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Principal Investigator: Henry M. Colecraft
Organization: COLUMBIA UNIVERSITY HEALTH SCIENCES
Fiscal Year: 2024
Award: $517,445
Funding agency: National Institute of Neurological Disorders and Stroke
SUMMARY
Mutations in CaV2.1 pore-forming 1A subunit cause a spectrum of neurological diseases including epileptic
encephalopathies (EE), familial hemiplegic migraine type 1 (FHM1), episodic ataxia type 2 (EA2), spinocerebellar
ataxia type 6 (SCA6), and intellectual disability (ID). The ClinVar database has entries for >1000 CACNA1A
mutations most of which (437) are classified as variants of unknown significance (VUS), pathogenic (137), or
likely pathogenic (61). There are several challenges for efforts to develop effective therapies for CACNA1A
channelopathies: 1) the large number of dmutations that give rise to disease make it unclear whether common
therapies can be found; 2) the full scope of functional alterations due to individual mutations and how these relate
to disease etiology are ambiguous; and 3) lack of novel therapeutics targeted to CaV2.1 functional deficiencies.
We hypothesize that the hundreds of distinct CACNA1A mutations fall into a few discrete functional groups that
can be targeted by novel bioengineered molecules tailored for each class. Our long-term objective is to gain an
in-depth perspective on how distinct CACNA1A mutations give rise to a spectrum of neurological disorders and
to develop molecules that can address the functional deficits as potential therapeutics. Here, we propose an
inter-disciplinary, multi-level proposal spanning single-channel and whole-cell Ca2+ channel biophysics, patient-
specific induced pluripotent stem cell neurons (hiPSC-neurons), mouse models of CACNA1A neurological
disease, and development of corrective molecules. The breadth of the proposal is enabled by collaboration and
combining resources between two labs− the Colecraft lab (Columbia University) has strong expertise in
molecular physiology and biophysics of CaV channels and developing innovative tools to regulate their functional
expression; the Rossignol lab (Montreal University) has expertise in generation and functional characterization
of CACNA1A mouse models of neurological disease. Dr. Rossignol is a clinician-scientist with a cohort of
CACNA1A patients who thus also brings a clinician’s perspective to the project. We propose three Aims all of
which are supported by strong preliminary data. 1) Determine holistic functional impact of distinct CACNA1A
mutations on recombinant CaV2.1 channels, and develop tailored approaches to correct different classes of
mutations. 2) Develop human ipsc-neurons to model and elucidate mechanisms of CACNA1A channelopathies
and to evaluate efficacy of novel potential therapeutic molecules. 3) Utilize mouse models to determine
mechanisms of disease and evaluate efficacy of novel tailored approaches to treat disease.
Terms: <Address><Assay><Behavioral Assay><Bioassay><Biological Assay><Biomedical Engineering><Biophysics><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Causality><Cav2.1><Cell Body><Cells><Clampings><Classification><ClinVar><Clinical><Closure by clamp><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Collaborations><Data><Data Bases><Databases><Development><Disease><Disorder><Dysfunction><Electrophysiology><Electrophysiology (science)><Engineering><Episodic ataxia><Etiology><Familial Hemiplegic Migraine><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Functional disorder><G-Proteins><GTP-Binding Proteins><GTP-Regulatory Proteins><Generations><Genes><Genetic Alteration><Genetic Change><Genetic defect><Guanine Nucleotide Coupling Protein><Guanine Nucleotide Regulatory Proteins><Hereditary><Histology><Image><Immunofluorescence><Immunofluorescence Immunologic><Individual><Induced DNA Alteration><Induced Mutation><Induced Sequence Alteration><Induced pluripotent stem cell derived neurons><Inherited><Intellectual disability><Intellectual functioning disability><Intellectual limitation><Ion Channel Gating><Ion Channel Gatings><Ions><Karyotype determination procedure><Karyotyping><Karyotyping Genetics><Lentivirus Infections><Lentivirus disease><Microelectrodes><Miniaturized Electrodes><Modeling><Molecular><Morphology><Mutation><Nerve Cells><Nerve Unit><Nervous System Diseases><Nervous System Disorder><Network Analysis><Neural Cell><Neurocyte><Neurologic Disorders><Neurological Disorders><Neuron from iPSC><Neuron from induced pluripotent stem cells><Neurons><Neurophysiology / Electrophysiology><P-Q type VDCC><P-Q type voltage-dependent calcium channel><Pathogenicity><Pathway Analysis><Patients><Personalized medical approach><Physiologic><Physiological><Physiology><Physiopathology><Recombinants><Regulation><Research Resources><Resources><Scientist><Slice><Spinocerebellar Ataxia-6><Stop Codon><Systematics><Termination Codon><Terminator Codon><Testing><Therapeutic><Transfer RNA><Translation Stop Signal><Triplet Codon-Amino Acid Adaptor><Type 6 Spinocerebellar Ataxia><Universities><Vesicle><bio-engineered><bio-engineers><bioengineering><biological engineering><biophysical foundation><biophysical principles><biophysical sciences><causation><clinical relevance><clinically relevant><cohort><data base><de novo mutation><de novo variant><de-ubiquitinase><de-ubiquitinating enzyme><determine efficacy><developmental><disease causation><effective therapy><effective treatment><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><efficacy testing><electrophysiological><epileptic encephalopathies><evaluate efficacy><examine efficacy><falls><flow cytophotometry><functional group><gain of function><genome editing><genome mutation><genomic editing><hiPSC><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><iPS neurons><iPSC derived-neurons><imaging><in vivo><individualized approach><induced human pluripotent stem cells><induced pluripotent stem cell neurons><innovate><innovation><innovative><intellectual and developmental disability><limited intellectual functioning><loss of function><mouse model><murine model><nanobodies><nanobody><neural network><neurological disease><neuron development><neuronal><neuronal development><neurons derived from induced pluripotent stem cells><neurotransmitter release><new drug target><new drug treatments><new druggable target><new drugs><new pharmacological therapeutic><new pharmacotherapy target><new therapeutic target><new therapeutics><new therapy><new therapy target><next generation therapeutics><novel><novel drug target><novel drug treatments><novel druggable target><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel pharmacotherapy target><novel therapeutic target><novel therapeutics><novel therapy><novel therapy target><omega-agatoxin-IVA-sensitive VDCC><pathophysiology><personalized approach><precision approach><premature><prematurity><presynaptic><screening><screenings><sdAb><single domain antibodies><tRNA><tailored approach><tool><trafficking><transfer Ribonucleic acids><ubiquitin isopeptidase><ubiquitin-specific isopeptidase><unclassified variant><variant of uncertain clinical significance><variant of uncertain significance><variant of undetermined significance><variant of unknown significance><voltage>