Novel bioengineering models to dissect cardiac cell-cell defects in arrhythmogenic cardiomyopathy

项目摘要 致心律失常性心肌病（ACM）的特征是进行性的纤维脂肪替代， 心肌心律失常和猝死致心肌病的纤维脂肪替代 通过产生非传导性基质，导致心肌梗死恶化， 导致心力衰竭的功能障碍。这种疾病的潜在机制仍不清楚;一个更好的方法是， 需要了解发病机制以找到更好的临床治疗方案。为了解决这个 挑战，需要可靠的物种特异性模型;在这里，我们提出开发一种新的人类模型， 这将作为一个系统来研究心脏纤维脂肪浸润的发病机制。这项研究整合了 工程和生物医学科学，应用组织工程，心脏生理学，生物信息学和干 细胞技术我们的长期目标是提供一种纤维脂肪心肌浸润模型， 潜在的疾病机制，这将导致患者急需的治疗方法的发展 患有与纤维脂肪浸润相关的心脏病的患者。其核心目标是 该提案旨在证明心肌的纤维脂肪浸润可以在3D工程中复制。 心脏组织，类似于缺乏收缩性和改变的电生理特性， 在患有ACM的患者中观察到。在我们的背景下，纤维脂肪浸润的分子特征 还将分析工程化的心脏组织模型。我们将从两个目标着手。在目标1中， 使用来自以下的hiPSC开发心肌纤维脂肪浸润的3D工程化心脏组织模型： ACM患者我们将联合收割机hiPSC-心肌细胞和hiPSC-心外膜细胞进行处理， 上皮-间充质转化;旨在类似ACM功能表型。在目标2中， 心外膜作为纤维脂肪浸润来源的作用;我们将开发3D工程心脏组织模型 使用来自健康供体的hiPSC的心肌纤维脂肪浸润。在这项研究中，我们提出了一个战略， 基于纤维脂肪浸润由心外膜激活诱导的证据; hiPSC衍生的心外膜细胞 将被处理以诱导它们进一步分化为成纤维细胞和脂肪细胞。我们将检查功能 和结构特性，沿着工程化心脏组织模型的单细胞转录组学。我们 我希望这项研究的结果将促进我们对特定线索的贡献的理解， ACM相关细胞在纤维脂肪重塑发病机制中的作用;我们的生理相关模型将有助于 阐明细胞间的相互作用和纤维脂肪浸润的发生和发展机制 心肌层该项目将通过领导开发一种 疾病的人体模型和加速应用生物医学技术来询问疾病 机制，这将有助于识别新的治疗靶点。

PROJECT SUMMARY Arrhythmogenic cardiomyopathy (ACM) is characterized by progressive fibrofatty replacement of the myocardium, arrhythmias, and sudden death. Fibrofatty substitution in arrhythmogenic cardiomyopathy contributes to worsening arrhythmogenesis by creating a non-conductive substrate, and causes ventricular dysfunction leading to heart failure. The mechanisms underlying this disease are still unclear; a better understanding of the pathogenesis is needed to find better options for clinical management. To address this challenge, reliable species-specific models are needed; here we propose to develop a novel human model, that will serve as a system to study the pathogenesis of cardiac fibrofatty infiltration. This study integrates engineering and biomedical sciences, applying tissue engineering, cardiac physiology, bioinformatics and stem cell technologies. Our long-term goal is to provide a model of fibrofatty myocardial infiltration to investigate underlying disease mechanisms, which will lead to the development of greatly needed therapies for patients who suffer from cardiac diseases related to the presence of fibrofatty infiltration. The central objective of this proposal is to demonstrate that fibrofatty infiltration of the myocardium can be replicated in a 3D engineered cardiac tissue, resembling deficient contractility and altered electrophysiological properties that mimic what is observed in patients that suffer from ACM. The molecular signatures of fibrofatty infiltration in the context of our engineered cardiac tissue model will also be analyzed. We will approach this in two aims. In Aim 1 we will develop a 3D engineered cardiac tissue model of fibrofatty infiltration of the myocardium using hiPSCs from patients with ACM. We will combine hiPSC-cardiomyocytes and hiPSC-epicardial cells treated to undergo epithelial-mesenchymal transition; aiming to resemble the ACM functional phenotype. In Aim 2, exploiting the role of the epicardium as source of fibrofatty infiltration; we will develop a 3D engineered cardiac tissue model of myocardial fibrofatty infiltration using hiPSCs from healthy donors. In this study, we propose a strategy based on evidence that fibrofatty infiltration is induced from epicardial activation; hiPSC-derived epicardial cells will be treated to induce their further differentiation into fibroblasts and adipocytes. We will examine functional and structural properties, along with single-cell transcriptomics of the engineered cardiac tissue models. We expect that results from this study will advance our understanding of the contribution of specific cues from ACM-related cells in the pathogenesis of fibrofatty remodeling; our physiologically relevant model will serve to unravel the cell-cell cross-talk and mechanisms responsible for initiation and progression of fibrofatty infiltration of the myocardium. This project will improve the health of patients with ACM by leading the development of a human model of the disease and accelerating the application of biomedical technologies to interrogate disease mechanisms, which will aid in the identification of novel therapeutic targets.