Towards Novel Therapies for CACNA1A Neurological Disorders

寻找 CACNA1A 神经系统疾病的新疗法

• 批准号: 10589799
• 负责人: Henry M. Colecraft
• 金额: $ 53.34万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-15 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10589799
• 关键词: Address; Behavioral Assay; Biological Assay; Biomedical Engineering; Biophysics; CRISPR/Cas technology; Cells; Classification; ClinVar; Clinical; Closure by clamp; Collaborations; Data; Databases; Development; Disease; Electrophysiology (science); Engineering; Episodic ataxia; Etiology; Familial Hemiplegic Migraine; Flow Cytometry; Functional disorder; GTP-Binding Proteins; Generations; Genes; Histology; Human; Image; Immunofluorescence Immunologic; Individual; Induced Mutation; Induced pluripotent stem cell derived neurons; Inherited; Intellectual functioning disability; Ion Channel Gating; Ions; Karyotype determination procedure; Lentivirus Infections; Microelectrodes; Modeling; Molecular; Morphology; Mutation; Neurons; P-Q type voltage-dependent calcium channel; Pathogenicity; Pathway Analysis; Patients; Physiological; Physiology; Recombinants; Regulation; Resources; Scientist; Slice; Terminator Codon; Testing; Therapeutic; Transfer RNA; Type 6 Spinocerebellar Ataxia; Universities; Vesicle; clinically relevant; cohort; de novo mutation; effective therapy; efficacy evaluation; efficacy testing; epileptic encephalopathies; falls; functional group; gain of function; genome editing; in vivo; induced pluripotent stem cell; innovation; loss of function; mouse model; nanobodies; nervous system disorder; neural network; neuron development; neurotransmitter release; new therapeutic target; novel; novel therapeutics; personalized approach; premature; presynaptic; screening; tool; trafficking; ubiquitin isopeptidase; variant of unknown significance; voltage

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.

概括 Cav2.1中的突变孔形成1a亚基引起一系列神经疾病，包括癫痫 脑病（EE），家族性偏瘫偏头痛1型（FHM1），情节性共济失调2（EA2），脊髓灰质 共济失调6型（SCA6）和近视残疾（ID）。 Clinvar数据库的条目> 1000 Cacna1a 突变大多数（437）分类为未知意义（VU），致病性（137）或 可能的致病性（61）。为开发有效疗法的cacna1a有一些挑战 通道病：1）引起疾病的大量疾病使它是否常见 可以找到疗法； 2）由于个体突变以及这些如何相关的功能变化范围 病因学是模棱两可的； 3）缺乏针对CAV2.1功能性缺陷的新型治疗。 我们假设数百个不同的Cacna1a突变属于几个离散的官能团 可以通过针对每个类别的新型生物工程分子来靶向。我们的长期目标是获得 关于不同的Cacna1a突变如何产生一系列神经系统疾病和 开发可以解决功能缺陷作为潜在治疗的分子。在这里，我们建议 跨学科的多层次提案，涵盖单通道和全细胞CA2+通道生物物理学，患者 - 特异性诱导的多能干细胞神经元（HIPSC-神经元），CACNA1A神经系统的小鼠模型 疾病，以及矫正分子的发展。该提案的广度是通过协作和 在两个实验室之间结合资源 - Colecraft Lab（哥伦比亚大学）在 CAV通道的分子生理和生物物理学以及开发创新的工具以调节其功能 表达; Rossignol Lab（蒙特利尔大学）在发电和功能表征方面具有专业知识 神经系统疾病的CACNA1A小鼠模型。 Rossignol博士是一位临床科学家 因此，CACNA1A患者也为该项目带来了临床观点。我们提出三个目标 由强大的初步数据支持。 1）确定不同CACNA1A的整体功能影响 重组CAV2.1通道上的突变，并开发量身定制的方法以纠正不同类别的类别 突变。 2）开发人IPSC-神经元以建模和阐明CACNA1A通道病的机制 并评估新型潜在治疗分子的有效性。 3）利用鼠标模型来确定 疾病的机制和评估新型量身定制方法治疗疾病的有效性。