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

项目摘要 非典冠状病毒2型大流行已感染2000多万人 截至2020年8月，全球人口数量大幅增加，极大地改变了我们的生活方式。尽管它的病死率相对较低 新的冠状病毒病(新冠肺炎)会在有既往疾病的人中引起严重疾病。 比如唐氏综合症(DS)患者。据预测，SARS-CoV-2会使患上SARS-CoV的风险增加8.9倍 新冠肺炎相关DS患者的住院和死亡，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。在这里我们 建议使用1)人诱导多能干细胞(IPSCs)，2)组织样有机物，3)活的SARS-CoV-2 病毒和iv)新冠肺炎患者的血清。我们将从DS和DS中生产40个IPSC衍生的有机化合物 非DS患者，并使他们感染SARS-CoV-2。我们将重点关注两个主要的临床表现 常见于新冠肺炎危重患者：病毒性心肌炎和重症肺 发炎。对于目标1，我们将产生由IPSC来源的心肌细胞组成的心脏器官， 评估SARS-CoV-2后内皮细胞和心脏成纤维细胞的功能和结构变化 感染。对于目标2，我们将产生由IPSC来源的肺上皮细胞、内皮细胞组成的肺器官 细胞、肺泡巨噬细胞，并鉴定SARS-CoV-2感染后的炎症征象。为了这两个目标， 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.