Discovery of small molecule regulators of atrial cardiomyocyte action potential duration to restore normal cardiac rhythm in atrial fibrillation

发现心房心肌细胞动作电位持续时间的小分子调节剂以恢复心房颤动的正常心律

• 批准号: 10643997
• 负责人: Alexandre Romain Colas
• 金额: $ 48.75万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-10 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10643997
• 关键词: Action Potentials; Affect; Aging; Animal Model; Anticoagulants; Arrhythmia; Atrial Fibrillation; Attention; Biological; Biological Assay; Biology; Cardiac; Cardiac Myocytes; Cardiac ablation; Cardiovascular Diseases; Cardiovascular system; Catalogs; Cells; Cellular Assay; Cellular biology; Chemicals; Clinical; Critical Pathways; Data; Developing Countries; Disease; Dose; Dryness; Electrophysiology (science); Exclusion; Future; Generations; Genetic; Goals; Grant; Heart Atrium; Heart Diseases; Heart failure; Human; Image; Kinetics; Lead; Mutation; Myocardial; Outcome; Output; Parents; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Physiological; Physiology; Powder dose form; Procedures; Proliferating; Property; Quality of life; Readiness; Reporting; Research; Resolution; Resources; Safety; Series; Stroke; Structure-Activity Relationship; Symptoms; Systems Analysis; Technology; Testing; Therapeutic; Thromboembolism; Tissues; Toxic effect; Treatment Cost; Validation; Ventricular; Work; analog; cheminformatics; cost; design; drug discovery; electrical property; experimental study; heart function; heart rhythm; high throughput screening; improved; in vivo evaluation; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; innovation; liquid chromatography mass spectrometry; mortality; novel; novel therapeutics; progenitor; response; scaffold; screening; side effect; small molecule; small molecule libraries; statistics; stem cells; stroke risk; tool; trait

PROJECT SUMMARY Atrial fibrillation (AF) is the most prevalent cardiac arrhythmia, afflicting over 33 million people worldwide and 6 million in the US. AF causes reduced quality of life, stroke and systemic thromboembolism, heart failure, and increased mortality. Treatment of AF and its complications with nonspecific drugs or procedures is characterized by unsatisfactory outcomes and significant cost. Acquired heart disease, cardiac remodeling, neurohormonal factors, aging, and genetic traits have all been correlated with presence of AF. Rapid, uncoordinated atrial chamber activity is due to shortened or prolonged cardiomyocyte action potential durations acting within a vulnerable myocardial substrate, causing persistent arrhythmia that features triggering or sustaining circuit re- entry or early and/or delayed after-depolarizations, respectively. We have recently developed a novel, high throughput kinetic imaging and analysis platform to characterize cardiomyocyte electrophysiological properties at single cell resolution, which can be used to conduct high throughput screening (HTS) on functional human atrial cardiomyocytes derived from Id1-programmed cardiac progenitors created from iPS cells. Our innovation is the use of this and related assays in a phenotypic screening cascade designed to discover previously unknown, atrial-specific modulators of cardiomyocyte electrical properties and rhythm. Our hypothesis is this approach will ultimately generate drug-like starting points for future disease-modifying cardiovascular therapeutics. The primary HTS assay has been fully optimized in a 384-well format, and as a demonstration of assay readiness, 400 compounds have been screened (Kolmogorov-Smirnov D-statistic >0.1). Multiple hits from pilot screens were identified and were confirmed and validated in concentration response experiments. A battery of downstream assays has been developed and piloted to establish a critical path-testing funnel. Several compounds identified from the pilot screen were tested to determine if they affected the action potential duration of atrial cardiomyocytes sensitized by the E375X mutation in KCNA5, and if they had effects on the action potential durations of wild type and primary atrial and ventricular cardiomyocytes. This proposal builds on data from the applicants, an established team from SBP (Drs. Colas and Larson) with basic biology and drug discovery expertise in the field and access to all necessary technologies. The overall goal of this proposal is to generate chemical biology research tools and starting points for new drugs. As the critical path assays are all in place, we anticipate we can rapidly obtain such probe molecules and start to explore their activity. Our future plans are to ultimately determine hits’ suitability for hit-to-lead activities, begin in vivo evaluation of lead compounds in animal models and eventually patients, and determine their cellular mechanism of action. This grant’s work product will serve as preliminary data for hit-to-lead (HTL) grant submissions and parent R01 grant submissions to pursue understanding of their biological mechanisms.

项目摘要 心房颤动（AF）是最常见的心律失常，全世界有超过3300万人受到影响， 百万在美国。AF导致生活质量下降、卒中和全身血栓栓塞、心力衰竭， 增加死亡率。使用非特异性药物或手术治疗AF及其并发症的特点是 不满意的结果和巨大的成本。获得性心脏病，心脏重塑，神经激素 因素、年龄和遗传特征都与AF的存在相关。 室活动是由于缩短或延长的心肌细胞动作电位持续时间内发挥作用， 脆弱的心肌基质，引起持续性心律失常，其特征是触发或维持电路重新激活， 分别为进入或早期和/或延迟后去极化。我们最近开发了一种新的，高 用于表征心肌细胞电生理特性的通量动力学成像和分析平台 在单细胞分辨率，可用于进行高通量筛选（HTS）功能的人 心房心肌细胞来源于由iPS细胞产生的Id 1编程的心脏祖细胞。我们的创新 是在表型筛选级联中使用这种和相关的测定， 未知的心房特异性心肌细胞电特性和节律调节剂。我们的假设是 这种方法最终将为未来改善心血管疾病的治疗提供类似药物的起点。 治疗学主要HTS测定已在384孔格式中进行了充分优化，并作为以下方面的证明： 在分析准备就绪的情况下，已经筛选了400种化合物（Kolmogorov-Smirnov D-统计>0.1）。多个匹配来自 鉴定了中试筛选，并在浓度响应实验中得到确认和验证。电池 已开发并试点了下游检测试剂盒，以建立关键路径测试漏斗。几 对从中试筛选中鉴定出的化合物进行测试，以确定它们是否影响动作电位时程 KCNA 5中E375 X突变致敏的心房心肌细胞，以及它们是否对 野生型和原代心房和心室心肌细胞的潜在持续时间。这一建议建立在数据基础上 从申请人，一个建立的团队从SBP（博士. Colas和拉森）与基础生物学和药物发现 在该领域的专业知识和获得所有必要的技术。本提案的总体目标是， 化学生物学研究工具和新药的起点。由于关键路径分析都已就绪，我们 预期我们可以快速获得这样的探针分子并开始探索它们的活性。我们未来的计划是 最终确定hits对hit-to-lead活性的适合性，开始在动物体内评价lead化合物 模型和最终患者，并确定其细胞作用机制。这项补助金的工作成果将 作为初步数据的命中铅（HTL）赠款提交和父R 01赠款提交，以追求 了解其生物学机制。