Human iPSC-derived atrial cardiomyocytes to model atrial fibrillation in a dish

人 iPSC 衍生的心房心肌细胞在培养皿中模拟心房颤动

• 批准号: 10549330
• 负责人: Dawood Darbar
• 金额: $ 58.78万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-17 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10549330
• 关键词: Action Potentials; Anti-Arrhythmia Agents; Arrhythmia; Atrial Fibrillation; CRISPR/Cas technology; Capsid Proteins; Cardiac; Cardiac Myocytes; Catheters; Cell Culture Techniques; Cell Line; Cell model; Cells; Cessation of life; Clinical; Collaborations; Critical Pathways; Data; Development; Dexamethasone; Disease; Electrophysiology (science); Engineering; Etiology; Extracellular Matrix; Extracellular Matrix Proteins; Failure; Familial atrial fibrillation; Fibroblasts; Functional disorder; Gap Junctions; Generations; Genetic; Genomics; Goals; Heart Atrium; Heart failure; Heritability; Heterogeneity; Human; Hydrogels; Insulin-Like Growth Factor I; Ion Channel; Link; Membrane; Mexiletine; Modeling; Molecular; Morbidity - disease rate; Mutation; Myofibroblast; Pathogenesis; Patient Care; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Play; Polymers; Potassium Channel; Protocols documentation; Research Personnel; Role; Signal Pathway; Signaling Molecule; Sodium; Structural Protein; System; Testing; Toxic effect; Translating; Translations; Tretinoin; Triiodothyronine; Variant; Ventricular; biobank; cardiovascular pharmacology; clinical care; common treatment; early onset; gain of function; gain of function mutation; genetic approach; human embryonic stem cell; imprint; improved; in vivo; indium arsenide; individual patient; induced pluripotent stem cell; innovation; insight; kindred; mortality; multi-ethnic; multidisciplinary; novel; paracrine; personalized medicine; polydimethylsiloxane; postnatal; ranolazine; response; skills; stem cell biology; stem cells; stroke risk; success; targeted treatment

Project Summary Atrial fibrillation (AF) is associated with significant morbidity and increased mortality. Despite recent advances in catheter-based treatments, antiarrhythmic drugs (AADs) are still commonly used to treat AF. However, response to membrane-active drugs is highly variable, in part, because of the failure to target therapy to the underlying mechanisms. Although genetic approaches have provided important insights into the underlying mechanisms of AF, the translation of these discoveries to the bedside care of patients has been limited due to the challenges of adequately recapitulating human AF in cellular models. The ability to derive patient-specific atrial cardiomyocytes (CMs) from human induced pluripotent stem cells (iPSC)-CMs holds great promise for modeling AF-linked mutations and developing cellular models of AF that are genetically-matched to specific patients. However, atrial iPSC-CMs have not been used to elucidate the underlying cellular mechanisms of AF- linked mutations and model heritable AF. Specific Aim 1 will create the UIC Multi-Ethnic Atrial HiPSC-CM Biorepository with the goal of generating atrial iPSC-CMs from familial AF kindreds to model AF-linked mutations. Our pilot data shows that atrial iPSC-CMs recapitulated the electrophysiologic (EP) phenotype of an AF-linked SCN5A mutation, and served as a platform for targeting the underlying cellular mechanism of the gain-of-function variant. Nonetheless, enhancing the maturity of iPSC-CMs remains important as modeling mature CMs will not only provide additional insights into the underlying cellular mechanisms of AF but also identify molecular signaling pathways important for atrial development. Specific Aim 2 will test the hypothesis that the EP and structural maturity of atrial iPSC-CMs can be significantly enhanced by precise microenvironmental engineering of in-vivo relevant cell-cell, cell-extracellular matrix, and cell-soluble factor interactions, such that these cells allow for optimal modeling of AF. We will also assess the role of cardiac fibroblasts in the pathogenesis of AF and determine if they impact EP maturity by co-culturing them with atrial iPSC-CMs. Specific Aim 3 will elucidate the underlying cellular mechanisms of AF-linked mutations using atrial iPSC-CMs. We will determine the EP phenotype of an SCN5A-E428K and KCNQ1-IAP54-56 mutation using patient-specific atrial iPSC-CMs. In addition, atrial iPSC-CMs from the 2 kindreds will be genetically corrected using CRISPR-Cas9 system to definitively establish causality. Thus, the overarching goals of this proposal are to harness the complementary skills of both Co-PIs (Drs. Darbar and Khetani) to establish the UIC Multi-Ethnic Atrial HiPSC Biorepository to serve as a platform for modeling AF-linked mutations, elucidate the underlying cellular mechanisms, and identify and assess novel mechanism-based therapies for AF. This platform will not only enable a more mechanism-based approach to the treatment of AF but will also `personalize' therapy with improved efficacy and reduced toxicities for individual patients.

项目摘要 心房颤动(房颤)与显著的发病率和增加死亡率有关。尽管最近取得了进展 在以导管为基础的治疗中，抗心律失常药物(AADS)仍然是治疗房颤的常用药物。然而， 对膜活性药物的反应是高度可变的，部分原因是未能针对 潜在的机制。尽管遗传方法提供了对潜在疾病的重要见解 房颤的机制，这些发现转化为患者的床边护理一直受到限制，因为 在细胞模型中充分概括人类房颤的挑战。能够派生出特定于患者的 从人诱导的多能干细胞(IPSC)-CMS中获得的心房肌细胞(CMS)有望在 建立房颤相关突变的模型并开发与特定基因相匹配的房颤细胞模型 病人。然而，心房IPSC-CMS还没有被用来阐明房颤的潜在细胞机制。 连锁突变和遗传性房颤模型。特定目标1将创建UIC多民族心房HiPSC-CM 生物库，目标是从家族性房颤家系中产生心房IPSC-CMS以建立房颤连锁模型 突变。我们的初步数据显示，心房IPSC-CMS重现了AN的电生理(EP)表型。 AF连锁的SCN5A突变，并作为靶向潜在的细胞机制的平台 函数增益变量。尽管如此，提高IPSC-CMS的成熟度作为建模仍然很重要 成熟的CMS不仅将为房颤的潜在细胞机制提供更多的见解，而且还将 确定对心房发育重要的分子信号通路。《特定目标2》将检验这一假设 精确度可显著提高心房IPSC-CMS的诱发电位和结构成熟度 体内相关细胞-细胞、细胞-细胞外基质和细胞可溶性因子的微环境工程 相互作用，使得这些细胞允许对房颤进行最佳建模。我们还将评估心脏的作用 成纤维细胞在房颤发病机制中的作用及与房颤共同培养对EP成熟度的影响 IPSC-CMS。特指目标3将阐明房颤相关突变的潜在细胞机制 心房IPSC-CMS。我们将确定SCN5A-E428K和KCNQ1-IAP54-56突变的EP表型 使用患者特定的心房IPSC-CMS。此外，这两个家族的房性IPSC-CMS将是遗传的 更正了使用CRISPR-CAS9系统确定因果关系的方法。因此，这一计划的首要目标是 建议是利用两个共同PIs(Darbar博士和Khetani博士)的互补技能来建立 UIC多民族房颤HiPSC生物库作为建模房颤相关突变的平台，阐明 潜在的细胞机制，并确定和评估新的基于机制的治疗房颤的方法。这 Platform不仅将使房颤的治疗更加基于机制，而且将 “个人化”治疗，提高疗效，减少个别患者的毒副作用。

项目成果

Genetics of atrial fibrillation-practical applications for clinical management: if not now, when and how?

• DOI: 10.1093/cvr/cvab153
• 发表时间: 2021-06-16
• 期刊: Cardiovascular research
• 影响因子: 10.8
• 作者: Kany S;Reissmann B;Metzner A;Kirchhof P;Darbar D;Schnabel RB
• 通讯作者: Schnabel RB

Human induced pluripotent stem cell-derived atrial cardiomyocytes carrying an SCN5A mutation identify nitric oxide signaling as a mediator of atrial fibrillation.

• DOI: 10.1016/j.stemcr.2021.04.019
• 发表时间: 2021-06-08
• 期刊: Stem cell reports
• 影响因子: 5.9
• 作者: Hong L;Zhang M;Ly OT;Chen H;Sridhar A;Lambers E;Chalazan B;Youn SW;Maienschein-Cline M;Feferman L;Ong SG;Wu JC;Rehman J;Darbar D
• 通讯作者: Darbar D