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

基本信息

批准号：
10548469
负责人：
Dawood Darbar
金额：
$ 2.6万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-17 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10548469
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 Coculture Techniques Collaborations Critical Pathways Data Development Dexamethasone Disease model 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 base 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

项目摘要

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.

项目概要心房颤动（AF）与显着的发病率和死亡率增加相关。尽管最近取得了进展在基于导管的治疗中，抗心律失常药物（AAD）仍然常用于治疗房颤。然而，对膜活性药物的反应变化很大，部分原因是未能针对膜活性药物进行靶向治疗。底层机制。尽管遗传学方法提供了对潜在的重要见解 AF 的机制，这些发现转化为患者的床边护理受到限制，因为在细胞模型中充分重现人类房颤的挑战。获得患者特异性的能力来自人类诱导多能干细胞 (iPSC) 的心房心肌细胞 (CM) - CM 具有巨大的应用前景模拟 AF 相关突变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。连锁突变和遗传性房颤模型。具体目标 1 将创建 UIC 多种族心房 HiPSC-CM 生物样本库的目标是从家族性 AF 亲属中生成心房 iPSC-CM，以建立 AF 相关模型突变。我们的试验数据显示，心房 iPSC-CM 重现了心房 iPSC-CM 的电生理 (EP) 表型。 AF 相关的 SCN5A 突变，并作为针对潜在细胞机制的平台功能获得变体。尽管如此，提高 iPSC-CM 的成熟度作为建模仍然很重要成熟的 CM 不仅可以为 AF 的潜在细胞机制提供更多见解，还可以确定对心房发育重要的分子信号传导途径。具体目标 2 将检验假设通过精确的方法可以显着增强心房 iPSC-CM 的 EP 和结构成熟度体内相关细胞-细胞、细胞-细胞外基质、细胞可溶性因子的微环境工程相互作用，使得这些细胞可以实现 AF 的最佳建模。我们还将评估心脏的作用成纤维细胞在 AF 发病机制中的作用，并通过与心房共培养来确定它们是否影响 EP 成熟度 iPSC-CM。具体目标 3 将使用以下方法阐明 AF 相关突变的潜在细胞机制：心房 iPSC-CM。我们将确定 SCN5A-E428K 和 KCNQ1-IAP54-56 突变的 EP 表型使用患者特异性心房 iPSC-CM。此外，来自 2 个家族的心房 iPSC-CM 将进行基因遗传使用 CRISPR-Cas9 系统进行纠正以明确建立因果关系。因此，本次会议的总体目标建议利用两位 Co-PI（Darbar 博士和 Khetani 博士）的互补技能来建立 UIC 多民族心房 HiPSC 生物样本库可作为 AF 相关突变建模平台，阐明潜在的细胞机制，并识别和评估新的基于机制的 AF 疗法。这平台不仅将实现一种更加基于机制的房颤治疗方法，而且还将 “个性化”治疗可以提高个体患者的疗效并降低毒性。