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Principal Investigator: Tulsi Patel
Organization: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
Fiscal Year: 2024
Award: $224,100
Funding agency: National Institute of Neurological Disorders and Stroke
PROJECT SUMMARY/ABSTRACT
In order to understand neurological diseases, it is essential to identify the affected neuronal cell types,
create model systems that accurately recapitulate normal function and disease phenotypes, and develop tools
that allow cellular manipulations. Motor neurons in the spinal cord control body movement by communicating
central motor commands with muscle targets. All spinal motor neurons are born and specified during embryonic
development, but their molecular identities and electrophysiological properties evolve for weeks in mice and
months in humans, until motor circuits and behavior become fully mature in post-natal life. In diseases like
Amyotrophic Lateral Sclerosis (ALS), sarcopenia, or spinal cord injury, the degeneration of specific subsets of
mature, adult motor neurons can lead to loss of muscle control, paralysis, and death. Although several studies
have mapped the mechanisms of motor neuron diversification in the embryonic spinal cord, our understanding
of motor neuron diversity in the adult spinal cord is in its infancy. This hinders the study of adult motor neuron
diseases as the affected motor neuron subtypes are not thoroughly defined. Furthermore, in vitro models that
faithfully recapitulate adult motor neuron identity do not exist, and adequate tools to access specific motor neuron
subtypes in vivo are lacking. This research plan aims to map the trajectory of post-mitotic motor neurons from
embryo to adulthood and use this data to both create viral tools that provide genetic access to specific subtypes
of motor neurons in vivo, and develop methods for generating adult-like motor neurons in vitro. This will be done
by first performing single cell transcriptome and chromatin profiling in mouse spinal motor neurons at various
embryonic to adult ages. The temporal chromatin profiles will be used to develop AAV tools that provide genetic
access to specific motor neuron subtypes at all ages, and to computationally identify candidate regulators of
subtype- and adult-specific identity. The identified regulators will then be used to mature the age of mouse stem
cell derived motor neurons in vitro. Finally, single cell transcriptomic and chromatin accessibility profiles of adult
human motor neurons will be generated and a combination of mouse and human-specific regulators will be used
to program the age of iPSC-derived motor neurons. This thorough approach will define the molecular features
of motor neuron subtypes that contribute to their differential susceptibility in disease, and establish tools
necessary for dissecting circuits, disease modeling, and the delivery of potential therapeutics.
The training phase of the award will be conducted in the laboratory of Dr. Hynek Wichterle at Columbia
University, and under the co- mentorship of Dr. David Gifford and Dr. Paola Arlotta. My career development plan
describes a detailed timeline for acquiring all the technical and professional skills necessary for the successful
transition into a career as an independent researcher. The completion of the proposed research plan will facilitate
future research in my lab aimed at understanding the temporal dynamics of neuronal identity, circuits, and
disease in motor neurons and other nervous system cells.
Terms: <21+ years old><AAV vector><AAV-based vector><ATAC sequencing><ATAC-seq><ATACseq><Adult><Adult Human><Affect><Age><Amyotrophic Lateral Sclerosis><Amyotrophic Lateral Sclerosis Motor Neuron Disease><Assay><Assay for Transposase-Accessible Chromatin using sequencing><Autopsy><Award><Basal Transcription Factor><Basal transcription factor genes><Behavior><Bio-Informatics><Bioassay><Bioinformatics><Biologic Models><Biological Assay><Biological Models><Brain region><Cell Body><Cell Communication and Signaling><Cell Line><Cell Nucleus><Cell Signaling><CellLine><Cells><Cessation of life><Chromatin><Communication><Data><Data Analyses><Data Analysis><Death><Development Plans><Dimensions><Disease><Disorder><Electrophysiology><Electrophysiology (science)><Embryo><Embryo Development><Embryogenesis><Embryonic><Embryonic Development><Gehrig's Disease><Gene Delivery><Gene Expression><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genetic><Genetic Transcription><Genomics><Human><In Vitro><Intracellular Communication and Signaling><Investigators><Laboratories><Lead><Life><Lou Gehrig Disease><Maps><Medulla Spinalis><Mentorship><Methods><Mice><Mice Mammals><Model System><Modeling><Modern Man><Molecular><Molecular Fingerprinting><Molecular Profiling><Motor><Motor Cell><Motor Neuron Disease><Motor Neurons><Movement><Murine><Mus><Muscle><Muscle Atrophy><Muscle Tissue><Muscular Atrophy><Nerve Cells><Nerve Unit><Nervous System><Nervous System Diseases><Nervous System Disorder><Neural Cell><Neurocyte><Neurologic Body System><Neurologic Disorders><Neurologic Organ System><Neurological Disorders><Neurons><Neurophysiology - biologic function><Neurophysiology / Electrophysiology><Nucleic Acid Regulator Regions><Nucleic Acid Regulatory Sequences><Nucleus><Palsy><Paralysed><Patients><Pb element><Phase><Plegia><Predisposition><Progenitor Cells><Property><RNA Expression><RNA Seq><RNA sequencing><RNAseq><Regulatory Regions><Research><Research Personnel><Researchers><Signal Transduction><Signal Transduction Systems><Signaling><Single-Nucleus Sequencing><Sorting><Specific qualifier value><Specified><Spinal><Spinal Cord><Spinal Cord Trauma><Spinal Trauma><Spinal cord injured><Spinal cord injury><Strains Cell Lines><Susceptibility><System><Testing><Therapeutic><Time><Training><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Traumatic Myelopathy><Universities><Viral><Virus><Work><adeno-associated viral vector><adeno-associated virus vector><adulthood><ages><assay for transposase accessible chromatin followed by sequencing><assay for transposase accessible chromatin seq><assay for transposase accessible chromatin sequencing><assay for transposase-accessible chromatin with sequencing><biological signal transduction><body movement><candidate identification><career><career development><cell type><cultured cell line><data interpretation><degenerative disorder of motor neurons><develop therapy><disease model><disease phenotype><disorder model><electrophysiological><genetic regulatory element><global gene expression><global transcription profile><heavy metal Pb><heavy metal lead><hiPSC><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><iPS><iPSC><iPSCs><in vitro Model><in vivo><induced human pluripotent stem cells><induced pluripotent cell><induced pluripotent stem cell><inducible pluripotent stem cell><infancy><infantile><intervention development><method development><molecular profile><molecular signature><motoneuron><motor neuron function><muscle breakdown><muscle degradation><muscle deterioration><muscle loss><muscle wasting><muscular><mutant><necropsy><neural circuit><neural circuitry><neural function><neurocircuitry><neurological disease><neuronal><neuronal circuit><neuronal circuitry><paralysis><paralytic><post-doctoral training><postdoctoral training><postmitotic><postmortem><postnatal><programs><sNuc-Seq><sarcopenia><sarcopenic><single nucleus RNA-sequencing><single nucleus seq><single-nucleus RNA-seq><skills><snRNA sequencing><snRNA-seq><stem cells><synaptic circuit><synaptic circuitry><therapy development><time use><timeline><tool><transcription factor><transcriptome><transcriptome sequencing><transcriptomic sequencing><transcriptomics><treatment development>