Roskamp Institute Inc

DELINEATING THE THERAPEUTIC EFFECT OF A NOVEL BBB-PENETRABLE PPAR AGONIST ON CHRONIC OUTCOMES FOLLOWING AGE-RELATED REPETITIVE MILD TBI

PATHOBIOLOGICAL STUDIES OF VESSEL BACE1 IN CEREBROVASCULAR AMYLOID ANGIOPATHY

DEVELOPMENT OF ULTRASELECTIVE DYRK1A INHIBITORS AS A NOVEL THERAPEUTIC MODALITY FOR TBI

A RANDOMIZED DOUBLE-BLIND PLACEBO-CONTROLLED CLINICAL TRIAL OF NICOTINAMIDE RIBOSIDE FOR RESTORING MITOCHONDRIAL BIOENERGETICS IN GULF WAR ILLNESS

A RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED CLINICAL TRIAL OF OLEOYLETHANOLAMIDE FOR TARGETING LIPID METABOLISM IN GULF WAR ILLNESS

INFLUENCE OF APOE ON LRP1 FUNCTION AND BETA-AMYLOID TRANSPORT ACROSS THE BBB

NEUROLOGICAL EFFECTS OF AEROSOLIZED RED TIDE NEUROTOXINS - THE OBSERVED INCREASED OCCURRENCES OF HARMFUL ALGAL BLOOMS (HABS) ARE LARGELY A CONSEQUENCE OF OCEAN WARMING DUE TO CLIMATE CHANGE, EUTROPHICATION, AND NUTRIENT POLLUTION, WHICH RAISE CONCERNS THAT HABS ARE NEGATIVELY IMPACTING AQUATIC ECOSYSTEMS, COASTAL RESOURCES AND THE HEALTH OF COASTAL COMMUNITIES. AMONG THESE HABS, KARENIA (K.) BREVIS AFFECTS MANY GULF COAST COMMUNITIES, PARTICULARLY SOUTHWEST FLORIDA. OVER THE LAST DECADE, K. BREVIS BLOOMS HAVE CAUSED MASSIVE DESTRUCTION TO MARINE LIFE AND MORBIDITY AND MORTALITY OF MARINE MAMMALS. THESE HARMFUL EFFECTS OF K. BREVIS ARE ATTRIBUTED TO THE RELEASE OF POTENT NEUROTOXINS, BREVETOXINS (PBTX). CURRENTLY, THE IMPACT OF AEROSOLIZED PBTX EXPOSURE ON THE HUMAN CENTRAL NERVOUS SYSTEM (CNS) IS NOT YET KNOWN. HUMAN CONSUMPTION OF PBTX CAUSES NEUROTOXIC SHELLFISH POISONING (NSP), INCLUDING A WIDE RANGE OF NEUROLOGICAL SYMPTOMS. WE RECENTLY SHOWED THAT SW FLORIDA RESIDENTS EXPOSED TO AEROSOLIZED PBTX ARE EXPERIENCING NSP-LIKE (NSPL) SYMPTOMS, AND THOSE WITH PAST MEDICAL HISTORIES (PMHX) OF MIGRAINE AND CHRONIC FATIGUE SYNDROME (CFS) WERE PARTICULARLY VULNERABLE TO THE NEUROLOGICAL EFFECTS OF AEROSOLIZED PBTX. CARRIERS OF THE APOLIPOPROTEIN E (APOE) E4 ALLELE WHO ARE GENETICALLY AT A HIGHER RISK OF COGNITIVE IMPAIRMENT MORE FREQUENTLY EXPERIENCE MEMORY PROBLEMS AND FATIGUE DURING RED TIDE BLOOMS. WE HAVE DETECTED PBTX IN BLOOD, AND ANTIBODIES AGAINST PBTX CORRESPOND WITH THE REPORTING OF NSPL SYMPTOMS FOLLOWING EXPOSURE TO AEROSOLIZED PBTX. HOWEVER, A DOSE-RESPONSE RELATIONSHIP BETWEEN AEROSOLIZED PBTX EXPOSURE AND THE EMERGENCE OF NSPL AND NEUROLOGICAL SYMPTOMS IS CURRENTLY UNKNOWN. THEREFORE, WE WILL DETERMINE A DOSE-RESPONSE RELATIONSHIP BETWEEN AEROSOLIZED PBTX EXPOSURE AND NSPL AND NEUROLOGICAL SYMPTOMS. WE WILL DETERMINE WHETHER INDIVIDUALS WITH PREEXISTING NEUROLOGICAL CONDITIONS OR E4 CARRIERS MORE FREQUENTLY EXPERIENCE NSPL COMPARED TO GENERAL RESIDENTS AND NON-CARRIERS. WE WILL ALSO DETERMINE WHETHER PBTX IN BIOSPECIMENS AND BLOOD PBTX ANTIBODIES CAN SERVE AS BIOMARKERS OF NSPL AND NEUROLOGICAL SYMPTOMS. CURRENTLY, THERE IS NO INFORMATION AVAILABLE ON PBTX PHARMACOKINETICS (PK) IN HUMANS. THEREFORE, WE WILL DETERMINE THE PK PARAMETERS USING PBTX IN BIOSPECIMENS AND DETERMINE THEIR RELATIONSHIP TO NSPL AND NEUROLOGICAL SYMPTOMS. USING PHYSIOLOGY-BASED PK MODELING, WE WILL ESTIMATE THE AMOUNTS OF PBTX THAT CAN REACH THE HUMAN BRAIN, WHICH WILL BE CRITICAL FOR UNDERSTANDING THE ADVERSE BRAIN HEALTH EFFECTS FROM AEROSOLIZED PBTX EXPOSURE. THIS WORK WILL INFORM FUTURE DECISION-MAKING FOR DEVELOPING STRATEGIES TO MINIMIZE THE RISKS OF NEUROLOGICAL SYMPTOMS IN HUMANS FOLLOWING AEROSOLIZED PBTX EXPOSURE. GIVEN THE ONGOING THREAT OF CLIMATE CHANGE ON THE FORMATION OF POWERFUL HURRICANES IN THE PACIFIC AND ATLANTIC OCEANS AND THE GULF OF MEXICO AND SUBSEQUENT OCCURRENCES OF INTENSE HABS IN COASTAL WATERS, THIS POPOSED WORK WILL PROVIDE A ROADMAP FOR FUTURE INVESTIGATIONS INTO STUDYING THE ADVERSE HUMAN HEALTH IMPACT FROM EXPOSURE TO HAB TOXINS.

GULF WAR PESTICIDE METABOLITE MEDIATED AUTOIMMUNE DYSFUNCTION IN GULF WAR ILLNESS

INVESTIGATION OF MITOCHONDRIAL EPIGENETIC CHANGES AS A BIOMARKER FOR GULF WAR ILLNESS AND AN OUTCOME MEASURE IN RESPONSE TO THERAPY

DEVELOPMENT OF OLEOYLETHANOLAMIDE FOR TREATING THE CNS SYMPTOMS OF GULF WAR ILLNESS

TREATMENT STRATEGIES IN A MOUSE MODEL OF CHRONIC GULF WAR ILLNESS

RESTORING THE BRAIN'S LIPID HOMEOSTASIS AS A THERAPEUTIC AVENUE FOR TREATING THE CNS SYMPTOMS OF GULF WAR ILLNESS

VALIDATING BLOOD BIOMARKERS OF BRAIN IMMUNE AND METABOLIC DYSFUNCTION IN GULF WAR ILLNESS.

TARGETING APOE BIOENERGETICS FOR REDUCING THE RISK OF NEURODEGENERATION AFTER CHRONIC TRAUMATIC BRAIN INJURY

AMNION CELL SECRETOME-MEDIATED THERAPY FOR TRAUMATIC BRAIN INJURY

AMNION-DERIVED MULTIPOTENT CELL SECRETOME-MEDICATED THERAPY FOR GULF WAR ILLNESS

ALTERED BRAIN FLUID DYNAMICS AND SOLUTE ELIMINATION FOLLOWING REPETITIVE MILD TRAUMATIC BRAIN INJURY

LIPIDOMICS FOR IDENTIFYING APOE4 ASSOCIATED BIOMARKERS OF AD RELATED COGNITIVE DECLINE IN TBI

THE INFLUENCE OF TRAUMATIC BRAIN INJURY ON GULF WAR ILLNESS PATHOGENESIS

INFLUENCE OF APOE GENOTYPE ON TAU ELIMINATION FROM THE BRAIN IN TRAUMATIC BRAIN INJURY AND ALZHEIMER'S DISEASE.

NOVEL THERAPEUTIC TARGET IDENTIFICATION THROUGH ANALYSIS OF CONVERGENT AD AND TBI...

TAU PROCESSING BY MURAL CELLS IN TRAUMATIC BRAIN INJURY AND ALZHEIMERS DISEASE

BRAIN AND PLASMA MOLECULAR CHARACTERIZATION OF THE PATHOGENIC TBI-AD INTERRELATIONSHIP IN MOUSE MODELS

CEREBROVASCULAR DEGENERATION FOLLOWING REPETITIVE MILD TRAUMATIC BRAIN INJURY CONTRIBUTES TO TAU DEPOSITION IN THE BRAIN

MICROGLIAL NOX2/NLRP3 INFLAMMASOME ACTIVATION DRIVES TAU PATHOLOGY AND COGNITIVE IMPAIRMENTS IN CHRONIC TRAUMATIC BRAIN INJURY

IMPACT OF APOE GENOTYPE ON ASTROGLIOPATHIES IN REPETITIVE MTBI.

IDENTIFICATION OF LIPID BIOMARKERS OF INFLAMMATION AND METABOLIC DISTRURBANCES IN GWI

ASTROGLIA PATHOBIOLOGY IN THE NEURODEGENERATIVE SEQUELAE OF REPETITIVE MILD TRAUMATIC BRAIN INJURY.

GLIAL CELL DYSFUNCTION IN NEURODEGENERATIVE SEQUELAE OF REPETITIVE MILD TRAUMATIC BRAIN INJURY

INFLUENCE OF APOE GENOTYPE ON THE NEURODEGENERATIVE SEQUELAE OF REPETITIVE MILD TBI IN AN HTAU MOUSE MODEL.

DELINEATING THE THERAPEUTIC EFFECT OF PTEN INHIBITION ON MITIGATING CHRONIC MICROGLIA ACTIVATION AND THE CHRONIC SEQUELAE OF REPETITIVE MTBI

EFFECT OF DELAYED MICROGLIAL SPECIFIC PPARΓ ACTIVATION ON AMELIORATING CHRONIC NEUROINFLAMMATION AND COGNITIVE DYSFUNCTION FOLLOWING R-MTBI - BRAIN INFLAMMATION IS A COMMON FEATURE OF REPETITIVE MILD TBI MEDIATED NEURODEGENERATION, AND THIS IS ASSOCIATED WITH THE MALADAPTIVE TRANSFORMATION OF MICROGLIA FROM AN EARLY NEURORESTORATIVE PHENOTYPE TO A PERSISTENTLY DYSFUNC- TIONAL AND CHRONICALLY ACTIVATED PRO-INFLAMMATORY STATE. YET, THE CRUCIAL FACTORS GOVERNING THE PROPAGATION AND PER- SISTENCE OF THESE DIFFUSE NEUROINFLAMMATORY RESPONSES IN THE CHRONIC SEQUELAE OF TBI REMAIN UNKNOWN. TO ADDRESS THIS, WE DEVELOPED A TRANSLATIONALLY RELEVANT PRECLINICAL MODEL OF R-MTBI. USING THIS MODEL, WE GENERATED A MOLECULAR LIBRARY OF MICROGLIA TRANSCRIPTOMIC PROFILES THAT PROVIDES A DETAILED TIME-COURSE OF THE MICROGLIA NEUROINFLAMMATORY RESPONSE TO TBI. IMPORTANTLY, WE OBSERVED DEFICITS IN GLUCOSE AND LIPID METABOLISM, OXIDATIVE STRESS, AND A PROINFLAM- MATORY SIGNATURE AT CHRONIC TIMEPOINTS, WHICH APPEARED TO BE MEDIATED BY THE LOSS OF CONSTITUTIVE PPAR𝛾 SIGNALING. PPAR𝛾 IS EXPRESSED IN MULTIPLE CELL TYPES AND PLAYS A CRITICAL ROLE IN REGULATING GLUCOSE AND LIPID METABOLISM, AND INFLAMMATION. TREATMENT WITH A PPAR𝛾 AGONIST HAS SHOWN EFFICACY IN MITIGATING COGNITIVE DEFICITS AND CHRONIC MI- CROGLIAL ACTIVATION IN OUR R-MTBI MODEL. HOWEVER, THE USE OF PPAR𝛾 AGONISTS CARRY SEVERAL CHALLENGES, SUCH AS THE OFTEN REPORTED OFF-TARGET EFFECTS AND LOW BBB PENETRATION. THESE DRUGS ALSO LACK THE SPECIFICITY NEEDED TO TARGET MI- CROGLIAL PPARΓ SIGNALING, AND IT REMAINS UNKNOWN WHETHER THE IMMUNOMODULATORY EFFECTS OF PPARΓ ARE CARRIED OUT BY MICROGLIA, OR OTHER CNS CELL-TYPES. FURTHER STUDIES ARE NEEDED TO CLARIFY THESE CELL-SPECIFIC ROLES TO OPTIMIZE THEIR THERAPEUTIC BENEFITS. WE RECENTLY DEVELOPED A CRE-LOXP MODEL THAT OVEREXPRESSES CONSTITUTIVELY ACTIVE PPARΓ IN MI- CROGLIA, AND IN PILOT STUDIES USING A PRE-INJURY TAMOXIFEN TREATMENT PARADIGM, WE REVEALED REPRESSION OF MICROGLIA INFLAMMATION AND ASTROGLIOSIS AT 30-DAYS POST INJURY (30DPI) AND IMPROVEMENT IN SPATIAL MEMORY. IN THIS NEW APPLI- CATION, WE WILL NOW CLARIFY THE CONSTITUTIVE ROLE OF PPARΓ IN THE CHRONIC STAGES OF TBI AND DEMONSTRATE WHETHER DELAYED PPARG OVEREXPRESSION IN ADULT MICROGLIA (AT 90DPI) COULD SERVE AS A NOVEL TARGET FOR MITIGATING CHRONIC NEUROINFLAMMATION AND COGNITIVE DEFICITS. IN THESE STUDIES, WE WILL EXAMINE FUNCTIONAL AND PATHOLOGICAL OUTCOMES AT 180- AND 270-DPI. FINALLY, WE WILL USE A FLOW CYTOMETRY AND MICROGLIAL TRANSCRIPTOMIC MINING APPROACH TO COMPARE TBI-DEPENDENT RESPONSES IN THE PRESENCE OR ABSENCE OF PPARG ACTIVATION TO REVEAL PHENOTYPIC CHANGES IN MICROGLIA SUBTYPES AND NEURORESTORATIVE MECHANISMS THAT CORRELATE WITH FAVORABLE OUTCOMES AND REPRESENT NOVEL THERAPEUTIC TARGETS TO BLOCK CHRONIC NEUROINFLAMMATION AND THE NEGATIVE CONSEQUENCES OF TBI AND RISK FOR NEURODEGENERATIVE DISEASES. BY OPTIMIZATION OF THE THERAPEUTIC EFFECT OF BOOSTING PPARΓ EXPRESSION IN MICROGLIA USING OUR NOVEL INDUC- IBLE PPARG MODEL WE AIM TO MAXIMIZE PPARΓ EFFECT AND REVEAL NOVEL TRACTABLE TARGETS THAT RESULT IN THE SAME POSITIVE DOWNSTREAM CONSEQUENCES OF MICROGLIAL SPECIFIC PPARΓ OVEREXPRESSION IN THE AFTERMATH OF TBI THAT CAN THEN BE EX- PLORED AS SAFER AND TOLERABLE NOVEL THERAPIES FOR TBI. OUR LONG-TERM GOAL IS TO IDENTIFY MOLECULAR MECHANISMS CAUSING POSTTRAUMATIC NEUROINFLAMMATION THAT CONTRIBUTE TO PROGRESSIVE BRAIN DEGENERATION AND COGNITIVE IMPAIRMENTS, IN ORDER TO DEVELOP EFFECTIVE DELAYED THERAPEUTIC INTERVENTIONS TO COMBAT NEUROLOGICAL COMPLICATIONS LINKED WITH TBI.

INFLUENCE OF MICROGLIAL CELL DEPLETION AND REPOPULATION ON ADUCANUMAB INDUCED ARIA AND VASCULAR LESIONS IN APOE4 CARRIERS WITH AD - AFTER MANY FAILED CLINICAL TRIALS, THE FDA RECENTLY APPROVED NEW BREAKTHROUGH ANTIBODY THERAPY FOR USE IN EARLY AD. BUT THESE THERAPIES HAVE BEEN REVEALED TO INCREASE THE FREQUENCY OF AMYLOID-RELATED IMAGING ABNORMALITIES (ARIAS). IMPORTANTLY, ARIA OCCURS MORE FREQUENTLY IN APOE4 CARRIERS. ALTHOUGH THE MECHANISM IS UNCLEAR, ARIA IS THOUGHT TO OCCUR AFTER REMOVAL OF AMYLOID FROM BLOOD VESSELS WITH CEREBRAL AMYLOID ANTIPATHY (CAA). APOE4 ALLELE IS ONE OF THE STRONGEST RISK FACTORS FOR AD AND HAS BEEN SHOWN TO PROMOTE CEREBROVASCULAR LESIONS SUCH AS CAA AND MICROHEMORRHAGES. BECAUSE CAA IS UNIVERSALLY OBSERVED IN AD, FURTHER INVESTIGATION OF APOE4-DEPENDENT MECHANISMS DRIVING CAA AND ARIA-EVENTS IS NEEDED. ACTIVATED MICROGLIA HAVE BEEN SHOWN TO DRIVE THE OUTCOME AND PACE OF APOE-MEDIATED NEURODEGENERATION. IMPORTANTLY, MICROGLIA CAN MODULATE CEREBROVASCULAR INTEGRITY AND CEREBRAL BLOOD FLOW, AND RELEASE FACTORS WHICH CONTRIBUTE TO BBB BREAKDOWN, CEREBROVASCULAR CELL DAMAGE AND NEUROVASCULAR INJURY. IN OUR STUDIES, WE REVEALED AN INCREASE IN INFLAMMATORY PATHWAYS IN AD VS CONTROL HUMAN CEREBROVESSELS, AND THIS WAS MORE PROMINENT IN APOE4 CARRIERS, WHO ALSO HAD HIGHER CAA SCORES. ANTI-AΒ ANTIBODY RELATED ARIA HAS ALSO BEEN ASSOCIATED WITH REACTIVE MICROGLIOSIS IN A PRIMATE MODEL OF CAA. IN HUMANS, IN VIVO PET IMAGING OF MICROGLIAL ACTIVATION HAS BEEN ASSOCIATED WITH THE MAGNITUDE AND SEVERITY OF ARIA. NO PRECLINICAL MODELS HAVE EVALUATED THIS ASSOCIATION BETWEEN ACTIVATED MICROGLIA RESPONSE AND ARIA-EVENTS, PARTICULARLY IN THE CONTEXT OF APOE4 GENOTYPE. IN THIS PROPOSAL, WE WILL ADDRESS THESE UNKNOWNS BY UTILIZING MICROGLIAL ABLATION AND REPOPULATION TECHNIQUES IN A PREVIOUSLY DESCRIBED CHRONIC ANTI-AΒ TREATED EFAD MOUSE MODEL OF AD EXPRESSING HUMAN APOE ISOFORMS AND AD MUTATIONS (I.E., ADUCANUMAB TREATED E3FAD AND E4FAD MICE). FROM THESE STUDIES, WE WILL GENERATE A DETAILED TIME-COURSE OF COGNITIVE FUNCTION, CEREBRAL BLOOD FLOW AND CEREBROVASCULAR REACTIVITY, AND CHARACTERIZE THE HISTOPATHOLOGICAL AND BIOCHEMICAL LEVEL CHANGES TO THE CEREBROVASCULATURE. NO STUDIES HAVE CHARACTERIZED THE CEREBROVASCULAR CELL PHENOTYPES IN ARIA PATIENTS. WE WILL THUS EMPLOY A SINGLE CELL TRANSCRIPTOMIC APPROACH TO CHARACTERIZE THE PHENOTYPIC BRAIN VASCULAR CELL TYPE RESPONSE(S) IN OUR EFAD MODELS FOLLOWING ADUCANUMAB TREATMENT WITH/WITHOUT MACROPHAGE DEPLETION AND REPOPULATION, AND REVEAL KEY UPSTREAM REGULATORS DRIVING THESE UNIQUE VASCULAR CELL RESPONSES AT THE SINGLE CELL LEVEL. FROM THIS PROPOSAL, OUR GOAL IS TO REVEAL WHETHER ACTIVATED MICROGLIA MEDIATED CEREBROVASCULAR INFLAMMATION CONTRIBUTES TO APOE4 MEDIATED ARIA EVENTS FOLLOWING ADUCANUMAB TREATMENT, AND II) IDENTIFY NOVEL TARGETS THROUGH WHICH APOE4 CONFERS THESE VASCULAR-SPECIFIC ABNORMALITIES, TO REVEAL NEW OPPORTUNITIES FOR IMPROVED ANTIBODIES AND ADJUNCT THERAPIES THAT CAN ONE DAY MINIMIZE THESE COMPLICATIONS IN APOE4 PATIENTS WITH AD.

ABERRANT PTEN SIGNALING IN CHRONICALLY ACTIVATED MICROGLIA DRIVES TAU PATHOLOGY, NEURODEGENERATION AND COGNITIVE DECLINE FOLLOWING REPETITIVE BRAIN INJURY - REPETITIVE MILD TBI (R-MTBI) CAN INDUCE NEUROLOGICAL IMPAIRMENT YEARS AFTER INJURY, INCREASING THE RISK FOR ALZHEIMER’S DISEASE (AD). NO SUITABLE TREATMENTS HAVE BEEN DEVELOPED TO RESCUE THE LONG-TERM CONSEQUENCES OF TBI. ADVANCING OUR UNDERSTANDING OF THE MECHANISMS DRIVING THE CHRONIC EFFECTS OF R-MTBI IS CRITICAL, AS THIS COULD LEAD TO THE IDENTIFICATION OF NOVEL THERAPIES. MICROGLIA MEDIATED NEUROINFLAMMATION IS A COMMON FEATURE OF TBI AND AD. EXPERIMENTAL EVIDENCE SUGGESTS THAT NEURODEGENERATION AND COGNITIVE DEFICITS ARE ASSOCIATED WITH THE MALADAPTIVE TRANSFORMATION OF MICROGLIA FROM AN EARLY NEURORESTORATIVE PHENOTYPE TO A DYSFUNCTIONAL AND CHRONICALLY ACTIVATED PROINFLAMMATORY STATE. FACTORS GOVERNING THE PERSISTENCE OF THESE NEUROINFLAMMATORY RESPONSES IN THE CHRONIC SEQUELAE OF TBI REMAIN ELUSIVE. WE HAVE ESTABLISHED A WT MOUSE MODEL THAT RECAPITULATES MANY OF THE FEATURES OF HUMAN R-MTBI AND THUS REPRESENTS A TRANSLATIONALLY RELEVANT PRECLINICAL PLATFORM TO INTERROGATE THESE FACTORS. HAVING ESTABLISHED THIS R-MTBI MODEL (AND GIVEN THE ROLE OF TAU AS A PATHOGNOMONIC LESION OF TBI), WE GENERATED MICROGLIAL TRANSCRIPTOMIC PROFILES IN HUMAN TAU KNOCK IN (TAUKI) MICE AT A RANGE OF TIMEPOINTS POST-INJURY. FIRSTLY, WHEN WE COMPARED MICROGLIAL RESPONSES IN TAUKI VS WT MICE, WE REVEALED AN INCREASE IN SIGNIFICANT GENE TRANSCRIPTS BETWEEN R-MTBI AND SHAM MICROGLIA BY 3 TO 9-FOLD AT CHRONIC TIMEPOINTS. IN THE TAUKI MODEL, WE REVEAL DEFICITS IN BIOENERGETICS, CYTOKINE SIGNALING, LIPID METABOLISM, AND A PRO- INFLAMMATORY SIGNATURE OF CHRONICALLY ACTIVATED MICROGLIA, WHICH APPEAR TO BE INFLUENCED BY THE ACTIVATION OF PHOSPHATASE AND TENSIN HOMOLOG (PTEN) SIGNALING, AND THIS CORRELATED WITH TBI DEPENDENT TAU PATHOLOGY. PTEN IS A LIPID PHOSPHATASE THAT ANTAGONIZES PI3K SIGNALING, A CRITICAL NODE VITAL FOR REGULATING CELL SURVIVAL, ENERGY BIOENERGETICS AND INFLAMMATION. PTEN IS HIGHLY EXPRESSED IN MYELOID CELLS, AND ITS DYSREGULATION CAN TRIGGER THE ACTIVATION OF NEUROINFLAMMATORY RESPONSES. PTEN ACTIVATION HAS BEEN REPORTED TO OCCUR EARLY IN TAUOPATHY MODELS, AND ITS INHIBITION, DAMPENS MICROGLIAL ACTIVATION, REDUCES SYNAPTIC PHAGOCYTOSIS BY MICROGLIA, AND PRESERVES NEURONAL INTEGRITY. WE THUS HYPOTHESIZE THAT PTEN ACTIVATION IS A MEDIATOR OF CHRONICALLY ACTIVATED MICROGLIA AND THEIR DYSFUNCTIONAL ACTIVITIES, AND A DRIVER OF TAU SEEDING AND PATHOGENESIS, SUCH THAT GENETIC DELETION OF PTEN IN MICROGLIA WILL MITIGATE THE DELETERIOUS EFFECTS OF CHRONIC MICROGLIAL ACTIVATION ON TAU PATHOLOGY AND LONG-TERM TBI OUTCOMES. TO BEGIN TO ADDRESS THIS, WE DEVELOPED A CRE-LOXP MODEL THAT SPECIFICALLY DELETES PTEN IN MYELOID CELLS BETWEEN 2.5 TO 3.5 MO POST- INJURY AND REVEALED RESCUE OF PRO-INFLAMMATORY MICROGLIAL RESPONSES AFTER R-MTBI. IN THIS NEW PROPOSAL, WE PLAN TO EXPAND ON THIS WORK, USING AN IMPROVED TAMOXIFEN INDUCIBLE MODEL ON A HUMAN TAUKI BACKGROUND THAT DELETES PTEN SIGNALING IN MICROGLIA, AND NOT OTHER MYELOID CELLS. WE WILL EXPLORE THE OPTIMAL TIME-WINDOW OF INTERVENTION (EARLY VS DELAYED) AND GENERATE A DETAILED LONGITUDINAL TIME-COURSE OF NEUROBEHAVIOR, MICROGLIAL PHENOTYPE AND FUNCTION, TAU SEEDING/PATHOGENESIS AND NEUROPATHOLOGICAL OUTCOMES AFTER R-MTBI. WE WILL ALSO USE A SINGLE CELL RNASEQ MINING APPROACH TO COMPARE TBI-DEPENDENT RESPONSES IN THE PRESENCE OR ABSENCE OF PTEN DELETION TO IDENTIFY THE DISTINCTIVE STATES OF MICROGLIA AND REPARATIVE MECHANISMS THAT DRIVES THEIR FAVORABLE OUTCOMES. OUR GOAL IS TO IDENTIFY MOLECULAR MECHANISMS DRIVING THE TRANSFORMATION OF CHRONIC MICROGLIA INTO A PROINFLAMMATORY AND NEUROTOXIC PHENOTYPE THAT CONTRIBUTE TO TAUOPATHY, NEURODEGENERATION AND COGNITIVE DEFICITS, IN ORDER TO DEVELOP NEW AND EFFECTIVE DELAYED THERAPEUTIC INTERVENTIONS FOR TBI AND AD.

CEREBROVASCULAR CONTRIBUTIONS TO APOE4-MEDIATED BRAIN BIOENERGETIC DEFICITS IN ALZHEIMER'S DISEASE - THE APOLIPOPROTEIN E (APOE) E4 ALLELE IS ONE OF THE MAJOR GENETIC RISK FACTORS FOR LATE-ONSET ALZHEIMER’S DISEASE (AD) AND AN IMPORTANT CONTRIBUTOR TO CEREBROVASCULAR (CV) DYSFUNCTION, WHICH IS NOW CONSIDERED A MAJOR COMPONENT OF AD PATHOLOGY. RECENT ADVANCES IN AD RESEARCH SUGGEST THAT E4 CARRIERS HAVE AN AGE-DEPENDENT VULNERABILITY IN SUPPLYING GLUCOSE TO THE BRAIN, WHICH CORRESPONDS WITH LOWER GLUCOSE METABOLISM AND PRECEDES BRAIN AMYLOID AND TAU PATHOLOGIES. THE CV SYSTEM REGULATES GLUCOSE TRANSPORT TO THE BRAIN TO SUPPORT NEURONAL BIOENERGETICS. WITH AGE, INDIVIDUALS WITH THE E4 ALLELE EXPERIENCE DEFICITS IN THEIR ABILITY TO TRANSPORT NUTRIENTS TO THE BRAIN, WHICH EVENTUALLY, FORCES NEURONS TO PERFORM FATTY ACID (FA) METABOLISM. FATTY ACID METABOLISM IS HARMFUL IF PERFORMED IN NEURONS AS IT CAN CONTRIBUTE TO OXIDATIVE STRESS. THE PROCESS OF FA METABOLISM REQUIRES L-CARNITINE FOR TRANSPORTING FA AS ACYLCARNITINES (CAR) INTO MITOCHONDRIA (L-CARNITINE BIOENERGETICS). THE IMPORTANCE OF THIS SYSTEM IN AD IS HIGHLIGHTED BY OUR RECENT STUDY SHOWING THAT L-CARNITINE-BIOENERGETIC DEFICITS ARE PRESENT IN E4 CARRIERS AND CORRELATE WITH CV PATHOLOGIES IN AD. OUR ANIMAL STUDIES HEREIN SHOW PATHWAYS THAT LINK GLUCOSE SENSING WITH ACETYL-COA CARBOXYLASE (ACC) FOR L-CARNITINE- BIOENERGETICS ARE ALTERED IN THE CEREBROVASCULATURE OF MICE WITH TARGETED REPLACEMENT OF MOUSE APOE WITH HUMAN APOE4 (E4-TR) AND AD MOUSE MODELS WITH THE HUMAN APOE4 ISOFORM. WE, THEREFORE, HYPOTHESIZE THAT E4 DISRUPTS L-CARNITINE-BIOENERGETICS WITHIN THE CV CELLS, WHICH CORRESPONDS WITH IMPAIRED TRANSPORT OF NUTRIENTS TO THE BRAIN PARENCHYMA. THIS INCREASES THE RELIANCE ON L-CARNITINE-BIOENERGETICS WITHIN NEURONS AND CONTRIBUTES TO OXIDATIVE STRESS AND INFLAMMATION IN THE BRAIN. TO TEST THIS HYPOTHESIS, WE WILL FIRST DETERMINE WHETHER BRAIN ENDOTHELIAL CELLS (BEC) OR MURAL CELLS EXPERIENCE ALTERED L-CARNITINE BIOENERGETICS AND DETERMINE THE DIFFERENTIAL IMPACT OF APOE GENOTYPES WITHIN BOTH CELLS WITH AGE. WE WILL THEN DETERMINE IF BOOSTING L-CARNITINE BIOENERGETICS PATHWAYS BY INHIBITING ACC IN BEC WILL HELP RESTORE NUTRIENT BALANCE IN THE BRAIN PARENCHYMA. THE PROPOSED STUDIES WILL PROVIDE NOVEL INSIGHTS INTO THE ROLE OF ACC-MEDIATED L-CARNITINE BIOENERGETICS FOR DEVELOPING THERAPEUTIC STRATEGIES SPECIFICALLY TARGETING E4 CARRIERS, WHO EXPERIENCE A SIGNIFICANTLY HIGHER CEREBROVASCULAR DISEASE BURDEN ASSOCIATED WITH AD PATHOGENESIS.

THE ROLE OF PPAR? IN ASTROCYTE PATHOBIOLOGY AFTER EXPOSURE TO REPETITIVE MILD TRAUMATIC BRAIN INJURY - REPETITIVE MILD TRAUMATIC BRAIN INJURY (R-MTBI) CAN INDUCE NEUROLOGICAL DAMAGE MANY YEARS AFTER THE CESSATION OF INJURY, INCREASING THE RISK FOR ADRD. NO DISEASE-MODIFYING TREATMENT STRATEGIES HAVE BEEN DEVELOPED TO MITIGATE THE LONG-TERM CONSEQUENCES OF R-MTBI. THERE IS AN URGENT NEED TO ADVANCE OUR CURRENT UNDERSTANDING OF THE CELLULAR MECHANISMS DRIVING THE CASCADE OF SECONDARY INJURY EVENTS, AS THIS COULD LEAD TO THE IDENTIFICATION AND DEVELOPMENT OF NOVEL THERAPEUTICS. ASTROCYTES PLAY AN IMPORTANT ROLE IN THESE SECONDARY INJURY EVENTS. AFTER ACUTE INSULT, THEY UNDERGO A DRAMATIC TRANSCRIPTOMIC AND MORPHOLOGICAL TRANSFORMATION. REACTIVE ASTROCYTES CAN BE POLARIZED INTO DIFFERENT STATES ADOPTING NEUROPROTECTIVE OR NEUROTOXIC PROPERTIES THAT CAN INFLUENCE BRAIN RECOVERY. NEUROPROTECTIVE ASTROCYTES CAN SERVE TO CREATE A PHYSICAL BARRIER TO LIMIT THE SPREAD OF DAMAGE, PREVENTEXCITOTOXICITY, BOOST METABOLIC SUPPORT FOR NEURONS, AND RELEASE TROPHIC FACTORS TO PROMOTE NEUROREPAIR. WHILE NEUROTOXIC ASTROCYTES CAN TAKE A DUAL ROLE OF NEUROINFLAMMATION AND GLIAL SCAR FORMATION THAT INHIBITS AXONAL REGENERATION AND PROMOTES NEURONAL DAMAGE, AND THIS CAN BE ACCOMPANIED BY LOSS OF THEIR CONSTITUTIVE SUPPORTIVE ROLES. OUR KNOWLEDGE OF THE MECHANISMS THAT REGULATE ASTROCYTE PHENOTYPES AND RESPONSES IN THE HEALTHY BRAIN OR AFTER BRAIN INJURY IS LACKING. TO ADDRESS THIS, WE ESTABLISHED A MOUSE MODEL OF R-MTBI THAT RECAPITULATES MANY OF THE FEATURES OF HUMAN TBI AND THUS REPRESENTS A TRANSLATIONALLY RELEVANT PRECLINICAL PLATFORM. USING THIS MODEL, WE GENERATED A MOLECULAR LIBRARY OF ASTROGLIA GENE PROFILES, AT A RANGE OF TIMEPOINTS POST-INJURY THAT PROVIDES A UNIQUE AND DETAILED TIME-COURSE OF THE ASTROGLIA RESPONSE TO TBI. PARTICULARLY, WE REVEAL DEFICITS IN CELLULAR METABOLISM, OXIDATIVE STRESS AND A PROINFLAMMATORY SIGNATURE OF ASTROGLIA, WHICH APPEARS TO BE INFLUENCED BY THE LOSS OF CONSTITUTIVE PPAR? SIGNALING IN ASTROCYTES. PPAR IS HIGHLY EXPRESSED IN GLIAL CELLS AND PLAYS A VITAL CONSTITUTIVE ROLE IN REGULATING CELL METABOLISM, BIOENERGETICS, CELL SURVIVAL AND IMMUNE FUNCTION. TREATMENT WITH A PPAR AGONIST HAS SHOWN EFFICACY IN RESTORING BEHAVIORAL OUTCOMES AND RESCUING ASTROGLIA PATHOBIOLOGY IN OUR R-MTBI MODEL. BECAUSE MULTIPLE CELL TYPES EXPRESS PPAR RECEPTORS, PPAR LIGANDS LACK THE SPECIFICITY NEEDED TO TARGET ASTROGLIA SPECIFIC PPAR SIGNALING IN VIVO. IN THIS PROPOSAL, WE PLAN TO CLARIFY THE CONSTITUTIVE ROLE OF PPAR IN REGULATING ASTROGLIAL RESPONSES IN THE HEALTHY BRAIN AND IN THE CONTEXT OF TBI, AND DEMONSTRATE WHETHER ASTROGLIA SPECIFIC PPAR ACTIVATION MITIGATES TBI MEDIATED ASTROGLIA ACTIVATION, INFLAMMATION, NEURODEGENERATION AND FUNCTIONAL OUTCOMES. WE WILL ACHIEVE THIS BY UTILIZING A TAMOXIFEN INDUCIBLE MOUSE MODEL THAT SPECIFICALLY TARGETS PPAR ACTIVATION IN ASTROCYTES. WE WILL INDUCE PPAR ACTIVATION INASTROCYTES USING THREE THERAPEUTIC TIME-WINDOWS (I.E., PRE-INJURY, EARLY AND DELAYED), AND EXAMINE FUNCTIONAL AND PATHOBIOLOGICAL OUTCOMES, SCRNASEQ PROFILES AND FUNCTIONAL ACTIVITIES OF ASTROGLIA AT 6 MO POST-INJURY. IN OUR SCRNASEQ STUDY, WE WILL COMPARE TBI-DEPENDENT RESPONSES IN THE PRESENCE OR ABSENCE OF PPAR ACTIVATION TO REVEAL ASTROGLIA-SPECIFIC TARGETS THAT CORRELATE WITH FAVORABLE OUTCOMES AT THE OPTIMALTIME-WINDOW OF TREATMENT, AND REPRESENT NOVEL THERAPEUTIC AND TRANSLATIONAL TARGETS. OUR GOAL IS TO CLARIFY THE ROLE OF PPAR AS A REGULATOR OF ASTROGLIA PATHOBIOLOGY IN THE CHRONIC SEQUELAE OF TBI AND IDENTIFY REPARATIVE MECHANISMS IN ASTROCYTES DRIVING FAVORABLE OUTCOMES THAT CAN BE EXPLORED AS NOVEL ASTROCYTE SPECIFIC TARGETS IN FUTURE WORK, NOT ONLY IN TBI BUT ADRD WHERE ASTROCYTE PATHOBIOLOGY IS A CRITICAL CONTRIBUTOR.

THE ROLE OF ADAPTIVE IMMUNITY IN ORGANOPHOSPHATE INDUCED CNS INJURY - CHLORPYRIFOS (CPF) IS AN ORGANOPHOSPHATE (OP) PESTICIDE, CLASSIFIED AS A CHEMICAL THREAT AGENT BY THE DEPARTMENT OF HOMELAND SECURITY BECAUSE IT CAN CAUSE NEUROTOXICITY IF RELEASED INTO THE CIVILIAN POPULATIONS. UNDER THESE CIRCUMSTANCES, CPF AND SIMILAR OP CHEMICALS HAVE A POTENTIAL TO CAUSE LONG-TERM CHRONIC MULTI SYMPTOM ILLNESSES (CMI), SUCH AS MYALGIC ENCEPHALOMYELITIS/CHRONIC FATIGUE SYNDROME (ME/CFS). TO DATE, MANY VICTIMS OF THE TOKYO SUBWAY SARIN GAS ATTACK ARE STILL EXPERIENCING CHRONIC HEALTH PROBLEMS CONSISTING OF COGNITIVE DIFFICULTIES, HEADACHES, MUSCLE WEAKNESSES AND FATIGUE, WHICH NEGATIVELY IMPACT THEIR QUALITY OF LIFE. SUCH SYMPTOMS ARE REPORTED BY INDIVIDUALS EXPOSED TO OP PESTICIDES AND ARE THOUGHT TO BE CAUSED BY MALADAPTIVE IMMUNE RESPONSES7. AS SUCH, THIS PROPOSAL WILL INVESTIGATE THE ROLE OF CPF IN MALADAPTIVE IMMUNE RESPONSES TO DEVELOP FUTURE APPROACHES THAT MITIGATE LONG-TERM MORBIDITY ASSOCIATED WITH CMI. ORDINARILY, A SMALL CHEMICAL IS NOT ANTIGENIC, BUT WHEN IT FORMS ADDUCTS WITH ENDOGENOUS PROTEINS, IT CAN BE RECOGNIZED BY THE IMMUNE SYSTEM AS A FOREIGN THREAT AGENT AND PROVOKE AN ADAPTIVE IMMUNE RESPONSE. OUR PRIOR WORK IN THIS AREA SHOWS THAT CERTAIN PESTICIDE METABOLITES CAN FORM ADDUCTS WITH PROTEINS AND THEN ELICIT AN ADAPTIVE IMMUNE RESPONSE, ACTIVATING T- AND B-CELLS THAT ULTIMATELY CONTRIBUTE TO THE PRODUCTION OF ANTIBODIES AGAINST THEM AND CORRESPONDING WITH BRAIN INFLAMMATION. ACCOUNTS OF ACUTE CPF POISONING IN HUMANS SUPPORT THIS NOTION BY SHOWING THAT CPF OR CPF-OXON (CPO) CAN FORM ADDUCTS ON SEVERAL AMINO ACID RESIDUES IN HUMAN ALBUMIN, WHICH ARE CONSIDERED BIOMARKERS OF OP EXPOSURE. HOWEVER, IT IS UNKNOWN WHETHER THESE CPF/CPO-PROTEIN ADDUCTS HAVE A ROLE IN ACTIVATING THE IMMUNE SYSTEM. WE THEREFORE HYPOTHESIZE THAT CPF/CPO-PROTEIN ADDUCTS FORMED IN VIVO AFTER CPF EXPOSURE CAN ACTIVATE T-CELL AND B-CELL RESPONSES, RESULTING IN ANTIBODY PRODUCTION. WE PROPOSE THAT CPF-PROTEIN SPECIFIC ANTIBODIES MAY CROSS-REACT WITH BRAIN PROTEINS AND CONTRIBUTE TO THE DEVELOPMENT OF CHRONIC AUTOIMMUNE DISORDERS. THE PROPOSED WORK BUILDS UPON AN EXISTING SCIENTIFIC PREMISE OF PESTICIDE-MEDIATED MALADAPTIVE IMMUNE RESPONSES. THESE STUDIES WILL CHARACTERIZE WHETHER ACUTE CPF ADMINISTRATION STIMULATES IMMUNE CELLS AND WHETHER THIS CORRESPONDS WITH ACTIVATION OF THE MICROGLIA AND ASTROGLIA AND NEUROINFLAMMATION AFTER CPF EXPOSURE. WE WILL DETERMINE WHETHER BRAIN IMMUNE ACTIVATION IS ASSOCIATED WITH FORMATION OF CPF/CPO-PROTEIN ADDUCTS. WE WILL EXAMINE THE PRESENCE OF IMMUNE CELLS THAT RECOGNIZE CPF/CPO-MODIFIED PROTEINS AND ANTIBODIES AGAINST THEM TO DETERMINE IF BLOOD ANTIBODIES CAN CROSS-REACT WITH BRAIN PROTEINS. UNDERSTANDING THE MECHANISMS OF OP- INDUCED CMI WILL FACILITATE THE DEVELOPMENT OF COUNTERMEASURE EFFORTS THAT TARGET THE IMMUNE SYSTEM IN ORDER TO MINIMIZE LONG-TERM MORBIDITY ASSOCIATED WITH SUCH ILLNESSES IN CIVILIAN POPULATIONS FOLLOWING A MASS CHEMICAL ATTACK WITH CPF OR SIMILAR CHEMICALS.

PROTEOMIC STUDIES OF PROTEIN PATHWAYS FOR THE ESTROGEN THERAPEUTIC WINDOW IN AD

AMNION CELL SECRETOME MEDIATED THERAPY FOR THE RESTORATION OF TRAUMATIC OPTIC NEUROPATHY AFTER BRAIN TRAUMA. NEW AWARD.

APPLICATION OF LIPIDOMICS TO IDENTIFY BIOMARKERS OF IMMUNE AND MITOCHONDRIAL DISTURBANCES IN CHRONIC FATIGUE SYNDROME.

TARGETING THE ASTROCYTE-CEREBROVASCULATURE SYSTEM TO CORRECT BRAIN BIOENERGETICS DEFECTS ASSOCIATED WITH APOE4 - THE APOLIPOPROTEIN E4 (APOE4) ALLELE IS THE MAJOR GENETIC RISK FACTOR FOR ALZHEIMER'S DISEASE (AD), A NEURODEGENERATIVE CONDITION MARKED BY IMPAIRMENT OF THE BRAIN BIOENERGETICS. ENERGY DYNAMICS IN THE BRAIN ARE TIGHTLY LINKED TO NEURONAL ACTIVITY AND HAVE A MAJOR IMPACT ON NEURONAL FUNCTION. GLUCOSE IS THE MAIN SOURCE OF ENERGY FOR THE BRAIN, AND ITS CEREBRAL FUELING IS GUARANTEED BY GLUCOSE TRANSPORTERS WITHIN THE BRAIN VASCULAR SYSTEM. GLUCOSE IS DIRECTLY CONSUMED BY NEURONS OR CAN BE METABOLIZED BY ASTROCYTES INTO LACTATE, WHICH IS THEN EXPORTED TO NEURONS AS A SUPPLEMENTARY ENERGY SOURCE. WHEN GLUCOSE SUPPLY TO THE BRAIN FROM THE BLOOD IS INSUFFICIENT, ASTROCYTES CAN ALSO GENERATE FATTY ACID (FA) METABOLITES THROUGH MITOCHONDRIAL FATTY ACID (FA) Β- OXIDATION (FAO) AND THESE METABOLITES SERVE AS ALTERNATIVE ENERGY SUBSTRATES FOR NEURONS. APOE4-MEDIATED DISTURBANCES OF THE FAO HAVE BEEN OBSERVED IN CELLS AT THE BLOOD BRAIN BARRIER (BBB) AND ASSOCIATED WITH ALTERATIONS IN THE BBB INTEGRITY SUGGESTING THAT PERTURBATIONS OF THE ENERGY METABOLISM OF THESE CELLS COULD LEAD TO DEFICITS OF NEURONAL ENERGY SUPPLY IN E4 CARRIERS. OUR PREVIOUS WORK SHOWED THE AMP-ACTIVATED PROTEIN KINASE (AMPK)-MEDIATED ACETYL-COA CARBOXYLASE ACTIVATION IS AFFECTED IN THE CEREBROVASCULATURE OF APOE4-TR AND E4FAD MICE, WITH CHANGES MORE PRONOUNCED IN THE PRESENCE OF AD PATHOLOGY IN EFAD MICE. AS THIS SYSTEM PLAYS A KEY ROLE IN LINKING GLUCOSE UPTAKE AND METABOLISM WITH FAO, WE HYPOTHESIZE THAT BRAIN BIOENERGETICS DEFICITS IN E4 CARRIERS, AND MORE IMPORTANTLY IN THE PRESENCE OF AD-RELATED INSULTS, RESULT FROM A COMBINATION OF LOW GLUCOSE AVAILABILITY DUE TO ITS POOR TRANSIT ACROSS THE BBB, PARTICULARLY THROUGH THE CEREBROVASCULATURE (CV), AND THEIR REDUCED ABILITY TO SWITCH TOWARDS COMPENSATORY MECHANISMS TO MEET BRAIN BIOENERGETIC NEEDS. THIS COULD BE MEDIATED BY AN INHIBITION OF THE GLP-1R-AMPK-ACC PATHWAY IN THE E4-CV/ASTROCYTE CROSSTALK SYSTEM, WHICH WOULD LOWER THE FAO IN THESE CELLS AND ALTER THEIR CAPACITY TO MAINTAIN BRAIN BIOENERGETIC HOMEOSTASIS. TARGETING THE GLP-1R-AMPK-ACC PATHWAY IN E4 CV/ASTROCYTES COULD SERVE AS A PROMOTER OF THE FAO TO CORRECT THE BIOENERGETICS IMBALANCE IN E4 CARRIERS. HOWEVER, THE IMPACT OF MODULATING THIS PATHWAY IN THE CV AND ASTROCYTES HAS NOT YET BEEN FULLY INVESTIGATED. HENCE, USING AN AD MOUSE MODEL, WE WILL EXAMINE THE INFLUENCE OF DIFFERENT APOE GENOTYPES ON THE GLP-1R-AMPK-ACC PATHWAY IN CV AND ASTROCYTES. WE WILL ALSO DETERMINE WHETHER TREATMENTS THAT INDUCE ACTIVATION OF THE GLP-1R-AMPK-ACC PATHWAY COULD REGULATE FAO IN E4 CARRIERS, WHICH COULD OPEN THE DOOR TO NEW WAYS OF TREATING AD. THUS, THESE PROPOSED STUDIES WILL PROVIDE CRITICAL INSIGHT INTO THE DEVELOPMENT OF APPROACHES FOR TARGETING BRAIN BIOENERGETIC PATHWAYS TO PREVENT OR REDUCE COGNITIVE DECLINE IN AD AMONG HIGH-RISK APOE E4 CARRIERS.

DELINEATING THE THERAPEUTIC EFFECT OF PTEN INHIBITION ON MITIGATING CHRONIC MICROGLIA ACTIVATION AND THE CHRONIC SEQUELAE OF REPETITIVE MTBI

INFLUENCE OF APOE4 GENOTYPE ON MICROGLIAL PATHOBIOLOGY AND TAU PATHOLOGY AFTER REPETITIVE MTBI - ONE OF THE HALLMARK, CHRONIC, FEATURES OF REPETITIVE MILD TBI (R-MTBI) IS THE DEPOSITION OF PHOSPHORYLATED TAU IN NEURONS. TAU LESIONS ARE ALSO ONE OF THE MAIN HALLMARK FEATURES OF ADRD. IT REMAINS UNKNOWN WHAT SPECIFIC MOLECULAR TRIGGERS PRECIPITATE THE PATH TOWARDS THIS DISTINCT TBI RELATED NEURODEGENERATIVE PHENOTYPE. THE E4 ALLELE IS A MAJOR GENETIC RISK FACTOR FOR AD; INDIVIDUALS CARRYING 1 COPY HAVE A 2-3 FOLD RISK FOR AD, WHILE THOSE WITH 2 COPIES HAVE A 15-FOLD RISK COMPARED TO E3 CARRIERS. DESPITE SOME CONTRADICTORY STUDIES, THE E4 ALLELE HAS LONG BEEN ASSOCIATED WITH A POOR OUTCOME AFTER TBI, BUT THE ROLE PLAYED BY APOE IN RESPONSE TO TBI IS STILL UNKNOWN AND WELL-DESIGNED LONGITUDINAL STUDIES ARE NEEDED. CONDUCTING SUCH STUDIES IN HUMANS REMAINS CHALLENGING AS EPIDEMIOLOGICAL AND PROSPECTIVE DATA ARE LACKING, PLUS THE HETEROGENEITY OF TBI ETIOLOGY, INCLUDING (BUT NOT LIMITED TO) SEVERITY LEVEL, AGE, COMORBIDITIES AND TIME POST-TBI, PRESENT AN ENORMOUSLY CONFOUNDING PROBLEM. THUS, THE BEST WAY TO ADDRESS THIS QUESTION IS IN TRANSLATIONALLY RELEVANT, WELL CHARACTERIZED AND CONTROLLED ANIMAL MODELS, WHEREIN KEY PREDISPOSING GENETIC FACTORS CAN BE TARGETED AND EXPRESSED, AND FINDINGS FROM LONGITUDINAL ANALYSES CAN BE RELATED TO THE LIMITED AUTOPSY INFORMATION FROM HUMAN TBI CASES WHO HAVE DIED AT DIFFERENT TIMEPOINTS AFTER THEIR INJURY. WE HAVE DEVELOPED AND CHARACTERIZED SUCH MOUSE MODELS OF R-MTBI, WHICH RECAPITULATE MANY FEATURES OF HUMAN TBI PATHOLOGY. IN RECENT WORK, WE HAVE EXPOSED HUMAN APOE-TARGETED REPLACEMENT MICE (APOE-TR), MICE HUMANIZED FOR TAU (TAUKI) AND CROSSES OF THESE MICE (E-TAU) TO OUR R-MTBI PARADIGM, AND OBSERVE TBI-DEPENDENT PTAU PATHOLOGY. FROM THESE STUDIES WE HAVE ALSO REVEALED THAT THE E4 ALLELE AUGMENTS THE PROINFLAMMATORY MICROGLIAL RESPONSE AND TAU PATHOLOGY IN INJURED MICE COMPARED TO E3. APOE IS UPREGULATED IN DISEASE ASSOCIATED MICROGLIA, WHICH HAS BEEN REPORTED IN AD BRAINS. IN OUR R-MTBI MODEL WE HAVE ALSO CONFIRMED AN INCREASE IN MICROGLIAL SPECIFIC APOE GENE EXPRESSION. DISEASE ASSOCIATED MICROGLIA HAVE BEEN REPORTED TO DRIVE THE OUTCOME AND PACE OF APOE4-DEPENDENT NEURODEGENERATION IN AD TRANSGENIC MODELS; YET VERY LITTLE IS KNOWN ABOUT THEIR CONTRIBUTION IN DRIVING APOE4 MEDIATED EFFECTS AFTER R-MTBI. WE WILL ADDRESS THESE UNKNOWNS USING MOUSE MODELS EXPRESSING HUMAN FORMS OF APOE/TAU, AND EXPOSE THEM TO OUR R-MTBI PARADIGM TO ADDRESS THESE TIMELY AND UNDER-STUDIED INTERACTIONS. WE WILL FIRST EXPOSE THESE MODELS TOPHARMACOLOGICAL MANIPULATION OF MICROGLIA USING DEPOPULATION/REPOPULATION PARADIGMS TO DELINEATE THEIR CONTRIBUTION TO THE APOE INFLUENCE ON R-MTBI PATHOGENESIS AND TAU PATHOLOGY. IN THE NEXT PART OF THE STUDY, WE WILL USE AN INDUCIBLE APOE-KI MODEL TO GENETICALLY MANIPULATE TO DELINEATE THEIR CONTRIBUTION TO THE APOE INFLUENCE ON MICROGLIAL TBI MEDIATED NEURODEGENERATION, TAU PATHOLOGY AND BEHAVIORAL OUTCOME. MICROGLIA SPECIFIC APOE EXPRESSION TRANSCRIPTOMIC PHENOTYPES AND EX VIVO FUNCTIONAL ACTIVITIES, WE WILL FINALLY COMPARE TBI-DEPENDENT MICROGLIAL TRANSCRIPTOMIC RESPONSES IN THE PRESENCE OR ABSENCE OF APOE DELETION IN THIS MODEL TO REVEAL MICROGLIAL SPECIFIC TARGETS THAT CORRELATE WITH FAVORABLE OUTCOMES AFTER R-MTBI AND REPRESENT NOVEL THERAPEUTIC TARGETS. WE WILL CONFIRM THE TRANSLATIONAL RELEVANCE OF OUR TARGETS IN R-MTBI/CONTROL AUTOPSY CASES FROM DIFFERENT APOE BACKGROUNDS. OUR FUTURE WORK WILL INTERROGATE THE FUNCTIONAL AND THERAPEUTIC ROLES OF THESE TARGETS. THIS STUDY IS MUCH NEEDED AS A FIRST STEP IN DECIPHERING THE ROLE OF APOE4 IN MICROGLIA PATHOBIOLOGY AFTER R-MTBI, WHICH IS CURRENTLY UNDER-INVESTIGATED.

THE ROLE OF PTEN IN MICROGLIAL PATHOBIOLOGY AFTER EXPOSURE TO REPETITIVE MILD TBI - REPETITIVE MILD TRAUMATIC BRAIN INJURY (R-MTBI) IS ONE OF THE STRONGEST RISK FACTORS FOR DEVELOPING NEURODEGENERATIVE DISEASES. TO DATE, NO DISEASE MODIFYING THERAPIES HAVE BEEN DEVELOPED TO PREVENT THE LONG- TERM CONSEQUENCES OF TBI. THERE IS A NEED TO ADVANCE OUR CURRENT UNDERSTANDING OF THE CELLULAR MECHANISMS DRIVING THE LONG-TERM NEUROLOGICAL DEFICITS AFTER TBI, AS THIS COULD LEAD TO THE IDENTIFICATION OF NOVEL THERAPEUTIC TARGETS. NEUROINFLAMMATION MEDIATED BY RESIDENT MICROGLIA IS A COMMON FEATURE OF HUMAN AND ANIMAL MODELS OF TBI. FACTORS GOVERNING THE PROPAGATION AND PERSISTENCE OF DISEASE ASSOCIATED MICROGLIAL RESPONSES IN THE CHRONIC SEQUELAE OF TBI REMAIN ELUSIVE. WE HAVE ESTABLISHED A MOUSE MODEL OF R-MTBI THAT RECAPITULATES MANY FEATURES OF HUMAN TBI AND THUS REPRESENTS A TRANSLATIONALLY RELEVANT PRECLINICAL PLATFORM. FROM THIS MODEL WE HAVE GENERATED A MOLECULAR LIBRARY OF MICROGLIA GENE PROFILES, AT A RANGE OF TIMEPOINTS POST-INJURY THAT PROVIDES A UNIQUE AND DETAILED TIME-COURSE OF THE MICROGLIAL NEUROINFLAMMATORY RESPONSE TO R-MTBI. PARTICULARLY, WE REVEAL DEFICITS IN ENERGY BIOENERGETICS, CYTOKINE SIGNALING, LIPID METABOLISM, AND A PRO-INFLAMMATORY SIGNATURE OF MICROGLIA AT CHRONIC TIMEPOINTS, WHICH APPEAR TO BE DRIVEN BY THE ACTIVATION OF PHOSPHATASE AND TENSIN HOMOLOG (PTEN) SIGNALING. PTEN IS A LIPID PHOSPHATASE THAT ANTAGONIZES PHOSPHATIDYLINOSITOL 3-KINASE SIGNALING, A CRITICAL NODE VITAL FOR REGULATING CELL SURVIVAL, ENERGY BIOENERGETICS, AUTOPHAGY AND INFLAMMATION. PTEN IS HIGHLY EXPRESSED IN MYELOID CELLS, AND ITS DYSREGULATION CAN TRIGGER THE ACTIVATION OF INFLAMMATORY RESPONSES. MULTIPLE CELL TYPES EXPRESS PTEN, THUS PTEN INHIBITORS LACK THE SPECIFICITY NEEDED TO TARGET PTEN SIGNALING IN MICROGLIA. IN A PILOT STUDY, WE HAVE SHOWN THAT PTEN DELETION IN MYELOID CELLS AFTER 1 MO DAMPENS DISEASE ASSOCIATED MICROGLIAL RESPONSES AND PROINFLAMMATORY SIGNATURE IN OUR MODEL. IN THIS NEW APPLICATION, WE PLAN TO EXTEND THESE STUDIES TO FURTHER CLARIFY THE ROLE OF PTEN IN REGULATING MICROGLIA RESPONSES IN THE CONTEXT OF TBI AND DEMONSTRATE WHETHER MICROGLIA SPECIFIC PTEN DELETION CAN MITIGATE TBI MEDIATED NEUROINFLAMMATION/NEURODEGENERATION AND CHRONIC FUNCTIONAL OUTCOMES. WE WILL COMPARE TBI- DEPENDENT RESPONSES IN THE PRESENCE OR ABSENCE OF PTEN DELETION TO REVEAL MICROGLIAL SPECIFIC TARGETS THAT CORRELATE WITH FAVORABLE OUTCOMES AFTER R-MTBI AND REPRESENT NOVEL THERAPEUTIC TARGETS. WE WILL ACHIEVE THIS BY UTILIZING A TAMOXIFEN INDUCIBLE MOUSE MODEL THAT WILL SPECIFICALLY TARGET PTEN DELETION IN MICROGLIA AND NOT OTHER MYELOID CELLS (HEXBCRE+/PTENFL/FL). WE WILL INDUCE PTEN DELETION USING THREE THERAPEUTIC TIME-WINDOWS (I.E., PRE-INJURY, EARLY AND DELAYED), AND EXAMINE FUNCTIONAL AND PATHOBIOLOGICAL OUTCOMES, SCRNASEQ PROFILES AND FUNCTIONAL ACTIVITIES OF MICROGLIA AT 6 MO POST-INJURY. OUR GOAL IS TO CLARIFY THE ROLE OF PTEN AS A NEGATIVE REGULATOR OF MICROGLIAL PATHOBIOLOGY IN THE CHRONIC SEQUELAE OF R-MTBI, AND TO IDENTIFY UNIQUE GENE SIGNATURES AND REPARATIVE MECHANISMS IN MICROGLIA INDUCED BY PTEN DELETION THAT CAN BE EXPLORED AS NOVEL MICROGLIAL SPECIFIC TARGETS IN TBI AND OTHER NEURODEGENERATIVE DISEASES WHERE NEUROINFLAMMATION IS A CRITICAL CONTRIBUTOR.

THE ROLE OF PPAR? IN MICROGLIA PATHOBIOLOGYAFTER EXPOSURE TO REPETITIVE MILD TRAUMATIC BRAIN INJURY - EXPOSURE TO REPETITIVE MILD TRAUMATIC BRAIN INJURY (R-MTBI) CAN INDUCE NEUROLOGICAL DAMAGE MANY YEARS FOLLOWING THE CESSATION OF INJURY, CONTRIBUTING TO AN INCREASED RISK FOR NEURODEGENERATIVE DISEASE IN LATER LIFE. TO DATE, NO SUITABLE TREATMENT STRATEGIES HAVE BEEN DEVELOPED TO RESCUE THE PERSISTENT AND LONG-TERM NEGATIVE CONSEQUENCES OF R-MTBI. A GREATER EMPHASIS SHOULD THEREFORE BE PLACED ON UNDERSTANDING THE UNDERLYING PATHOBIOLOGICAL MECHANISMS DRIVING THE LONG-TERM NEUROLOGICAL DEFICITS AFTER R-MTBI, AS THIS COULD LEAD TO THE IDENTIFICATION OF NOVEL THERAPEUTIC TARGETS. NEUROINFLAMMATION IS A COMMON FEATURE OF HUMAN AND PRECLINICAL ANIMAL MODELS OF TBI. THE FACTORS GOVERNING THE PROPAGATION AND PERSISTENCE OF NEUROINFLAMMATORY RESPONSES IN THE CHRONIC SEQUELAE OF TBI REMAIN ELUSIVE. WE HAVE ESTABLISHED A MOUSE MODEL OF R-MTBI THAT RECAPITULATES MANY OF THE FEATURES OF HUMAN TBI AND THUS REPRESENTS A TRANSLATIONALLY RELEVANT PRECLINICAL PLATFORM FOR SUCH STUDIES. FROM THIS MODEL WE HAVE GENERATED A MOLECULAR LIBRARY OF MICROGLIA GENE PROFILES AT A RANGE OF TIMEPOINTS POST-INJURY THAT PROVIDES A UNIQUE AND DETAILED TIME-COURSE OF THE MICROGLIAL NEUROINFLAMMATORY RESPONSE TO R-MTBI. PARTICULARLY, WE OBSERVED DEFICITS IN ENERGY BIOENERGETICS, ALTERED GLUCOSE AND LIPID METABOLISM, AND A PRO-INFLAMMATORY SIGNATURE AT CHRONIC TIMEPOINTS, WHICH APPEARED TO BE DRIVEN BY THE LOSS OF CONSTITUTIVE PPAR SIGNALING IN MICROGLIA. PPAR IS EXPRESSED IN MULTIPLE CELL TYPES AND PLAYS A CRITICAL ROLE IN REGULATING GLUCOSE AND LIPID METABOLISM, ENERGY BIOENERGETICS AND INFLAMMATION. TREATMENT WITH A PPAR AGONIST HAS SHOWN EFFICACY IN RESTORING BEHAVIORAL AND MICROGLIAL PATHOBIOLOGICAL CONSEQUENCES IN OUR R- MTBI MODEL. HOWEVER, BECAUSE MULTIPLE CELL TYPES EXPRESS PPAR RECEPTORS, PHARMACOLOGICAL PPAR LIGANDS LACK THE SPECIFICITY NEEDED TO TARGET MICROGLIAL PPAR SIGNALING IN VIVO. IN THIS NEW APPLICATION, WE PLAN TO CLARIFY THE CONSTITUTIVE ROLE OF PPAR IN REGULATING BRAIN MICROGLIAL CELL RESPONSES IN THE CONTEXT OF TBI AND DEMONSTRATE WHETHER MICROGLIA SPECIFIC PPAR ACTIVATION MITIGATES TBI MEDIATED NEUROINFLAMMATION AND SUBSEQUENT NEURODEGENERATION IN OUR R-MTBI MODEL. WE WILL COMPARE TBI-DEPENDENT RESPONSES IN THE PRESENCE OR ABSENCE OF PPAR ACTIVATION TO REVEAL MICROGLIAL SPECIFIC TARGETS THAT CORRELATE WITH FAVORABLE OUTCOMES AFTER R-MTBI AND REPRESENT NOVEL THERAPEUTIC TARGETS. WE WILL ACHIEVE THIS BY UTILIZING A TAMOXIFEN INDUCIBLE MOUSE MODEL THAT SPECIFICALLY TARGETS PPAR ACTIVATION IN MICROGLIA. WE WILL INDUCE PPAR ACTIVATION IN MICROGLIA USING A PRE-INJURY TAMOXIFEN TREATMENT PARADIGM, AND EXAMINE FUNCTIONAL AND PATHOBIOLOGICAL OUTCOMES, AND GLIAL CELL TRANSCRIPTOMIC PROFILES AT 3 AND 6 MO POST-INJURY. OUR GOAL IS TO CLARIFY THE ROLE OF PPAR AS A MASTER REGULATOR OF MICROGLIAL PATHOBIOLOGY IN THE CHRONIC SEQUELAE OF R-MTBI, AND TO IDENTIFY UNIQUE GENE SIGNATURES AND REPARATIVE MECHANISMS IN MICROGLIA INDUCED BY PPAR ACTIVATION THAT CAN BE EXPLORED AS NOVEL MICROGLIAL SPECIFIC TARGETS, NOT ONLY IN TBI BUT OTHER NEURODEGENERATIVE DISEASES WHERE NEUROINFLAMMATION IS A CRITICAL CONTRIBUTOR.

THE ROLE OF NEUROIMMUNE-NEUROENDOCRINE INTEGRATION AND PREMATURE HYPOTHALAMIC AGING IN TBI-RELATED NEURODEGENERATION

AMNION CELL SECRETOME MEDIATED THERAPY FOR GULF WAR ILLNESS.

LONG TERM ASSESSMENT OF NEUROLOGICAL EFFECTS AFTER RED TIDE EXPOSURE

EFFECT OF MILD TBI ON FEMALE MICE WITH A CONTROLLED ESTROUS CYCLE

THE EFFECTS OF APOE4 ON CARNITINE/ACYLCARNITINE MEDIATED BIOENERGETIC DEFICITS IN ALZHEIMER'S DISEASE - ALZHEIMER’S DISEASE IS A NEURODEGENERATIVE CONDITION WHICH AFFECTS 50 MILLION PEOPLE WORLDWIDE. IT IS INCREASING BEING RECOGNIZED THAT, IN ADDITION TO AMYLOID AND TAU, THERE ARE OTHER FACTORS WHICH CONTRIBUTE TO THE COMPLEX AND HETEROGENOUS ETIOLOGY OF AD. IN THAT REGARD, RECENT INVESTIGATIONS SHOW THAT BRAIN BIOENERGETIC DEFICITS AND ABNORMAL LIPID METABOLISM ALTER THE BRAIN’S MILIEU, MAKING IT MORE SUSCEPTIBLE TO AGE-RELATED INSULTS AND THEREBY INCREASING THE BRAIN’S VULNERABILITY TO DEVELOPING AD. CARNITINE AND ACYLCARNITINES ARE CRITICAL FOR CENTRAL NERVOUS SYSTEM (CNS) BIOENERGETICS DUE TO THEIR ROLE IN FATTY ACIDS OXIDATION IN MITOCHONDRIA FOR THE BRAIN’S ENERGY REQUIREMENTS. BIOENERGETIC DEFICITS ARE OBSERVED AT AN EARLY AGE IN SUBJECTS WITH THE APOLIPOPROTEIN E (APOE) E4 ALLELE, WHICH IS THE MOST IMPORTANT GENETIC RISK FACTOR FOR MAJORITY OF LATE-ONSET AD CASES. THE PROPOSED WORK WILL EXPLORE THE INFLUENCE OF APOE E4 IN CARNITINE- AND ACYLCARNITINE-MEDIATED BIOENERGETIC DEFICITS IN THE PATHOGENESIS OF AD. WE RECENTLY OBSERVED THAT CARNITINE, A CARNITINE METABOLITE TRIMETHYLAMINE N-OXIDE (TMAO) AND ACYLCARNITINE LEVELS ARE ALTERED IN THE BLOOD AND BRAINS OF AD PATIENTS. WE ALSO SHOW THAT E4 CARRIERS AT PRECLINICAL OR EARLY STAGES OF AD HAVE ELEVATED LEVELS OF TMAO. ELEVATED MEDIUM CHAIN ACYLCARNITINE (MCA) WERE DETECTED IN THE BRAIN AND BLOOD OF E4 CARRIERS, REFLECTING AN INCOMPLETE FATTY ACID OXIDATION. THESE DATA, TOGETHER WITH THE EXISTING EVIDENCE OF GLUCOSE HYPOMETABOLISM IN THE BRAINS OF E4 CARRIERS, SUGGEST THAT BIOENERGETIC DEFICIT MAY BE AN EARLY EVENT IN AD. WE THEREFORE HYPOTHESIZE THAT APOE E4 DEPENDENT DEFICITS IN THE TRANSPORT OF PERIPHERAL CARNITINE AND ACYLCARNITINES TO THE BRAIN AND IN THEIR METABOLISM IN THE PERIPHERY OR IN THE BRAIN CONTRIBUTE TO THE BRAIN BIOENERGETIC DEFICITS WHICH MAKE THE BRAIN VULNERABLE TO AD PATHOLOGY. WE WILL FIRST CHARACTERIZE ABNORMAL CARNITINE AND ACYLCARNITINE PROFILES IN THE BRAIN AND BLOOD OF NEUROPATHOLOGICALLY DIAGNOSED AD PATIENTS AND IN AD MOUSE MODELS TO EXAMINE THE INFLUENCE OF APOE GENOTYPES ON THE RELATIONSHIP BETWEEN CARNITINE/ACYLCARNITINES AND AD. WE WILL EXAMINE IF PERIPHERALLY ADMINISTERED STABLE ISOTOPE LABELED CARNITINE AND ACYLCARNITINES ARE DIFFERENTIALLY TRANSPORTED AND METABOLIZED IN AN AD MOUSE MODEL WITH 5 X AD MUTATIONS AND WITH GENETIC TARGETED REPLACEMENT OF MURINE APOE WITH HUMAN APOE ISOFORMS (EFAD MICE). WE WILL ALSO DETERMINE WHETHER THESE CHANGES OCCUR PRIOR TO OR WITH THE ONSET OF AMYLOID AND TAU PATHOLOGIES. THESE STUDIES WILL CLARIFY WHETHER APOE E4 CONTRIBUTES TO BOTH TRANSPORT AND METABOLISM DEFICIENCIES OF PERIPHERAL CARNITINE AND ACYLCARNITINES AND IDENTIFY NEW AVENUES FOR DEVELOPING TREATMENT STRATEGIES FOR AD, PARTICULARLY FOR E4 POSITIVE INDIVIDUALS WHO ARE AT A HIGH RISK OF DEVELOPING AD.

INFLUENCE OF APOE GENOTYPE ON CEREBROVASCULAR CELL PATHOBIOLOGY IN AD, AND THE CONTRIBUTION OF MICROGLIA INFLAMMATION - ALZHEIMER’S DISEASE (AD) IS AN AGE-RELATED NEURODEGENERATIVE DISORDER AND THE PREDOMINANT TYPE OF DEMENTIA, MARKED BY BRAIN DEPOSITS OF AMYLOID PLAQUES AND NEUROFIBRILLARY TANGLES. AN OFTEN OVERLOOKED PATHOGNOMONIC LESION OF AD IS CEREBROVASCULAR DEGENERATION. ABNORMALITIES IN CEREBRAL BLOOD FLOW ARE A PRECLINICAL FEATURE OF AD THAT MANIFEST MANY YEARS BEFORE SYMPTOM ONSET. VASCULAR LESIONS ARE ALSO ROUTINELY OBSERVED IN AD BRAINS AT DIFFERENT STAGES OF THE DISEASE. DISRUPTIONS TO CEREBROVASCULAR INTEGRITY CAN IMPACT ON NEURONAL FUNCTION, HOWEVER, THE MAIN DRIVER OF THESE VASCULAR CHANGES IN AD REMAINS ELUSIVE. THE APOLIPOPROTEIN E4 (APOE4) ALLELE IS ONE OF THE STRONGEST RISK FACTORS FOR AD, AND HAS BEEN SHOWN TO SIGNIFICANTLY IMPACT ON CEREBROVASCULAR HEALTH, PRECIPITATING DEFICITS IN CEREBRAL PERFUSION, VASCULAR LESIONS AND DAMAGE TO THE BLOOD BRAIN BARRIER. YET, MORE LONGITUDINAL STUDIES ARE NEEDED TO BETTER UNDERSTAND HOW APOE4 CAN SIGNIFICANTLY IMPACT ON CEREBROVASCULAR CELL PATHOBIOLOGY IN THE SEQUELAE OF AD PATHOGENESIS. IN THE BRAIN, APOE IS PRODUCED PRIMARILY BY GLIAL CELLS, AND IN AD, DISEASE ASSOCIATED MICROGLIA (DAM’S) ARE KNOWN TO UPREGULATE APOE EXPRESSION, WHICH MEDIATES THE TRANSCRIPTIONAL PROINFLAMMATORY PHENOTYPE OF THESE CELLS. DAM’S HAVE BEEN SHOWN TO DRIVE THE OUTCOME AND PACE OF APOE-MEDIATED NEURONAL DYSFUNCTION, BUT VERY LITTLE IS CURRENTLY KNOWN ABOUT THE ROLE THEY PLAY IN CONTRIBUTING TO CEREBROVASCULAR CHANGES IN AD, PARTICULARLY, HOW THIS IS INFLUENCED BY APOE GENOTYPE. TO ADDRESS THESE QUESTIONS, WE PLAN TO USE MOUSE MODELS EXPRESSING HUMAN FORMS OF APOE AND A TO CLARIFY THE ROLE PLAYED BY APOE4 AS A MOLECULAR DRIVER OF CEREBROVASCULAR CELL PATHOBIOLOGY, AND HOW THIS IS INFLUENCED BY DAM’S. WE WILL EXPLORE LONGITUDINAL CEREBROVASCULAR CELL PATHOBIOLOGY AT TIMEPOINTS REPRESENTING PRE, PERI AND POST ONSET OF A PATHOLOGY USING HISTOPATHOLOGICAL AND ULTRASTRUCTURAL ANALYSES. WE WILL ALSO CONFIRM THE CONTRIBUTION OF DAMS ON CEREBROVASCULAR PATHOBIOLOGY BY UTILIZING MICROGLIA ABLATION TECHNIQUES TO DEPOPULATE DAMS AND ALSO REPOPULATE NEWBORN MICROGLIA IN OUR MOUSE MODELS. TO DATE, NO STUDIES HAVE CHARACTERIZED THE MOLECULAR TRANSCRIPTS OF REACTIVE CEREBROVASCULAR CELL PHENOTYPES IN THESE COMBINED MODELS. THUS, WE WILL CONCLUDE OUR STUDIES USING SINGLE CELL GENE ANALYSES OF CEREBROVASCULAR CELLS (ENDOTHELIAL CELLS, PERICYTES, SMOOTH MUSCLE CELLS) TO REVEAL THEIR UNIQUE AND DETAILED TIME-COURSE OF RESPONSES. WE WILL CONFIRM DIFFERENTIALLY EXPRESSED GENES IN AUTOPSIED CEREBROVASCULAR TISSUE FROM STAGED AD CASES TO VALIDATE THE TRANSLATIONAL RELEVANCE OF OUR FINDINGS AND MAP AGAINST MOUSE AD-RELATED PATHOGENIC TIMELINES. FROM THIS PROPOSAL, WE AIM TO IDENTIFY NOVEL (CEREBROVASCULAR) CELL SPECIFIC TARGETS IN THE EARLY STAGES OF DISEASE THROUGH WHICH THE APOE4 ALLELE CONFERS NEUROLOGICAL RISK. OUR FUTURE GOAL WILL BE TO EXPLORE THE FUNCTIONAL EFFECTS OF THESE TARGETS, AND TO PROVIDE NEW THERAPEUTIC OPPORTUNITIES TO PROMOTE CEREBROVASCULAR HEALTH AND RESTORATION OF COGNITIVE FUNCTION IN AD.

DEVELOPMENT OF NEURAL STEM CELL MEDIATED THERAPY FOR REPETITIVE MILD TBI.

A RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED CROSSOVER STUDY OF THE ANTI-INFLAMMATORY COMPOUND ANATABINE TO TREAT PAIN IN GWI PATIENTS