Regenerative Research Foundation

INVESTIGATING THE FUNCTIONAL IMPACT OF AD RISK GENES ON NEURO-VASCULAR INTERACTIONS - PROJECT SUMMARY/ABSTRACT CEREBROVASCULAR PATHOLOGY IS PRESENT THROUGHOUT STAGES OF ALZHEIMER’S DISEASE AND IS CORRELATED WITH COGNITIVE CHANGES. THERE IS STRONG EVIDENCE THAT VASCULAR DYSFUNCTION IS A SIGNIFICANT DRIVER OF NEUROPATHOLOGY. OUR LONG- TERM OBJECTIVE IS TO UNDERSTAND THE FUNCTION OF ALZHEIMER’S DISEASE-ASSOCIATED RISK GENES IN VASCULAR CELLS, THEIR CONTRIBUTION TO THE DEVELOPMENT OF CEREBROVASCULAR PATHOLOGY AND THE OPPORTUNITIES TO USE THIS INFORMATION IN THERAPEUTIC DEVELOPMENT. THERE ARE OVER 27 ALZHEIMER’S DISEASE-ASSOCIATED RISK (AD-RISK) LOCI ENCOMPASSING NUMEROUS GENETIC VARIANTS IN NON-CODING AND CODING REGIONS AND HUNDREDS OF LINKED GENES. OUR OVERARCHING HYPOTHESIS IS THAT A SUBSET OF AD-RISK GENES IMPAIRS VASCULAR FUNCTION, CAUSING RELEASE OF INFLAMMATORY FACTORS, BLOOD BRAIN BARRIER (BBB) IMPAIRMENT, AND REDUCED PERFUSION, THUS CONTRIBUTING TO NEURODEGENERATION. TO ADDRESS THIS, WE HAVE ASSEMBLED A MULTI-DISCIPLINARY TEAM WITH A PROVEN TRACK RECORD OF COLLABORATION, INCLUDING WITH ADSP AND ADGP MEMBERS, WHO BRING EXPERTISE IN VASCULAR PATHOLOGY IN DEMENTIA, ENDOTHELIAL CELL (EC) SIGNALING AND EC FUNCTIONAL TESTING, ALZHEIMER’S DISEASE GENOMICS, SINGLE CELL AND NUCLEAR TRANSCRIPTOMICS, BIOINFORMATICS, CRISPR-BASED GENE EDITING FOR LARGE SCALE SCREENING AND AD MOUSE MODELS FOR IN-DEPTH FUNCTIONAL ASSESSMENT IN VIVO. NOTABLY, WE WILL ADDRESS DIFFERENCES IN GENE EFFECTS RELATED TO THE IMPORTANT BIOLOGICAL VARIABLES, SEX AND METABOLIC DISEASE. MEN AND WOMEN DIFFER IN THEIR GENETIC RISK FOR ALZHEIMER’S DISEASE, WITH SEX-SPECIFIC POLYGENIC RISK SCORES PROVIDING BETTER PREDICTION OF ONSET, PROGRESSION, AND PATHOLOGY THAN POOLED-SEX SCORES. OVER 80% OF INDIVIDUALS WITH ALZHEIMER’S DISEASE HAVE CO-MORBID METABOLIC DISEASE, WHICH EXACERBATES VASCULAR PATHOLOGY. WE HAVE IDENTIFIED THE TOP 50 AD-RISK SNPS AND 600 AD-ASSOCIATED GENES, AND THESE WILL BE TARGETED FOR INDUCED PLURIPOTENT STEM CELL (IPSC)-DERIVED ENDOTHELIAL CELL (EC) SCREENS BY PRIME EDITING AND CRISPR-BASED GENE INHIBITION AND ACTIVATION APPROACHES RESPECTIVELY. IPSC-BASED PRODUCTION OF HUMAN ECS AND MURAL CELLS IN 2D AND 3D MODELS HAS BEEN OPTIMIZED AND SCALED TO ENABLE EFFICIENT FUNCTIONAL TESTING OF THE IMPACT OF GENE CHANGES, INCLUDING ON NEURO-VASCULAR INTERACTIONS IN CEREBRAL ORGANOIDS. DISCOVERIES MADE IN THESE HUMAN CELL SYSTEMS WILL BE VALIDATED BY AN IN-DEPTH INVESTIGATION OF GENE EXPRESSION CHANGES IN INDIVIDUAL ECS AND MURAL CELLS ACROSS A LARGE COLLECTION OF ALZHEIMER’S DISEASE BRAIN SAMPLES USING SINGLE NUCLEAR SEQUENCING. THE EC TRANSLATOME WILL ALSO BE OBTAINED FROM MOUSE ALZHEIMER’S DISEASE MODELS THAT INCORPORATE SEX AND METABOLIC DISEASE. THESE DIVERSE DATASETS WILL BE HARMONIZED AND INTEGRATED IN ORDER TO MAP VASCULAR PHENOTYPES OF AD-RISK GENES AND IDENTIFY CRITICAL MOLECULAR PATHWAYS THAT ARE TARGETABLE DRIVERS OF AD CEREBROVASCULAR PATHOLOGY. THESE DATA WILL ADD TO THE BREADTH OF KNOWLEDGE BEING GATHERED BY OTHER GROUPS TO FURTHER ELUCIDATE UNDERLYING NEURONAL, GLIAL, MICROGLIAL, ENDOTHELIAL AND MURAL CELL-CELL INTERACTIONS THAT CONTRIBUTE IN A SUBSTANTIAL WAY TO THE COMPLEX ARCHITECTURE OF ALZHEIMER DISEASE PATHOLOGY. 1

DEFINING CHARACTERISTICS OF CORTICAL PROGENITOR CELLS OVER TIME IN MOUSE AND HUMAN

PHASE1/2A CLINICAL TRIAL OF RPESC-DERIVED RPE TRANSPLANTATION AS THERAPY FOR NON-EXUDATIVE AGE-RELATED MACULAR DEGENERATION - PROJECT SUMMARY/ABSTRACT AGE-RELATED MACULAR DEGENERATION (AMD) IS A MAJOR CAUSE OF BLINDNESS IN OUR AGING POPULATION. EARLY AMD PATHOGENESIS INVOLVES ATROPHY OF THE RETINAL PIGMENT EPITHELIUM (RPE) WITH ACCOMPANYING LOSS OF RETINAL FUNCTION AND VISION. ALTHOUGH THERAPY IS AVAILABLE FOR EXUDATIVE (WET) AMD, AN EFFECTIVE TREATMENT IS NOT AVAILABLE FOR THE MORE COMMON NON- EXUDATIVE (DRY) AMD FORM. PLURIPOTENT STEM CELL (PSC)-DERIVED RPE (PSC-RPE) TRANSPLANTATION HAS SHOWN PROMISE FOR AMD IN EARLY CLINICAL TRIALS. DUE TO THE HIGHLY PROLIFERATIVE AND PLASTIC NATURE OF PSC, HOWEVER, EXTENSIVE DIFFERENTIATION TO RPE IS NEEDED PRIOR TO TRANSPLANTATION TO AVOID TUMOR GROWTH AND GENOTYPE INSTABILITY INHERENT TO THE PSC SOURCE. WE DISCOVERED AN ADULT RPE STEM CELL (RPESC) WITH RESTRICTED PROLIFERATIVE AND LINEAGE POTENTIAL. RPESC-DERIVED RPE (RPESC- RPE) DO NOT FORM TUMOR ENABLING TRANSPLANTATION OF LESS DIFFERENTIATED RPE AT THE PROGENITOR STAGE. WE FOUND THAT TRANSPLANTED RPE PROGENITOR CELLS WERE MORE EFFECTIVE THAN HIGHLY DIFFERENTIATED PROGENY AT VISION RESCUE IN THE ROYAL COLLEGE OF SURGEONS RAT MODEL OF AMD. RPESC-RPE DIFFERENTIATED FOR 4 WEEKS INTO AN INTERMEDIATE RPE PROGENITOR STAGE RESCUED VISION MORE EFFECTIVELY THAN CELLS DIFFERENTIATED FOR 8 WEEKS INTO THE MATURE RPE PHENOTYPE. THIS IMPROVED EFFICACY COMBINED WITH LACK OF TUMORGENICITY PROVIDES COMPELLING RATIONALE FOR THE PROPOSED PHASE 1/2A CLINICAL TRIAL OF RPESC-RPE TRANSPLANTATION AS THERAPY FOR DRY AMD. EXPERIENCED CLINICAL TRIAL TEAMS HAVE BEEN ASSEMBLED AT THE UNIVERSITY OF MICHIGAN KELLOGG EYE CENTER (KEC) AND STANFORD UNIVERSITY. THE KEC TEAM INCLUDES A VITREORETINAL SURGEON EXPERIENCED IN STEM CELL RESEARCH WHO WILL RECRUIT, PERFORM INTERVENTIONS, AND MANAGE PARTICIPANT CARE. ANOTHER RETINAL SPECIALIST WILL DIRECT POST-INTERVENTION ASSESSMENT AT THE KEC CLINICAL RESEARCH CENTER, AND A SENIOR KEC RETINAL SPECIALIST WILL SERVE AS ON-SITE MEDICAL MONITOR. A CLINICAL TRIALIST HIGHLY EXPERIENCED IN EARLY PHASE OPHTHALMIC TRIALS WILL PROVIDE REGULATORY, DESIGN AND STATISTICAL SUPPORT. THE NEURAL STEM CELL INSTITUTE (NSCI) AND THE STANFORD UNIVERSITY BYERS EYE INSTITUTE WILL WORK WITH KEC TO PROVIDE SCIENTIFIC AND CLINICAL GUIDANCE FOR THE PROPOSED TRIAL. SINGLE CELL TRANSCRIPTOMIC ANALYSES GENERATED AT NSCI WILL BE CORRELATED WITH CLINICAL OUTCOMES, WHICH WILL CONTRIBUTE TO THE RECOGNIZED NEED FOR IMPROVED CELL PRODUCT IDENTITY AND POTENCY MEASURES IN REGENERATIVE MEDICINE GENERALLY. WE PROPOSE TO COMBINE A STRONG PROGRAM IN STEM CELL BIOLOGY WITH AN EXPERIENCED TEAM IN THE CONDUCT OF OPHTHALMIC CLINICAL TRIALS SOUND CLINICAL TRIAL CONDUCT AIMS TO PRODUCE RELIABLE OUTCOMES RESULTS TO EVALUATE RPESC-RPE PROGENITOR CELL TRANSPLANTATION AS THERAPY FOR DRY AMD. OUTCOMES WILL BE CORRELATED WITH PRODUCT IDENTITY AND POTENCY MEASURES AT THE SINGLE CELL LEVEL. COMPLETION OF THE PROPOSED WORK WILL IMPROVE UNDERSTANDING OF RM PRODUCT CHARACTERIZATION AND ADVANCE A UNIQUE TYPE OF ADULT STEM CELL TO REPLACE RPE FOR DRY AMD PATIENT BENEFIT.

IPSC MODELING OF AD USING PROGERIN

CHARACTERIZING HUMAN RPE CELL PROLIFERATION TO ADVANCE ENDOGENOUS REGENERATION - PROJECT SUMMARY / ABSTRACT DECADES OF STUDY HAVE DEMONSTRATED THAT ADULT HUMAN RETINAL PIGMENT EPITHELIUM (RPE) CELLS HAVE STRONG PROLIFERATIVE CAPACITY IN VITRO, WHICH INDICATES THAT THE RPE LAYER HAS THE POSSIBILITY OF SELF-REPAIR. WE HAVE SHOWN THAT ADULT RPE FROM ELDERLY DONORS OR DONORS WITH AGE-RELATED MACULAR DEGENERATION (AMD) CAN PROLIFERATE IN CULTURE AND PRODUCE A NEAR-NATIVE, RENEWED RPE MONOLAYER. DESPITE THIS, RPE CELLS IN VIVO DO NOT REGENERATE THE DAMAGED COBBLESTONE RPE LAYER IN PATIENTS WITH DEGENERATING CELLS, SUCH AS THOSE WITH DRY AMD. THE ENVIRONMENT IN VIVO MUST EFFECTIVELY PREVENT REPAIR OF THE COBBLESTONE RPE MONOLAYER, EITHER THROUGH LACK OF MITOGENS OR INHIBITORY MOLECULES OR A COMBINATION OF BOTH. THE OVERARCHING GOAL OF THESE STUDIES IS TO ACHIEVE SAFE, CONTROLLED PROLIFERATION OF ENDOGENOUS RPE CELLS TO ENABLE SELF-REPAIR OF THE RPE LAYER IN PATIENTS WITH AMD. THE OBJECTIVE OF THIS PROPOSAL IS TO CHARACTERIZE THE ENVIRONMENTAL FACTORS THAT POSITIVELY AND NEGATIVELY CONTROL THE PROLIFERATION OF ADULT HUMAN RPE CELLS. THE FIRST SPECIFIC AIM IS TO CHARACTERIZE THE ADULT HUMAN RPE CELL SURFACEOME ON DIVIDING AND NON-DIVIDING CELLS USING AN INNOVATIVE MASS SPECTROMETRY AND BIOINFORMATIC PLATFORM. THIS WILL PROVIDE THE FIRST COMPREHENSIVE ANALYSIS OF THE MOLECULES ON THE RPE PLASMA MEMBRANE AND REVEAL CELL SURFACE RECEPTORS AND SECRETED PROTEINS THAT RESPOND TO ENVIRONMENTAL FACTORS IMPACTING CELL DIVISION. INVESTIGATING BOTH NORMAL AND AMD RPE WILL PROVIDE A GREATER UNDERSTANDING OF HOW RPE CELLS CHANGE WITH DISEASE. THE SECOND SPECIFIC AIM IS TO EXAMINE A TRANSCRIPTIONAL NETWORK WE HAVE IDENTIFIED THAT IS ASSOCIATED WITH ADULT HUMAN RPE CELL PROLIFERATION TO DETERMINE WHICH MOLECULES ARE CRITICAL. TO DO THIS EFFICIENTLY, WE WILL FIRST EMPLOY CRISPRI, THEN ADDITIONAL FUNCTIONAL SCREENS. THE THIRD SPECIFIC AIM WILL EXAMINE WHETHER EXOGENOUS FACTORS CAN ACTIVATE PROLIFERATION OF QUIESCENT, COBBLESTONE HUMAN RPE, INCLUDING THOSE IN SITU ON BRUCH’S MEMBRANE EXPLANTS, AND THOSE FROM PATIENTS WITH AMD. IN ADDITION TO THE MAIN OBJECTIVE, THIS STUDY WILL GENERATE NEW KNOWLEDGE ABOUT RPE MOLECULES THAT CAN BE USED TO TARGET RPE IN VIVO. THE PROPOSED WORK ALSO HAS THE POTENTIAL TO IMPROVE RPE CELL PROLIFERATION EX VIVO FOR MORE EFFICIENT CELL MANUFACTURING. MOST IMPORTANTLY, THIS STUDY WILL CREATE A FOUNDATION FOR SAFELY STIMULATING RPE CELL PROLIFERATION IN VIVO. ENDOGENOUS ACTIVATION OF RPE CELL PROLIFERATION TO COUNTERACT RPE CELL LOSS IN AMD HAS THE POTENTIAL TO AVOID SURGERY AND IMMUNOSUPPRESSION INVOLVED IN RPE CELL TRANSPLANTATION, WHICH WOULD GREATLY BENEFIT THE ELDERLY AMD PATIENT POPULATION.

DEFINING THE MOLECULAR MECHANISMS UNDERLYING HUMAN RPE PLASTICITY

ASSESSMENT OF SELECTIVE INHIBITORS OF NUCLEAR EXPORT (SINE) FOLLOWING SPINAL CORD INJURY.

CELL TYPE AND REGIONAL VULNERABILITY IN FRONTOTEMPORAL DEMENTIA - PROJECT SUMMARY/ABSTRACT A KEY QUESTION IN TAUOPATHY RESEARCH IS WHY SOME BRAIN CELL POPULATIONS ARE SIGNIFICANTLY AFFECTED WHILE OTHERS ARE RELATIVELY SPARED. THE LONG-TERM GOAL OF THIS STUDY IS TO UNDERSTAND THIS DIFFERENTIAL CELL VULNERABILITY AND USE THE KNOWLEDGE TO DEVELOP THERAPIES THAT PROTECT NEURAL CELLS FROM TAUOPATHY-RELATED DEGENERATION. MUTATIONS IN THE MAPT GENE THAT ENCODES TAU COMMONLY CAUSE EXTENSIVE PATHOLOGY IN THE FOREBRAIN, WITH SIGNIFICANT LOSS OF FRONTAL AND TEMPORAL LOBE CEREBRAL CORTICAL CELLS LEADING TO BEHAVIORAL, LANGUAGE AND COGNITIVE DEFICITS. HOWEVER, A SUBSET OF MAPT MUTATIONS ALSO CAUSE SIGNIFICANT DEGENERATION OF MIDBRAIN DOPAMINERGIC NEURONS IN THE SUBSTANTIA NIGRA CONTRIBUTING TO A PARKINSONISM PHENOTYPE. FURTHERMORE, THESE CELLS ARE CONNECTED: MIDBRAIN DOPAMINERGIC NEURONS WIDELY INNERVATE THE PREFRONTAL CORTEX AND ARE RECIPROCALLY INNERVATED VIA CORTICO-STRIATAL-NIGRAL CIRCUITS. WHY SPECIFIC MAPT MUTATIONS SIGNIFICANTLY AFFECT THE MIDBRAIN IN ADDITION TO CORTEX WHILE OTHERS DO NOT, AND HOW CONNECTIVITY BETWEEN THESE REGIONS WITH THE POTENTIAL FOR PATHOLOGICAL TAU SPREAD MAY BE INVOLVED, REPRESENT SIGNIFICANT GAPS IN KNOWLEDGE THAT WE WILL ADDRESS IN THIS PROPOSED STUDY. OVER THE PAST FEW YEARS, WE HAVE HELPED CREATE A LARGE IPSC LINE COLLECTION FROM PATIENTS WITH FAMILIAL DEMENTIA DUE TO MUTATIONS IN THE MAPT GENE, INCLUDING ISOGENIC CONTROLS. PHENOTYPIC ANALYSES SHOW MAPT MUTANT AND CONTROL IPSC-DERIVED CEREBRAL CORTICAL CELLS ARE INITIALLY PHENOTYPICALLY SIMILAR BUT DEVELOP DIFFERENCES WITH MATURATION THAT INCLUDE INCREASED TAU AGGREGATION, TAU HYPERPHOSPHORYLATION AND VULNERABILITY TO SEVERAL STRESSORS, ASSOCIATED WITH THE MUTATION. HOWEVER, TO DATE, STUDIES COMPARING FOREBRAIN AND MIDBRAIN CELL POPULATION RESPONSES TO MAPT MUTATIONS THAT DIFFERENTIALLY AFFECT THESE BRAIN REGIONS HAVE NOT BEEN DONE. SUCH COMPARISONS HAVE THE POTENTIAL TO REVEAL COMMON AND UNIQUE MOLECULAR MECHANISMS THAT UNDERLIE CELL VULNERABILITY. OUR APPROACH IS TO USE HUMAN IPSC-DERIVED 3D ORGANOIDS, WHICH RECAPITULATE COMPLEX CELL-CELL INTERACTIONS IN A HUMAN CELL SYSTEM AND ENABLE LONG-TERM CULTURE OVER SEVERAL MONTHS. WE WILL CREATE CORTICAL AND MIDBRAIN ORGANOIDS FROM TWO MAPT MUTATIONS THAT PRIMARILY AFFECT CORTEX AND TWO THAT AFFECT BOTH CORTEX AND MIDBRAIN, VERSUS RESPECTIVE ISOGENIC CONTROLS. IN AIM 1 WE WILL EXAMINE THE IMPACT OF THESE MAPT MUTATIONS ON CELL POPULATIONS AND GENE EXPRESSION OVER TIME USING SINGLE CELL TRANSCRIPTOMICS TO DEFINE HOW DIVERSE CELL TYPES RESPOND TO EACH MUTATION. IN AIM 2, WE WILL CREATE ASSEMBLOIDS OF CORTICAL AND MIDBRAIN ORGANOIDS TO MODEL THE CIRCUITRY BETWEEN THE REGIONS AND DETERMINE WHETHER THIS CONNECTIVITY ALTERS PATTERNS OF CELL VULNERABILITY AND ENABLES THE SPREAD OF PATHOLOGICAL TAU FROM ONE REGION TO ANOTHER, DEPENDING ON SPECIFIC MAPT MUTATION. IN AIM 3 WE WILL PROBE THE IMPACT OF STIMULATING TAU DEGRADATION ON DIFFERENTIAL CELL VULNERABILITY IN CEREBRAL CORTEX AND MIDBRAIN.

QUANTIFYING CHANGES IN NEURAL STEM CELL LINEAGES IN THE AGING NICHE

ASYMMETRIC CELL DIVISION OF CNS PROGENITOR CELLS

HUMAN IPSC MODEL OF CEREBRO-VASCULAR INTERACTIONS IN ADRD - WE PROPOSE TO GENERATE AND VALIDATE A NOVEL MODEL THAT INCORPORATES HUMAN CEREBRAL CORTICAL NEURONS AND GLIA, VASCULAR ENDOTHELIAL CELLS, NEURAL CREST PERICYTES AND MICROGLIA FROM HUMAN INDUCED PLURIPOTENT STEM CELLS (IPSCS). THE GOAL IS A MODEL WITH IMPROVED CONSTRUCT, FACE, AND PREDICTIVE VALIDITY TO STUDY THE CEREBRO-VASCULAR INTERACTIONS COMMON TO ALZHEIMER’S DISEASE-RELATED DEMENTIAS (ADRD). THE NINDS 2022 ADRD SUMMIT REPORT DESCRIBES THE NEED FOR SUCH A MODEL. IN PRIOR STUDIES WE DEVELOPED A REPRODUCIBLE HUMAN 3D CEREBRAL CORTEX ORGANOID MODEL TESTED ACROSS 63 IPSC LINES WITH 90% SUCCESS. IT IMPROVED EFFICIENCY IN ORGANOID PRODUCTION FROM 10% TO APPROACHING 100%. ORGANOIDS GROWN WITH THE PROTOCOL HAVE UNIFORM SIZE, SHAPE, PREDICTABLE GROWTH CURVES AND CELL COMPOSITION. SINGLE CELL TRANSCRIPTOME STUDIES DEMONSTRATE HIGH OVERLAP BETWEEN IPSC LINES FROM DIFFERENT DONORS, AN IMPORTANT ADVANCE FOR STUDIES OF BOTH GENETIC AND SPORADIC DISEASE. WE ALSO RECENTLY DEVELOPED AN IPSC-DERIVED 3D VASCULATURE MODEL FORMING ROBUST, VESSEL-LIKE STRUCTURES OF ENDOTHELIAL CELLS AND PERICYTES IN AN INERT HYDROGEL. PRELIMINARY STUDIES DEMONSTRATE THAT THESE TWO ADVANCED MODELS CAN BE INTEGRATED TO GENERATE A HUMAN CELL 3D CEREBRO-VASCULAR MODEL. IN AIM 1, THE R61 PHASE, WE WILL OPTIMIZE THE INCORPORATION OF THESE TWO MODELS TO ACHIEVE GREATEST REPRODUCIBILITY OF CELL COMPOSITION REFLECTING VASCULARIZED CEREBRAL CORTICAL TISSUE IN VIVO. THE MODEL WILL BE GENERATED FROM AT LEAST THREE DIFFERENT PSEN1 MUTATION IPSC LINES VERSUS HEALTHY CONTROLS. THERE IS SIGNIFICANT NEED FOR THIS MODEL GIVEN THAT CURRENT PSEN1 MUTATION TRANSGENIC AND KNOCK-IN MOUSE MODELS LACK SEVERAL KEY FEATURES OF NEURO- AND VASCULAR PATHOLOGY SEEN IN HUMAN DISEASE. QUANTITATIVE METRICS WILL BE USED TO ASSESS EFFICIENCY AND REPRODUCIBILITY OF THE MODEL AND ITS ABILITY TO REPRODUCE PSEN1 MUTATION CORTICAL PATHOLOGY, INCLUDING CEREBRAL AMYLOID ANGIOPATHY (CAA). WITH THE ACHIEVEMENT OF PROSPECTIVELY DETERMINED QUANTITATIVE CRITERIA, WE WILL ADVANCE TO THE R33 PHASE OF THE PROJECT. IN AIM 2, THE PSEN1 MUTATION MODELS WILL BE EXAMINED FOR FACE, CONSTRUCT, AND PREDICTIVE VALIDITY. THIS PHASE WILL INCLUDE COLLABORATORS WITH EXPERTISE IN EXAMINING PSEN1 MUTATION CARRIER TISSUE PATHOLOGY, BRAIN SINGLE NUCLEAR TRANSCRIPTOMES, METABOLOMICS, LIPIDOMICS, AND CEREBROSPINAL FLUID (CSF) COMPOSITION. WE WILL ASSESS ESTABLISHED CHARACTERISTICS OF PSEN1 MUTATION-INDUCED CEREBRAL PATHOLOGY, SUCH AS ALTERED EXPRESSION OF A40,42 AND 43, AND TAU. VALIDATION TESTS WILL INCLUDE THOSE USED FOR CLINICAL EVALUATION WHERE TEST SENSITIVITY, DYNAMIC RANGE AND PREDICTIVE VALUE ARE KNOWN. THE PREDICTIVE VALIDITY OF THE MODEL WILL BE ASSESSED BY COMPARING PSEN1 MUTATION TO GENETIC CONTROLS, INCLUDING CRISPR ENGINEERED ISOGENIC CONTROL LINES. IN ADDITION, WE WILL ASSESS THE IMPACT OF ANTIBODY THERAPIES THAT ARE APPROVED FOR PSEN1 MUTATION CARRIERS, TO DETERMINE WHETHER THEY CAN BIND A SPECIES, OFFSET OTHER SIGNS OF PATHOLOGY, AND OFFER INSIGHT PERTINENT TO DRUG-RELATED VASCULAR SIDE-EFFECTS. WE WILL DEMONSTRATE INDEPENDENT REPLICATION OF THE MODEL IN A DIFFERENT INSTITUTION LAB TO DEMONSTRATE ROBUSTNESS. TRANSPARENT SHARING OF DETAILED PROTOCOLS AND DATA WILL AID ADOPTION OF THE MODEL.

NEUROVASCULAR STEM CELL NICHE IN THE ADULT SVZ

CHARACTERIZATION OF HUMAN RPE SUBPOPULATIONS AT THE SINGLE CELL LEVEL

IPSC-BASED MODEL TO STUDY FRONTOTEMPORAL DEMENTIA DUE TO MAPT MUTATION - OUR GOAL IS TO GENERATE AND VALIDATE A NOVEL, CELLULARLY COMPLEX MODEL OF FRONTOTEMPORAL DEMENTIA (FTD) DUE TO MUTATIONS IN THE MAPT GENE THAT ENCODES TAU (FTD-TAU). THE NINDS 2022 ADRD SUMMIT REPORT DESCRIBES THE NEED FOR SUCH A MODEL. OVER 50 MUTATIONS IN MAPT CAUSE FTD-TAU, WHICH MANIFESTS BY ABNORMAL TAU ACCUMULATION IN SUBSETS OF CEREBRAL CORTICAL NEURONS, ASTROCYTES, AND OLIGODENDROCYTES. THIS LEADS TO CELLULAR DYSFUNCTION AND DEATH, PRIMARILY IN THE FRONTAL AND TEMPORAL CORTICAL REGIONS, LEADING TO DEFICITS IN EXECUTIVE FUNCTION, LANGUAGE, AND BEHAVIORAL RESPONSES. MAPT MUTATION CARRIERS FREQUENTLY ALSO HAVE SIGNIFICANT DEGENERATION OF SUBCORTICAL AREAS, INCLUDING THE STRIATUM AND THE MIDBRAIN DOPAMINERGIC SUBSTANTIA NIGRA (SNC) AND MIDBRAIN VENTRAL TEGMENTAL AREA (VTA) NEURONS, CONTRIBUTING TO PARKINSONISM AND BEHAVIORAL CHANGES. HENCE THERE IS NEED TO MODEL MULTIPLE BRAIN AREAS TO MORE FULLY CAPTURE BRAIN CELL VULNERABILITY. FURTHERMORE, THERE ARE EXTENSIVE CONNECTIONS BETWEEN THESE MIDBRAIN AND FOREBRAIN AREA NEURONS THAT ARE AFFECTED IN FTD- TAU, WHICH IMPLICATES THIS CIRCUITRY IN PATHOLOGY DEVELOPMENT AND SPREAD. PRIOR 3D HUMAN CELL MODELS OF FTD- TAU HAVE FOCUSED ON THE CEREBRAL CORTEX NEURONS AND GLIA, WHICH DEMONSTRATE KEY FEATURES OF CORTICAL TAUOPATHY, INCLUDING ACCUMULATION OF ABNORMALLY PHOSPHORYLATED TAU AND DEGENERATION OF GLUTAMATERGIC NEURONS. HERE, WE PROPOSE TO BUILD A MODEL OF AXON-CONNECTED 3D CEREBRAL CORTEX, STRIATUM, AND MIDBRAIN ORGANOIDS THAT INCORPORATES THE MAJOR CELL TYPES AND RECONSTRUCTS KEY ELEMENTS OF THE FOREBRAIN-MIDBRAIN CIRCUITY AFFECTED IN FTD-TAU. TO FURTHER ADVANCE THE MODEL AND INCLUDE CRITICAL CELL TYPES IMPLICATED IN FTD-TAU, IT WILL INCORPORATE MICROGLIA. THE GOAL IS A HUMAN CELL MODEL OF AFFECTED BRAIN REGIONS THAT EXHIBITS IMPROVED CONSTRUCT AND FACE VALIDITY OVER EXISTING MODELS. IN AIM 1 WE WILL DEVELOP AND OPTIMIZE PRODUCTION AND CONNECTIVITY OF THE CORTICAL- STRIATAL-MIDBRAIN (COST-MB) MODEL. IN AIM 2 WE WILL GENERATE COST-MB MODELS FROM DIFFERENT MAPT MUTATIONS VS ISOGENIC CONTROL LINES. WE WILL DEEPLY PHENOTYPE THE MODELS AND COMPARE THEM TO PATIENT BRAIN SAMPLES WITH MAPT MUTATIONS IN FACE AND CONSTRUCT VALIDATION STUDIES. QUANTITATIVE METRICS WILL BE USED TO ASSESS REPRODUCIBILITY OF THE MODEL AND ITS PHENOTYPIC RESEMBLANCE TO PATIENT FOREBRAIN AND MIDBRAIN PATHOLOGY. WE WILL DEMONSTRATE INDEPENDENT REPLICATION OF THE MODEL IN MORE THAN ONE INSTITUTION LAB TO DEMONSTRATE ROBUSTNESS. TRANSPARENT SHARING OF DETAILED PROTOCOLS AND DATA WILL AID ADOPTION OF THE COST-MB FTD-TAU MODEL. THE MODEL WILL BE GENERATED USING OUR COLLECTION OF HUMAN INDUCED PLURIPOTENT STEM CELL (IPSC) LINES WITH MAPT MUTATIONS IN CODING AND NON-CODING REGIONS THAT AFFECT BOTH FOREBRAIN AND MIDBRAIN PATHOLOGY AND CRISPR-CORRECTED ISOGENIC CONTROL LINES; THESE LINES ARE FULLY AVAILABLE TO OTHER RESEARCHERS. THE COST-MB MODEL CAN BE USED IN THE FUTURE TO STUDY DISEASE MECHANISMS AND DEVELOP THERAPEUTICS TO COMBAT FTD-TAU AND POTENTIALLY OTHER TAUOPATHIES OR RELATED NEURODEGENERATIVE DISEASES THAT CAUSE MULTI-REGION BRAIN CELL LOSS.

CHARACTERIZING THE MOLECULAR DETERMINANTS OF HUMAN NEURAL SPECIFICATION

CELL TYPE AND REGIONAL VULNERABILITY IN FRONTOTEMPORAL DEMENTIA - PROJECT SUMMARY/ABSTRACT A KEY QUESTION IN TAUOPATHY RESEARCH IS WHY SOME BRAIN CELL POPULATIONS ARE SIGNIFICANTLY AFFECTED WHILE OTHERS ARE RELATIVELY SPARED. THE LONG-TERM GOAL OF THIS STUDY IS TO UNDERSTAND THIS DIFFERENTIAL CELL VULNERABILITY AND USE THE KNOWLEDGE TO DEVELOP THERAPIES THAT PROTECT NEURAL CELLS FROM TAUOPATHY-RELATED DEGENERATION. MUTATIONS IN THE MAPT GENE THAT ENCODES TAU COMMONLY CAUSE EXTENSIVE PATHOLOGY IN THE FOREBRAIN, WITH SIGNIFICANT LOSS OF FRONTAL AND TEMPORAL LOBE CEREBRAL CORTICAL CELLS LEADING TO BEHAVIORAL, LANGUAGE AND COGNITIVE DEFICITS. HOWEVER, A SUBSET OF MAPT MUTATIONS ALSO CAUSE SIGNIFICANT DEGENERATION OF MIDBRAIN DOPAMINERGIC NEURONS IN THE SUBSTANTIA NIGRA CONTRIBUTING TO A PARKINSONISM PHENOTYPE. FURTHERMORE, THESE CELLS ARE CONNECTED: MIDBRAIN DOPAMINERGIC NEURONS WIDELY INNERVATE THE PREFRONTAL CORTEX AND ARE RECIPROCALLY INNERVATED VIA CORTICO-STRIATAL-NIGRAL CIRCUITS. WHY SPECIFIC MAPT MUTATIONS SIGNIFICANTLY AFFECT THE MIDBRAIN IN ADDITION TO CORTEX WHILE OTHERS DO NOT, AND HOW CONNECTIVITY BETWEEN THESE REGIONS WITH THE POTENTIAL FOR PATHOLOGICAL TAU SPREAD MAY BE INVOLVED, REPRESENT SIGNIFICANT GAPS IN KNOWLEDGE THAT WE WILL ADDRESS IN THIS PROPOSED STUDY. OVER THE PAST FEW YEARS, WE HAVE HELPED CREATE A LARGE IPSC LINE COLLECTION FROM PATIENTS WITH FAMILIAL DEMENTIA DUE TO MUTATIONS IN THE MAPT GENE, INCLUDING ISOGENIC CONTROLS. PHENOTYPIC ANALYSES SHOW MAPT MUTANT AND CONTROL IPSC-DERIVED CEREBRAL CORTICAL CELLS ARE INITIALLY PHENOTYPICALLY SIMILAR BUT DEVELOP DIFFERENCES WITH MATURATION THAT INCLUDE INCREASED TAU AGGREGATION, TAU HYPERPHOSPHORYLATION AND VULNERABILITY TO SEVERAL STRESSORS, ASSOCIATED WITH THE MUTATION. HOWEVER, TO DATE, STUDIES COMPARING FOREBRAIN AND MIDBRAIN CELL POPULATION RESPONSES TO MAPT MUTATIONS THAT DIFFERENTIALLY AFFECT THESE BRAIN REGIONS HAVE NOT BEEN DONE. SUCH COMPARISONS HAVE THE POTENTIAL TO REVEAL COMMON AND UNIQUE MOLECULAR MECHANISMS THAT UNDERLIE CELL VULNERABILITY. OUR APPROACH IS TO USE HUMAN IPSC-DERIVED 3D ORGANOIDS, WHICH RECAPITULATE COMPLEX CELL-CELL INTERACTIONS IN A HUMAN CELL SYSTEM AND ENABLE LONG-TERM CULTURE OVER SEVERAL MONTHS. WE WILL CREATE CORTICAL AND MIDBRAIN ORGANOIDS FROM TWO MAPT MUTATIONS THAT PRIMARILY AFFECT CORTEX AND TWO THAT AFFECT BOTH CORTEX AND MIDBRAIN, VERSUS RESPECTIVE ISOGENIC CONTROLS. IN AIM 1 WE WILL EXAMINE THE IMPACT OF THESE MAPT MUTATIONS ON CELL POPULATIONS AND GENE EXPRESSION OVER TIME USING SINGLE CELL TRANSCRIPTOMICS TO DEFINE HOW DIVERSE CELL TYPES RESPOND TO EACH MUTATION. IN AIM 2, WE WILL CREATE ASSEMBLOIDS OF CORTICAL AND MIDBRAIN ORGANOIDS TO MODEL THE CIRCUITRY BETWEEN THE REGIONS AND DETERMINE WHETHER THIS CONNECTIVITY ALTERS PATTERNS OF CELL VULNERABILITY AND ENABLES THE SPREAD OF PATHOLOGICAL TAU FROM ONE REGION TO ANOTHER, DEPENDING ON SPECIFIC MAPT MUTATION. IN AIM 3 WE WILL PROBE THE IMPACT OF STIMULATING TAU DEGRADATION ON DIFFERENTIAL CELL VULNERABILITY IN CEREBRAL CORTEX AND MIDBRAIN.

TRANSPLANTATION OF ADULT, TISSUE-SPECIFIC RPE STEM CELLS AS THERAPY FOR NON-EXUDATIVE AGE-RELATED MACULAR DEGENERATION AMD

STEM CELLS IN THE CNS

MODELING NF2 TUMORS FOR DRUG SCREENING USING INDUCED PLURIPOTENT STEM CELLS

IPSC LINES FOR MODELING AGE-RELATED MACULAR DEGENERATION

EFFECT OF NF1 MUTATION ON CHOROID PLEXUS FUNCTION

DEVELOPING ANTIBODY-OLIGONUCLEOTIDE BRIDGES TO SIMPLIFY SINGLE CELL SPATIAL TRANSCRIPTOMICS - PROJECT SUMMARY SINGLE CELL RNA-SEQUENCING (SCRNA-SEQ) HAS ENABLED RESEARCHERS TO INVESTIGATE A WIDE ARRAY OF BIOLOGICAL PROCESSES AND HOW TISSUE HETEROGENEITY CONTRIBUTES TO FUNCTION. THESE TECHNOLOGIES HAVE LED TO THE DEVELOPMENT OF PROGRAMS, SUCH AS THE NIH HUMAN BIOMOLECULAR ATLAS PROGRAM, TO UNDERSTAND HOW CELLS INTERACT IN AN ORGANISM TO DRIVE ITS FUNCTION. THE DEVELOPMENT OF SPATIAL TRANSCRIPTOMIC TECHNOLOGIES HAS ALLOWED RESEARCHERS TO INVESTIGATE HOW CELL-CELL INTERACTIONS AFFECT CELLULAR GENE EXPRESSION WITHIN A TISSUE. CURRENT APPROACHES GENERALLY RELY IN SITU RNA BASED METHODOLGIES OR SLIDE-BASED SEQUENCING IN COMBINATION WITH SCRNA-SEQ (E.G. SLIDE-SEQ) TO GENERATE SPATIAL MAPS OF INTERACTING CELLS. THESE TECHNOLOGIES HAVE HELPED DEMONSTRATE THE POWER OF COMBINING THE TISSUES STRUCTURAL DATA WITH TRANSCRIPTIONAL DATA TO BETTER UNDERSTAND CELL BEHAVIOR IN A TISSUE AND GOING FORWARD IMPROVING THE INTERFACE BETWEEN THESE TECHNOLOGIES WILL BE KEY TO UNCOVERING HOW STRUCTURE AND CELL TRANSCRIPTION DRIVES FUNCTION. HOWEVER, MULTIPLE BARRIERS IMPEDE THE WIDESPREAD ADOPTION OF THE CURRENT METHODS FOR PERFORMING SPATIAL TRANSCRIPTOMICS, INCLUDING SIGNIFICANT FINANCIAL COST, TIME REQUIREMENTS, AND A LACK OF SUFFICIENT EXPERTISE. HEREIN, WE PROPOSE TO DEVELOP A SIMPLE REAGENT COMPATIBLE WITH CURRENT PROTOCOLS FOR DROP-BASED AND PIPETTE-BASED SCRNA-SEQ TECHNOLOGIES. THIS REAGENT WILL BE ABLE TO GENERATE SPATIAL DATA IN THE NORMAL COURSE OF SCRNA-SEQ EXPERIMENTS WITHOUT SIGNIFICANTLY INCREASING COST OR TIME COMMITMENTS. WE PROPOSE TO INVENT “ANTIBODY-OLIGONUCLEOTIDE BRIDGES” AND BUILD SOFTWARE CAPABLE OF BIOINFORMATICALLY GENERATING A NETWORK OF CELL-CELL CONTACTS ACROSS THE TISSUE TO RECAPITULATE THE SPATIAL RELATIONSHIPS OF THE CELLS IN A TISSUE. IN THIS PROPOSAL, WE WILL CARRY OUT A PROOF-OF- PRINCIPAL EXPERIMENT FOR THE AO BRIDGE SCRNA-SEQ APPROACH FOR GENERATING THE SPATIAL RELATIONSHIPS BETWEEN CELLS.

A HUMAN IPSC-DERIVED MICROPHYSIOLOGICAL 3D MODEL TO STUDY CAA/AD - SUMMARY/ABSTRACT VASCULAR PATHOLOGY HAS BEEN IDENTIFIED AS A CRITICAL DRIVER OF ALZHEIMER’S DISEASE. THIS PROPOSAL AIMS TO ESTABLISH A HUMAN INDUCED PLURIPOTENT STEM CELL (IPSC)-DERIVED MODEL TO STUDY THE IMPACT OF GENETIC AND ENVIRONMENTAL FACTORS ON THE DEVELOPMENT OF ALZHEIMER’S DISEASE-RELATED VASCULAR PATHOLOGY. MOST ALZHEIMER’S DISEASE PATIENTS HAVE CEREBRAL AMYLOID ANGIOPATHY (CAA), THE SYMPTOMS OF WHICH INCLUDE BUILD-UP OF AMYLOID AROUND VESSELS, CEREBRAL HEMORRHAGES, MICROBLEEDS, AND INFLAMMATION. STUDIES IN MULTIPLE MOUSE MODELS OF ALZHEIMER’S DISEASE HAVE DEMONSTRATED THAT HEMOPOIETIC-DERIVED BRAIN-RESIDENT MICROGLIA AND CIRCULATING MONOCYTES AND PERIVASCULAR MACROPHAGES CAN BE PROTECTIVE AGAINST CAA. THE APOE44 GENOTYPE IS THE STRONGEST GENETIC RISK FACTOR FOR ALZHEIMER’S DISEASE AND CAA. DESPITE THE CLEAR INTERPLAY BETWEEN ALZHEIMER’S DISEASE, CAA, AND APOE GENOTYPE, THE MOLECULAR MECHANISMS UNDERLYING THE VASCULAR CHANGES ARE NOT FULLY UNDERSTOOD. THESE OBSERVATIONS MOTIVATE US TO CREATE A MODEL USING HUMAN CELLS TO STUDY THE UNDERLYING MECHANISMS OF MICROGLIA- VASCULAR INTERACTIONS IN CELLS OF DIFFERENT APOE GENOTYPES. IN PRIOR PUBLISHED STUDIES, WE CREATED A 3D HUMAN IPSC-DERIVED VASCULAR MODEL IN A HYDROGEL SCAFFOLD. IT UNDERGOES VASCULOGENIC AND ANGIOGENIC EVENTS AND FORMS A MODEL PLEXUS (THE VAMP MODEL). WE SHOWED THAT THE VAMP MODEL CAN BE BUILT INSIDE A MICROFLUIDIC DEVICE TO ENABLE REGULATABLE PERFUSION, MIMICKING BLOOD FLOW. WE HAVE ADVANCED THE MODEL BY INCORPORATING MICROGLIA (VAMP-MG) TO REFLECT THE KEY CELL-CELL INTERACTIONS IN CAA BETTER. THE WORK PROPOSED IN TWO SPECIFIC AIMS WILL INCREASE THE UTILITY OF THE VAMP-MG MODEL IN FIVE SIGNIFICANT WAYS: WE WILL 1) BUILD THE MODEL FROM A RECENTLY ESTABLISHED COLLECTION OF APOE44 VERSUS APOE33 ISOGENIC IPSC LINES TO INCORPORATE GENETIC RISK FACTORS; 2) INCORPORATE LIVE REPORTERS TO MONITOR THE ACTIVATION STATES OF MICROGLIA AND VASCULAR CELLS IN REAL-TIME; 3) CREATE THE VAMP-MG MODEL IN MICROFLUIDIC CHIPS TO ACHIEVE PERFUSABLE VASCULATURE; 4) FLOW HUMAN PLASMA FROM ALZHEIMER’S DISEASE, AGED-MATCHED HEALTHY, OR YOUNG HEALTHY DONORS THROUGH THE VESSELS TO IMPROVE MODELING OF DISEASE-RELEVANT ENVIRONMENTAL FACTORS; 5) USE RIBO-TAG TECHNOLOGY TO IDENTIFY TRANSCRIPTOMIC CHANGES IN THE VASCULAR ENDOTHELIAL CELLS AND THE MICROGLIA. THE MODEL WILL BE DEEPLY CHARACTERIZED AND VALIDATED AT CELLULAR AND MOLECULAR LEVELS. ALZHEIMER’S DISEASE-RELEVANT PHENOTYPES, INCLUDING AMYLOID DEPOSITION AND CLEARANCE, A SECRETION, AND INFLAMMATORY FACTOR PRODUCTION, WILL BE ASSESSED. WE WILL COMPARE TRANSCRIPTOMIC DATA COLLECTED FROM THE MODEL TO PUBLISHED HUMAN ALZHEIMER’S DISEASE BRAIN VERSUS HEALTHY CONTROL BRAIN TRANSCRIPTOMIC DATA, INCLUDING SINGLE NUCLEAR RNA-SEQ DATASETS. COMPLETING THE PROPOSED WORK WILL ENHANCE OUR ABILITY TO STUDY VASCULAR AND MICROGLIAL RESPONSES TO INFLAMMATORY SIGNALS IN REAL TIME, GENERATE TRANSCRIPTOMIC DATA AT CELL TYPE LEVELS, AND ESTABLISH APOE44 VERSUS APOE33 PHENOTYPES IN MICROGLIA-VASCULAR MODELS TO A NEW LEVEL OF COMPLEXITY. IN THE FUTURE, THIS VAMP-MG MODEL CAN POTENTIALLY DEFINE KEY GENETIC AND ENVIRONMENTAL FACTORS THAT EXACERBATE OR ALLEVIATE ALZHEIMER’S DISEASE-VASCULAR PHENOTYPES.

HIGH-DENSITY MICROELECTRODE ARRAY - PI: KIEHL, THOMAS R. , REGENERATIVE RESEARCH FOUNDATION, HIGH-DENSITY MICROELECTRODE ARRAY SUMMARY/ABSTRACT WITH THIS APPLICATION, WE REQUEST FUNDING TO PROCURE THE HYPERCAM ALPHA PLATFORM FROM 3BRAIN AG (ZÜRICH SWITZERLAND). THE HYPERCAM ALPHA IS A MULTI-WELL, HIGH-DENSITY MICROELECTRODE ARRAY SYSTEM FOR OBSERVING AND STIMULATING IN VITRO ELECTROPHYSIOLOGICAL ACTIVITY IN ELECTRO-ACTIVE CELL CULTURES, CELL CONSTRUCTS, ORGANOIDS, AND PRIMARY TISSUE SAMPLES. THIS INSTRUMENT PROVIDES SEVERAL KEY FEATURES: 1) HIGH-RESOLUTION RECORDING CAPABILITY; 2) DYNAMIC STIMULATION FUNCTIONALITY, 3) A MULTI-WELL FORMAT; 4) 3D SENSOR OPTIONS, AND 5) SOPHISTICATED ANALYSIS SOFTWARE. THESE FEATURES ENSURE ITS UTILITY ACROSS A RANGE OF BIOLOGICAL INVESTIGATIONS TO MEET THE HIGH-THROUGHPUT AND HIGH-FIDELITY DEMANDS OF OUR USERS AND COLLABORATORS. THIS SUBMISSION ADDRESSES A SIGNIFICANT UNMET NEED IN OUR INSTITUTE AND REGION, NAMELY THE ABILITY TO TEST THE FUNCTIONAL IMPACT OF CELLULAR INTERACTIONS, GENETIC CHANGES OR OF PERTURBAGENS ON NEURAL SYSTEMS AT SCALE. THE RESEARCHERS IDENTIFIED IN THIS SUBMISSION (SPANNING FIVE INSTITUTIONS) ARE ENGAGED IN THE STUDY OF A WIDE RANGE OF NEUROLOGICAL DISFUNCTION FROM DISEASE, AGING, INJURY AND DEVELOPMENTAL DISORDERS. PLACEMENT OF THIS INSTRUMENT WILL DIRECTLY ADVANCE WORK IN FRONTOTEMPORAL DEMENTIA, ALZHEIMER’S, VASCULAR CONTRIBUTIONS TO DEMENTIA, SEX- SPECIFIC AGING IMPACT ON COGNITIVE FUNCTION, TISSUE REGENERATION, SPINAL CORD INJURY, BRAIN DEVELOPMENT, AND DISEASE IMPACT ON NEUROLOGICAL DEVELOPMENT. THIS INSTRUMENT WILL HELP TO QUANTIFY MODEL SYSTEMS, SUCH AS BRAIN ORGANOIDS, THAT HAVE FAST BECOME THE WORKHORSES OF THE FIELD. PROJECTS DESCRIBED HERE ALSO ADVANCE THE DEVELOPMENT OF NEW, MORE CAPABLE, MODEL SYSTEMS AND EVEN OF NEXT GENERATION MEA-RELATED DEVICES. THE PRIMARY, CONCRETE, LONG-RANGE BENEFIT OF THIS INSTRUMENT TO THE BIOMEDICAL RESEARCH GOALS OF RRF/NSCI/NEURACELL IS TO PROVIDE ROUTINE, CLEAR, HIGH-RESOLUTION, EASILY CAPTURED, CHARACTERIZATION AND QUANTIFICATION OF THE BEHAVIOR AND FUNCTION OF THE NUMEROUS NEUROLOGICAL SYSTEMS UNDER STUDY AT OUR INSTITUTION. THE USE OF THIS INSTRUMENT WILL BE INTEGRATED INTO NEARLY EVERY RESEARCH PROGRAM WITHIN OUR WALLS. BEYOND OUR ORGANIZATION, THIS INSTRUMENT WILL: AMPLIFY THE EXISTING PROGRAMS OF OUR MAJOR USERS; HELP OUR MINOR USERS ESTABLISH AND GROW THEIR RELATED RESEARCH PROGRAMS; AND ENDOW A FOUNDATION FOR FUTURE TECHNOLOGY DEVELOPMENT.

ROLE OF STAUFEN2 IN ASYMMETRIC DIVISION OF MOUSE CORTICAL PROGENITOR CELLS

ENDOTHELIAL AROMATASE IN SEX-SPECIFIC CEREBROVASCULAR DYSFUNCTION AFTER ISCHEMIA

PHASE1/2A CLINICAL TRIAL OF RPESC-DERIVED RPE TRANSPLANTATION AS THERAPY FOR NON-EXUDATIVE AGE-RELATED MACULAR DEGENERATION

HUMAN IPSC-BASED MODELING OF RPE MELANOGENESIS: REACTIVATION AND PROTECTION