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：
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
项目类别：
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

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.

项目摘要心房颤动（AF）与显着的发病率和死亡率增加有关。尽管最近进步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部分是高度变化的，部分原因是未能靶向治疗基本机制。尽管遗传方法为基础提供了重要的见解 AF的机制，由于在细胞模型中充分概括人AF的挑战。得出特定于患者的能力来自人类诱导的多能干细胞（IPSC）-CMS的心房心肌细胞（CMS）对建模与AF相关的突变和开发与特异性匹配的AF的细胞模型患者。但是，心房IPSC-CMS尚未用于阐明AF-的潜在细胞机制连接的突变和模型可遗传的AF。特定目标1将创建UIC多民族心房HIPSC-CM 生物座位的目的是从家族型AF产生心房IPSC-CM，以模拟AF链接突变。我们的试验数据表明，心房IPSC-CMS概括了电生理（EP）表型与AF连接的SCN5A突变，并作为靶向基础细胞机制的平台功能收益变体。尽管如此，增强IPSC-CM的成熟度仍然很重要成熟的CMS不仅将提供有关AF的潜在细胞机制的其他见解，还将提供确定对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-CMS的EP和结构成熟度可以通过精确地显着增强体内相关的细胞细胞，细胞 - 细胞基质和细胞溶因子的微环境工程相互作用，使这些细胞允许对AF的最佳建模。我们还将评估心脏的作用成纤维细胞在AF的发病机理中，并确定它们是否通过与房屋共同培养影响EP成熟 IPSC-CMS。特定的目标3将使用AF连接突变的基本细胞机制心房IPSC-CMS。我们将确定SCN5A-E428K和KCNQ1-IAP54-56突变的EP表型使用患者特异性心房IPSC-CMS。此外，来自两个亲属的心房IPSC-CM将是遗传的使用CRISPR-CAS9系统纠正以确定建立因果关系。因此，这个总体目标提案是利用Co-Pis（Drs。Darbar和Khetani）的互补技能来建立 UIC多种族心房HIPSC生物座席可作为建模AF链接突变的平台，阐明潜在的细胞机制，并识别和评估基于机制的新型AF疗法。这平台不仅将启用一种基于机制的方法来治疗AF，而且还将 “个性化”治疗，具有提高功效和对个别患者的毒性降低的毒性。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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