Human organoid model for COVID-19 myocarditis

COVID-19 心肌炎的人体类器官模型

• 批准号: 10746509
• 负责人: Ying Mei
• 金额: $ 23.15万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-17 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10746509
• 关键词: 3-Dimensional; Actinin; Acute; Adult; Autopsy; Biopsy; COVID-19; COVID-19 cytokine storm; COVID-19 impact; COVID-19 patient; Calcium; Cardiac; Cardiac Myocytes; Cells; Circulation; Clinical; Coculture Techniques; Complication; Data; Disease model; Dissociation; Endothelial Cells; Exclusion; Fibroblasts; Foundations; Goals; Harvest; Heart; Heart Diseases; Heart Injuries; Hospitalization; Human; Hyperactivity; Immune; Immune system; In Vitro; Infiltration; Inflammation; Inflammatory; Interleukins; Investigation; Lung; Macrophage; Metabolic; Modeling; Morbidity - disease rate; Myocarditis; Myocardium; Netherlands; Organ; Organoids; PECAM1 gene; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phenotype; Research; Risk; Role; Sampling; Serum; Stains; Stimulus; Stromal Cells; Structure; Time; Tissues; VWF gene; Vascular Endothelium; Vimentin; comparative; cytokine; cytokine release syndrome; drug testing; experimental study; fabrication; follow-up; human model; human pluripotent stem cell; immune cell infiltrate; immune modulating agents; innovation; insight; migration; monocyte; mortality; peripheral blood; public health relevance; recruit; severe COVID-19; transcriptome; transcriptome sequencing; viral RNA; 3-Dimensional; Actinin; Acute; Adult; Autopsy; Biopsy; COVID-19; COVID-19 cytokine storm; COVID-19 impact; COVID-19 patient; Calcium; Cardiac; Cardiac Myocytes; Cells; Circulation; Clinical; Coculture Techniques; Complication; Data; Disease model; Dissociation; Endothelial Cells; Exclusion; Fibroblasts; Foundations; Goals; Harvest; Heart; Heart Diseases; Heart Injuries; Hospitalization; Human; Hyperactivity; Immune; Immune system; In Vitro; Infiltration; Inflammation; Inflammatory; Interleukins; Investigation; Lung; Macrophage; Metabolic; Modeling; Morbidity - disease rate; Myocarditis; Myocardium; Netherlands; Organ; Organoids; PECAM1 gene; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phenotype; Research; Risk; Role; Sampling; Serum; Stains; Stimulus; Stromal Cells; Structure; Time; Tissues; VWF gene; Vascular Endothelium; Vimentin; comparative; cytokine; cytokine release syndrome; drug testing; experimental study; fabrication; follow-up; human model; human pluripotent stem cell; immune cell infiltrate; immune modulating agents; innovation; insight; migration; monocyte; mortality; peripheral blood; public health relevance; recruit; severe COVID-19; transcriptome; transcriptome sequencing; viral RNA

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.

项目摘要：急性心脏损伤（ACI）发生在高达20-25％的住院Covid-19患者和 与发病率和死亡率的风险增加有关。 Covid-19已显示诱导免疫 系统“过度火”和“失火”，导致造影症发炎。这样的细胞因子风暴已经 通过直接细胞因子损伤和间接机制造成器官损伤（例如，招募 促炎性免疫细胞进入器官）。心肌炎（例如，单核细胞浸润）是常见的并发症 在COVID-19 ACI心中。但是，浸润的单核细胞对Covid-19 ACI心脏的影响仍然 定义不足。由于先前的研究仅限于任何一种临床样本（例如，外周血液， 尸检/活检组织）或体外2D共培养实验，这些研究产生了有限的功能见解。 3D 有机模型提供了一个强大的平台，以研究心脏和心脏之间的功能相互作用 循环中的免疫细胞。为了开发人类心脏的有机模型进行疾病建模，我们 开发的3D人类心脏器官由人多能干细胞衍生的心肌细胞组成 （HPSC-CMS），人类心脏成纤维细胞，内皮细胞和基质细胞。我们的初步数据显示了 白介素（IL）-1b处理的类器官概括了转录组，结构和功能的关键特征 COVID-19 ACI心。由于IL-1B不是19个特定刺激，因此这些结果为使用的基础奠定了基础 来自COVID-19 ACI患者的血清将Covid-19细胞因子损伤概括为心脏。另外， 类型器中的免疫细胞排除限制了COVID-19中超炎性的全部概括 ACI心。该建议的目的是利用血清和外周血单核细胞（PMBC） 从COVID-19 ACI患者收获到1）模拟COVID-19细胞因子损伤对人心脏的影响， 2）开发第一个体外共互联-19肌动炎器官模型，以模拟免疫球浸润至 COVID-19 ACI心脏并评估浸润的COVID-19。该提议的中心假设 是浸润的COVID-199免疫细胞（例如，单核细胞）加剧了COVID-19 ACI的过度炎症 心。该提案具有创新性，因为它将首次开发出心肌炎的有机模型 模拟免疫细胞浸润到心肌中。根据，我们将追求以下两个目标：1） 确定COVID-19 ACI血清对人心脏器官的影响，2）确定 COVID-19 ACI血清治疗的类器官，从COVID-19 ACI患者中募集免疫细胞。提议 研究将导致R01的随访，重点是渗透的效果和机制 COVID-19 ACI心脏中的单核细胞。此外，心肌炎的3D有机模型可用于 免疫调节药物对心脏的影响的机理研究。