RF1NS144660

MECHANISM OF ALS/FTD DETERMINED BY MULTIMODAL SINGLE CELL SPATIAL TRANSCRIPTOMICS - PROJECT SUMMARY ABNORMAL CYTOPLASMIC ACCUMULATION AND NUCLEAR DEPLETION OF THE RNA-BINDING PROTEIN TDP-43, COLLECTIVELY REFERRED TO AS TDP-43 PROTEINOPATHY, HAS BEEN REPORTED IN 40% OF FRONTAL TEMPORAL DEMENTIA (FTD), 97% OF INSTANCES OF AMYOTROPHIC LATERAL SCLEROSIS (ALS), 50% OF ALZHEIMER'S DISEASE (AD), AND 100% OF LIMBIC- PREDOMINANT AGE-RELATED TDP-43 ENCEPHALOPATHY (LATE). ACROSS THIS DISEASE SPECTRUM, NEURONS DEVELOP DISTINCT STAGES OF TDP-43 MISLOCALIZATION AND AGGREGATION. A MAJOR GENETIC CAUSE OF FTD AND ALS IS A GGGGCC (G4C2) HEXANUCLEOTIDE EXPANSION IN THE C9ORF72 GENE (C9ORF72 IN MICE). WHILE A NORMAL HUMAN C9ORF72 ALLELE HAS LESS THAN 20 G4C2 REPEATS, PATHOGENIC REPEATS IN DISEASE ARE IN THE HUNDREDS, WITH SOMATIC EXPANSION UP TO 4000 REPEATS. THERE IS CONSENSUS THAT TDP-43 NUCLEAR LOSS AND CYTOPLASMIC AGGREGATION ARE HALLMARKS OF C9ORF72 REPEAT EXPANSION-MEDIATED NEURODEGENERATIVE DISEASE. HOWEVER, THE PRECISE MECHANISM BY WHICH AN EXPANDED G4C2 REPEAT INITIATES PATHOGENESIS IS NOT ESTABLISHED. THREE PRIMARY MECHANISMS HAVE BEEN PROPOSED: 1) HAPLOINSUFFICIENCY OF C9ORF72 PROTEIN, 2) TOXIC RNA FOCI GENERATED FROM SENSE AND/OR ANTISENSE REPEAT-CONTAINING TRANSCRIPTS, AND/OR 3) ACCUMULATION OF DIPEPTIDE REPEAT (DPR) PROTEIN PRODUCTS. TO IDENTIFY MECHANISMS OF PATHOGENESIS IN TDP-43 PROTEINOPATHIES, WE HAVE DEVELOPED A MULTIMODAL MERFISH (MULTIPLEXED ERROR-ROBUST FLUORESCENCE IN SITU HYBRIDIZATION) APPROACH, ENABLING THE MEASUREMENT OF KEY CANDIDATE PROTEINS (INCLUDING TDP-43 AND DPRS) TO DETERMINE CELLULAR PATHOLOGIC STATE WHILE SIMULTANEOUSLY MEASURING ANY EFFECT ON THE CORRESPONDING TRANSCRIPTOME. BY INTEGRATING THIS APPROACH WITH SINGLE-NUCLEAR RNA SEQUENCING, OUR EXTENSIONS ALLOW FOR THE DETERMINATION OF NEARLY COMPLETE TRANSCRIPTOMES IN UP TO 80,000 INDIVIDUAL CELLS WITHIN A SINGLE INTACT TISSUE SLICE. THIS APPROACH PRESERVES THE CRITICAL SPATIAL CONTEXT WITHIN INTACT TISSUE AND ENABLES CORRELATING PATHOLOGIC PROTEIN MISLOCALIZATION WITH MEASUREMENTS OF SOMATIC REPEAT EXPANSION AND SINGLE-CELL TRANSCRIPTOMIC RNA PROFILES. WITH THIS TECHNOLOGY, WE WILL DETERMINE WHICH CELL TYPES AND GENES ARE AFFECTED BY TDP-43 PROTEINOPATHY, FIRST IN SPORADIC ALS. NEXT, WE WILL EXTEND THIS ANALYSIS TO C9ORF72-ALS/FTD TO DETERMINE WHICH CELL TYPES AND GENES ARE AFFECTED BY REPEAT EXPANSION-MEDIATED C9ORF72 PATHOLOGICAL HALLMARKS, TDP-43 PROTEINOPATHY, AS WELL AS THE PRESENCE AND EXTENT OF SOMATIC REPEAT EXPANSION. FINALLY, WE WILL DETERMINE WHETHER ANY GENES AND/OR DISEASE MECHANISMS ARE EXACERBATED BY REDUCED C9ORF72 ACTIVITY. BY GENERATING AND EXAMINING COHORTS OF MICE WITH ALS-LIKE MOTOR NEURON DEGENERATION FROM EXPRESSION OF A G4C2 HEXANUCLEOTIDE REPEAT WITH NORMAL, REDUCED, OR COMPLETE ABSENCE OF C9ORF72 PROTEIN, WE WILL LEVERAGE MULTIMODAL MERFISH TO DETERMINE RNA TRANSCRIPTOMES IN ALL CELLS AS WELL AS THE PRESENCE OF TDP-43 PROTEINOPATHY, SENSE/ANTISENSE RNA FOCI, AND DPR ACCUMULATION FROM EARLY TO LATE DISEASE STAGES. THIS ANALYSIS WILL DETERMINE WHETHER, AND IF SO HOW, C9ORF72 LOSS OF FUNCTION AFFECTS REPEAT EXPANSION-LINKED TOXICITY.

San Diego University Of California

(1) TO SUPPORT EXTRAMURAL RESEARCH FUNDED BY THE NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE (NINDS) INCLUDING: BASIC RESEARCH THAT EXPLORES THE FUNDAMENTAL STRUCTURE AND FUNCTION OF THE BRAIN AND THE NERVOUS SYSTEM, RESEARCH TO UNDERSTAND THE CAUSES AND ORIGINS OF PATHOLOGICAL CONDITIONS OF THE NERVOUS SYSTEM WITH THE GOAL OF PREVENTING THESE DISORDERS, RESEARCH ON THE NATURAL COURSE OF NEUROLOGICAL DISORDERS, IMPROVED METHODS OF DISEASE PREVENTION, NEW METHODS OF DIAGNOSIS AND TREATMENT, DRUG DEVELOPMENT, DEVELOPMENT OF NEURAL DEVICES, CLINICAL TRIALS, AND RESEARCH TRAINING IN BASIC, TRANSLATIONAL AND CLINICAL NEUROSCIENCE. THE INSTITUTE IS THE LARGEST FUNDER OF BASIC NEUROSCIENCE IN THE US AND SUPPORTS RESEARCH ON TOPICS INCLUDING BUT NOT LIMITED TO: DEVELOPMENT OF THE NERVOUS SYSTEM, INCLUDING NEUROGENESIS AND PROGENITOR CELL BIOLOGY, SIGNAL TRANSDUCTION IN DEVELOPMENT AND PLASTICITY, AND PROGRAMMED CELL DEATH, SYNAPSE FORMATION, FUNCTION, AND PLASTICITY, LEARNING AND MEMORY, CHANNELS, TRANSPORTERS, AND PUMPS, CIRCUIT FORMATION AND MODULATION, BEHAVIORAL AND COGNITIVE NEUROSCIENCE, SENSORIMOTOR LEARNING, INTEGRATION AND EXECUTIVE FUNCTION, NEUROENDOCRINE SYSTEMS, SLEEP AND CIRCADIAN RHYTHMS, AND SENSORY AND MOTOR SYSTEMS. IN ADDITION, THE INSTITUTE SUPPORTS BASIC, TRANSLATIONAL AND CLINICAL STUDIES ON A NUMBER OF DISORDERS OF THE NERVOUS SYSTEM INCLUDING (BUT NOT LIMITED TO): STROKE, TRAUMATIC INJURY TO THE BRAIN, SPINAL CORD AND PERIPHERAL NERVOUS SYSTEM, NEURODEGENERATIVE DISORDERS, MOVEMENT DISORDERS, BRAIN TUMORS, CONVULSIVE DISORDERS, INFECTIOUS DISORDERS OF THE BRAIN AND NERVOUS SYSTEM, IMMUNE DISORDERS OF THE BRAIN AND NERVOUS SYSTEM, INCLUDING MULTIPLE SCLEROSIS, DISORDERS RELATED TO SLEEP, AND PAIN. PROGRAMMATIC AREAS, WHICH ARE PRIMARILY SUPPORTED BY THE DIVISION OF NEUROSCIENCE, ARE ALSO SUPPORTED BY THE DIVISION OF EXTRAMURAL ACTIVITIES, THE DIVISION OF TRANSLATIONAL RESEARCH, THE DIVISION OF CLINICAL RESEARCH, THE OFFICE OF TRAINING AND WORKFORCE DEVELOPMENT, THE OFFICE OF PROGRAMS TO ENHANCE NEUROSCIENCE WORKFORCE DEVELOPMENT, AND THE OFFICE OF INTERNATIONAL ACTIVITIES. (2) TO EXPAND AND IMPROVE THE SMALL BUSINESS INNOVATION RESEARCH (SBIR) PROGRAM, TO INCREASE PRIVATE SECTOR COMMERCIALIZATION OF INNOVATIONS DERIVED FROM FEDERAL RESEARCH AND DEVELOPMENT, TO INCREASE SMALL BUSINESS PARTICIPATION IN FEDERAL RESEARCH AND DEVELOPMENT, AND TO FOSTER AND ENCOURAGE PARTICIPATION OF SOCIALLY AND ECONOMICALLY DISADVANTAGED SMALL BUSINESS CONCERNS AND WOMEN-OWNED SMALL BUSINESS CONCERNS IN TECHNOLOGICAL INNOVATION. TO UTILIZE THE SMALL BUSINESS TECHNOLOGY TRANSFER (STTR) PROGRAM, TO STIMULATE AND FOSTER SCIENTIFIC AND TECHNOLOGICAL INNOVATION THROUGH COOPERATIVE RESEARCH AND DEVELOPMENT CARRIED OUT BETWEEN SMALL BUSINESS CONCERNS AND RESEARCH INSTITUTIONS, TO FOSTER TECHNOLOGY TRANSFER BETWEEN SMALL BUSINESS CONCERNS AND RESEARCH INSTITUTIONS, TO INCREASE PRIVATE SECTOR COMMERCIALIZATION OF INNOVATIONS DERIVED FROM FEDERAL RESEARCH AND DEVELOPMENT, AND TO FOSTER AND ENCOURAGE PARTICIPATION OF SOCIALLY AND ECONOMICALLY DISADVANTAGED SMALL BUSINESS CONCERNS AND WOMEN-OWNED SMALL BUSINESS CONCERNS IN TECHNOLOGICAL INNOVATION.

93.853 - Extramural Research Programs in the Neurosciences and Neurological Disorders

National Institute of Neurological Disorders and Stroke (NNDS) [HHS - NIH]

La Jolla, California 92093 United States

Single Zip Code

NIH Research Project Grant (Parent R01 Clinical Trial Not Allowed) (PA-20-185)