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.

总结 CaV2.1孔形成蛋白1A亚基的突变导致一系列神经系统疾病，包括癫痫 脑病（EE）、家族性偏瘫偏头痛1型（FHM 1）、发作性共济失调2型（EA 2）、脊髓小脑性 共济失调6型（SCA 6）和智力残疾（ID）。ClinVar数据库有>1000 CACNA 1A的条目 突变，其中大部分（437）被归类为未知意义（VUS），致病性（137），或 可能是致病的（61）。开发CACNA 1A有效疗法的努力面临着几个挑战 通道病：1）引起疾病的大量突变使其不清楚是否常见 可以找到治疗方法; 2）由于个体突变引起的功能改变的全部范围以及这些变化如何相互关联 疾病病因不明确; 3）缺乏针对CaV2.1功能缺陷的新疗法。 我们假设数百种不同的CACNA 1A突变属于几个离散的功能组， 可以被针对每一类的新型生物工程分子靶向。我们的长远目标是 深入探讨不同的CACNA 1A突变如何引起一系列神经系统疾病， 来开发能够解决功能缺陷的分子，作为潜在的治疗方法。在这里，我们提出一个 跨学科，多层次的建议，跨越单通道和全细胞钙通道生物物理学，病人- 特异性诱导多能干细胞神经元（hiPSC-神经元），CACNA 1A神经系统的小鼠模型 疾病和纠正分子的发展。提案的广度是通过协作实现的， 结合两个实验室之间的资源-Colecraft实验室（哥伦比亚大学）在以下方面拥有强大的专业知识： CaV通道的分子生理学和生物物理学，并开发创新工具来调节其功能 表达; Rossignol实验室（蒙特利尔大学）在生成和功能表征方面具有专业知识 CACNA 1A小鼠神经系统疾病模型。Rossignol博士是一名临床科学家， CACNA 1A患者因此也为该项目带来了临床医生的观点。我们提出三个目标， 这得到了强有力的初步数据的支持。1)确定不同CACNA 1A的整体功能影响 突变的重组CaV2.1通道，并开发量身定制的方法来纠正不同类型的 突变。2)开发人ipsc神经元以模拟和阐明CACNA 1A通道病的机制 并评价新的潜在治疗分子的功效。3)利用小鼠模型来确定 疾病的机制，并评估新的定制方法来治疗疾病的疗效。