ConProject-001

ConProject-001

• 批准号: 10316591
• 负责人: Ilya B Bezprozvanny
• 金额: $ 72.31万
• 依托单位: CHAPMAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10316591
• 关键词: Affect; Ataxia; Behavioral; Binding; Biological Process; Calcium; Calcium Signaling; Cells; Cerebellar Ataxia; Cessation of life; Clinical; Clinical Research; Collaborations; Complex; Computational Biology; Crystallization; Development; Disease; Dominant Genetic Conditions; Drug Targeting; Electrophysiology (science); Equilibrium; Exposure to; Fluorescence; Functional disorder; Future; Genetic; Goals; Health; ITPR1 gene; Industrialization; Inositol; Knowledge; Laboratories; Letters; Metabolic; Methods; Movement; Mus; Muscle; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Online Mendelian Inheritance In Man; Pathogenesis; Pathology; Patients; Pattern; Phase; Phenotype; Play; Population; Potassium; Pre-Clinical Model; Proteins; Purkinje Cells; Research; Riluzole; Role; SCA2 protein; Signal Transduction; Slice; Specificity; Spinocerebellar Ataxias; Structure; Symptoms; Testing; Therapeutic; Therapeutic Agents; Transgenic Mice; Type 2 Spinocerebellar Ataxia; base; biophysical techniques; clinical development; effective therapy; high throughput screening; interest; mouse model; mutant; nervous system disorder; novel; novel therapeutics; polyglutamine; positive allosteric modulator; pre-clinical; preclinical development; prototype; receptor; skills; structural biology; therapeutically effective; tool; translational neuroscience; virtual

The broad, long-term objective of the project is to develop novel therapeutic agents for spinocerebellar ataxias (SCAs). There is no effective treatment or cure for SCAs, in large part due to insufficient understanding of the disease pathogenesis. In this project, we will focus on spinocerebellar ataxia type 2 (SCA2). SCA2 is an autosomal dominant genetic neurodegenerative disorder caused by polyQ expansion in ataxin-2 (Atxn2) protein. Cerebellar Purkinje cells (PCs) are the main locus of pathology in SCA2 patients. Previously, our laboratory discovered abnormal neuronal calcium signaling in PCs, which play an important role in the pathogenesis of SCAs. Previous studies also suggest that neuronal activity of PCs is abnormal in ataxic mouse models. We demonstrated that positive allosteric modulators (PAMs) of small conductance calcium-activated potassium (SK) channels can normalize the abnormal firing pattern of PCs in SCA2 mice and alleviate behavioral and neuropathological phenotypes. These results suggest that SK PAMs can be potentially useful as therapeutic agents for the treatment of SCA2 and possibly for other ataxias. This project brings together the complementary skills and expertise of two laboratories for the discovery of such agents. To achieve this goal, we will focus on the following specific aims: R21 Phase Aim 1. To identify novel subtype-selective SK PAM candidates. We will determine the crystal structure of SK2 channels in complex with the prototype subtype-selective PAMs. We will obtain structural information that will be used to perform v irtual high throughput screening for subtype-selective SK PAMs. R21 Phase Aim 2. To validate functional activity of novel SK PAMs. We will use fluorescence-based high throughput assay and electrophysiological methods to test SK PAM candidates. R33 Phase Aim 3. To investigate the specificity of novel SK PAMs for SK channels. We will evaluate the specificity of the SK PAM candidates for SK channels over other K+, Na+ and Ca2+ channels using electrophysiological recordings. R33 Phase Aim 4. To investigate the effects of novel SK PAMs on the activity of cerebellar PCs in SCA2 mice. We will perform ex. vivo. cerebellar slice electrophysiological recordings of PC activity in these studies. The resulting subtype-selective SK PAMs will provide critical leads for future preclinical and clinical development, fulfilling the critical and unmet needs in ataxia patients. These molecules will also offer novel and unique research tools for understanding the biological function of SK channels in health and disease. Thus, the proposed project will have significant impact in basic and translational neuroscience.

该项目广泛的长期目标是开发脊髓小脑共济失调的新型治疗药物 （管制计划协议）。SCA没有有效的治疗或治愈方法，很大程度上是由于对 发病机理在这个项目中，我们将重点关注脊髓小脑共济失调2型（SCA 2）。SCA 2是一个 由共济失调蛋白-2（Atxn 2）中polyQ扩增引起的常染色体显性遗传神经变性疾病 蛋白小脑浦肯野细胞（PC）是SCA 2患者的主要病理部位。此前，我们 一个实验室在PC中发现了异常的神经元钙信号，这在神经元的钙代谢中起着重要作用。 SCA的发病机制。以往的研究也提示共济失调小鼠PC的神经元活动异常 模型我们证明了小电导钙激活的正变构调节剂（PAM）， 钾通道可使SCA 2小鼠PC异常放电模式正常化， 行为和神经病理表型。这些结果表明，SK PAM可能是有用的 作为治疗SCA 2和可能用于其它共济失调的治疗剂。该项目汇集了 两个实验室在发现这类制剂方面的技能和专门知识互补。为了实现这一目标， 我们会致力达致以下的具体目标： R21阶段目标1.鉴定新的亚型选择性SK PAM候选物。我们将确定水晶 SK2通道与原型亚型选择性PAM的复合结构。我们将获得结构 将用于对亚型选择性SK PAM进行虚拟高通量筛选的信息。 第二阶段目标。验证新型SK PAM的功能活性。我们将使用基于荧光的高 通量测定和电生理学方法来测试SK PAM候选物。 第三章.研究新型SK PAM对SK通道的特异性。我们将评估 SK PAM候选物对于SK通道相对于其他K+、Na+和Ca 2+通道的特异性， 电生理记录。 第四章.研究新型SK PAM对SCA 2小脑PC活性的影响 小鼠我们将执行前。vivo.小脑切片电生理记录PC活动在这些研究中。 由此产生的亚型选择性SK PAM将为未来的临床前和临床 发展，满足共济失调患者的关键和未满足的需求。这些分子还将提供新的 以及独特的研究工具，用于了解SK通道在健康和疾病中的生物学功能。 因此，该项目将对基础和转化神经科学产生重大影响。