Human organoid model for COVID-19 myocarditis

项目概述：急性心脏损伤(ACI)发生在高达20-25%的住院新冠肺炎患者和 与发病和死亡风险增加有关。新冠肺炎已被证明可诱导免疫 系统“过火”和“失火”，导致生理上的炎症。这样的细胞因子风暴一直是 被证明通过直接细胞因子侮辱和间接机制(例如，招募)导致器官损害 促炎症免疫细胞进入器官)。心肌炎(如单核细胞浸润)是一种常见的并发症。 在新冠肺炎ACI的心中。然而，渗透的单核细胞对新冠肺炎脑梗塞心脏的影响仍然存在 定义不足。由于先前的研究仅限于临床样本(例如，外周血， 无论是身体解剖/活组织检查)还是体外2D共培养实验，这些研究都产生了有限的功能见解。3D 器官模型为研究心脏和心脏之间的功能相互作用提供了一个强大的平台 循环中的免疫细胞。为了开发一种用于疾病建模的人体心脏器官模型，我们 由人多能干细胞来源的心肌细胞组成的三维人体心脏器官 (HPSC-CMS)、人心脏成纤维细胞、内皮细胞和基质细胞。我们的初步数据显示 白介素1b处理的有机化合物概述了转录组、结构和功能的关键特征。 新冠肺炎ACI红心。由于IL-1b不是新冠肺炎的特异性刺激因子，这些结果为使用奠定了基础 来自新冠肺炎急性脑梗塞患者的血清概括了新冠肺炎细胞因子对心脏的侮辱。此外， 排除器官中的免疫细胞限制了新冠肺炎炎症的全面重演 ACI红心。这项提议的目标是利用血清和外周血单核细胞(PMBC) 从新冠肺炎脑梗塞患者身上采集的数据用于1)模拟新冠肺炎细胞因子侮辱对人心脏的影响， 2)建立了第一个新冠肺炎心肌炎器官体外模型，模拟免疫细胞的渗透 并评估新冠肺炎急性心肌梗死后新冠肺炎单核细胞的浸润情况。这项提议的核心假设是 浸润的新冠肺炎免疫细胞(如单核细胞)加剧了新冠肺炎急性脑梗塞的炎症反应 红心。这项提议具有创新性，因为它将首次开发心肌炎的器官类型模型 模拟免疫细胞向心肌的渗透。因此，我们将追求以下两个目标：1) 2)测定新冠肺炎脑梗塞血清对人心脏脏器的影响。 新冠肺炎脑梗塞患者血清经有机化合物处理后，可从患者体内招募免疫细胞。建议数 研究将导致R01的后续应用，重点是渗透的效果和机制 新冠肺炎急性心肌梗死患者心脏的单核细胞。此外，心肌炎的3D器官模型可用于 免疫调节药物对心脏影响的机制研究。

Project Summary: Acute Cardiac Injuries (ACIs) occur in up to 20-25% of hospitalized COVID-19 patients and are associated with increased risks of morbidity and mortality. COVID-19 has been shown to induce immune system “overfiring” and “misfiring”, resulting in supraphysiological inflammation. Such cytokine storms have been shown to lead to organ damage through both direct cytokine insults and indirect mechanisms (e.g., recruitment of proinflammatory immune cells into organs). Myocarditis (e.g., monocyte infiltration) is a common complication in COVID-19 ACI hearts. However, the impacts of the infiltrated monocytes on COVID-19 ACI hearts remain under-defined. As previous studies have been limited to either clinical samples (e.g., peripheral blood, autopsy/biopsy tissues) or in vitro 2D co-culture experiments, these studies yield limited functional insights. 3D organotypic models provide a powerful platform to study the functional interactions between hearts and immune cells in circulation. To develop an organotypic model of human hearts for disease modeling, we developed 3D human cardiac organoids composed of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs), human cardiac fibroblasts, endothelial cells, and stromal cells. Our preliminary data showed the Interleukin (IL)-1b treated organoids recapitulated key features of transcriptome, structure, and function of COVID-19 ACI hearts. As IL-1b is not a COVID-19 specific stimulus, these results laid the foundation to use serum from COVID-19 ACI patients to recapitulate the COVID-19 cytokine insults to the hearts. In addition, the exclusion of immune cells in the organoids has limited the full recapitulation of hyperinflammation in COVID-19 ACI hearts. The goal of this proposal is to leverage the serum and peripheral blood mononuclear cells (PMBCs) harvested from COVID-19 ACI patients to 1) simulate the effects of COVID-19 cytokine insults on human hearts, 2) develop the first in vitro COVID-19 myocarditis organoid model to simulate immune cell infiltration to COVID-19 ACI hearts and assess the infiltrated COVID-19 monocytes. The central hypothesis of this proposal is that infiltrated COVID-19 immune cells (e.g., monocytes) exacerbate the hyperinflammation of COVID-19 ACI hearts. This proposal is innovative in that it will, for the first time, develop an organotypic model of myocarditis to simulate immune cell infiltration into myocardium. Accordingly, we will pursue the following 2 Aims: 1) Determine the effects of COVID-19 ACI serum on human cardiac organoids, and 2) Determine the ability of COVID-19 ACI serum treated organoids to recruit immune cells from COVID-19 ACI patients. The proposed studies will lead to follow-up R01 applications focusing on the effects and mechanisms of the infiltrated monocytes in COVID-19 ACI hearts. In addition, the 3D organotypic model of myocarditis can be used for mechanistic studies of the effects of immunomodulatory drugs on hearts.