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（AF）与显著的发病率和死亡率增加相关。尽管最近取得了一些进展 在基于导管的治疗中，抗心律失常药物（AAD）仍然通常用于治疗AF。然而， 对膜活性药物的反应是高度可变的，部分原因是未能靶向治疗， 基本机制。尽管遗传学方法为了解潜在的 由于AF的发病机制，这些发现对患者床边护理的转化受到限制， 在细胞模型中充分再现人类AF的挑战。获得患者特异性 来自人类诱导多能干细胞（iPSC）的心房心肌细胞（CM）-CM具有很大的希望， 对AF相关突变进行建模，并开发与特定基因匹配的AF细胞模型， 患者然而，心房iPSC-CM尚未用于阐明AF的潜在细胞机制。 特定目标1将创建UIC多种族心房HiPSC-CM 生物储存库，目标是从家族性AF激动剂产生心房iPSC-CM，以模拟AF相关 突变。我们的初步数据显示，心房iPSC-CM重现了心房肌细胞的电生理（EP）表型。 AF连锁的SCN 5A突变，并作为靶向潜在的细胞机制的平台， 功能增益变体。尽管如此，提高iPSC-CM的成熟度仍然很重要， 成熟的CM不仅可以为AF的潜在细胞机制提供更多的见解， 鉴定心房发育重要的分子信号通路。具体目标2将检验假设 心房iPSC-CM的EP和结构成熟度可以通过精确的 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，这样这些细胞允许AF的最佳建模。 成纤维细胞在房颤发病机制中的作用，并通过将它们与心房肌细胞共培养来确定它们是否影响EP成熟。 iPSC-CM。具体目标3将阐明AF连锁突变的潜在细胞机制， 心房iPSC-CM。我们将确定SCN 5A-E428 K和KCNQ 1-IAP 54 -56突变的EP表型 使用患者特异性心房iPSC-CM。此外，来自2种激酶的心房iPSC-CM将在遗传上与来自2种激酶的心房iPSC-CM相似。 使用CRISPR-Cas9系统进行纠正，以明确建立因果关系。因此，本项目的总体目标 建议是利用两个共同PI（Darbar博士和Khetani博士）的互补技能， UIC多种族心房HiPSC生物储存库作为AF相关突变建模的平台，阐明 潜在的细胞机制，并确定和评估新的机制为基础的治疗AF。这 该平台不仅能够实现更基于机制的AF治疗方法， “个性化”治疗，提高疗效，降低个体患者的毒性。

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