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

项目总结 致心律失常性心肌病(ACM)的特点是进行性纤维脂肪替代 心肌、心律失常和猝死。致心律失常性心肌病的纤维脂肪替代 通过产生不导电的底物而加剧心律失常，并导致室性心动过速 功能障碍会导致心力衰竭。这种疾病背后的机制仍不清楚；一种更好的 需要了解发病机制，以便为临床治疗找到更好的选择。要解决这个问题 挑战，需要可靠的特定物种模型；在这里，我们建议开发一种新的人类模型， 这将为研究心肌纤维脂肪渗出的发病机制提供一个系统。这项研究整合了 工程和生物医学科学，应用组织工程学、心脏生理学、生物信息学和干细胞 细胞技术。我们的长期目标是提供一种用于研究纤维脂肪心肌浸润的模型。 潜在的疾病机制，这将导致为患者开发急需的治疗方法 患有与心脏疾病相关的纤维脂肪浸润者。这样做的中心目标是 建议证明心肌的纤维脂肪渗透可以在3D工程中复制 心脏组织，类似于收缩能力不足和改变的电生理特性，模拟 在患有ACM的患者中观察。在我们的研究背景下纤维脂肪渗透的分子特征 工程心脏组织模型也将被分析。我们将通过两个目标来实现这一点。在目标1中，我们将 利用hPSCs建立心肌纤维脂肪渗出的三维工程化心肌组织模型 ACM患者。我们将结合HiPSC-心肌细胞和HiPSC-心外膜细胞进行治疗 上皮-间充质转化；旨在类似ACM的功能表型。在目标2中，利用 心外膜作为纤维脂肪渗出来源的作用；我们将建立一个3D工程化心脏组织模型 使用来自健康供者的hPSCs研究心肌纤维脂肪的渗透。在本研究中，我们提出了一种策略 基于纤维脂肪浸润是由心外膜激活引起的证据；HiPSC来源的心外膜细胞 将被诱导进一步分化为成纤维细胞和脂肪细胞。我们将检查功能 以及结构特性，以及工程心脏组织模型的单细胞转录。我们 期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；我们的生理学相关模型将有助于 解开细胞间的相互作用和纤维脂肪渗透的发生和发展的机制 对心肌的影响。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审问疾病 机制，这将有助于确定新的治疗靶点。

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.