Arizona Veterans Research And Education Foundation

VASCULAR METABOLIC MEMORY IN TYPE 2 DIABETES

MOLECULAR TOOL DEVELOPMENT TO IDENTIFY, ISOLATE, AND INTERROGATE THE ROD MICROGLIA PHENOTYPE IN NEUROLOGICAL DISEASE AND INJURY - OUR PATHOLOGICAL EXAMINATION OF ACQUIRED NEUROLOGICAL INJURY UNCOVERED THE ROD MICROGLIA VARIANT OF ACTIVATED IMMUNE CELLS IN THE BRAIN. OVER A CENTURY AGO, SIMILAR ROD MICROGLIA WERE HAND DRAWN TO INDICATE NEUROPATHOLOGY IN GENERAL PARESIS, TYPHOID, AND CHEMICAL EXPOSURE, AND THEN LARGELY IGNORED. FOLLOWING OUR 2012 PUBLICATION, ROD MICROGLIA HAVE BEEN VISUALIZED IN POST-MORTEM TISSUE WITH CLINICAL AND NEUROPATHOLOGICAL CONFIRMATION OF ALZHEIMER’S DISEASE AND RELATED DEMENTIAS (ADRD), IN ADDITION TO EPILEPSY, MENTAL HEALTH DISORDERS, AND OTHERS. YET NO MOLECULAR TOOLS EXIST FOR PRECISE MOLECULAR INVESTIGATION OF ROD MICROGLIA. AS CELLULAR MECHANISMS OF ADRD AND INJURY INCLUDE NEUROINFLAMMATION AND CONCOMITANT MICROGLIA ACTIVATION, A CRITICAL PROBLEM EMERGES FROM OUR INABILITY TO DETECT AND ANALYZE ROD MICROGLIA WITHIN THE CONTEXT OF ADRD/BRAIN INJURY NEUROINFLAMMATION. THE BODY OF KNOWLEDGE ON ROD MICROGLIA RELIES SOLELY ON HISTOLOGICAL STAINS AND NON-SPECIFIC MICROGLIA ANTIBODIES (E.G., IBA1, CD68, CX3CR1) FOR POST-MORTEM MICROSCOPIC IDENTIFICATION. OUR MOST RECENT SINGLE-CELL RNASEQ DATA FROM 100 ALZHEIMER’S DISEASE (AD) AND CONTROL SUBJECTS IDENTIFIED A UNIQUE CLUSTER OF MICROGLIA DEFINED BY GENES FOR MOTILITY, CYTOSKELETAL ACTIN DYNAMICS, AND MOVEMENT OF MICROGLIAL PROCESSES; CONFIRMATION OF THIS CLUSTER AS ROD MICROGLIA CANNOT OCCUR WITHOUT NEW MOLECULAR TOOLS TO PHENOTYPE THIS ELUSIVE MICROGLIA VARIANT. FOR THIS BIOENGINEERING RESEARCH GRANT (PAR 19-158), COLLECTIONS OF MOLECULAR TOOLS AND MARKERS UNIQUE TO THE ROD MICROGLIA VARIANT WILL BE DEVELOPED, VALIDATED, AND VERIFIED TO ENABLE FUTURE DIAGNOSTIC, PROGNOSTIC, AND MECHANISTIC STUDIES OF NEUROINFLAMMATION PROGRESSION IN CASES OF ADRD AND BRAIN INJURY. TO DEVELOP THESE TOOLS, WE PROPOSE A DIFFUSE BRAIN-INJURY MODEL THAT GENERATES FIELDS OF ROD MICROGLIA ADJACENT TO NEURONAL DENDRITES. FROM THIS TISSUE, WE WILL CAPTURE (LCM) CX3CR1-EGFP ROD MICROGLIA AND NON-ROD MICROGLIA FOR PHAGE DISPLAY BIOPANNING FROM A LIBRARY OF HUMAN-DERIVED VARIABLE HEAVY-CHAIN BINDING DOMAIN ANTIBODIES (DAB). SYNTHETIC CYCLIZED HCDR3 PEPTIDES WILL BE DEVELOPED AS DAB MIMETICS UNIQUE TO ROD MICROGLIA (OBJECTIVE 1). SPATIAL TRANSCRIPTOMICS OF THE ROD MICROGLIA VARIANT USING THE NANOSTRING GEOMX DIGITAL SPATIAL PROFILER WILL DETERMINE DIFFERENTIALLY EXPRESSED GENES BETWEEN ROD AND NON-ROD MICROGLIA (OBJECTIVE 2). PATHWAY ANALYSIS WILL IDENTIFY CELLULAR PROCESSES ASSOCIATED WITH THE STRUCTURE, FUNCTION, MOTILITY, AND ORIGIN OF ROD MICROGLIA. FOR EACH OBJECTIVE, NEW MOLECULAR TOOLS ARE VALIDATED AND VERIFIED ACROSS TIME POST-INJURY, BETWEEN SEXES, AND IN EXPERIMENTAL MODELS OF MIXED PATHOLOGY TBI, STROKE, AND AD (FAMILIAL AND LATE-ONSET AMYLOIDOSIS), AS WELL AS POST-MORTEM HUMAN AD/ADRD TISSUE. THE NEW TOOLS PROPOSED IN THIS APPLICATION WILL BE ABLE TO IDENTIFY, ISOLATE, AND INTERROGATE THE ROD MICROGLIA VARIANT, WHILE EXPANDING THE CONCEPT OF NEUROINFLAMMATION IN AGING, INJURY, AND DISEASE TO INCLUDE THE OVERLOOKED ROD MICROGLIA VARIANT THAT PRESENTS ADJACENT TO NEURONAL DENDRITES. WITH REFINED MOLECULAR TOOLS, ROD MICROGLIA CAN BE INVESTIGATED ACROSS MANY NEUROLOGICAL CONDITIONS AND INCORPORATED INTO UNIFYING PRINCIPLES THAT EXPLAIN MICROGLIAL ACTIVATION AND NEUROINFLAMMATION. 33

PROBING THE MECHANISTIC ROLE OF VASCULAR DYSFUNCTION AND VASCULAR INFLAMMATION IN TBI MEDIATED COGNITIVE DYSFUNCTION

VCR - VIRTUAL COGNITIVE REHABILITATION USING A VIRTUAL REALITY SPATIAL NAVIGATION APPLICATION FOR VETERANS WITH A HISTORY OF TBI

IDENTIFICATION OF RECESSIVE PROSTATE CANCER RISK ALLELES IN THE MILLION VETERANS PROGRAM

NEUROGENIC TREMORS TRAINING (TRE) FOR STRESS AND PTSD: A CONTROLLED CLINICAL TRIAL

UNDERSTANDING VASCULAR AGING-RELATED DEMENTIA THROUGH MEDIN SIGNALING - ABSTRACT AGE IS THE MOST IMPORTANT RISK FACTOR FOR CARDIO-CEREBROVASCULAR DISEASES AND DEMENTIA DISORDERS. EPIDEMIOLOGIC, PRECLINICAL AND CLINICAL DATA SHOW THAT VASCULAR DISEASE IS STRONGLY ASSOCIATED WITH DEMENTIA DISORDERS, INCLUDING ALZHEIMER'S DISEASE (AD) AND AD-RELATED DISORDERS SUCH AS VASCULAR DEMENTIA (VAD). THE MECHANISTIC LINKS AMONG VASCULAR DISEASE, AGING AND DEMENTIA REMAIN POORLY UNDERSTOOD. THERE IS GROWING EVIDENCE THAT MEDIN, A 50-AMINO ACID PEPTIDE THAT FORMS ONE OF THE MOST COMMON YET LEAST STUDIED HUMAN AMYLOIDOSES, IS AN IMPORTANT DRIVER OF VASCULAR AGING PATHOLOGIES. MEDIN ACCUMULATES IN THE VASCULATURE WITH AGING AND IS IMPLICATED IN AD, VAD AND AORTIC DISEASE. LITTLE IS KNOWN AS TO THE MECHANISMS BY WHICH MEDIN INDUCES CELL AND TISSUE INJURY, AND NO ANIMAL MODEL OF MEDIN PATHOLOGY CURRENTLY EXISTS. WE SHOWED THAT MEDIN IMPAIRED ENDOTHELIAL FUNCTION AND CELL VIABILITY AND INDUCED PRO-INFLAMMATORY ACTIVATION, IN PART THROUGH RECEPTOR FOR ADVANCED GLYCATION ENDPRODUCTS (RAGE). OUR GOALS ARE TO DISCOVER BIOLOGICAL PATHWAYS OF MEDIN TOXICITY USING NOVEL GENOME-WIDE CRISPR/CAS9 KNOCKOUT GENETIC SCREENING, TEST F(BA)S PEPTIDE AND RAGE KNOCKOUT TO REVERSE MEDIN TOXICITY AND TEST THE IN VIVO ROLE OF MEDIN IN VASCULO-NEUROPATHOLOGY BY CREATING A MOUSE MODEL WITH ENDOTHELIAL OVEREXPRESSION OF MEDIN. IN AIM 1, WE WILL PROBE TOXIC SIGNALING MECHANISMS USING A SYNTHETIC LETHALITY-BASED GENOME-WIDE CRISPR/CAS9 SCREENING IN ENDOTHELIAL CELLS. THIS WILL GENERATE A LIST OF CANDIDATE GENES/PATHWAYS THAT FACILITATE OR PROTECT AGAINST MEDIN TOXICITY AND COULD BE POTENTIAL DRUG TARGETS. IN AIM 2, WE WILL TEST IF PEPTIDE F(BA)S CAN REVERSE MEDIN TOXICITY IN CULTURED ENDOTHELIAL CELLS AND EX VIVO IN ISOLATED HUMAN DONOR HUMAN CEREBRAL ARTERIES. IN AIM 3, WE WILL TEST IN VIVO THE ROLE OF MEDIN IN AGING-INDUCED VASCULAR AND COGNITIVE DYSFUNCTION AND ASSESS WHETHER RAGE KNOCKOUT WILL BE PROTECTIVE. IN AN EXPLORATORY SUBAIM, WE WILL CREATE A TRANSGENIC MOUSE MODEL OF ENDOTHELIUM-SPECIFIC MEDIN OVEREXPRESSION. ONCE SUCCESSFULLY IMPLEMENTED, THE PROPOSAL WILL SHED LIGHT ON THE MECHANISMS UNDERLYING MEDIN VASCULO-NEUROPATHOLOGY AND CREATE A VALUABLE AND NOVEL PRECLINICAL ANIMAL MODEL OF VASCULAR AGING THAT CAN BE USED TO IDENTIFY AND TEST NEW DRUG TARGETS.

THE HARVEST FOR HEROES PRODUCE PRESCRIPTION PROGRAM ADDRESSES THE CRITICAL ISSUE OF FOOD INSECURITY AMONG VETERANS, A GROWING CONCERN THAT IMPACTS THEIR HEALTH AND WELL-BEING. LIMITED OR UNCERTAIN ACCESS TO ADEQUATE NUTRITIOUS FOOD, A CONDITION CALLED FOOD INSECURITY, IS MORE PREVALENT AMONG VETERANS AND IS TIED TO WORSE MANAGEMENT OF CHRONIC ILLNESSES LIKE DIABETES, HYPERTENSION, AND OBESITY. IT CAN ALSO LEAD TO MORE FREQUENT VISITS TO EMERGENCY MEDICAL SERVICES, WHICH IS COSTLY FOR THE MEDICAL SYSTEM. ADDITIONALLY A LACK OF NUTRITION EDUCATION CAN INDEPENDENTLY CONTRIBUTE TO POOR HEALTH OUTCOMES. THIS PROGRAM WILL PROVIDE ENROLLED VETERANS WHO MEET SPECIFIC CRITERIA WITH NUTRITION EDUCATION FROM A REGISTERED DIETITIAN AND VOUCHERS FOR FRESH FRUITS AND VEGETABLES FROM LOCAL FARMERS MARKETS. BY OFFERING THESE VETERANS THE ABILITY TO ADD MORE FRESH FRUITS AND VEGETABLES TO THEIR DIET USING PRODUCE VOUCHERS AND LEARNING HOW TO CREATE NUTRITIOUS MEALS AND PREPARE FRUITS AND VEGETABLES WITH THE HELP OF THEIR DIETITIAN, WE AIM TO SHOW IMPROVEMENT IN FOOD INSECURITY, CHRONIC DISEASE MANAGEMENT, AND AGGREGATE REDUCTION OF HEALTHCARE COSTS.PARTICIPATING VETERANS WILL BE GIVEN 5 VISITS WITH A DIETITIAN FOR EDUCATION AND COUNSELING ON MEAL PLANNING, EATING HEALTHY ON A BUDGET, AND INCREASING INTAKE OF FRUITS AND VEGETABLES. ACTIVE PARTICIPANTS WILL ALSO RECEIVE VOUCHERS WEEKLY, REDEEMABLE FOR FRESH PRODUCE AT LOCAL FARMERS MARKETS. OVER THE COURSE OF THE 20-24 WEEKS THAT EACH VETERAN IS ENROLLED, DATA WILL BE COLLECTED ON FRUIT AND VEGETABLE INTAKE, FOOD INSECURITY, MARKERS OF CHRONIC DISEASE, AND EMERGENCY HEALTHCARE USE. THE AIMS OF THIS STUDY ARE TO SHOW AN INCREASED INTAKE OF FRESH PRODUCE DUE TO IMPROVED NUTRITION KNOWLEDGE, SKILLS, AND CONFIDENCE IN THESE VETERANS. ADDITIONALLY, WE AIM TO SHOW THAT THESE NUTRITION CHANGES WILL LEAD TO SUSTAINABLE BEHAVIOR CHANGE, DECREASES IN FOOD INSECURITY, AND IMPROVEMENT IN VETERANS' CHRONIC HEALTH CONDITIONS. ULTIMATELY, HARVEST FOR HEROES AIMS TO DEMONSTRATE THAT CORRECTING FOOD INSECURITY WITH A COMBINATION OF PRODUCE PRESCRIPTIONS AND NUTRITION EDUCATION WILL REDUCE HEALTHCARE COSTS AND PROMOTE THE LONG-TERM HEALTH OF OUR VETERANS.

ANTI-MEDIN IMMUNOTHERAPY FOR VASCULAR AGING AND RELATED DEMENTIAS - ABSTRACT AGE IS THE MOST IMPORTANT RISK FACTOR FOR CARDIO-CEREBROVASCULAR DISEASES (CVD) AND DEMENTIA DISORDERS. EPIDEMIOLOGIC, PRECLINICAL AND CLINICAL DATA SHOW THAT VASCULAR DISEASE IS STRONGLY ASSOCIATED WITH DEMENTIA DISORDERS, INCLUDING ALZHEIMER'S DISEASE (AD) AND AD-RELATED DEMENTIAS (ADRD) SUCH AS VASCULAR DEMENTIA (VAD). INDEED, UNBIASED DATA-DRIVEN ANALYSES SHOWED THAT VASCULAR DYSFUNCTION IS THE EARLIEST AND STRONGEST BRAIN PATHOLOGY ASSOCIATED WITH LATE ONSET AD. THERE IS GROWING EVIDENCE THAT MEDIN, A 50-AMINO ACID PEPTIDE THAT FORMS ONE OF THE MOST COMMON YET LEAST STUDIED HUMAN AMYLOIDOSES, IS AN IMPORTANT DRIVER OF VASCULAR AGING PATHOLOGIES. WE AND OTHERS SHOWED THAT VASCULAR MEDIN BURDEN WAS ASSOCIATED WITH VAD, AD AND AORTIC ANEURYSMS. MEDIN, A CLEAVAGE PRODUCT OF MILK FAT GLOBULE-EGF FACTOR 8 PROTEIN (MFGE8), ACCUMULATES IN VESSELS WITH AGING, CAUSES ENDOTHELIAL DYSFUNCTION THROUGH OXIDATIVE AND NITRATIVE STRESS AND INDUCES ENDOTHELIAL PRO- INFLAMMATORY ACTIVATION THAT COULD INITIATE NEUROINFLAMMATION. KNOCKOUT OF MFGE8 DOMAIN CONTAINING MEDIN PREVENTED CEREBROVASCULAR MEDIN AMYLOID FORMATION AND RESTORED CEREBROVASCULAR FUNCTION. WE DESIGNED AND PRODUCED AN IMMUNOGEN MODIFIED FROM THE AGGREGATION-PRONE C TERMINUS OF MEDIN TO BE USED AS ANTI-MEDIN VACCINE AND 3 MONOCLONAL ANTIBODIES (MABS) THAT HAVE HIGH MEDIN AFFINITY. OUR OVERALL GOAL IS TO EVALUATE ANTI- MEDIN IMMUNOTHERAPY USING MEDIN VACCINATION (AIM 1) AND ANTI-MEDIN MABS (AIM 2) TO ATTENUATE OR PREVENT MEDIN-INDUCED VASCULAR DYSFUNCTION AND NEUROVASCULAR PATHOLOGY IN AGING C57BL/6 MICE AND IN SENESCENCE ACCELERATED MOUSE-PRONE 8 (SAMP8) MICE. AIM 1 IS TO DEVELOP AND TEST THE BENEFIT OF ANTI-MEDIN VACCINATION IN PREVENTING CEREBROVASCULAR AND COGNITIVE DYSFUNCTION AND NEUROVASCULAR PATHOLOGY IN C57BL/6 AGED AND SAMP8 MICE. WE WILL FIRST DETERMINE AN OPTIMAL VACCINATION REGIMEN AND THEN TEST THE EFFICACY OF THAT REGIMEN. AIM 2 IS TO DEVELOP AND PILOT TEST ANTI-MEDIN MAB IMMUNOTHERAPY TO ATTENUATE MEDIN-INDUCED CEREBROVASCULAR AND COGNITIVE DYSFUNCTION AND NEUROVASCULAR PATHOLOGY IN C57BL/6 AGED AND SAMP8 MICE. FIRST, WE WILL DETERMINE WHICH MAB (19H1, 6F2 AND 13B7) WILL HAVE THE BEST EFFECT IN REVERSING MEDIN-INDUCED ENDOTHELIAL CYTOTOXICITY. NEXT, WE WILL DO PILOT FEASIBILITY IN VIVO TESTING OF THE IDEAL DOSING OF ANTI-MEDIN MABS TO REDUCE CEREBROVASCULAR MEDIN AND PRESERVE CEREBROVASCULAR AND COGNITIVE FUNCTION. THE PROPOSAL IS INNOVATIVE AND PARADIGM-CHANGING IN POTENTIALLY ESTABLISHING MEDIN AS A NEW TARGET FOR VASCULAR DISEASE AND DEMENTIA, AND ANTI-MEDIN IMMUNOTHERAPY AS A POTENTIAL NEW TREATMENT APPROACH FOR VASCULAR AGING AND RESULTING PATHOLOGIES.

SEVERE ALCOHOLIC PANCREATITIS-ASSOCIATED ACUTE LUNG INJURY IN VETERANS: RISKS, MECHANISMS, PREDICTION, AND THERAPEUTIC RELEVANCE

DISCOVERING THE ORIGIN OF VASCULAR AGING AMYLOID PROTEIN MEDIN - ABSTRACT AGE IS THE MOST IMPORTANT RISK FACTOR FOR CARDIOVASCULAR AND CEREBROVASCULAR DISEASES (CVD), THE LEADING CAUSES OF MORTALITY AND MORBIDITY IN THE US AND WORLDWIDE. AGING CAUSES VASCULAR IMPAIRMENT INDEPENDENT OF THE TRADITIONAL CARDIOMETABOLIC RISK FACTORS, WHILE MAGNIFYING THE LATTER’S DAMAGE. MEDIN IS A 50-AMINO ACID AMYLOID PRECURSOR DERIVED FROM ITS PARENT PROTEIN MILK FAT GLOBULE-EGF FACTOR 8 PROTEIN (MFGE8). IT ACCUMULATES IN THE VASCULATURE WITH AGING AND CONTRIBUTES TO THE MOST COMMON FORM OF HUMAN AMYLOIDOSIS. MEDIN IS IMPLICATED IN VASCULAR AGING, AORTIC DISEASE, ALZHEIMER’S DISEASE AND VASCULAR DEMENTIA. LITTLE IS KNOWN AS TO THE MECHANISM BY WHICH MEDIN IS GENERATED AND WE DO NOT KNOW THE PROTEASE(S) RESPONSIBLE FOR MFGE8 CLEAVAGE. THE MAIN GOAL OF THE PROPOSAL IS TO ADDRESS THIS MAJOR KNOWLEDGE GAP THROUGH USE OF NOVEL GENOMIC AND BIOINFORMATICS TOOLS. IN AIM 1, WE WILL DEVELOP IN HUMAN ENDOTHELIAL CELLS A RELIABLE AND QUANTITATIVE REPORTER SYSTEM FOR THE PROCESSING ACTIVITY FOR MFGE8, AND USE IT AS A SELECTABLE PHENOTYPE TO CONDUCT GENOME WIDE CRISPR/CAS9 KNOCKOUT SCREENING TO IDENTIFY GENES INVOLVED IN MEDIN GENERATION EITHER DIRECTLY OR INDIRECTLY, BUT WITH PARTICULAR FOCUS ON GENES FOR PROTEASE/S. IN AIM 2, WE WILL CONDUCT IN SILICO MOLECULAR DYNAMICS STUDY TO UNDERSTAND MFGE8 CONFORMATION CHANGES REQUIRED TO EXPOSE THE MEDIN CLEAVAGE SITES AND, INFORMED BY GENE TARGETS IDENTIFIED IN AIM 1, USE IN SILICO PROTEIN BIOINFORMATICS TO SELECT POTENTIAL PROTEASE/S THAT ARE STRUCTURALLY PREDICTED TO INTERACT WITH MFGE8 IN APPROPRIATE CONFORMATION. IN AIM 3, WE WILL FUNCTIONALLY INTERROGATE THE CANDIDATE GENES/PROTEINS FROM AIMS 1 AND 2 BY OVER-EXPRESSING OR KNOCKING THEM OUT INDIVIDUALLY IN REPORTER ENDOTHELIAL CELLS AND IN NAÏVE ENDOTHELIAL CELLS, TO ASSESS PREDICTED CHANGES IN MEDIN GENERATION AND CONFIRM THEIR BIOLOGIC RELEVANCE. IDENTIFYING ENZYMES, PROTEINS AND MOLECULAR STRUCTURAL DETERMINANTS OF MEDIN GENERATION IS A CRITICAL STEP IN UNDERSTANDING AND REVERSING MEDIN PATHOPHYSIOLOGY THAT WOULD BE USEFUL IN ADDRESSING VASCULAR AGING, VASCULAR DEMENTIA, AD AND AORTIC DISEASE.

HUMAN VASCULAR MODEL TO STUDY ALZHEIMER'S DISEASE

ROLE OF METHIONINE SULFOXIDE AND ADVANCED GLYCATION ENDPRODUCTS IN CARDIOVASCULAR AND RENAL COMPLICATIONS OF TYPE 2 DIABETES