Examining COVID-19 in Down Syndrome Patients Using Human iPSC-Derived Organoids

项目摘要 严重急性呼吸系统综合征冠状病毒2型（SARS-CoV-2）大流行已感染2000多万人 截至2020年8月，全球范围内的人们都在改变我们的生活方式。尽管其病死率相对较低 率，新型冠状病毒病（COVID-19）可导致患有既存疾病的人患严重疾病 就像唐氏综合症（DS）患者一样。据预测，SARS-CoV-2可导致8.9倍的风险增加， DS患者中与COVID-19相关的住院和死亡，但我们的实验资源有限， 了解不同疾病易感性和进展的基本机制。这里我们 建议使用i）人类诱导多能干细胞（iPSC），ii）组织样类器官，iii）活SARS-CoV-2 iv）从COVID-19患者收集的血清。我们将从DS产生40个iPSC衍生的类器官， 非DS个体并感染SARS-CoV-2。我们将重点关注两大临床表现 常见于COVID-19危重患者：病毒性心肌炎和严重肺 炎症对于目标1，我们将产生由iPSC衍生的心肌细胞组成的心脏类器官， 内皮细胞和心脏成纤维细胞，以评估SARS-CoV-2后它们的功能和结构变化 感染对于目标2，我们将产生由iPSC衍生的肺上皮细胞、内皮细胞和内皮细胞组成的肺类器官。 细胞，肺泡巨噬细胞，并确定SARS-CoV-2感染后的炎症特征。对于这两个目标， 我们将进行单细胞RNA测序，以确定不同细胞类型之间的差异基因反应， 有助于SARS-CoV-2感染后的表型变化。

Project Summary Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic has infected more than 20 million people worldwide as of August 2020 and significantly altered our way of life. Despite its relative low case-fatality rate, the new coronavirus disease (COVID-19) can cause serious illness in people with pre-existing conditions like individuals with Down Syndrome (DS). It is projected that SARS-CoV-2 can cause 8.9-fold increased risk of COVID-19 related hospitalization and deaths in DS patients, yet we have limited experimental resource to understand the basic mechanisms underlying differential disease susceptibility and progression. Here we propose to use i) human induced pluripotent stem cells (iPSCs), ii) tissue-like organoids, iii) live SARS-CoV-2 virus and iv) serum collected from COVID-19 patients. We will generate 40 iPSC-derived organoids from DS and non-DS individuals and infect them with SARS-CoV-2. We will focus on two major clinical manifestations commonly observed in COVID-19 patients with critical conditions: viral myocarditis and severe pulmonary inflammation. For Aim 1, we will generate cardiac organoids composing of iPSC-derived cardiomyocytes, endothelial cells, and cardiac fibroblasts to assess their functional and structural changes after SARS-CoV-2 infection. For Aim 2, we will generate lung organoids composing of iPSC-derived lung epithelial cells, endothelial cells, alveolar macrophages, and identify an inflammatory signature after SARS-CoV-2 infection. For both Aims, we will perform single cell RNA sequencing to identify differential gene response among different cell types which contribute to phenotypic changes following SARS-CoV-2 infection.