCC2) PHOSPHORYLATION AND ISOFORM EXPRESSION, RENAL FUNCTION, AND BP. PRELIMINARY DATA SUGGEST THAT TNF, VIA ACTIVATION OF TNF RECEPTOR 1 (TNFR1), INHIBITS PHOSPHO-NKCC2 (PNKCC2) EXPRESSION BY A MECHANISM INVOLVING ACTIVATION OF THE SERINE/THREONINE PHOSPHATASE, CALCINEURIN (CN). THE EFFECTS OF TNF ON CN HAVE NOT BEEN EXPLORED IN THE KIDNEY, THUS EXPERIMENTS WILL ADDRESS TNF-DEPENDENT INCREASES IN CN ACTIVITY AS WELL AS EXPRESSION OF THE CATALYTIC SUBUNIT CNAB AND REGULATORY SUBUNIT CNB. THE GENETIC AND LENTIVIRUS STRATEGIES WILL BE ADAPTED TO DETERMINE THE EFFECTS OF SALT INTAKE ON TNFR1-DEPENDENT CN-MEDIATED INHIBITION OF PNKCC2 EXPRESSION, ELECTROLYTE EXCRETION, AND THE BP RESPONSE TO HS INTAKE. THE NKCC2A AND NKCC2B ISOFORMS ARE STRATEGICALLY LOCALIZED ALONG THE MAMMALIAN TAL AND CONTRIBUTE TO REGULATORY FUNCTIONS IN RESPONSE TO HIGH AND LOW SALT CONDITIONS, RESPECTIVELY. TNF INHIBITS THE EXPRESSION OF BOTH ISOFORMS SUGGESTING A ROLE FOR THIS CYTOKINE IN BOTH THE MTAL AND CTAL/MD SEGMENTS OF THE TAL. WE PREVIOUSLY SHOWED THAT IN EACH INSTANCE, TNF REGULATES RENAL FUNCTION INVOLVING THESE ISOFORMS IN A MANNER THAT LIMITS REABSORPTION OF NACL. HOWEVER, THE MOLECULAR MECHANISM BY WHICH TNF SUPPRESSES BOTH NKCC2A AND NKCC2B MRNA IN RESPONSE TO HIGH AND LOW SALT INTAKE, RESPECTIVELY, HAS NOT BEEN DETERMINED. PREVIOUS MIRNA PROFILING OF THE TAL IN COMBINATION WITH NEW PRELIMINARY DATA HAVE IDENTIFIED 3 CANDIDATE MIRNAS THAT REGULATE NKCC2 ISOFORM MRNA ABUNDANCE. FOR INSTANCE, MIRNA-195 EXPRESSION IS INDUCED BY TNF DERIVED FROM THE TAL AND INHIBITS NKCC2A MRNA ACCUMULATION AND PNKCC2 EXPRESSION IN MICE INGESTING HS. COLLECTIVELY, THE STUDIES WILL DEFINE A NOVEL INTRATUBULAR REGULATORY SYSTEM IN WHICH TNF PRODUCTION BY RENAL TUBULAR EPITHELIAL CELLS, IN RESPONSE TO INCREASES IN SALT INTAKE, REGULATES NKCC2 ISOFORM EXPRESSION AND FUNCTION AND CONTRIBUTES TO BP HOMEOSTASIS.

NOVEL MECHANISM-BASED TREATMENTS FOR INFANTILE SPASMS

PROTECTIVE ANTIBODY IN STREPTOCOCCAL INFECTION MODELS - ABSTRACT THE INITIAL CONTACT BETWEEN NEWLY TRANSMITTED GROUP A STREPTOCOCCI (GAS) AND ITS HUMAN HOST CAN BE READILY THWARTED BY ANTIBODY (AB) DIRECTED TO THE APPROPRIATE GAS ANTIGENS (AGS). TO BEST SIMULATE THIS MICROENVIRONMENT, PHYSIOLOGICAL (I.E., LOW) INOCULUM DOSES OF GAS ARE ESSENTIAL. THE GOAL OF THIS PROPOSAL IS TO IDENTIFY AGS, ALONE AND IN COMBINATIONS REPRESENTING MULTICOMPONENT VACCINES, THAT ARE THE MOST EFFECTIVE TARGETS OF AB-MEDIATED IMMUNITY AND COLLECTIVELY CAN PROVIDE WORLDWIDE COVERAGE AGAINST ALL GAS STRAINS. THE UNDERLYING RATIONALE FOR THE PROPOSED APPROACH RESTS ON TWO PRINCIPLES: THAT SERUM OBTAINED FROM CHILDREN JUST PRIOR TO A NEW GAS INFECTION LACKS PROTECTIVE AB, AND THAT SERUM OBTAINED FROM MOST ADULTS HAS AT LEAST LOW LEVELS OF PERSISTING PROTECTIVE AB. THE LATTER IS BASED ON THE VERY LOW INCIDENCE OF GAS INFECTION IN ADULTS, LIKELY THE RESULT OF PROTECTIVE IMMUNITY THAT DEVELOPED FOLLOWING REPEATED INFECTIONS EARLIER IN LIFE. HYPOTHESES ON PROTECTIVE AG-SPECIFIC AB GENERATED FROM ANALYSIS OF PEDIATRIC SERUM (AIM 1) ARE EXPERIMENTALLY TESTED IN MICE (AIM 2). PRE-INFECTION PEDIATRIC SERUM IS ANALYZED FOR LACK OF PROTECTIVE AB TO LEADING VACCINE CANDIDATES. ADULT SERUM IS USED AS A SOURCE FOR PURIFIED IMMUNOGLOBULIN (IG) SPECIFIC TO THE VACCINE TARGET AGS MISSING IN SUSCEPTIBLE CHILDREN. IMMUNODEFICIENT MICE ARE PASSIVELY IMMUNIZED WITH THE AG-SPECIFIC IG PURIFIED FROM ADULTS AND CHALLENGED WITH LOW INFECTIVE DOSES OF GAS. THE AG-SPECIFIC IG THAT CONFERS PROTECTION IN MICE EQUATES TO A HUMAN SERUM-BASED CORRELATE OF PROTECTION (COP). FOUR MAJOR FORMS OF GAS DISEASE ARE MODELED IN MICE AND EVALUATED FOR PROTECTIVE IMMUNITY: UPPER RESPIRATORY TRACT INFECTION, IMPETIGO, SKIN AND SOFT TISSUE INFECTION AND INVASIVE DISEASE. GAS FROM ALL THREE MAJOR SUBPOPULATIONS OF STRAINS ARE TESTED: THROAT SPECIALISTS, SKIN SPECIALISTS AND GENERALISTS. THE AG TARGETS EVALUATED ARE LEADING VACCINE CANDIDATES AND INCLUDE BROADLY AND SEMI-CONSERVED AGS AS WELL AS SURFACE AND SECRETED AGS. IF SUCCESSFUL, THIS WORK WILL IDENTIFY A COLLECTION OF AG TARGETS THAT CAN SERVE AS THE BASIS FOR GLOBAL PROTECTION. IT WILL ALSO DELIVER TWO NEW STANDARDIZED PLATFORMS – SCREENING OF PRE-INFECTION HUMAN SERUM AB TO GAS AGS AND IMPROVED MOUSE MODELS - FOR FUTURE IDENTIFICATION AND STRAIGHTFORWARD COMPARISON OF ADDITIONAL GAS VACCINE CANDIDATES.

WEIBEL - PALADE BODIES - SENTINELS OF ACUTE ISCHEMIA

CMV, ADV AND EBV VIRAL CYTOTOXIC T-LYMPHOCYTES GENERATED BY A NOVEL CYTOKINE CAPTURE SYSTEM IN CHILDREN, ADOLESCENTS AND YOUNG ADULTS WITH REFRACTORY VIRAL INFECTION AND T-CELL IMMUNODEFICIENCY

NOX4-ASSOCIATED OXIDATIVE STRESS MEDIATES VASCULAR AND KIDNEY IMPAIRMENT IN THE LOW BIRTH WEIGHT ADULT

HYPERTENSION AND INFLAMMATION: NOVEL INSIGHTS FROM HUMAN ANGIOTENSIN TYPE 1 RECEPTOR VARIANTS

THICK ASCENDING LIMB-DERIVED TNF, SALT SENSITIVITY, AND BLOOD PRESSURE REGULATION

FERROCHELATASE & GUANYLATE CYCLASE REGULATION IN VASCULAR DYSFUNCTION

INTERVENTION MODELS FOR THYROID PROLIFERATIVE DISEASE USING A BIOACTIVE FOOD COMP

IDENTIFICATION AND CIRCUMVENTION OF RESISTANCE AFTER DONOR-DERIVED EX VIVO EXPANDED NK CELLS AND HAPLO-HCT IN CHILDREN AND YOUNG ADULTS WITH HIGH-RISK AML

REGULATION OF RENAL TNF PRODUCTION AND FUNCTION

ECSOD DERIVED PEROXIDE IN PULMONARY ADAPTATION TO HYPOXIA

VASCULAR DYSFUNCTION IN CFS

"CORNEAL ARACHIDONATE METABOLITES VIA CYTOCHROME P450"

INTRANASAL PEPTIDE DELIVERY TO REDUCE PSYCHOLOGICAL STRESS INJURY

SUBJECTIVE SYMPTOMS AFTER TREATMENT OF LYME DISEASE

TOXOPLASMA GONDII EVASION OF IMMUNE EFFECTER MECHANISMS

KIR5.1 REGULATES KIR4.1 UBIQUITINATION BY NEDD4-2 IN DCT

PRESYNAPTIC LONG-TERM SYNAPTIC DEPRESSION

NOVEL COMBINATORIAL TARGETED CHIMERIC ANTIGEN RECEPTOR NATURAL KILLER CELL-BASED CELLULAR IMMUNOTHERAPY WITH ADJUVANT IMMUNOMODULATION IN PATIENTS WITH AML

ION CHANNELS REGULATING REM-RELATED CHOLINERGIC NEURONS

REGULATION OF KIR4.1/KIR5.1 AND RENAL POTASSIUM EXCRETION - MAINTAINING K+ HOMEOSTASIS IS ESSENTIAL FOR THE FUNCTION OF HEART, SKELETAL MUSCLES AND NEURONS BECAUSE HYPOKALEMIA OR HYPERKALEMIA COULD CAUSE LIFE-THREATENING CONSEQUENCE SUCH AS CARDIAC ARRHYTHMIA. A RECENT DEVELOPMENT IN THE FIELD HAS FIRMLY ESTABLISHED THE ROLE OF THIAZIDE-SENSITIVE NA-CL COTRANSPORTER (NCC) IN THE REGULATION OF K+ HOMEOSTASIS BECAUSE THE COORDINATED ACTION AMONG NCC, EPITHELIAL NA+ CHANNEL (ENAC) AND ROMK IS ESSENTIAL FOR MAXIMALLY ENHANCING RENAL K+ EXCRETION (EK) DURING HIGH DIETARY K+ INTAKE (HK) AND FOR EFFECTIVELY PREVENTING K+ WASTING DURING LOW DIETARY K+ INTAKE (LK). THE BASOLATERAL KIR4.1 AND KIR5.1 CHANNELS IN THE DISTAL CONVOLUTED TUBULE (DCT) PLAY AN IMPORTANT ROLE IN CONTROLLING NCC EXPRESSION AND ACTIVITY. OUR PREVIOUS STUDIES HAVE DEMONSTRATED THAT HK INDUCED INHIBITION OF KIR4.1/KIR5.1 IS AN ESSENTIAL STEP FOR HK-INDUCED INHIBITION OF NCC. CONVERSELY, LK-INDUCED STIMULATION OF KIR4.1/KIR5.1 OF THE DCT IS AN ESSENTIAL STEP FOR LK-INDUCED STIMULATION OF NCC. THE MECHANISM BY WHICH THE BASOLATERAL KIR4.1/KIR5.1 IN THE DCT REGULATES NCC ACTIVITY DEPENDS ON CL--SENSITIVE WITH-NO-LYSINE KINASE (WNK). THE ROLE OF KIR4.1 AND KIR5.1 IN THE REGULATION OF RENAL K+ EXCRETION HAS BEEN DEMONSTRATED IN THE MOUSE MODELS: THE DELETION OF KIR4.1 INHIBITS NCC ACTIVITY AND CAUSES RENAL K+ WASTING, WHEREAS THE DELETION OF KIR5.1 INCREASES NCC ACTIVITY AND REDUCES RENAL K+ EXCRETION ABILITY DURING HK INTAKE. THUS, KIR4.1 AND KIR5.1 IN THE DCT SERVE AS TWO IMPORTANT MEMBERS OF “POTASSIUM-SENSOR” MECHANISM. ALTHOUGH THE ROLE OF KIR4.1/KIR5.1 IN REGULATING NCC AND RENAL K+ EXCRETION IS WELL ESTABLISHED, THE REGULATORY MECHANISM BY WHICH DIETARY K+ INTAKE MODULATES KIR4.1 AND KIR5.1 IS NOT COMPLETELY UNDERSTOOD. WE NOW PROPOSE TO EXAMINE THE ROLE OF MECHANISTIC TARGET OF RAPAMYCIN (MTOR) COMPLEX 1 (MTORC1) AND MTOR COMPLEX 2 (MTORC2) IN MEDIATING THE EFFECT OF DIETARY K+ INTAKE ON KIR4.1/KIR5.1 OF THE DCT. WE HYPOTHESIZE 1) ACTIVATION OF MTORC1, PARTIALLY VIA INSULIN-LIKE-GROWTH FACTOR 1 (IGF-1), IS CRITICALLY INVOLVED IN MEDIATING LK-INTAKE-INDUCED STIMULATION OF KIR4.1/KIR5.1 OF THE DCT AND NCC; 2) ACTIVATION OF MTORC2 INHIBITS KIR4.1/KIR5.1 BY PKC DURING HK AND IS INVOLVED IN HK-INTAKE-INDUCED INHIBITION OF NCC. THE PROPOSAL HAS THREE SPECIFIC AIMS: 1)TO TEST THE HYPOTHESIS THAT MTORC1 REGULATES KIR4.1/KIR5.1 OF DCT AND NCC UNDER CONTROL CONDITIONS (NORMAL K+) AND PLAYS A ROLE IN DETERMINING THE BASELINE RENAL K+ EXCRETION; 2) TO TEST THE HYPOTHESIS THAT ACTIVATION OF IGF1-MTORC1 PLAYS A KEY ROLE IN MEDIATING LK-INDUCED STIMULATION OF KIR4.1/KIR5.1 IN THE DCT AND NCC AND IN SUPPRESSING RENAL K+ EXCRETION; 3) TEST THE HYPOTHESIS THAT ACTIVATION OF MTORC2 MEDIATES HK-INDUCED INHIBITION OF KIR4.1/KIR5.1 VIA PKC AND NCC IN THE DCT THEREBY ENHANCING ENAC-DEPENDENT RENAL EK. THE SIGNIFICANCE OF OUR PROPOSAL IS TO EXPLORE THE NOVEL ROLE OF MTORC1/MTORC2 PATHWAYS IN MEDIATING EFFECTS OF LK OR HK ON KIR4.1/KIR5.1, NCC, ENAC AND ROMK IN THE DCT AND CONNECTING TUBULE.

NEUROPROTECTIVE STRATEGIES IN SEIZURE-INDUCED DAMAGE

COMBINED SODIUM AND CALCIUM IMAGING OF DENDRITIC FUNCTION

NEURAL MECHANISMS OF ULTRASOUND-INDUCED ENHANCEMENT OF SENSORY ACUITY

WHITE MATTER INJURY IN GERMINAL MATRIX HEMORRHAGE

A RANDOMIZED STUDY OF MATERNAL DONOR DERIVED CMV CYTOTOXIC T-LYMPHOCYTES (CTLS) AND VALGANCICLOVIR VS VALGANCICLOVIR IN NEONATES WITH MODERATE/SEVERE MATERNAL ACQUIRED CMV INFECTION - PROJECT SUMMARY CONGENITAL CYTOMEGALOVIRUS (CCMV) INFECTION THAT IS ACQUIRED IN-UTERO, ALTHOUGH AN ORPHAN DISEASE (20-80,000 CASES/YR IN U.S.) IS THE MOST COMMON CONGENITAL INFECTION IN THE U.S. AND THE LEADING CAUSE OF LONG-TERM NEURODEVELOPMENTAL DISABILITIES. CCMV INFECTION REPRESENTS 41% OF ALL CMV ASSOCIATED CHILDHOOD DEATHS. THIS HIGH DEGREE OF MORBIDITY AND MORTALITY IS IN LARGE PART SECONDARY TO THE LARGE CMV GENOME WHOSE GENES ARE DESIGNED TO FACILITATE EVASION, BLOCKING, IMPAIRING AND MODULATION OF HOST ANTI-VIRAL IMMUNE RESPONSES AND AN IMMATURITY AND DEFICIT IN NEONATAL T-CELL ADAPTIVE IMMUNITY AT BIRTH. THIS MALADAPTIVE T-CELL IMMUNE RESPONSE AT BIRTH RESULTS FROM TEMPORAL AND DEVELOPMENTAL IMMUNE DYSREGULATION IN-UTERO WITH A RESULTING BIAS TOWARDS TOLERANCE AND PREVENTING MATERNAL IMMUNOLOGICAL REJECTION, THEREBY SUPPRESSING NORMAL T-CELL MATURATION AND DEVELOPMENT OF MEMORY T-CELLS TO FOREIGN MICROBIAL ANTIGENS. IMPORTANTLY, A POOR CMV SPECIFIC T-CELL ADAPTIVE IMMUNE RESPONSE PREDISPOSES NEONATES WITH CCMV ACQUIRED IN-UTERO TO A SIGNIFICANT INCREASED RISK OF SERIOUS LONG-TERM MORBIDITY AND INCREASED MORTALITY. RESTORING, AT LEAST TEMPORARILY, BOTH CD4 AND CD8 SPECIFIC CMV T- CELL MEDIATED IMMUNITY IN NEONATES WITH MATERNAL ACQUIRED CCMV IS THEREFORE CRITICAL TO ERADICATING PERSISTENT, REFRACTORY AND/OR RESISTANT CMV INFECTION AND UNDESIRABLE LONG-TERM SEQUELAE AND REPRESENTS AN UNMET NEED. WE HAVE PREVIOUSLY ESTABLISHED A VIRAL CYTOTOXIC T-CELL LYMPHOCYTE CONSORTIUM (VIRCTLC) FUNDED UNDER AN FDA ORPHAN GRANT TO MANUFACTURE UNDER GMP CONDITIONS AND ENRICH FOR HAPLOIDENTICAL (MATERNAL) DONOR-DERIVED CMV SPECIFIC CTLS UTILIZING A CMV SPECIFIC PEPTIVATOR® AND THE CYTOKINE CAPTURE SYSTEM (CCS®) ON THE MILTENYI CLINIMACS® DEVICE FOR TREATMENT OF INFANTS AND CHILDREN WITH REFRACTORY/PERSISTENT CMV INFECTION WITH PRIMARY OR SECONDARY T-CELL DEFICIENCIES. WE, THEREFORE, HYPOTHESIZE THAT MANUFACTURING MATERNAL DONOR-DERIVED CMV CTLS AND SUBSEQUENT ADMINISTRATION IN NEONATES WITH MATERNAL ACQUIRED CCMV WILL BE FEASIBLE AND SAFE AND EFFECTIVE IN COMBINATION WITH STANDARD OF CARE (SOC) (VALGANCICLOVIR [VALGCV]) COMPARED TO THOSE TREATED WITH SOC ALONE. THE THREE OVERARCHING AIMS OF THIS FDA ORPHAN GRANT PROPOSAL INCLUDES: 1) TO DETERMINE FEASIBILITY, SAFETY AND COMPARE THE EFFICACY OF MATERNAL DONOR-DERIVED CMV SPECIFIC CTLS WITH SOC VS SOC ALONE; 2) TO CHARACTERIZE THE GENOMIC AND IMMUNOMIC SIGNATURES AND PERSISTENCE OF MATERNAL DONOR-DERIVED CMV CTLS AND COMPARISON OF INNATE AND ADAPTIVE IMMUNE LANDSCAPE POST CMV CTL ADMINISTRATION; AND 3) TO QUANTIFY THE SEVERITY AND INCIDENCE OF LONG-TERM NEUROLOGICAL SEQUELAE AND DEVELOPMENTAL DISABILITIES BETWEEN THE TWO TREATMENT GROUPS. CMV CTLS WILL BE MANUFACTURED UNDER GMP CONDITIONS UTILIZING THE CCS® AND ADMINISTERED EVERY 2 WEEKS UP TO 5 DOSES WITH VALGCV X 6 MONTHS OR VALGCV ALONE. GENOMIC AND IMMUNOMIC STUDIES WILL BE PERFORMED BY SCRNASEQ, MASS CYTOMETRY, HIGH DIMENSIONAL FLOW CYTOMETRY AND NANOSTRING IMMUNOPROFILING. LONG-TERM NEUROLOGICAL SEQUELAE WILL BE MEASURED BY AUDIOGRAMS, BRAIN MRIS, DEVELOPMENTAL TESTING, AMONG OTHERS.

INTRANASAL NEUROPEPTIDE Y FOR PTSD AND OTHER STRESS TRIGGERED NEUROPSYCHIATRIC DISORDERS

RESIDENCY TRAINING IN PRIMARY CARE

TRANSCRIPTIONAL REGULATION OF HUMAN ANGIOTENSIN RECEPTOR

PULMONARY VASODILATION AND GUANYLATE CYCLASE REGULATION

GENOTYPIC VARIATION AND B. BURGDORFERI PATHOGENESIS

CALCIUM WAVES IN PYRAMIDAL NEURONS

CHARACTERIZING THE PHYSICOCHEMICAL PROPERTIES OF MEMBRANELESS CONDENSATES AND ITS REGULATION BY DELTA-9-TETRAHYDROCANNABINOL IN HIV/SIV INFECTION. - ABSTRACT THE `CENTRAL DOGMA' OF MOLECULAR BIOLOGY IS THAT IN CELLS, THE FUNCTIONS OF PROTEINS AND NUCLEIC ACIDS ARE PRIMARILY DETERMINED BY THEIR SEQUENCE. HOWEVER, THIS DOGMA HAS BEEN CHALLENGED BY STUDIES OF SECRETED MOLECULES INCLUDING THE MEMBRANE ENCASED EXTRACELLULAR VESICLES (EVS) AND THE MEMBRANELESS STRUCTURES KNOWN AS BMCS, WHICH INDICATE THAT CELLS CREATE DISCRETE CHEMICAL ENVIRONMENTS WHERE THEY ORGANIZE COMPLEX BIOMOLECULES. BMCS FORM IN MANY COMPARTMENTS OF THE BODY, INCLUDING THE NUCLEUS (NUCLEOLUS, CENTROSOME, CAJAL BODY, STRESS GRANULES), THE CYTOPLASM (P-BODIES, STRESS GRANULES), AND EVEN THE NEURONAL CYTOPLASM (NEURONAL TRANSPORT GRANULES). WHILE THE DETAILS OF BMC BIOLOGY IS STILL EVOLVING, IT IS KNOWN THAT RNA AND PROTEINS WITH INTRINSICALLY DISORDERED REGIONS (IDRS) ARE KEY FACTORS THAT INFLUENCE INTRACELLULAR COMPOSITION, SIZE, AND MORPHOLOGY OF BMCS. THE SIZE AND COMPOSITION OF BMCS ARE ALSO THOUGHT TO BE DIFFERENT. BMCS PARTICIPATE IN THE REGULATION OF VARIOUS PHYSIOLOGICAL PROCESSES, SUCH AS MODULATION OF HOST TRANSCRIPTION AND STRESS RESPONSE. BMCS ALSO PLAY IMPORTANT ROLES IN PATHOPHYSIOLOGICAL CONDITIONS, INCLUDING CANCER, NEURODEGENERATIVE DISEASES, AND VIRAL PATHOGENESIS, INCLUDING MODULATION OF HIV REPLICATION. INDEED, IT IS KNOWN THAT HIV INFECTION RESULTS IN PERIPHERAL AND NEURO INFLAMMATION, AND THAT CHRONIC EXPOSURE TO THC IN THE CONTEXT OF HIV IN HUMANS OR SIV INFECTION IN RHESUS MACAQUES RESULTS IN AMELIORATION OF DISEASE OR SYMPTOMS. HOWEVER, THE REGULATORS OF HIV DISEASE AND THC RESPONSE IS NOT COMPLETELY UNDERSTOOD. DESPITE THE REPORTED ROLE OF BMCS IN VARIOUS CONDITIONS, WHETHER OR NOT BMCS REGULATE THE PATHOGENESIS OF HIV INFECTION AND THE INTERSECTION OF HIV AND THC USE IS UNKNOWN. IT IS ALSO UNKNOWN WHETHER BMCS ASSEMBLE IN VIVO, WHERE THEY MAY PLAY RELEVANT BIOLOGICAL ROLES. THERE IS PAUCITY OF INFORMATION ON THE PRESENCE OF BMCS IN BODY FLUIDS (IN VIVO) AND THEIR ROLE IN HIV PATHOGENESIS. THE REASON FOR THIS SCARCITY OF INFORMATION IS PARTLY DUE TO LACK OF TOOLS FOR ISOLATION OF BMCS FROM BODY FLUIDS. IN PRELIMINARY STUDIES, WE DEVELOPED A NOVEL SIZE-GUIDED PARTICLE PURIFICATION LIQUID CHROMATOGRAPHY (PPLC). WE USED PPLC TO ISOLATE AND RETRIEVE DYE-FREE BMCS FROM HUMAN SEMEN, RHESUS MACAQUE BLOOD PLASMA AND CEREBROSPINAL FLUID (CSF). USING STANDARD ELECTRON MICROSCOPY AND ENERGY DISPERSIVE X-RAY EQUIPPED TRANSMISSION ELECTRON MICROSCOPE (TEM-EDX) ANALYSES, WE OBTAINED STRUCTURAL AND ELEMENTAL INFORMATION OF THE BMCS. PROTEOMICS ANALYSIS AND RNA BIOANALYZER STUDY WERE USED TO SHOW THAT THE BMCS CONTAIN PROTEIN AND RNA, WHILE CELL BIOLOGY ASSAY INDICATE THAT BMCS ARE CYTOTOXIC TO CELLS. THESE DATA PROVIDE COMPELLING EVIDENCE THAT BIOACTIVE BMCS ARE PRESENT IN VIVO. THUS, IN THIS PROPOSAL, WE WILL USE PPLC TO ISOLATE BMCS FROM RHESUS MACAQUE BLOOD, CSF, SALIVA, AND INTESTINAL CONTENTS. WE WILL THEN USE VARIOUS STRUCTURAL, BIOCHEMICAL, AND CELL BIOLOGY ASSAYS TO PROVIDE STRUCTURAL, MOLECULAR, AND FUNCTIONAL CLASSIFICATION OF BMCS FROM SIV INFECTED AND THC TREATED MACAQUES, WHICH WILL PROVIDE A FRAMEWORK TO DISSECT BMC FUNCTIONS IN HIV/AIDS AND THC USE.

THE HIPPOCAMPUS AND DENTATE GYRUS ARE CRITICALLY IMPORTANT BRAIN REGIONS NECESSARY FOR THE FORMATION OF LONG-TERM MEMORIES, AND DAMAGE TO THESE REGIO

PREVENTION OF VASCULOPATHY AND NEPHROPATHY IN METABOLIC SYNDROME

ADENOSINE AND EPOXYEICOSATRIENOIC ACIDS

REACTIVE OXYGEN SPECIES AND VASCULAR O2 SENSING

PROSTAGLANDINS: KININS AND ANGIOTENSIN INTERACTIONS

AWARD DESCRIPTION: THE WOMEN'S INSTITUTE FOR SCIENCE ENTREPRENEURSHIP (WISE) AIMS TO OFFER UNIQUE OPPORTUNITIES TO WOMEN COLLEGE STUDENTS TO LEVERAGE THEIR EDUCATION IN STEM FIELDS TO EXPLORE NEW SCIENTIFIC CONCEPTS AND LAUNCH NEW BUSINESSES. BY OFFERING A MULTIDISCIPLINARY ENTREPRENEURSHIP EDUCATIONAL OPPORTUNITY, THE PROJECT PROMISES TO OPEN AND EXPAND CAREER TRAJECTORIES FOR PROFESSIONALLY UPWARD MOBILE WOMEN. THEY WOULD HAVE THE POTENTIAL TO POSITION THEMSELVES AS RESPECTED CONTRIBUTORS WHO HAVE OPPORTUNITIES FOR SUBSTANTIVE DIFFERENCES TO THEIR RESPECTIVE FIELDS, THEIR COMMUNITIES, AND WORLD-WIDE.PURPOSE: THE PROJECTS AIMS TO ADDRESS THE FOLLOWING : 1) DESPITE PROGRESS, WOMEN REMAIN UNDERREPRESENTED IN THE BIOMEDICAL SCIENCES; 2. THE UNDERGRADUATE YEARS ARE IDEAL GROUND FOR ENCOURAGEMENT AND RECRUITMENT OF WOMEN INTO THE BIOMEDICAL SCIENCES; AND 3) LONGITUDINAL MENTORING, PARTICIPATION IN ACTUAL RESEARCH, AND SHARING THE JOYS OF DISCOVERY ARE PROVEN METHODS TO GROOM TOMORROW'S SCIENTIFIC LEADERS IN THE HEALTH CARE FIELD.ACTIVITIES TO BE PERFORMED: WISE WILL CONNECT INTERDISCIPLINARY TEAMS OF STUDENTS, RESEARCHERS, FACULTY, AND BIOTECH ENTREPRENEURS TO WORK COLLABORATIVELY TO DEVELOP SYNERGY. WISE WILL WORK CLOSELY WITH THE FACULTY OF LCW, NYMC AND THE TOURO COLLEGE & UNIVERSITY SYSTEM, ALONG WITH THE ENTREPRENEURIAL, BUSINESS, AND SOCIAL VENTURE COMMUNITIES, FOR OFFERED PROGRAMS. THE PROJECT WILL BE IMPLEMENTED IN TWO PHASES: PHASE 1: LCW ENTREPRENEURSHIP EDUCATION AND NETWORKING: STUDENTS TO TAKE ADVANTAGE OF A MENU OF PLANNED ACTIVITIES BEGINNING IN THEIR FRESHMAN YEAR, INCLUDING: SPEAKER'S SERIES, ENTREPRENEURSHIP ROUNDTABLES, RESEARCH PAPERS, INTERNSHIPS, STARTUPNATION (AN ENTREPRENEURIAL-FOCUSED TRIP TO ISRAEL, WITH AN EMPHASIS ON THE STARTUP AND INVESTMENT CLIMATE IN ISRAEL AND SURROUNDING COUNTRIES), MENTORING, E-TREK (IN-PERSON AND VIRTUAL VISITS TO ENTREPRENEURIAL SETTINGS IN THE GREATER NYC AREA TO INTRODUCE STUDENTS TO STARTUP ENVIRONMENTS, ALUMNAE AND RESOURCES), SEED MONEY GRANT PROGRAM AND NEW VENTURE COMPETITION, AND STUDENT CLUBS FOCUSED ON ENTREPRENEURSHIP. PHASE 2: NYMC SCIENTIFIC TRAINING AND BIOTECH INTERNSHIPS: INTERNSHIP FOR 10 PROGRAM PARTICIPANTS, SUMMER RESEARCH (INTENSIVE 8-WEEK RESEARCH PROJECT UNDER THE SUPERVISION OF A FACULTY RESEARCHER), A YEAR-LONG UNDERGRADUATE EXPERIENCE, RESEARCH AND SEMINARS, REAL-WORLD EXPERIENCE, M.S. IN BIOMEDICAL SCIENCE & MANAGEMENT EXPECTED OUTCOMES: THE WOMEN'S INSTITUTE FOR SCIENCE ENTREPRENEURSHIP (WISE) IS A TRANSFORMATIVE PROGRAM THAT, IF SUCCESSFUL, WILL ALLOW FEMALE UNDERGRADUATES TO APPLY THEIR STEM EDUCATION TO CULTIVATE NEW SCIENTIFIC CONCEPTS AND LAUNCH NEW BUSINESSES. WISE AIMS TO CREATE A MULTIDISCIPLINARY ENTREPRENEURSHIP EDUCATIONAL OPPORTUNITY FOR STUDENTS AND LAUNCH CAREERS FOR ALUMNI WHO HAVE THE PROMISE TO MAKE SIGNIFICANT DIFFERENCES TO THEIR COMMUNITIES AND AROUND THE WORLD.INTENDED BENEFICIARIES: STEM TECHNOLOGY, WHICH IS A MAJOR DRIVER OF ECONOMIC DEVELOPMENT IN NEW YORK, BENEFITS DIRECTLY FROM STIMULATION OF NEW COHORTS OF TALENTED WOMEN TO ENTER THE FIELD. THIS PROJECT DRAWS WOMEN INTO NEW YORK MEDICAL COLLEGE AND OTHER ACADEMIC MEDICAL CENTERS IN NEW YORK, ENABLING IT TO OUT-COMPETE OTHER REGIONS OF THE COUNTRY WITH THE UNIQUE PROFESSIONAL OPPORTUNITIES AFFORDED BY THIS PROJECT. IN ADDITION, NEW YORK MEDICAL COLLEGE, WHICH ALREADY IS A MAJOR CONTRIBUTOR TO THE LOCAL ECONOMY, IS UTILIZING THIS PROJECT TO RECRUIT SIGNIFICANT NEW NUMBERS OF PROFESSIONAL WOMEN WHO WILL SERVE AS RESEARCH FELLOWS AND ENTREPRENEURS, FURTHER STIMULATING ECONOMIC DEVELOPMENT IN WESTCHESTER AND THE MID-HUDSON REGION. THROUGH OFFERING A MULTIDISCIPLINARY ENTREPRENEURSHIP EDUCATIONAL OPPORTUNITY, THE PROJECT PROMISES TO OPEN AND EXPAND CAREER TRAJECTORIES FOR NUMEROUS PROFESSIONALLY UPWARD MOBILE WOMEN. SUBRECIPIENT: N/A

EMERGENCY FINANCIAL AID GRANTS TO STUDENTS UNDER THE CORONAVIRUS AID, RELIEF, AND ECONOMIC SECURITY (CARES) ACT

POPULATION GENOMICS OF STREPTOCOCCUS PYOGENES

TISSUE-SPECIFIC INFECTION BY GROUP A STREPTOCOCCI

DEVELOPING AND EVALUATING COUNTERMEASURES AGAINST TETRAMETHYLENEDISULFOTETRAMINE

EXAMINING THE CELLULAR HETEROGENEITY OF ADULT MYOCARDIUM USING TRANSGENIC MICE

REGULATED EXPRESSION OF B. BURGDORFERI VIRULENCE GENES

DISSECTING THE MECHANISMS THAT CONTROL FGF23

GENE EXPRESSION BIOMARKERS FOR DIAGNOSIS OF LYME DISEASE

MECHANISM OF CHEMOREFLEX AND BAROREFLEX ALTERATIONS CAUSING POSTURAL TACHYCARDIA SYNDROME IN POTS PATIENTS WITH ORTHOSTATIC HYPERPNEA AND HYPOCAPNIA. - PROJECT SUMMARY/ABSTRACT POSTURAL TACHYCARDIA SYNDROME (POTS) IS CHRONIC ORTHOSTATIC INTOLERANCE WITH EXCESSIVE UPRIGHT TACHYCARDIA WITHOUT HYPOTENSION AND OCCURS MOSTLY IN YOUNG FEMALES (>85%). SYMPTOMS ARE LIGHTHEADEDNESS, FATIGUE, COGNITIVE LOSS, AND DYSPNEA WITH HYPERPNEIC HYPOCAPNIA AND VENTILATORY INSTABILITY IN APPROXIMATELY 50% OF OUR NEW POTS PATIENTS. OUR RECENT STUDIES SUPPORT AN INCREASED HYPOXIC VENTILATORY RESPONSE, SYMPATHOEXCITATION WITH SENSITIZATION OF THE CAROTID BODY PERIPHERAL CHEMOREFLEX. HYPOCAPNIA ALONE PRODUCES TACHYCARDIA AND REDUCES CEREBRAL BLOOD FLOW (CBF) ENGENDERING MANY POTS SYMPTOMS. UNLIKE VOLUNTARY HYPERVENTILATION, HYPERPNEIC POTS IS RELATED TO DECREASED CENTRAL BLOOD VOLUME AND CARDIAC OUTPUT, INCREASED SYSTEMIC VASCULAR RESISTANCE AND BP, SPLANCHNIC BLOOD POOLING, AND A SHIFT IN THE SIGMOIDAL BAROREFLEX RELATION THAT FAVORS TACHYCARDIA EVEN WHILE SUPINE. CAROTID BODY SENSITIVITY IS HIGHLY PLASTIC AND CAN BE RAPIDLY CONDITIONED BY CHRONIC INTERMITTENT HYPOXIA OR BY “STAGNANT HYPOXIA” - RECURRENT ISCHEMIA OF THE CAROTID BODY. STAGNANT HYPOXIA CAN BE PRODUCED BY “INITIAL ORTHOSTATIC HYPOTENSION” (IOH) COMPRISING A TRANSIENT FALL IN BP AND CBF ON STANDING. IOH IS ABNORMAL IN POTS WITH A PARADOXICAL DECREASE IN CEREBRAL CONDUCTANCE INDICATING IMPAIRED CEREBRAL AUTOREGULATION. WE HYPOTHESIZE THAT A MECHANISM FOR THE GENESIS OF POTS INVOLVES CAROTID BODY SENSITIZATION INITIATED BY RECURRENT IOH RESULTS IN HYPERPNEIC HYPOCAPNIA DRIVING TACHYCARDIA DIRECTLY AND INDIRECTLY BY RESETTING ARTERIAL BAROREFLEXES. WE WILL COMPARE FEMALE POTS PATIENTS AGED 15 TO 39 YEARS WITH (N=40) AND WITHOUT (N=40) ORTHOSTATIC HYPERPNEA, TO HEALTHY VOLUNTEERS (N=40) WITH THE FOLLOWING AIMS: 1. TO TEST ORTHOSTATIC CARDIORESPIRATORY RESPONSES TO DETERMINE WHETHER PROLONGED IOH PRECEDES UPRIGHT HYPOCAPNIA IN HYPERPNEIC POTS BUT NOT IN CONTROLS OR NON-HYPERPNEIC POTS. CEREBRAL BLOOD FLOW, RESPIRATORY AND HEMODYNAMIC MEASUREMENTS, INVESTIGATING SPLANCHNIC BLOOD FLOW AND MEASURING CHANGES OF CBV, REGIONAL BLOOD VOLUMES, AND CARDIAC OUTPUT DURING A 10 MIN STAND TO QUANTIFY IOH, AND A 10 MIN TILT TEST TO 70° TO QUANTITATE CARDIORESPIRATORY CHANGES WILL ALLOW US TO STRATIFY HYPERPNEIC AND NON-HYPERPNEIC POTS PATIENTS. 2.TO TEST IF CHEMOREFLEX SENSITIZATION OF VENTILATION AND SYMPATHETIC ACTIVITY ARE ABNORMAL WHEN SUPINE AND UPRIGHT (AT 45O) AND HOW THAT INTERACTS WITH OXFORD MEASURED CARDIOVAGAL AND SYMPATHETIC BAROREFLEXES UNDER CONTROLLED GAS CONDITIONS: ISOCAPNIC HYPOXIA AND ISOCAPNIC HYPEROXIA TO MEASURE CAROTID BODY REFLEX; HYPEROXIC ISOCAPNIA AND HYPEROXIC HYPERCAPNIA TO MEASURE CENTRAL CHEMOREFLEXES. HYPEROXIA SILENCES PERIPHERAL CHEMORECEPTORS AND WILL NORMALIZE BAROREFLEX AND TILT RESPONSES. 3. TO EMPLOY PHARMACEUTICAL MODULATION OF THE CHEMOREFLEX TO ELUCIDATE MECHANISMS OF HYPERPNEIC POTS, WITH THE POTENTIAL OF DETERMINING TREATMENT MODALITIES TO REDUCE CHEMOREFLEX SENSITIVITY AND HYPERPNEA – USING AN ANGIOTENSIN TYPE 1 RECEPTOR BLOCKER, DIETARY NITRATE TO DONATE NO, AND AN ADENOSINE RECEPTOR ANTAGONIST.

HEMODYNAMIC FORCES REGULATE BMPS IN CORONARY ARTERIES

NON-CANONICAL WNT SIGNAL TRANSDUCTION AND CARDIOGENESIS

ZEISS LSM 980 PLUS AIRYSCAN 2 FOR NYMC IMAGING CORE

TARGETS FOR SELENIUM IN PROSTATE CANCER PREVENTION

STEM CELL-DERIVED DEVELOPMENTAL HUMAN CORTICAL INTERNEURONS TO TREAT INTRACTABLE EPILEPSY - ABSTRACT EPILEPSY IS A SEVERE NEUROLOGICAL DISEASE AFFECTING MORE THAN 65 MILLION PEOPLE WORLDWIDE AND IS CHARACTERIZED BY UNPREDICTABLE ABNORMAL ELECTRICAL DISCHARGES RESULTING IN RECURRENT SEIZURES. ABOUT ONE THIRD OF PATIENTS WITH EPILEPSY SUFFER FROM INTRACTABLE SEIZURES THAT DO NOT RESPOND TO ANTIEPILEPTIC DRUGS (AEDS). NEUROSURGICAL INTERVENTIONS AND NEUROSTIMULATOR DEVICES ARE USEFUL OPTIONS FOR ONLY A FRACTION OF PATIENTS WITH DRUG-REFRACTORY SEIZURES, UNDERSCORING THE URGENT NEED TO DEVELOP NEW THERAPIES. ONE STRATEGY WITH CONSIDERABLE PROMISE IS TO ENGRAFT NEW NEURONS TO PROVIDE ENHANCED GABAERGIC INHIBITION IN AN ACTIVITY-DEPENDENT MANNER. HOWEVER, USE OF FETAL NEURONS FOR CELL THERAPY IS ASSOCIATED WITH PRACTICAL AND ETHICAL ISSUES. THEREFORE, TO OVERCOME SUCH HURDLES, IN OUR PREVIOUS STUDIES, WE PIONEERED THE TRANSPLANTATION OF HUMAN PLURIPOTENT STEM CELLS (HPSCS)- DERIVED MEDIAL GANGLIONIC EMINENCE (MGE)-TYPE HUMAN DEVELOPMENTAL CORTICAL INTERNEURONS (CINS) INTO EPILEPTIC MOUSE BRAINS AND DEMONSTRATED THEIR INTEGRATION INTO DYSFUNCTIONAL CIRCUITRY, ACCOMPANIED BY THE SUPPRESSION OF SEIZURES AND COMORBID BEHAVIORAL ABNORMALITIES. FURTHERMORE, WE HAVE ALSO DETERMINED THE OPTIMAL STAGE OF HUMAN CIN DIFFERENTIATION TO ENSURE MAXIMAL INTEGRATION INTO HOST CIRCUITRY AS WELL AS SAFETY WITHOUT RISK OF TUMOR FORMATION, AND DEVELOPED A METHOD TO EFFICIENTLY GENERATE THESE SAFE AND HIGHLY MIGRATORY POPULATIONS OF CINS FROM HPSCS IN LARGE QUANTITIES, BRINGING CELL THERAPY FOR EPILEPSY ONE STEP CLOSER TO REALITY. HOWEVER, THERE ARE STILL IMPORTANT ISSUES TO ADDRESS PRIOR TO THE CLINICAL TRANSLATION OF THIS PROMISING RESTORATIVE THERAPY; 1) WHAT IS THE SYNAPTIC CONNECTION SPECIFICITY OF HUMAN DEVELOPMENTAL CINS IN ADULT EPILEPTIC CIRCUITRY? 2) WHAT ARE SAFE AND OPTIMAL DENSITIES OF HUMAN CIN GRAFTS FOR INHIBITION OF EPILEPTIC HOST CIRCUITRY? 3) DO HUMAN DEVELOPMENTAL CIN GRAFTS MAINTAIN LONG-TERM EFFICACY AND SAFETY IN EPILEPTIC BRAINS? TO TACKLE THESE ISSUES, WE WILL TEST OUR HYPOTHESIS THAT HUMAN IPSC-DERIVED DEVELOPMENTAL CINS WITH OPTIMAL GRAFTING DENSITIES PREFERENTIALLY INNERVATE HOST EXCITATORY NEURONS AND AMELIORATE SEIZURE ACTIVITY WITH LONG-TERM EFFICACY AND SAFETY. WE WILL TRANSPLANT MIGRATORY HUMAN CINS INTO NOD SCID GAMMA (NSG) MICE WITH INTRAHIPPOCAMPAL KAINIC ACID-INDUCED TEMPORAL LOBE EPILEPSY (KA-TLE), A MODEL OF HUMAN HIPPOCAMPAL SCLEROSIS, THE MOST COMMON CAUSE OF DRUG-RESISTANT EPILEPSY, AND ANALYZE GRAFTED CINS’ SYNAPTIC INTEGRATION SPECIFICITY AND HOST INHIBITION IN THE EPILEPTIC BRAINS. THE LONG-TERM MAINTENANCE OF ANTI-EPILEPTIC EFFICACY WILL BE EXTENSIVELY ANALYZED BY 24/7 VIDEO-EEG RECORDINGS 3 MONTHS, 6 MONTHS AND 9 MONTHS AFTER TRANSPLANTATION. WE WILL ANALYZE THE GRAFTS IMMUNOHISTOCHEMICALLY TO DETERMINE THE EXTENT OF CELL SURVIVAL, MATURATION, INTEGRATION, AND MOST IMPORTANTLY, CELL PROLIFERATION AS A MEASURE OF GRAFT SAFETY WITHOUT RISK OF UNCONTROLLED GROWTH. COMPLETION OF THESE STUDIES IS PIVOTAL FOR TRANSLATING THIS EXPERIMENTAL THERAPY INTO A VIABLE THERAPEUTIC STRATEGY FOR INTRACTABLE EPILEPSY.

IMPACT OF HEALTH DEPARTMENT WORKER SAFETY TRAINING ON HEALTH IMPACTS AFTER SANDY

ADIPOCYTE EET-PGC1ALPHA-HO-1 IN OBESITY-DRIVEN HYPERTENSION

FACSARIA II CELL SORTER

OVERCOMING IMMUNE ESCAPE MECHANISMS IN IMMUNOTHERAPY OF NEUROBLASTOMA

POSTDOCTORAL TRAINING IN GENERAL, PEDIATRIC AND PUBLIC HEALTH DENTISTRY AND DENTAL HYGIENE

MACROLIDE RESISTANCE TRANSFER IN STREPTOCOCCUS PYOGENES - PROJECT SUMMARY GROUP A STREPTOCOCCUS (GAS) IS A STRICT HUMAN PATHOGEN THAT PRIMARILY INFECTS THE EPITHELIA AT THE THROAT OR SKIN, LEADING TO ~750 MILLION INFECTIONS PER YEAR. HIGH RATES OF MORBIDITY AND MORTALITY RESULT FROM INVASIVE GAS (IGAS) DISEASE. DESPITE ITS IMPORTANCE AS A GLOBAL PATHOGEN, THERE IS NO VACCINE AVAILABLE FOR GAS. IN THE C.D.C.'S ANTIBIOTIC RESISTANCE THREATS REPORT OF 2019, ERYTHROMYCIN-RESISTANT GAS ARE LISTED AS A “CONCERNING THREAT” AND THE % OF INVASIVE GAS (IGAS) ISOLATES RESISTANT TO ERYTHROMYCIN HAS RECENTLY TRIPLED. MACROLIDES ARE COMMONLY PRESCRIBED FOR PATIENTS WITH SS-LACTAM ALLERGIES, AND LINCOSAMIDES ARE HIGHLY EFFECTIVE AGAINST IGAS DISEASE BECAUSE EXOTOXIN PRODUCTION IS HALTED. THE PROBLEM OF ANTIBIOTIC-RESISTANCE IN GAS IS FURTHER COMPOUNDED BY 2019-2020 REPORTS ON THE EMERGENCE OF STABLE SS-LACTAM RESISTANCE DUE TO ALTERED PENICILLIN- BINDING PROTEINS; THE POTENTIAL FOR LATERAL SPREAD OF RESISTANCE GENES TO OTHER GAS STRAINS IS VERY HIGH. THE PROPOSED STUDY SEEKS A DEEPER UNDERSTANDING OF THE BIOLOGICAL CAUSES AND CLINICAL CONSEQUENCES OF THE ACQUISITION BY GAS OF MOBILE GENETIC ELEMENTS (MGES) HARBORING MACROLIDE-RESISTANCE GENES (R-GENES). AIM 1 SEEKS TO DEFINE THE GENETIC ARCHITECTURE OF THE (NEAR) COMPLETE REPERTOIRE OF MGES THAT HARBOR MACROLIDE- RESISTANCE GENES IN GAS. AIM 2 USES EXPERIMENTAL MODELS OF HORIZONTAL TRANSFER OF R-GENE-MGES BETWEEN GAS STRAINS, TO OPTIMIZE MICROENVIRONMENTAL CONDITIONS AND TO GENERATE ISOGENIC PAIRS OF PARENTAL-RECIPIENT AND NEW RECOMBINANT STRAINS. AIM 3 EVALUATES THE EFFECT OF MGE ACQUISITION ON HOST CELL PHENOTYPES THAT ARE INDEPENDENT OF DRUG-RESISTANCE; SEVERAL CARGO GENES ARE PREDICTED TO ALTER GLOBAL GENE EXPRESSION AND/OR CONTRIBUTE TO VIRULENCE. TRANSCRIPTOMES AND FITNESS WILL BE COMPARED FOR THE ISOGENIC PAIRS. A MOUSE MODEL FOR IGAS DISEASE WILL TEST THE HYPOTHESIS THAT MGE ACQUISITION LEADS TO AN INCREASE IN THE INTRINSIC VIRULENCE OF THE NEW RECOMBINANT. IF CORRECT, DATA MAY EXPLAIN THE EPIDEMIOLOGICAL FINDINGS ON THE HIGH ASSOCIATION OF MACROLIDE-RESISTANCE WITH IGAS DISEASE AND THEREBY, PROVIDE A PLATFORM FOR FUTURE STUDIES THAT PROBE MOLECULAR MECHANISMS. TOOLS TO BE DEVELOPED FROM THE PROPOSED WORK INCLUDE A CONSOLIDATED STRUCTURAL ORGANIZATION FOR THE MACROLIDE- RESISTANCE MGES, TO BE POSTED ON THE INTERACTIVE USER-FRIENDLY WWW.PUBMLST.ORG WEBSITE (AIM 1), AND IMPROVED EXPERIMENTAL PROTOCOLS FOR HORIZONTAL GENE TRANSFER BY FILTER-MATING (AIM 2). IN ADDITION TO TESTING THE HYPOTHESIS THAT MGES IMPART PHENOTYPIC CHANGES IN AN ANTIBIOTIC-FREE ENVIRONMENT, TRANSCRIPTOME ANALYSIS (AIM 3) IS EXPLORATORY AND MAY PROVIDE A WINDOW INTO CRITICAL MOLECULAR MECHANISMS.

POPULATION ANALYSIS OF GROUP A STREPTOCOCCAL PHENOTYPES

THE PRESENT APPLICATION SEEKS TO STUDY THE LONG-TERM CONSEQUENCES OF COSMIC RADIATION EXPOSURE. SPACE TRAVEL INCREASES SOLAR PARTICLE RADIATION EXPOSURE WHICH IS SIGNIFICANTLY ELEVATED ONCE TRAVEL MOVES BEYOND LOW EARTH ORBIT. THIS INCLUDES A COMBINATION OF HIGH ENERGY PROTONS AND HEAVY IONS SUCH AS 56FE 28SI AND 16O. THE ADVERSE RISK OF RADIATION-INDUCED HEART FAILURE IS ALSO EVIDENT AS A LONG-TERM CONSEQUENCE OF ACCIDENTAL RADIATION EXPOSURE OR CANCER TREATMENT. SURVIVORS OF CANCER ARE ALSO AT RISK FOR OTHER ADVERSE HEALTH OUTCOMES INCLUDING ABNORMAL PULMONARY FUNCTION ENDOCRINE DISORDERS NEUROCOGNITIVE IMPAIRMENT AND OSTEOPOROSIS. ALL OF THESE ORGAN SYSTEMS ARE CHARACTERIZED BY A LOW TURNOVER OF CELLS AND IT IS LIKELY THAT AN ACCELERATED CELL DEATH AND/OR THE FAILURE OF REGENERATION BY THE PLURIPOTENT CELLS MAY BE THE UNDERLYING CAUSE OF ORGAN FAILURE. ALTHOUGH THIS APPLICATION WILL FOCUS ON HEART FAILURE OUR FINDINGS WILL HAVE IMPLICATIONS FOR MANY ORGAN SYSTEMS. OUR PRELIMINARY STUDIES OBSERVED DEGRADATION OF CARDIOVASCULAR FUNCTION IN A MODEL OF COSMIC RADIATION (HIGH-LET) EXPOSURE. MICE WERE EXPOSED TO 50 CGY (FE56) AT 3 MONTHS OF AGE AND THEN STUDIED AT 24 MONTHS OF AGE. DEGRADATION OF CARDIAC FUNCTION WAS EVIDENT BY SIGNIFICANT DECREASES IN MYOCARDIAL CONTRACTILITY AND RELAXATION. CONCOMITANT WITH THIS WERE SIGNIFICANT CHANGES IN MITOCHONDRIAL AND STEM/PROGENITOR CELL FUNCTION. IN THE PRESENT GROUND BASED APPLICATION WE PROPOSE TO EVALUATE THE IMPACT OF HIGH-LET; TO IDENTIFY THE PATHWAY TO HEART FAILURE AND EVALUATE THREE DISTINCT PROTOCOLS AS POTENTIAL COUNTERMEASURES. HYPOTHESIS: RADIATION-INDUCED CARDIOMYOPATHY IS THE RESULT OF A CELL SPECIFIC FAILURE. CELL SPECIFIC FAILURE IS THE RESULT OF AN INABILITY TO MAINTAIN THE BALANCE OF REPAIR/REPLACEMENT THAT ULTIMATELY LEADS TO ACTIVATION OF DEGRADATION PATHWAYS AND ACCELERATED CELL LOSS. THIS STUDY WILL UTILIZE BOTH CULTURED CELLS AS WELL AS SWISS WEBSTER MICE TO BE RANDOMLY ASSIGNED TO CONTROL OR SINGLE HEAVY ION HIGH- LET EXPOSURE GROUPS. PHASE 1 WILL USE SINGLE EXPOSURES TO HIGH-LET EXPOSURE OF 1) 56FE (50 CGY) OR 2) PROTON (200 CGY). PHASE 2 WILL UTILIZE A MIXED FIELD EXPOSURE FOLLOWING THE GUIDELINES OF THE HERO ANNOUNCEMENT (80JSC017N0001-BPBA; APPENDIX C). ALL EXPOSURES WILL BE PERFORMED AT THE NASA SPACE RADIATION LABORATORY AT BROOKHAVEN NATIONAL LABORATORY. WE PLAN TO STUDY THREE DISTINCT COUNTERMEASURES INCLUDING 1) MITOTEMPO 2) METFORMIN 3) LISINOPRIL. MITOTEMPO IS AN ANTIOXIDANT THAT PARTITIONS TO THE MITOCHONDRIA. METFORMIN AN ANTIDIABETIC MAINSTAY HAS MORE RECENTLY BEEN SHOWN TO HAVE SIGNIFICANT ANTICANCER PROPERTIES. LISINOPRIL AN INHIBITOR OF THE ANGIOTENSIN SYSTEM HAS BEEN SHOWN TO MITIGATE RADIATION INJURIES FROM LOW-LET EXPOSURE. EVALUATIONS WILL BE MADE ON SEVERAL LEVELS TO INCLUDE 1) DETERMINATION OF CARDIOVASCULAR FUNCTION 2) IDENTIFICATION OF CELL SPECIFIC FAILURE 3) INTRACELLULAR DETERMINATION OF DAMAGE FOCUSING ON GENOMIC AND MITOCHONDRIAL DNA DAMAGE. CELL FAILURE IN TERMS OF METABOLIC FAILURE ACCELERATED SENESCENCE AS WELL AS DNA DAMAGE WILL BE DETERMINED. A NUMBER OF INVESTIGATIONS IN SPACE BIOLOGY ARE CURRENTLY ONGOING AT NYMC AND COLLABORATIONS AND SHARING OF RESOURCES WILL ENHANCE THE RESEARCH PRODUCTS DERIVED FROM FUNDING THIS APPLICATION. RADIATION INDUCED CARDIOMYOPATHIES ARE OBSERVED MONTHS OR YEARS AFTER EXPOSURE. THE PRESENT APPLICATION WILL SEPARATE THE INSULT FROM THE CONSEQUENCES. OUR PRELIMINARY FINDINGS DEMONSTRATED SIGNIFICANT DEGRADATION OF CARDIAC FUNCTION SIMILAR TO THE ACCELERATED AGING PHENOTYPE OBSERVED WITH CHEMOTHERAPY. IT MAY NOT BE RELEVANT THAT ANY SINGLE CELL DIES BUT THAT THE BALANCE OF CELL DEATH AND CELL REPLACEMENT IS UPSET. ALTHOUGH FOCUSED ON THE HEART THESE INVESTIGATIONS WILL HAVE WIDESPREAD APPLICATION TO OTHER ORGAN SYSTEMS AND COLLECTIVELY SERVE THE LONG TERM HEALTH AND WELL BEING OF FLIGHT CREWS.

TARGETING EBV'S DEUBIQUITINATING ACTIVITY FOR THERAPY - PROJECT SUMMARY/ABSTRACT EPSTEIN-BARR VIRUS (EBV), A HUMAN TUMOR VIRUS, IS THE CAUSATIVE AGENT OF MONONUCLEOSIS AND IMMUNOBLASTIC LYMPHOMAS. EBV IS STRONGLY ASSOCIATED WITH BURKITT’S LYMPHOMA, HODGKIN’S LYMPHOMA, AND NASOPHARYNGEAL CARCINOMA (NPC). ADDITIONALLY, RECENT STUDIES SUGGEST THAT EBV HAS A PATHOGENIC ROLE WITH THE ONSET OF MULTIPLE SCLEROSIS (MS). APPROXIMATELY 90% OF THE WORLD’S POPULATION IS INFECTED WITH EBV, BUT MOST DO NOT PRESENT DISEASE. HOWEVER, FOR THOSE THAT DO DEVELOP EBV-RELATED ILLNESS THERE REMAINS NO DIRECTED SMALL MOLECULE THERAPY. EBV-TRIGGERED DISEASE, CAUSING DEBILITATING ILLNESS AND DEATH, REMAINS A WORLD-WIDE PROBLEM. INTERESTINGLY, EBV EXPRESSES A UNIQUE PROTEIN (BPLF1) THAT POSSESSES DEUBIQUITINATING ACTIVITY. BPFL1 IS KNOWN TO REGULATE BOTH CELLULAR AND VIRAL TARGET ACTIVITIES, YET IT REMAINS LARGELY UNSTUDIED. OUR WORK HAS IMPLICATED BPLF1 IN A WIDE RANGE OF VIRAL AND CELLULAR PROCESSES INCLUDING INFECTIVITY (90% REDUCTION WITH KNOCKOUT OF BPLF1), VIRAL DNA REPLICATION, AND DNA REPAIR. ALSO WE RECENTLY REPORTED SURPRISING NEW FINDINGS THAT KNOCKOUT OF BPLF1 DELAYS AND REDUCES HUMAN B-CELL IMMORTALIZATION AND LYMPHOMA FORMATION IN HUMANIZED MICE. THE AIM OF THIS PROPOSAL IS TO DISCOVER AND CHARACTERIZE THE FIRST-IN-WORLD SMALL MOLECULE INHIBITORS OF BPLF1 DEUBIQUITINATING ACTIVITY. THE GOALS OF THIS PROPOSAL ARE TO 1) USE HIGH THROUGHPUT SCREENING TO IDENTIFY NOVEL CHEMOTYPES FOR A LEAD OPTIMIZATION EFFORT AND 2) PRODUCE FURTHER VALIDATION THAT SMALL MOLECULE INHIBITION OF BPLF1 DEUBIQUITINATING ACTIVITY IS A PROMISING AVENUE FOR TREATING DISEASES CAUSED BY EBV INFECTION. THIS WORK WILL LAY THE FOUNDATION FOR A CHEMICAL PROBE AND DRUG DISCOVERY EFFORT TO COMBAT EBV-ASSOCIATED DISEASE.

THE ROLE OF DNA POLYMERASE DELTA 4 IN LUNG CARCINOGENESIS INDUCED BY GENOTOXIC CARCINOGENS - ABSTRACT LUNG CANCER IS THE LEADING CAUSE OF CANCER-RELATED DEATH IN THE WORLD. EXPOSURES TO AIRBORNE CARCINOGENS CONTRIBUTE SIGNIFICANTLY TO LUNG CARCINOGENESIS. GENOTOXIC CARCINOGENS, WHICH CAN COME FROM ENVIRONMENTAL AND/OR OCCUPATIONAL SETTINGS, ARE THE MOST IMPORTANT CAUSES OF LUNG MALIGNANCY IN HUMANS. THESE INCLUDE HEXAVALENT CHROMIUM (CR(VI)), NICOTINE-DERIVED NITROSAMINE KETONE (NNK), BENZO(A)PYRENE (BAP), AND URETHANE. EXPOSURE TO THESE CARCINOGENS CAN LEAD TO FORMATION OF DNA ADDUCTS THAT INDUCE DNA DOUBLE-STRAND BREAKS. DNA DAMAGE IS A KEY EARLY EVENT AND THE DRIVING FORCE OF GENETIC ABNORMALITIES THAT LEAD TO MALIGNANT TRANSFORMATION OF NORMAL LUNG EPITHELIAL CELLS. IN HUMANS, THERE ARE TWO FORMS OF POLΔ: POLΔ4 AND POLΔ3. POLΔ4 IS A TETRAMER CONTAINING THE P12 SUBUNIT WHICH IS ABSENT IN YEAST. THE DEGRADATION OF P12 IN RESPONSE TO DNA DAMAGE LEADS TO THE CONVERSION OF POLΔ4 TO POLΔ3. THIS IS A NOVEL CELLULAR REGULATORY SYSTEM THAT ORCHESTRATES THE FORMATION OF POLΔ3 WHICH PLAYS A KEY ROLE IN DNA REPAIR AND REPLICATION. OUR RECENT WORK SHOWS THAT POLΔ4 IS THE ENZYME FORM THAT IS INVOLVED IN THE KEY STEP IN HOMOLOGY DIRECTED REPAIR (HDR) OF DSBS. LOSS OF POLΔ4 LEADS TO HDR DEFICIENCY AND IS A NOVEL POTENTIAL CONTRIBUTOR TO TUMORIGENESIS. OUR GOALS ARE DIRECTED TO THE UNEXPLORED ROLES OF POLΔ4 LOSS IN LUNG TUMORIGENESIS. OUR FIRST AIM IS TO CHARACTERIZE THE EFFECTS OF THE LUNG CARCINOGENS (CR(VI), BAP AND NNK ON P12 DEGRADATION IN LUNG EPITHELIAL CELL LINES AS WELL AS THE MECHANISMS INVOLVED IN THIS PROCESS. THESE STUDIES HAVE NOT BEEN DONE IN NON-NEOPLASTICALLY TRANSFORMED CELLS. WE WILL STUDY THE POTENTIAL OF CHRONIC EXPOSURE TO THE ABOVE-MENTIONED CARCINOGENS ON P12 DEGRADATION, AS WELL AS THEIR CAPACITY TO LEAD TO EPIGENETIC CHANGES THAT SUPPRESS P12 EXPRESSION AFTER CHRONIC EXPOSURE. OUR SECOND AIM IS TO TEST THE HYPOTHESIS THAT P12 DEPLETION FACILITATES NEOPLASTIC TRANSFORMATION OF LUNG EPITHELIAL CELLS. WE WILL EXAMINE WHETHER P12 DELETION PROMOTES NEOPLASTIC TRANSFORMATION OF LUNG EPITHELIAL CELLS INDUCED BY THESE CARCINOGENS. WE WILL ALSO INVESTIGATE WHETHER P12 DELETION ENHANCES THE ABILITY OF URETHANE TO INDUCE LUNG TUMORS TO PROVIDE AN IN VIVO READOUT OF THE EFFECTS OF P12 ON LUNG TUMORIGENESIS. AS DELETION OF P12 ELIMINATES THE POSSIBILITY OF FORMING THE POLΔ4 ENZYME, THE P12 KNOCKOUT CELL AND MOUSE MODELS ARE ABSENT OF POLΔ4. THESE MODELS THUS HAVE A UNIQUE AND DEFINED LOCUS IN THE CONVERGENT STEP OF D-LOOP EXTENSION, A CRUCIAL STEP IN HDR. THE P12 KNOCKOUT MOUSE MODEL PROVIDES A UNIQUE OPPORTUNITY TO EXAMINE THE ROLE OF P12 IN LUNG CARCINOGENESIS INDUCED BY GENOTOXIC CARCINOGENS IN VIVO IN A CLEAN BACKGROUND. THESE STUDIES WILL LEAD TO EVIDENCE FOR A ROLE OF POLΔ4 IN LUNG TUMORIGENESIS THAT ORIGINATES FROM A NOVEL ROUTE TO HR-DEFICIENCY AND HAVE THE POTENTIAL TO OPEN UP FURTHER AVENUES OF RESEARCH THAT COULD CONTRIBUTE TO FUTURE THERAPEUTIC APPROACHES IN LUNG CANCER.

SYSTEMATIC SEARCH FOR NOVEL TREATMENTS OF INFANTILE SPASMS AMONG SIGMA-1 RECEPTOR LIGANDS - EVERY YEAR IN THE US ~1700 CHILDREN ARE NEWLY DIAGNOSED WITH EPILEPTIC SPASMS DURING INFANCY (INFANTILE SPASMS; IS). IS DEVELOP BETWEEN 3-12 MONTHS OF AGE WITH A PREDOMINANCE (60%) IN MALES. IS ARE ASSOCIATED WITH SIGNIFICANT MORTALITY AND MORBIDITY. MEDICAL TREATMENT OPTIONS FOR IS ARE DIFFERENT THAN FOR ANY OTHER TYPES OF EPILEPSY. THERE ARE TWO DRUGS WITH A REASONABLE EVIDENCE OF EFFICACY, BOTH APPROVED BY FDA: ACTH (ADRENOCORTICOTROPIN) AND VIGABATRIN, ELIMINATING SPASMS IN 50-55% OF PATIENTS IN LONG TERM. HOWEVER, ACTH CARRIES ENORMOUS COST BURDEN AND, IN UP TO 43% OF CASES HAS SIGNIFICANT AND SERIOUS ADVERSE EFFECTS, WHICH INCLUDE OBESITY, ARTERIAL HYPERTENSION, ELECTROLYTE IMBALANCE, GASTRIC ULCER, GROWTH RETARDATION, CARDIOMYOPATHY, AND IMMUNOSUPPRESSION AS WELL AS BRAIN ATROPHY. VIGABATRIN IS ALMOST AS EFFECTIVE AS ACTH SHORT-TERM BUT IT LAGS IN EFFECTS AFTER ONE YEAR. VIGABATRIN HAS A SIGNIFICANT RISK FOR CONCENTRIC VISUAL FIELD DEFICITS DUE TO PERIPHERAL RETINOPATHY, WHICH DEVELOPS IN AN UNPREDICTABLE MANNER. DESPITE THE TREATMENTS, UP TO 85% OF PATIENTS WITH IS HAVE DEVELOPMENTAL REGRESSION AND 67% SUFFER FROM INTRACTABLE EPILEPSY LATER. WE IDENTIFIED THE FOLLOWING GAPS: THERE IS NO SYSTEMATIC RIGOROUS APPROACH IN THE PRECLINICAL SEARCH FOR NOVEL IS TREATMENTS. CURRENT TREATMENTS OF IS (EVEN IF IN COMBINATION) ARE INSUFFICIENT AND MAY HAVE SERIOUS ADVERSE EFFECTS. MOST OF THE DEVELOPED MODELS OF IS LACK VALIDATION USING THE ACTH EFFICACY. SIGMA-1 RECEPTOR LIGANDS HAVE NOT BEEN EXAMINED FOR EFFICACY AGAINST IS. OUR PROPOSAL USES A VALIDATED RAT MODEL OF IS CONSISTING OF PRENATAL PRIMING AND POSTNATAL TRIGGER OF SPASMS DURING DEVELOPMENTALLY APPROPRIATE PERIOD. THE SPASMS IN OUR MODEL ARE SENSITIVE TO TREATMENT WITH ACTH AS WELL AS VIGABATRIN. THERE IS GOOD ICTAL AND INTERICTAL EEG CORRELATE OF THIS MODEL WITH IS. THERE ARE ALSO DELAYED SPASMS OR SEIZURES WITH DELAYED EEG EPILEPTIFORM ACTIVITY. THE MODEL, INCLUDING ACTH EFFICACY, HAS BEEN USED AND REPRODUCED IN INDEPENDENT LABORATORIES. IN THIS PROPOSAL WE WILL INITIATE SYSTEMATIC SEARCH FOR COMPOUNDS POTENTIALLY EFFECTIVE AGAINST IS. OUR PRELIMINARY STUDIES SHOW THAT SIGMA RECEPTOR LIGANDS (SIGMA-1 RECEPTOR ALLOSTERIC MODULATORS) HAVE ROBUST EFFECTS AGAINST THE SPASMS IN OUR MODEL. THEREFORE, THIS CLASS OF COMPOUNDS MAY PRODUCE NOVEL TARGETS FOR IS TREATMENT. OUR PROPOSAL WILL TEST THE TREATMENT CANDIDATES AMONG SIGMA RECEPTOR LIGANDS IN THE THREE-TIER SYSTEM. IN TIER 1, ALL POTENTIAL TREATMENT COMPOUNDS WILL BE TESTED AGAINST THE EXPRESSION OF SPASMS IN THE RANDOMIZED PROSPECTIVE TRIAL. IN TIER 2, THOSE DRUGS WITH >50% EFFICACY WILL BE FORWARDED TO THE EEG STUDY INVESTIGATING THEIR EFFECTS ON BOTH ACUTE AND DELAYED EEG CHANGES. TIER 3 WILL INVESTIGATE COGNITIVE IMPROVEMENTS AS WELL AS RULE OUT BEHAVIORAL ADVERSE EFFECTS AFFORDED BY THE CANDIDATE DRUGS SUCCESSFUL IN TIERS 1 AND 2. SPECIFIC AIM IS TO DETERMINE EFFICACY OF SIGMA RECEPTOR LIGANDS AGAINST THE SPASMS IN PRENATALLY PRIMED RATS IN THREE TIERS. THE PROPOSAL WILL TAKE ADVANTAGE OF UNIQUE FEATURES OF OUR RODENT MODEL OF IS AND DELIVER AT LEAST ONE PROSPECTIVE TREATMENT CANDIDATE WITH EFFICACY COMPARABLE TO OR BETTER THAN ACTH AND WITH FEWER ADVERSE EFFECTS FOR AN IND STUDY.

REDUCING ORTHOSTATIC INTOLERANCE WITH ORAL REHYDRATION IN PATIENTS WITH MYALGIC ENCEPHALOMYELITIS/CHRONIC FATIGUE SYNDROME

TAFENOQUINE AS A POTENTIAL REVOLUTIONARY TREATMENT FOR BABESIOSIS

NEUROMOTOR MODELING OF ADDUCTOR SPASMODIC DYSPHONIA

ADVANCEMENT OF A MUCOSAL SUBUNIT VACCINE IN AN OUTBRED MODEL OF RESPIRATORY TULAREMIA

A NOVEL MUCOSAL VACCINE FOR PSEUDOMONAS AERUGINOSA INFECTION - PROJECT SUMMARY PSEUDOMONAS AERUGINOSA (PA) IS AN OPPORTUNISTIC PATHOGEN THAT CAUSES A DIVERSE ARRAY OF DISEASE MANIFESTATIONS. IT IS A MAJOR CAUSE OF HEALTHCARE ASSOCIATED INFECTIONS WORLDWIDE, CHRONIC LUNG INFECTION IN PATIENTS WITH CYSTIC FIBROSIS (CF), AND BURN WOUND INFECTIONS. THERE IS A HIGH RATE OF ANTIMICROBIAL RESISTANCE IN PA, LEADING TO SIGNIFICANT MORBIDITY AND MORTALITY FROM INFECTION. THE WORLD HEALTH ORGANIZATION HAS CLASSIFIED MULTIDRUG RESISTANT (MDR) PA AS A PRIORITY 1 PATHOGEN FOR RESEARCH. AS INFECTIONS WITH MDR STRAINS OF PA HAVE BECOME COMMONPLACE, TREATMENT OPTIONS HAVE BECOME LIMITED. WE PROPOSE TO DEVELOP A VACCINE TO PREVENT PA INFECTION IN HIGH-RISK INDIVIDUALS. PRIOR ATTEMPTS TO DEVELOP A PA VACCINE HAVE FOCUSED ON PROTECTION FROM RESPIRATORY INFECTION, MOST NOTABLY VENTILATOR-ASSOCIATED PNEUMONIA AND LUNG INFECTION IN CF PATIENTS. THESE ATTEMPTS HAVE BEEN UNSUCCESSFUL DESPITE THE INDUCTION OF DETECTABLE VACCINE-SPECIFIC ANTIBODY RESPONSES IN IMMUNIZED PATIENTS. WE AND OTHERS HYPOTHESIZE THAT THE PARENTERAL IMMUNIZATION SCHEME AND ADJUVANTS USED IN THESE STUDIES DO NOT PRODUCE THE FULL SPECTRUM OF BALANCED HUMORAL AND CELLULAR IMMUNITY NECESSARY FOR EFFECTIVE PROTECTION FROM PA IN THE RESPIRATORY TRACT. THIS CAN BE OVERCOME THROUGH DIRECT IMMUNIZATION AT THE MUCOSAL SURFACE WITH AN ADJUVANT THAT CAN INDUCE TH17 IMMUNITY, AS TH17 IMMUNITY HAS BEEN SHOWN TO BE A CRITICAL COMPONENT FOR PROTECTION TO PA. FURTHERMORE, VACCINES ADMINISTERED AT MUCOSAL SURFACES HAVE ALSO BEEN SHOWN TO GENERATE PROTECTIVE SYSTEMIC IMMUNE RESPONSES. WE ARE DEVELOPING A VACCINE THAT CAN BE ADMINISTERED MUCOSALLY, PROVIDING COMPLETE PROTECTION NOT ONLY AGAINST RESPIRATORY INFECTION WITH PA, BUT ALSO AGAINST NON-MUCOSAL DISEASE MANIFESTATIONS; THUS, PROVIDING COMPLETE IMMUNITY TO THE PATHOGEN. THE VACCINE WILL CONSIST OF A MINIMUM OF FOUR VIRULENCE FACTORS TO PROVIDE BROAD PROTECTION AGAINST THE LARGE NUMBER OF PA SEROTYPES PRESENT IN NATURE. IN THIS APPLICATION, WE WILL EVALUATE PCRV, OPRF, OPRI, AND EXOTOXIN A TOXOID AS VACCINE TARGETS. THESE ANTIGENS WILL BE CONJUGATED TO TOBACCO MOSAIC VIRUS, WHICH WE HAVE PREVIOUSLY DEMONSTRATED TO BE AN EFFECTIVE DELIVERY PLATFORM FOR THE MUCOSAL DELIVERY OF SUBUNIT VACCINE ANTIGENS. IN A PILOT STUDY, WE DEMONSTRATED THAT IN DELIVERY OF TMV-PCRV PROTECTED 66% OF MICE FROM LETHAL CHALLENGE WITH 10XLD50 OF PA IN AN ACUTE LUNG INFECTION MODEL, WHEREAS ALL UNINFECTED MICE SUCCUMBED TO INFECTION. IN THE PRESENT STUDY WE WILL OPTIMIZE VACCINE IMMUNOGENICITY FOR EACH OF THE FOUR TMV CONJUGATES, EVALUATING FUNCTIONAL ANTIBODY AND T CELL RESPONSES FOLLOWING IN VACCINATION, AND TESTING PROTECTIVE EFFICACY IN AN ACUTE LUNG INFECTION MODEL OF PA. WE WILL THEN TEST THE ABILITY OF A COMBINED MULTIVALENT VACCINE AGAINST FIVE DIFFERENT STRAINS OF PA USING BOTH THE LUNG INFECTION MODEL, AND A FOREIGN IMPLANT BIOFILM MODEL. USING THIS WE WILL ESTABLISH PROOF OF PRINCIPLE FOR OUR APPROACH AND DEVELOP A PROTOTYPE VACCINE TO MOVE INTO PRECLINICAL DEVELOPMENT IN A SUBSEQUENT R01 APPLICATION.

SODIUM IMAGING OF SYNAPTIC FUNCTION

GABAERGIC NEUROGENESIS IN HUMANS AND THE EFFECT OF PREMATURITY

IMPROVING PRIMORDIAL FOLLICLE SURVIVAL AFTER TRANSPLANTATION OF CRYOPRESERVED HUM

A CRITICAL ROLE OF NLRP6 IN WEST NILE VIRUS PATHOGENESIS IN MICE

PRECLINICAL DEVELOPMENT OF A MULTIVALENT TULAREMIA VACCINE

HYPERPNEA IN ORTHOSTATIC INTOLERANCE

CHONDROITIN SULFATE PROTEOGLYCAN AND MYELINATION IN INTRAVENTRICULAR HEMORRHAGE

PLK3 IN NICKEL-INDUCED LUNG CARCINOGENESIS

ANALYSIS OF THE PERIPHERAL BLOOD TRANSCRIPTOME TO IDENTIFY CLINICAL CORRELATES OF PATHOLOGY IN PATIENTS WITH BABESIOSIS

MICRORNAS IN DENDRITIC CELL DIFFERENTIATION AND FUNCTION

PRESYNAPTIC LONG-TERM SYNAPTIC DEPRESSION

ROLE OF SIN3A IN THE EPIGENETIC REGULATION OF THE BONE MORPHOGENETIC PROTEIN RECEPTOR TYPE 2 IN PULMONARY ARTERIAL HYPERTENSION - PROJECT SUMMARY PAH IS A FATAL DISEASE CHARACTERIZED BY THE PROGRESSIVE REMODELING OF THE DISTAL PULMONARY VASCULAR ARTERIES, RESULTING IN ELEVATED PULMONARY VASCULAR RESISTANCE AND PULMONARY ARTERY PRESSURE, WHICH MAY LEAD TO SERIOUS COMPLICATIONS, SUCH AS RIGHT HEART FAILURE, AND ULTIMATELY DEATH. ACCUMULATING EVIDENCE INDICATES THAT ENDOTHELIAL DYSFUNCTION IS ONE OF THE FIRST TRIGGERS IN PAH THAT LEADS TO UNCONTROLLED PROLIFERATION OF VASCULAR CELLS, VASCULAR REMODELING, AND OCCLUSION OF THE PULMONARY BLOOD VESSELS. ONE OF THE MOST COMMON PATHOMECHANISMS IN PAH IS THE ALTERATION OF THE BONE MORPHOGENETIC PROTEIN TYPE 2 RECEPTOR (BMPR2) SIGNALING IN VASCULAR CELLS. THE LOSS OF BMPR2 FUNCTION, INDUCED BY MUTATION OR A LOSS OF EXPRESSION, IS ASSOCIATED WITH A SEVERE HEMODYNAMIC PROFILE AND POOR OUTCOMES IN PAH PATIENTS. MULTIPLE STUDIES POINT TO THE PULMONARY ENDOTHELIUM AS THE CELL TYPE THAT IS MOST CRITICALLY IMPACTED BY BMPR2 LOSS IN PAH. HOWEVER, THE MOLECULAR MECHANISMS UNDERLYING THE REGULATION OF BMPR2 EXPRESSION AND ITS ASSOCIATED PAH-LIKE PHENOTYPE REMAIN LARGELY UNKNOWN. OUR GROUP HAS RECENTLY SHOWN THAT SIN3A PLAYS A CENTRAL ROLE IN THE DNA AND HISTONE METHYLATION OF THE BMPR2 PROMOTER IN PULMONARY ARTERY SMOOTH MUSCLE CELLS AND THE PATHOGENESIS OF PAH. HOWEVER, THE EPIGENETIC AND TRANSCRIPTIONAL MECHANISMS BY WHICH SIN3A REGULATES THE BMPR2 GENE IN PAEC REMAIN TO BE ELUCIDATED. PROPOSAL ARE TO UNCOVER THE ROLE OF SIN3A IN THE PULMONARY ENDOTHELIAL DYSFUNCTION OUR OBJECTIVES IN THIS IN PAH, IDENTIFY THE DOWNSTREAM MECHANISMS UNDERLYING THE REGULATION OF BMPR2 EXPRESSION IN PAEC, AND EVALUATE THE THERAPEUTIC EFFECTS OF MODIFIED MRNA ENCODING SIN3A IN ANIMAL MODELS OF PAH. OUR PRELIMINARY DATA SHOWED THAT SIN3A IS SIGNIFICANTLY DOWNREGULATED IN HPAEC ISOLATED FROM PAH PATIENTS. IN VITRO, WE OBSERVED THAT SIN3A SILENCING DOWNREGULATES BMPR2 EXPRESSION AND SIGNALING WHILE POTENTIATING PAEC PROLIFERATION AND MIGRATION. MECHANISTICALLY, WE DISCOVERED A NOVEL MOLECULAR PATHWAY BY WHICH SIN3A MODULATES BMPR2 LEVELS IN HPAECS. OUR DATA SHOWED THAT SIN3A OVEREXPRESSION UPREGULATES FOXK2 BY REPRESSING ENHANCER OF ZESTE HOMOLOG 2 (EZH2)-MEDIATED HISTONE METHYLATION IN THE FOXK2 PROMOTER. ULTIMATELY, SIN3A OVEREXPRESSION INCREASES FOXK2 BINDING TO THE BMPR2 PROMOTER AND UPREGULATES BMPR2 LEVELS IN HPAECS. BASED UPON THESE FINDINGS, WE HYPOTHESIZE THAT THE LOSS OF SIN3A IMPAIRS BMPR2 EXPRESSION IN PAEC, TRIGGERS ENDOTHELIAL DYSFUNCTION AND VASCULAR REMODELING IN PAH. IN THIS PROPOSAL, OUR HYPOTHESIS WILL BE TESTED BY PURSUING THE FOLLOWING THREE SPECIFIC AIMS: AIM 1) TO INVESTIGATE THE ROLE OF SIN3A IN ENDOTHELIAL CELL DYSFUNCTION AND DECIPHER THE MOLECULAR MECHANISM UNDERLYING THE REGULATION OF BMPR2 IN PAH-HPAEC. AIM 2) TO ELUCIDATE THE EFFECTS OF SIN3A DEFICIENCY IN THE ONSET OF PAH IN SMOOTH MUSCLE CELLS AND ENDOTHELIAL CELLS USING A DUAL APPROACH. AIM 3) TO EVALUATE THE THERAPEUTIC EFFICACY OF SIN3A MODRNA IN PRECLINICAL MODELS OF PAH. DEFINING THE REGULATORY MECHANISMS UNDERLYING THE LOSS OF BMPR2 EXPRESSION IN PAH WILL BE OF GREAT RELEVANCE. FINALLY, RESTORING THE EXPRESSION OF SIN3A IN THE LUNGS USING SIN3A MODIFIED RNA MIGHT BE A NEW PROMISING STRATEGY FOR TREATING PAH.

NEW TREATMENTS FOR DRUG RESISTANT EPILEPSY THAT TARGET PRESYNPATIC TRANSMITTER RELEASE

MODELING LENNOX-GASTAUT SYNDROME FOR IMPROVED CLINICAL AND NEUROBEHAVIORAL OUTCOMES - LENNOX-GASTAUT SYNDROME (LGS) IS A DEVELOPMENTAL EPILEPTIC ENCEPHALOPATHY (SEIZURES FURTHER CONTRIBUTE TO COGNITIVE DETERIORATION). LGS IS RELATIVELY FREQUENT (1-2 MILLION PEOPLE WORLDWIDE), ACCOUNTING FOR 1-2% OF ALL PATIENTS WITH EPILEPSY, MORE IN MALES. THOUGH DESCRIBED IN THE SIXTIES, INTERNATIONAL LEAGUE AGAINST EPILEPSY (ILAE) RECENTLY UPDATED AND FOCUSED LGS DEFINITION. LGS PRESENTS WITH: (1) VARIABLE SEIZURE TYPES STARTING BEFORE 18 YEARS OF AGE, HOWEVER TONIC SEIZURES MUST BE INCLUDED PLUS ONE ADDITIONAL TYPE. (2) COGNITIVE AND BEHAVIORAL IMPAIRMENTS (MAY NOT BE PRESENT AT SEIZURE ONSET). (3) EEG MUST SHOW DIFFUSE SLOW SPIKE-AND-WAVE AS WELL AS GENERALIZED FAST PAROXYSMAL ACTIVITY. LGS COMMONLY DEVELOPS FROM THE SEVERE INFANTILE EPILEPSY, VERY OFTEN (IN 20-30%) FROM INFANTILE EPILEPSY SPASM SYNDROME (IESS). CONVERSELY, 20-50% OF INFANTS WITH IESS TRANSITION TO LGS, ESPECIALLY THOSE WITH STRUCTURAL- METABOLIC ETIOLOGY OF IESS. LAMOTRIGINE, FELBAMATE AND POSSIBLY TOPIRAMATE MAY BE HELPFUL IN TREATMENT OF DROP ATTACKS (ATONIC SEIZURES) ASSOCIATED WITH LGS. RECENTLY, FENFLURAMINE APPEARS TO BE PROMISING IN THE REDUCTION OF DROP ATTACKS AND TONIC-CLONIC SEIZURES IN PATIENTS WITH LGS. LGS IS OTHERWISE VERY REFRACTORY, PERSISTS TILL ADULTHOOD, AND REMAINS RESISTANT TO THERAPY RESULTING IN INTELLECTUAL DISABILITY IN MORE THAN 90% OF PATIENTS. THERE IS NO ANIMAL MODEL OF LGS THAT WOULD ASSIST IN TESTING NEW MORE EFFECTIVE TREATMENTS TO IMPROVE DIRE NEUROBEHAVIORAL PROGNOSIS IN MOST PATIENTS WITH LGS. THE GOAL OF THIS PROPOSAL IS TO DEVELOP MODEL OF LGS FOR TESTING NOVEL TREATMENTS. IN THE R61 PHASE, THE MODEL WILL BE DEVELOPED AND QUANTIFIED. R61 SPECIFIC AIM IS TO FULLY CHARACTERIZE SPONTANEOUS ELECTROCLINICAL SIGNS OF THE LGS MODEL IN LARGE COHORTS OF PREPUBERTAL RATS BASED ON TWO COMPLEMENTARY ETIOLOGIES FOUND IN HUMAN CONDITION (TWO DIFFERENT MODELS OF IESS = TWO EXPERIMENTAL ARMS). THE MILESTONE DETERMINING GO/NO GO TRANSITION TO THE R33 PHASE IS THE 40% OCCURRENCE OF ELECTROCLINICAL FEATURES OF LGS (BOTH EEG AND BEHAVIORAL SIGNS IN A SINGLE SUBJECT) IN MODEL ANIMALS. R61 SPECIFIC AIMS ARE TO PROVIDE EXTERNAL VALIDATION TO THE DEVELOPED LGS MODEL. IF SUCCESSFUL, THE R33 PHASE WILL COMPLETE EXTERNAL VALIDATION IN TERMS OF FACE VALIDITY (ADDING BEHAVIORAL TESTING ON TOP OF ELECTROCLINICAL FEATURES), COMPARING ETIOLOGICAL EFFICACY FOR CONSTRUCT VALIDITY, AND TESTING TWO DIFFERENT TREATMENT PARADIGMS USED FOR LGS IN HUMANS AS PREDICTIVE VALIDITY. METHODS USE PREPARATION OF THE IESS MODELS AND CONTROLS, INTRACRANIAL SURGERY, EEG ELECTRODE IMPLANTATION, 24/7 VIDEO- EEG RECORDINGS AND ANALYSIS, DRUG ADMINISTRATION, AS WELL AS SEPARATE COHORT BEHAVIORAL TESTING FOR MOTOR BEHAVIORS, ANXIETY, AUTISTIC TRAITS AND COGNITION. PURPOSE OF THE LGS MODEL DEVELOPMENT IS TO PROVIDE A TOOL FOR TESTING NEW TREATMENT APPROACHES FOR THIS DEVASTATING DEVELOPMENTAL EPILEPTIC ENCEPHALOPATHY.

MODULATION OF HOST INNATE IMMUNE RESPONSE BY FRANCISELLA TULARENSIS

CALCIUM WAVES IN PYRAMIDAL NEURONS

ROLE OF KIR4.1 IN REGULATING NCC AND ROMK IN DCT - PROJECT SUMMARY/ABSTRACT KIR4.1/KIR5.1 IN DISTAL CONVOLUTED TUBULE (DCT) PLAYS A KEY ROLE IN MEDIATING THE EFFECT OF LOW K (LK) OR HIGH K (HK) AND LOW SODIUM (LS) OR HIGH SODIUM (HS) INTAKE ON NCC THROUGH CL--SENSITIVE WITH-NO-LYSINE KINASE (WNK) WHICH REGULATES STE20-PROLINE-ALANINE RICH KINASE (SPAK), A KEY KINASE FOR CONTROLLING NCC ACTIVITY. ALTHOUGH MANY DETAILS OF THIS SIGNALING MECHANISM EXPLAINING THE EFFECTS OF CHRONIC DIETARY K+ OR NA+ INTAKE HAVE BEEN ELUCIDATED, WE HAVE DEMONSTRATED THAT CHRONIC PERTURBATIONS REMODEL THE DISTAL NEPHRON. THUS, THE OBSERVED CHANGES MAY NOT REFLECT THE INITIAL DYNAMIC RESPONSE OF KIR4.1/KIR5.1 AND NCC IN THE DCT. IMPORTANTLY, DIETARY INTAKE IS TYPICALLY EPISODIC, LEADING TO HIGHLY DYNAMIC ELECTROLYTE EXCRETORY PATTERNS. THE DCT COMPRISES AN EARLY PART (DCT1) AND A LATE PART (DCT2). WHILE BASOLATERAL KIR4.1/KIR5.1 AND APICAL NCC ARE EXPRESSED ALONG BOTH DCT1 AND DCT2, THE EPITHELIAL SODIUM CHANNEL (ENAC) AND ROMK ARE ONLY DETECTED IN THE DCT2. THIS RAISES THE POSSIBILITY THAT THE RESPONSE OF KIR4.1/KIR5.1 AND NCC TO SHORT-TERM DIETARY K+ OR NA+ MAY DIFFER BETWEEN DCT1 AND DCT2, CONTRIBUTING TO RAPID HOMEOSTATIC EFFECTS. WE NOW HYPOTHESIZE THAT OVERNIGHT BUT NOT CHRONIC DIETARY K+-INDUCED REGULATION OF KIR4.1/KIR5.1 AND NCC OCCURS PREDOMINANTLY IN THE DCT1, WHEREAS OVERNIGHT BUT NOT CHRONIC DIETARY NA+ OR ANGII PERFUSION- INDUCED REGULATION OF KIR4.1/KIR5.1 AND NCC OCCURS PREDOMINANTLY IN THE DCT2 VIA AT1AR. THIS HYPOTHESIS IS SUPPORTED BY SEVERAL LINES OF PRELIMINARY DATA: 1) OVERNIGHT LK INTAKE INCREASES KIR4.1/KIR5.1 CURRENTS ONLY IN THE DCT1 BUT NOT IN DCT2; 2) OVERNIGHT LS INTAKE INCREASES KIR4.1/KIR5.1 CURRENTS PREDOMINANTLY IN THE DCT2 BUT TO A LESS DEGREE IN DCT1; 3) ACUTE APPLICATION OF ANGII STIMULATES KIR4.1/KIR5.1 ONLY IN THE DCT2 AND ENHANCES NCC FUNCTION; 4) OVERNIGHT ANGII INFUSION ROBUSTLY STIMULATES KIR4.1/KIR5.1 CHANNEL ACTIVITY AND INCREASES WHOLE-CELL K+ CURRENTS IN THE DCT2 ; 5) OVERNIGHT ANGII INFUSION STIMULATES NCC EXPRESSION/ACTIVITY IN KCNJ10+/+ MICE BUT NOT IN INDUCIBLE KIDNEY-SPECIFIC KIR4.1 KNOCKOUT MICE. THE PHYSIOLOGICAL SIGNIFICANCE OF THE PROPOSAL IS TO ELUCIDATE THE MECHANISMS UNDERLYING DUAL REGULATION OF THE DCT, SUCH THAT DCT1 BEHAVES LIKE A “K+-SENSOR” AND DCT2 BEHAVES LIKE A “NA+-SENSOR”. THIS DIFFERENTIAL RESPONSE IS ESSENTIAL FOR NORMAL PHYSIOLOGICAL HOMEOSTASIS DURING SHORT-TERM ALTERATIONS IN K+ OR NA+ INTAKE. THE APPLICATION HAS THREE AIMS:1) TEST HYPOTHESIS THAT THE EFFECT OF OVERNIGHT DIETARY K+ ON NCC EXPRESSION/ACTIVITY IS INITIATED BY MODULATING KIR4.1/KIR5.1 PREDOMINATELY IN THE DCT1, WHEREAS THE EFFECT OF OVERNIGHT DIETARY NA+ ON NCC EXPRESSION/ACTIVITY IS INITIATED THROUGH REGULATING KIR4.1/KIR5.1 PREDOMINANTLY IN THE DCT2; 2)TEST THE HYPOTHESIS THAT ACUTE OR OVERNIGHT ANGII TREATMENT ACTIVATES KIR4.1/KIR5.1 AND NCC PREDOMINATELY IN THE DCT2 BUT NOT IN THE DCT1 AND THAT THE ACTIVATION OF KIR4.1/KIR5.1 IS ESSENTIAL FOR ACUTE BUT NOT CHRONIC ANGII-INDUCED STIMULATION OF WNK4 AND NCC.; 3)TEST THE HYPOTHESIS THAT DELETION OF AT1AR IMPAIRS THE STIMULATORY EFFECT OF LS BUT NOT LK ON KIR4.1/KIR5.1, WNK4 AND PNCC.

CHARACTERIZING THE PHYSICOCHEMICAL PROPERTIES OF MEMBRANELESS CONDENSATES AND ITS REGULATION BY DELTA-9-TETRAHYDROCANNABINOL IN HIV/SIV INFECTION. - ABSTRACT THE `CENTRAL DOGMA' OF MOLECULAR BIOLOGY IS THAT IN CELLS, THE FUNCTIONS OF PROTEINS AND NUCLEIC ACIDS ARE PRIMARILY DETERMINED BY THEIR SEQUENCE. HOWEVER, THIS DOGMA HAS BEEN CHALLENGED BY STUDIES OF SECRETED MOLECULES INCLUDING THE MEMBRANE ENCASED EXTRACELLULAR VESICLES (EVS) AND THE MEMBRANELESS STRUCTURES KNOWN AS BMCS, WHICH INDICATE THAT CELLS CREATE DISCRETE CHEMICAL ENVIRONMENTS WHERE THEY ORGANIZE COMPLEX BIOMOLECULES. BMCS FORM IN MANY COMPARTMENTS OF THE BODY, INCLUDING THE NUCLEUS (NUCLEOLUS, CENTROSOME, CAJAL BODY, STRESS GRANULES), THE CYTOPLASM (P-BODIES, STRESS GRANULES), AND EVEN THE NEURONAL CYTOPLASM (NEURONAL TRANSPORT GRANULES). WHILE THE DETAILS OF BMC BIOLOGY IS STILL EVOLVING, IT IS KNOWN THAT RNA AND PROTEINS WITH INTRINSICALLY DISORDERED REGIONS (IDRS) ARE KEY FACTORS THAT INFLUENCE INTRACELLULAR COMPOSITION, SIZE, AND MORPHOLOGY OF BMCS. THE SIZE AND COMPOSITION OF BMCS ARE ALSO THOUGHT TO BE DIFFERENT. BMCS PARTICIPATE IN THE REGULATION OF VARIOUS PHYSIOLOGICAL PROCESSES, SUCH AS MODULATION OF HOST TRANSCRIPTION AND STRESS RESPONSE. BMCS ALSO PLAY IMPORTANT ROLES IN PATHOPHYSIOLOGICAL CONDITIONS, INCLUDING CANCER, NEURODEGENERATIVE DISEASES, AND VIRAL PATHOGENESIS, INCLUDING MODULATION OF HIV REPLICATION. INDEED, IT IS KNOWN THAT HIV INFECTION RESULTS IN PERIPHERAL AND NEURO INFLAMMATION, AND THAT CHRONIC EXPOSURE TO THC IN THE CONTEXT OF HIV IN HUMANS OR SIV INFECTION IN RHESUS MACAQUES RESULTS IN AMELIORATION OF DISEASE OR SYMPTOMS. HOWEVER, THE REGULATORS OF HIV DISEASE AND THC RESPONSE IS NOT COMPLETELY UNDERSTOOD. DESPITE THE REPORTED ROLE OF BMCS IN VARIOUS CONDITIONS, WHETHER OR NOT BMCS REGULATE THE PATHOGENESIS OF HIV INFECTION AND THE INTERSECTION OF HIV AND THC USE IS UNKNOWN. IT IS ALSO UNKNOWN WHETHER BMCS ASSEMBLE IN VIVO, WHERE THEY MAY PLAY RELEVANT BIOLOGICAL ROLES. THERE IS PAUCITY OF INFORMATION ON THE PRESENCE OF BMCS IN BODY FLUIDS (IN VIVO) AND THEIR ROLE IN HIV PATHOGENESIS. THE REASON FOR THIS SCARCITY OF INFORMATION IS PARTLY DUE TO LACK OF TOOLS FOR ISOLATION OF BMCS FROM BODY FLUIDS. IN PRELIMINARY STUDIES, WE DEVELOPED A NOVEL SIZE-GUIDED PARTICLE PURIFICATION LIQUID CHROMATOGRAPHY (PPLC). WE USED PPLC TO ISOLATE AND RETRIEVE DYE-FREE BMCS FROM HUMAN SEMEN, RHESUS MACAQUE BLOOD PLASMA AND CEREBROSPINAL FLUID (CSF). USING STANDARD ELECTRON MICROSCOPY AND ENERGY DISPERSIVE X-RAY EQUIPPED TRANSMISSION ELECTRON MICROSCOPE (TEM-EDX) ANALYSES, WE OBTAINED STRUCTURAL AND ELEMENTAL INFORMATION OF THE BMCS. PROTEOMICS ANALYSIS AND RNA BIOANALYZER STUDY WERE USED TO SHOW THAT THE BMCS CONTAIN PROTEIN AND RNA, WHILE CELL BIOLOGY ASSAY INDICATE THAT BMCS ARE CYTOTOXIC TO CELLS. THESE DATA PROVIDE COMPELLING EVIDENCE THAT BIOACTIVE BMCS ARE PRESENT IN VIVO. THUS, IN THIS PROPOSAL, WE WILL USE PPLC TO ISOLATE BMCS FROM RHESUS MACAQUE BLOOD, CSF, SALIVA, AND INTESTINAL CONTENTS. WE WILL THEN USE VARIOUS STRUCTURAL, BIOCHEMICAL, AND CELL BIOLOGY ASSAYS TO PROVIDE STRUCTURAL, MOLECULAR, AND FUNCTIONAL CLASSIFICATION OF BMCS FROM SIV INFECTED AND THC TREATED MACAQUES, WHICH WILL PROVIDE A FRAMEWORK TO DISSECT BMC FUNCTIONS IN HIV/AIDS AND THC USE.

VACULOGENESIS AND RENAL MESODERMAL PROGENITOR CELLS

MODULATION OF MACROPHAGE FUNCTION BY FRANCISELLA TULARENSIS

STRUCTURE/FUNCTION STUDIES OF QUINONE REDUCTASE 2

TOBACCO MOSAIC VIRUS AS A MUCOSAL DELIVERY PLATFORM FOR AN INTRANASAL VACCINE AGAINST PSEUDOMONAS AERUGINOSA - PSEUDOMONAS AERUGINOSA (PA) IS A GRAM-NEGATIVE OPPORTUNISTIC PATHOGEN THAT IS ONE OF THE MOST PROMINENT CAUSES OF HEALTHCARE ACQUIRED INFECTIONS WORLDWIDE. IT IS ALSO A SIGNIFICANT CAUSE OF MORBIDITY AND MORTALITY ASSOCIATED WITH BURN WOUNDS, AND IN CHRONIC LUNG INFECTIONS ASSOCIATED WITH CYSTIC FIBROSIS (CF). PA HAS A WIDE VARIETY OF VIRULENCE MECHANISMS AND CAN CAUSE MANY DIFFERENT TYPES OF MUCOSAL AND SOFT TISSUE INFECTIONS. IT IS ONE OF THE MOST COMMON CAUSES OF VENTILATOR ASSOCIATED PNEUMONIA, AND A TOP TEN CAUSE OF SURGICAL SITE INFECTIONS, CATHETER ASSOCIATED URINARY TRACT INFECTIONS, AND BLOOD INFECTIONS FROM CENTRAL LINES. THE SPREAD OF ANTIMICROBIAL RESISTANCE HAS MADE PA INFECTIONS INCREASINGLY DIFFICULT TO TREAT, AND MULTI-DRUG RESISTANT (MDR) PA IS CONSIDERED A CRITICAL THREAT TO PUBLIC HEALTH BY THE WORLD HEALTH ORGANIZATION AND THE CENTERS FOR DISEASE CONTROL AND PREVENTION. NEW APPROACHES FOR THE TREATMENT AND/OR PREVENTION OF PA ARE NEEDED. VACCINES ARE ONE OF THE MOST SUCCESSFUL MEDICAL INTERVENTIONS IN HISTORY. MOST VACCINES ARE GIVING PARENTERALLY VIA INJECTION, A ROUTE OF ADMINISTRATION THAT DOES NOT PROVIDE STRONG PROTECTION AT MUCOSAL SURFACES, WHICH IS THE SITE OF ENTRY FOR MOST PATHOGENS. MUCOSAL VACCINATION PROVIDES STRONG PROTECTION AT MUCOSAL SITES, SUCH AS THE RESPIRATORY TRACT, AS WELL AS SYSTEMICALLY. THIS IS IDEAL FOR A PATHOGEN LIKE PA WHICH CAN CAUSE DISEASE AT DIFFERENT SITES. A CHALLENGE FOR MUCOSAL VACCINATION IS THE NEED FOR DELIVERY METHODS THAT CAN OVERCOME THE NATURAL TOLEROGENIC MECHANISMS THAT OCCUR AT THESE SITES, WHILE NOT INDUCING OVERT INFLAMMATORY REACTIONS. WE ARE DEVELOPING AN INTRANASAL PROTEIN SUBUNIT VACCINE AGAINST PA, USING TOBACCO MOSAIC VIRUS (TMV) AS A DELIVERY PLATFORM. TMV IS A PLANT PATHOGEN THAT DOES NOT INFECT ANIMAL CELLS AND IS HIGHLY IMMUNOGENIC, BEING AN OPTIMAL SIZE AND COMPOSITION FOR RECOGNITION BY THE IMMUNE SYSTEM. WE HAVE DEVELOPED A RECOMBINANT TMV THAT EXPRESSES A SURFACE EXPOSED LYSINE ON THE COAT PROTEIN ALLOWING FOR THE CONJUGATION OF VACCINE ANTIGENS TO THE VIRAL SURFACE. IN PRELIMINARY STUDIES WE DEMONSTRATED THAT IN VACCINATION WITH TMV CONJUGATED TO PCRV, THE TIP PROTEIN OF THE PA TYPE III SECRETION SYSTEM, IN THE PRESENCE OF THE TH17 SKEWING ADJUVANT CURDLAN, PROTECTED MICE AGAINST LETHAL RESPIRATORY CHALLENGE WITH PA. PROTECTION WAS ASSOCIATED WITH PRODUCTION OF ANTI-PCRV ANTIBODY AND ELEVATED IL-17 AND IFNΓ IN THE LUNGS AND SPLEENS OF MICE FOLLOWING INFECTION. IN THE PRESENT STUDY WE WILL DEFINE ADDITIONAL VACCINE ANTIGENS TO CREATE A MULTI-VALENT VACCINE THAT WILL PROVIDE BROAD PROTECTION AGAINST MULTIPLE STRAINS OF PA. WE WILL ALSO IDENTIFY THE MECHANISM(S) OF VACCINE MEDIATED PROTECTION TO PROVIDE ADDITIONAL PATHWAYS THAT CAN BE TARGETED FOR IMPROVING VACCINE EFFICACY. PA IS A MAJOR PUBLIC HEALTH THREAT THAT REQUIRES NEW AND INNOVATIVE METHODS TO PROTECT PATIENTS IN HEALTH CARE SETTINGS, FIRST RESPONDERS AND OUR MILITARY, AND CF PATIENTS FROM DISEASE AND DEATH. THE GOAL OF THIS PROJECT IS AN EFFECTIVE MUCOSAL VACCINE THAT WILL PROVIDE BROAD COVERAGE AGAINST DIFFERENT TYPES OF PA INFECTIONS FOR USE IN AT RISK PATIENTS TO LIMIT THE IMPACT OF MDR PA AND IMPROVE PATIENT OUTCOMES.

MODIFICATION OF DNA POLYMERASE D BY A NOVEL MECHANISM DURING REPLICATION STRESS

CHARACTERIZATION OF ESTROGEN-MTORC1 SIGNALING NETWORK IN TSC/LAM

TNF-UMOD INTERACTIONS AND SUSCEPTIBILITY TO SALT-INDUCED HYPERTENSION

MCH DISTANCE LEARNING

HTS ASSAY DEVELOPMENT TARGETING YERSINIA PESTIS TOPOISOMERASE I

CHARACTERIZATION OF FRANCISELLA TULARENSIS-SPECIFIC BACTERIOPHAGES - BACTERIOPHAGES ARE VIRUSES THAT INFECT AND KILL BACTERIA EITHER BY DISRUPTING THEIR METABOLISM OR BY CAUSING BACTERIAL LYSIS. THESE UNIQUE PROPERTIES OF BACTERIOPHAGES HAVE BEEN EXPLOITED AS ANTIMICROBIAL AGENTS FOR OVER 100 YEARS. THE EMERGENCE OF ANTIBIOTIC RESISTANCE AND THE LACK OF INTRODUCTION OF NEW ANTIBIOTICS IN RECENT YEARS, HAS RENEWED INTEREST IN THE DEVELOPMENT OF BACTERIOPHAGE-BASED THERAPIES IN CLINICAL SITUATIONS WHERE ANTIBIOTICS ARE NO LONGER EFFECTIVE. THE OVERALL GOAL OF THIS PROPOSAL IS TO CHARACTERIZE A PANEL OF BACTERIOPHAGES THAT COULD FURTHER BE DEVELOPED AS A THERAPY TO TREAT FATAL HUMAN DISEASE TULAREMIA CAUSED BY A GRAM-NEGATIVE BACTERIUM, FRANCISELLA TULARENSIS (FT). FT IS A HIGHLY UBIQUITOUS, INFECTIOUS, INTRACELLULAR BACTERIUM PREVALENT THROUGHOUT NORTH AMERICA. FT IS CLASSIFIED AS A TIER 1 CATEGORY A SELECT AGENT BY THE CDC DUE TO ITS HISTORY OF WEAPONIZATION BY SEVERAL COUNTRIES, AND ITS POTENTIAL TO BE USED AS A BIOTERROR AGENT. ANTHRAX POWDER ATTACKS AFTER 9/11 INCIDENCE HAVE RENEWED THE MEDICAL AND SCIENTIFIC INTEREST IN FT RESEARCH. TULAREMIA IS A NOTIFIABLE DISEASE IN THE USA. NATURALLY OCCURRING TULAREMIA IS REPORTED FROM ALL THE STATES OF THE USA, EXCEPT HAWAII. CURRENTLY, NO VACCINE IS AVAILABLE FOR TULAREMIA PROPHYLAXIS. UNFORTUNATELY, ANTIBIOTIC TREATMENT IS NOT PARTICULARLY EFFECTIVE AGAINST SEVERE RESPIRATORY TULAREMIA AND MAY NOT BE EFFECTIVE AT ALL, IF A WEAPONIZED ANTIBIOTIC-RESISTANT STRAIN USED IN A BIOTERROR ATTACK CAUSES AN OUTBREAK. TOGETHER, THESE INDICATE THAT THE DEVELOPMENT OF EFFECTIVE THERAPEUTIC MEASURES AGAINST THIS DREADED BIOTHREAT AGENT IS NECESSARY. BACTERIOPHAGES MAY PROVIDE AN EXCELLENT ALTERNATIVE TO THE CONVENTIONAL ANTIBIOTIC THERAPY FOR THE TREATMENT OF TULAREMIA WITH THE ADDED ADVANTAGE BEING THAT THEY ARE INNOCUOUS, SELF-REPLICATING, AMPLIFY AT THE SITE OF INFECTION, AND DO NOT PERTURB MICROBIOTA. HOWEVER, BACTERIOPHAGES AGAINST FRANCISELLA HAVE NOT BEEN REPORTED IN THE LITERATURE TO-DATE. AS A PROOF-OF-CONCEPT, WE HAVE ISOLATED A PANEL OF FRANCISELLA-SPECIFIC LYTIC PHAGE POPULATIONS FROM ENVIRONMENTAL SAMPLES. IN SPECIFIC AIM 1 OF THIS PROPOSAL, WE WILL ISOLATE A COLLECTION OF INDIVIDUAL FT-SPECIFIC PHAGES FROM THESE SAMPLES AND CHARACTERIZE THEIR GENETIC, BIOCHEMICAL, AND PHYSICAL PROPERTIES. IN SPECIFIC AIM 2, WE WILL CHARACTERIZE THEIR BIOLOGICAL PROPERTIES. SPECIFICALLY, WE WILL DETERMINE THEIR GROWTH CHARACTERISTICS, HOST-RANGE, AND POTENTIAL CELL SURFACE RECEPTORS. WE WILL FORMULATE AN FT-SPECIFIC PHAGE COCKTAIL AND DEVELOP ASSAYS TO TEST THE ABILITY OF PHAGE COCKTAIL(S) TO KILL INTRACELLULAR FRANCISELLA. THIS IS A FIRST COMPREHENSIVE STUDY DIRECTED AT THE CHARACTERIZATION OF FT-SPECIFIC BACTERIOPHAGES. ISOLATION AND CHARACTERIZATION OF FT-SPECIFIC PHAGES AGAINST CATEGORY A TIER-1 SELECT AGENT FT SCHUS4 AND FORMULATION OF NOVEL FT-SPECIFIC BACTERIOPHAGE COCKTAIL CAPABLE OF KILLING INTRACELLULAR BACTERIA CONSTITUTE THE MOST INNOVATIVE ASPECTS OF THIS PROPOSAL. WE EXPECT THAT THE OUTCOME OF THE PROPOSED STUDIES WILL PAVE THE WAY FOR THE DEVELOPMENT OF A NOVEL THERAPEUTIC APPROACH FOR THE TREATMENT OF TULAREMIA IN HUMANS.

UNCOVERING THE ROLE OF GPR75 AS AN ACTIVATOR OF FATTY ACID TRANSPORTERS IN NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD) - THE PREVALENCE OF OBESITY IS A GLOBAL CONCERN WITH NEARLY 2 IN 5 ADULTS (42.4%) IN THE US HAVING OBESITY. RAPID AND SUSTAINED INCREASES IN WEIGHT GAIN RESULT IN VARIOUS OBESITY-DRIVEN COMPLICATIONS AND HEALTH OUTCOMES INCLUDING NON- ALCOHOLIC FATTY LIVER DISEASE (NAFLD). RECENTLY, SEVERAL PREDICTED LOSS OF FUNCTION VARIANTS OF THE UNDERSTUDIED ORPHAN G PROTEIN- COUPLED RECEPTOR (GPCR), GPR75 WERE IDENTIFIED TO BE ASSOCIATED WITH LEANNESS AND A PROTECTIVE PHENOTYPE AGAINST OBESITY IN A WORLD-WIDE MULTI-ETHNIC EXOME SEQUENCING OF OVER 640,000 INDIVIDUALS. OUR GROUP WAS INSTRUMENTAL IN THESE STUDIES AND OUR PROPOSAL SEEKS TO EXAMINE THE ROLE OF GPR75 IN OBESITY AND NAFLD. SPECIFICALLY, WE HYPOTHESIZE THAT THE PAIRING OF GPR75 TO ITS HIGH-AFFINITY LIGAND, 20-HETE, A VASOACTIVE AND PROINFLAMMATORY LIPID, EXACERBATING DIET-INDUCED OBESITY, DRIVING NAFL AND NONALCOHOLIC STEATOHEPATITIS (NASH) WHICH IS CHARACTERIZED BY PRONOUNCED INFLAMMATION, CELL DAMAGE AND FIBROSIS. TO TEST THIS HYPOTHESIS, WE PROPOSE TWO SPECIFIC AIMS. AIM 1 SEEKS TO DETERMINE THE DEGREE TO WHICH THE PAIRING OF 20-HETE/GPR75 CONTRIBUTES TO THE PATHOGENESIS OF OBESITY-DRIVEN NAFLD/NASH. THIS AIM WILL EVALUATE THE SEVERITY OF OBESITY, DIABETES/INSULIN RESISTANCE AND LIVER DAMAGE IN MICE DEFICIENT IN GPR75 AND EXPOSED TO ELEVATIONS IN 20-HETE AND A HIGH-FAT DIET FEEDING PROTOCOL ACROSS TIME. A NOVEL WATER-SOLUBLE GPR75 RECEPTOR BLOCKER, AAA, WILL BE USED TO ASSESS THE DEPENDENCY OF THE DISEASE DEVELOPMENT AND PROGRESSION ON 20-HETE-GPR75 PAIRING. AIM 2 LOOKS TO IDENTIFY THE CELLULAR MECHANISM BY WHICH THE 20-HETE-GPR75 PAIRING DRIVES INCREASES IN FATTY ACID UPTAKE AND INFLAMMATION WHICH CONTRIBUTE TO NAFLD. SPECIFICALLY, IT WILL DETERMINE HOW THE ACTIVATION OF GPR75 VIA 20-HETE STIMULATES THE ACTIVITY OF THE FATTY ACID TRANSPORTER 2 (FATP2) IN HEPATOCYTES. WE WILL ALSO EVALUATE HOW THE COMBINATION OF 20-HETE AND FATTY ACID INFLUX DRIVE VARIOUS PROINFLAMMATORY AND PROFIBROTIC SIGNALS. THIS PARTICULAR AIM WILL ALSO INCORPORATE THE USE OF AAA (GPR75 RECEPTOR BLOCKER) AND HEPATOCYTES DEFICIENT IN GPR75. THE IMPLICATIONS BEHIND THE PROPOSED RESEARCH ARE HIGHLY INNOVATIVE AS THEY WILL LAY THE FOUNDATION AND FUNDAMENTALS AS WE CONTINUE TO LEARN MORE ABOUT THE ROLE OF GPR75 IN OBESITY AND NAFLD/NASH. OUR PRELIMINARY DATA STRONGLY SUGGEST THAT GPR75 IS A DRUGGABLE TARGET WITH TRANSLATIONAL APPLICATIONS FOR THE PREVENTION AND TREATMENT OF OBESITY AND NAFLD/NASH, DISEASES THAT PRESENTLY PLAGUE THE GLOBAL HEALTHCARE SYSTEM. THEREFORE, WE BELIEVE THAT THIS APPLICATION FITS STRONGLY WITH THE IDG’S INITIATIVE TO SUPPORT STUDIES PERTAINING TO POORLY CHARACTERIZED GPCRS IN HUMAN HEALTH AND DISEASE.

EVALUATING THE EFFICACY OF TARGETED DEEP BRAIN STIMULATION (DBS) IN ATTENUATING REFRACTORY INFANTILE EPILEPSY SPASM SYNDROME (IESS) IN A RODENT MODEL - INFANTILE EPILEPTIC SPASMS SYNDROME (IESS) IS A DEVASTATING EARLY-ONSET EPILEPSY SYNDROME CHARACTERIZED BY SPASMS AND INTERICTAL HYPSARRHYTHMIA ON EEG OCCURRING DURING INFANCY. CURRENT TREATMENTS, INCLUDING ACTH AND VIGABATRIN, FREQUENTLY FAIL AND CARRY SERIOUS SIDE EFFECTS SUCH AS IMMUNE SUPPRESSION AND VISION LOSS. THERE IS A PRESSING NEED FOR SAFER, MORE EFFECTIVE THERAPIES, PARTICULARLY FOR DRUG-RESISTANT CASES. THIS PROPOSAL EXPLORES DEEP BRAIN STIMULATION (DBS) TARGETING SPECIFIC HYPOTHALAMIC NUCLEI AS A NOVEL STRATEGY TO SUPPRESS SEIZURES AND NORMALIZE EEG IN IESS. OUR CENTRAL HYPOTHESIS IS THAT FREQUENCY-SPECIFIC DBS OF THE PARAVENTRICULAR HYPOTHALAMIC NUCLEUS (PVH) AND ARCUATE NUCLEUS (ARC) WILL MODULATE PATHOLOGICAL NETWORK EXCITABILITY, SUPPRESS SPASMS AND HYPSARRHYTHMIA, AND PREVENT SUBSEQUENT SPONTANEOUS SEIZURES. PRELIMINARY MICROINFUSION DATA STRONGLY SUPPORT A CENTRAL ROLE FOR THE PVH-ARC NETWORK IN REGULATION OF SPASMS. DBS TARGETING OF THIS CIRCUITRY OFFERS A MECHANISTICALLY GUIDED, NEUROMODULATORY STRATEGY THAT MAY OVERCOME LIMITATIONS OF CURRENT THERAPIES. USING A VALIDATED RAT MODEL THAT RECAPITULATES BOTH THE CLINICAL AND ELECTROGRAPHIC FEATURES OF IESS (VIA PRENATAL BETAMETHASONE EXPOSURE FOLLOWED BY POSTNATAL NMDA-INDUCED SPASMS) WE WILL TEST THE FOLLOWING AIMS: AIM 1: EVALUATE PROOF-OF-PRINCIPLE THERAPEUTIC EFFICACY OF DBS IN PVH OR ARC USING HIGH- FREQUENCY (HF) STIMULATION IN ARC AND LOW-FREQUENCY (LF) STIMULATION IN PVH, MIMICKING INHIBITORY AND EXCITATORY NEUROMODULATION, RESPECTIVELY. AIM 2: OPTIMIZE DBS STIMULATION PARADIGMS BY (A) ASSESSING HOW LF VS HF STIMULATION OF EACH TARGET MODULATES SEIZURES AND EEG FEATURES, AND (B) EVALUATING THE EFFECTS OF CONCURRENT STIMULATION IN BOTH PVH AND ARC. KEY OUTCOME MEASURES INCLUDE SPASM FREQUENCY AND LATENCY, EEG QUANTIFICATION OF HYPSARRHYTHMIA AND ELECTRO-DECREMENT, AND ASSESSMENT OF SPONTANEOUS SEIZURES LATER IN DEVELOPMENT. DATA WILL BE BENCHMARKED AGAINST PHARMACOLOGIC INTERVENTIONS INCLUDING ACTH AND ANAVEX®2-73 [BLARCAMESINE]. BY LEVERAGING TECHNIQUES IN DEVELOPMENTAL EEG MONITORING AND DBS IN A RIGOROUSLY VALIDATED ANIMAL MODEL, THIS STUDY HAS THE POTENTIAL TO TRANSFORM THERAPEUTIC APPROACHES FOR IESS AND OPEN NEW AVENUES FOR TREATING DRUG-RESISTANT PEDIATRIC EPILEPSIES.

PILIN GENOTYPING FOR GROUP A STREPTOCOCCI

T.GONDII RESISTANCE TO REACTIVE NITROGEN SPECIES

SECOND-HIT AND SEXUAL DIMORPHISM EFFECTS IN PAH

MOLECULAR MECHANISM OF CELLULAR ACTIVATION OF THE HEPATITIS C VIRUS POLYMERASE

POLD4 GENE EXPRESSION AS A PROGNOSTIC INDICATOR FOR SYNTHETIC LETHALITY WITH PARP INHIBITORS IN LUNG CANCER CELLS

20-HETE REGULATION OF ANDROGEN-INDUCED HYPERTENSION

INHIBITION OF NO DEPENDENT SPLANCHNIC HYPEREMIA IMPROVES CFS/POTS

CARES – HIGHER EDUCATION EMERGENCY RELIEF FUND (FIPSE)""

A NOVEL HUMAN DNA DAMAGE RESPONSE WHICH DIRECTLY ALTERS DNA POLYMERASE DELTA

NEW YORK MEDICAL COLLEGE BIOTERRORISM: CDM DISASTER MEDICINE AND EMERGING INFECTIONS

PROTEINSIMPLE JESS SYSTEM FOR PROTEIN ANALYSIS AT NYMC - PROJECT SUMMARY THIS IS A REQUEST FOR JESS PROTEIN ANALYSIS SYSTEM (PROTEIN SIMPLE) TO BE INTEGRATED INTO THE NEW YORK MEDICAL COLLEGE (NYMC) FOR PROTEIN QUANTITATION AND CHARACTERIZATION. NYMC IS DEDICATED TO BUILDING BIOMEDICAL RESEARCH. A KEY PART OF NYMC STRATEGIC PLAN IS IMPROVING INVESTIGATOR-IDENTIFIED CRITICAL CORE RESEARCH FACILITIES. SURVEYS OF NYMC FUNDED INVESTIGATORS, INCLUDING THOSE FUNDED BY NIH AND THOSE FUNDED FROM OTHER SOURCES, CONSISTENTLY RANK A PROTEIN ANALYSIS SYSTEM THAT MAY QUANTITATIVE AND REPRODUCIBLE RESULTS AS THE MOST ESSENTIAL CORE PIECE OF EQUIPMENT FOR THEIR CURRENT AND FUTURE RESEARCH, ESPECIALLY IN THE ABSENCE OF A FUNCTIONAL PROTEOMICS CORE. THE PROPOSED PROTEINSIMPLE JESS SYSTEM INTEGRATED INTO THE COLLEGE HISTOLOGICAL LABORATORY AND IMAGING CORE WILL MEET A TREMENDOUS UNMET NEED FOR INVESTIGATORS FOR STATE-OF-THE ART AUTOMATED SIZE-BASED SEPARATION AND NANO-IMMUNOASSAY PLATFORM FOR THE DETECTION AND CHARACTERIZATION OF PROTEINS MOLECULAR WEIGHTS IN DENATURED PROTEIN FROM TISSUES, CELLS, AND EXTRACELLULAR PARTICLES (EXTRACELLULAR VESICLES AND EXTRACELLULAR CONDENSATES). THE RESEARCH OF MANY OF THE MAJOR AND MINOR USERS AT NYMC INVOLVES ANALYSIS OF GENE EXPRESSION AT THE PROTEIN LEVEL AND THE ENHANCED SIGNAL TO NOISE RATIO PROVIDED BY THE PROTEINSIMPLE JESS SYSTEM IS ESSENTIAL. CURRENTLY THERE IS NOT A FUNCTIONAL PROTEOMICS CORE AT NYMC. THE PROPOSAL INCLUDES 4 NIH-FUNDED MAJOR USERS FROM VARIOUS DEPARTMENTS AND DISCIPLINES AT NYMC. NIH FUNDED USERS COMPRISE 100% OF THE ESTIMATED ACCESSIBLE USER TIME (AUT) FOR THE INSTRUMENT. THE OTHER USERS COMPRISE ADDITIONAL INVESTIGATORS WITH CRITICAL NEED FOR QUANTITATIVE AND REPRODUCIBLE RESULTS. THERE IS NO FUNCTIONAL PROTEIN ANALYSIS CORE OR EQUIPMENT AT NYMC. ALL INVESTIGATORS DEPEND ON TRADITIONAL WESTERN BLOTTING THAT IS PLAGUED BY POOR REPRODUCIBILITY, LACK OF ACCURATE QUANTITATION, EXTENSIVE TIME TO RESULT AND RELIABILITY ISSUES. ALL MAJOR/MINOR USERS HAVE AGREED THAT THE PROTEINSIMPLE JESS SYSTEM WILL BEST FIT THE CURRENT/FUTURE NEEDS OF INVESTIGATORS TO PROVIDE MORE QUANTITATIVE AND REPRODUCIBLE RESULTS, THUS, MEETING ESTABLISHED GUIDELINES ON NIH’S REPRODUCIBILITY AND RIGOR. IN ADDITION TO THE AUTOMATION, SENSITIVITY, AND THROUGHPUT ADVANTAGES OF THE PROTEINSIMPLE JESS SYSTEM, THE SYSTEM CREATES ELECTRONIC DATA FILES THAT ARE IMPERVIOUS TO MANIPULATION, AS IT CONTAINS THE RAW AND ANALYZED DATA, WHICH CAN EASILY BE UPLOADED TO A DATA REPOSITORY WHEN IT COMES TIME TO PUBLISH. IN SUMMARY, THE PROTEINSIMPLE JESS SYSTEM PROMOTES DATA REPRODUCIBILITY BECAUSE IT PRECISELY CONTROLS SAMPLE LOADING, ANTIBODY ADDITIONS, INCUBATIONS, AND ALL WASHES, THEREBY ELIMINATING THE INCONSISTENCIES AND USER-DEPENDENT VARIABILITY THAT MAY BE INTRODUCED DURING TRADITIONAL WESTERN BLOTTING, CULMINATING IN INTRA-ASSAY COEFFICIENTS OF VARIABILITY (CVS) <15%.

DISSECTING MICROVASCULAR UNDERPINNINGS OF INJURY-INDUCED ALTERATIONS IN FUNCTIONAL CEREBRAL BLOOD FLOW

DECODING DYNAMIC ABNORMALITIES IN CEREBRAL HEMODYNAMIC RESPONSES OBTAINED FROM NEAR-INFRARED OPTICAL DIAGNOSTICS

ROLE OF GPROTEIN AND SNAP-25 IN NMDA VS MGLUR PRESYNAPTIC LONG-TERM PLASTICITY

CONTRIBUTION OF BONE TO FGF23 ELEVATION IN KIDNEY DISEASE

MULTISCALE, MULTIMODAL OPTICAL INVESTIGATION OF LONG-TERM, INJURY-INDUCED ALTERATIONS IN NEUROVASCULAR COUPLING

NSF/FDA SIR: MICRO- AND MACRO-SCALE VALIDATION OF DIFFUSE CORRELATION SPECTROSCOPY FOR MONITORING FUNCTIONAL HEMODYNAMICS IN THE MICROVASCULATURE OF

NSF/FDA SIR: MULTIMODAL CHARACTERIZATION OF QUANTITATIVE BIOMARKERS FOR TRAUMATIC BRAIN INJURY MEASURED VIA PORTABLE DEVICE TECHNOLOGY

DOXORUBICIN-INDUCED CARDIAC PROGENITOR CELL DEATH IS ANTECEDENT TO HEART FAILURE.

SUBCELLULAR MECHANISMS IN PATHOGENESIS OF PULMONARY ARTERIAL HYPERTENSION

20-HETE INCREASES LARGE ARTERY STIFFNESS AND SYSTOLIC BLOOD PRESSURE IN THE METABOLIC SYNDROME

HEPATITIS C VIRUS NS3 HELICASE INHIBITOR DISCOVERY

FOURTH INTERNATIONAL SYMPOSIUM ON CHILDHOOD, ADOLESCENT AND YOUNG ADULT NON-HODGK

WORKSHOP ON PHENOTYPIC AND DEVELOPMENTAL PLASTICITY, DECEMBER 15-20, 2007; TRIVANDRUM & KERALA, INDIA

SIXTH INTERNATIONAL SYMPOSIUM ON CHILDHOOD, ADOLESCENT AND YOUNG ADULT NON-HODGKIN LYMPHOMA

1ST ANNUAL TANDEM ASPHO/PBMTC EDUCATIONAL MTG: NEW FRONTIERS IN PEDIATRIC ALLOSCT

SEVENTH INTERNATIONAL SYMPOSIUM ON CHILDHOOD, ADOLESCENT AND YOUNG ADULT NON-HODGKIN LYMPHOMA - PROJECT SUMMARY WE PROPOSE TO ORGANIZE AND CONDUCT THE SEVENTH INTERNATIONAL SYMPOSIUM ON CHILDHOOD, ADOLESCENT AND YOUNG ADULT (CAYA) NON-HODGKIN LYMPHOMA (NHL) AT THE INTERCONTINENTAL HOTEL IN NEW YORK CITY, NY, ON OCTOBER 20-23, 2022. THE FIRST, SECOND AND FOURTH SYMPOSIA WERE HELD AT THE NEW YORK SHERATON HOTEL AND TOWERS IN NYC, NY, IN 2003, 2006 AND 2012, THE THIRD IN FRANKFURT, GERMANY IN 2009, THE FIFTH IN VARESE, ITALY IN 2015 AND THE SIXTH IN 2018, IN ROTTERDAM, THE NETHERLANDS. CURRENT PROBLEMS THAT STILL AFFECT CAYA PATIENTS WITH NHL INCLUDE POOR PROGNOSIS OF CAYA PATIENTS FROM LOW SOCIOECONOMIC RESOURCED COUNTRIES, POOR OUTCOME IN PATIENTS WHO RELAPSE/PROGRESS, UNACCEPTABLE SHORT AND LONG-TERM MORBIDITY AND A PAUCITY OF INFORMATION ON GENOMIC, EPIGENETIC AND IMMUNOLOGICAL ABERRATIONS ASSOCIATED WITH CAYA NHL. UNFORTUNATELY, THERE HAVE BEEN LIMITED EDUCATIONAL RESOURCES AVAILABLE TO INCREASING OUR UNDERSTANDING OF THE BASIC BIOLOGY, MECHANISMS OF DRUG RESISTANCE, DEVELOPMENT OF NOVEL TRANSLATIONAL AND TARGETED THERAPIES AND LONG TERM EFFECTS OF TREATMENT OF THIS ORPHAN DISEASE AND ALMOST NEGLIGIBLE EDUCATIONAL CONFERENCE SUPPORT FROM THE NIH AND PHARMACEUTICAL/BIOTECHNOLOGY COMPANIES SUPPORTING INTERNATIONAL CONFERENCES. THE PRIMARY OBJECTIVES:1) IDENTIFY NEW GENOMIC, EPIGENETIC AND IMMUNOMIC ABERRATIONS IN THE PATHOLOGY OF MATURE B-NHL, ALCL, MATURE NK/T, LYMPHOBLASTIC LYMPHOMA, AND RARE NHL IN CAYA; 2) TO DETERMINE THE SAFETY AND EFFICACY OF NOVEL TARGETED CELLULAR AND HUMORAL IMMUNOTHERAPY IN CAYA PATIENTS WITH NHL; 3) TO REVIEW THE ‘STATE OF THE SCIENCE” IN AYA WITH NHL AND HL; 4) TO ENHANCE THE UNDERSTANDING OF WELLNESS AND SURVIVORSHIP OF CAYA WITH NHL; AND 5) TO PROMOTE COLLABORATION OF RESEARCH AND TREATMENT PROTOCOLS IN CAYA WITH NHL IN LOW SOCIOECONOMIC RESOURCED COUNTRIES. SECONDARY OBJECTIVES: 1) TO CREATE AN OPPORTUNITY FOR INVESTIGATORS IN TRAINING AND YOUNG FACULTY TO CHAIR SESSIONS, PRESENT THEIR RESEARCH AND INTERFACE WITH MORE SENIOR INVESTIGATORS; 2) TO ENHANCE COMMUNICATION AND COLLABORATION WITH A MULTIDISCIPLINARY GROUP OF INTERNATIONAL INVESTIGATORS IN BASIC, TRANSLATIONAL AND CLINICAL RESEARCH IN CAYA NHL; 3) TO PROVIDE AN OPPORTUNITY TO CONDUCT A STATE OF THE ART EDUCATIONAL PROGRAM IN CAYA NHL FOR INVESTIGATORS FROM LESS RESOURCED SOCIOECONOMIC ENVIRONMENTS; AND 4) TO FACILITATE AND BRIDGE THE CLINICAL AND SCIENTIFIC GAP BETWEEN CHILDHOOD VS AYA NHL AND PROMOTE COLLABORATION BETWEEN PEDIATRIC AND MEDICAL ONCOLOGISTS. THE RESULTS OF THE SEVENTH INTERNATIONAL SYMPOSIUM WILL MOST LIKELY INCREASE OUR UNDERSTANDING OF THE BASIC, TRANSLATIONAL AND CLINICAL STATE OF THE SCIENTIFIC RESEARCH IN THIS GROUP OF RARE DISEASES AND PROVIDE AN OPPORTUNITY AND PLATFORM TO ENGAGE YOUNG AND SENIOR MULTIDISCIPLINARY AND MULTINATIONAL INVESTIGATORS TO DEVELOP INTERNATIONAL COLLABORATIVE RESEARCH THAT WILL ULTIMATELY LEAD TO THE DEVELOPMENT OF LESS TOXIC AND MORE SPECIFIC MOLECULAR, IMMUNE AND SMALL MOLECULE TARGETED BASED THERAPIES FOR FUTURE GENERATIONS OF PATIENTS WITH CAYA NHL.

TARGETING DNA REPAIR IN OVARIAN AGING

PCL - ALLOPATHIC MEDICINE - CORRECTIONS IN BUDGET PERIOD/PROJECT PERIOD