SARS-CoV-2 Infection of Pluripotent Stem Cell-Derived Human Lung Alveolar Type 2 Cells Elicits a Rapid Epithelial-Intrinsic Inflammatory Response.
SARS-CoV-2 Infection of Pluripotent Stem Cell-Derived Human Lung Alveolar Type 2 Cells Elicits a Rapid Epithelial-Intrinsic Inflammatory Response.
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DOI：
10.1101/2020.06.30.175695
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发表时间：
2020-12-03
期刊：
影响因子：
23.9
通讯作者：
Kotton, Darrell N
中科院分区：
文献类型：
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作者：
Huang, Jessie;Hume, Adam J;Kotton, Darrell N
最严重和致命的SARS-CoV-2感染导致急性呼吸窘迫综合征，这是2019冠状病毒病（COVID-19）的一种临床表型，与靶向远端肺上皮的病毒粒子有关，特别是该组织的兼性祖细胞，肺泡上皮2型细胞（AT 2）。人们对人类肺泡对SARS-CoV-2感染的最初反应知之甚少，部分原因是无法从患者身上获得这些细胞，特别是在疾病的早期阶段。在这里，我们提出了一个体外人体模型，模拟与SARS-CoV-2的远端肺上皮细胞的初始顶端感染，使用AT 2已适应气液界面培养后，其衍生自诱导多能干细胞（iAT 2）。我们发现，SARS-CoV-2诱导感染的iAT 2的快速全球转录组学变化，其特征在于转变为主要由NF-kB靶基因编码的细胞因子分泌的炎性表型，延迟的上皮干扰素反应，以及成熟的肺泡上皮程序的快速丧失。随着时间的推移，感染的iAT 2表现出细胞毒性，可导致这些关键肺泡兼性祖细胞死亡，如在COVID-19肺尸检中体内观察到的那样。重要的是，使用iAT 2的药物测试证实了对remdesivir的抗病毒剂量反应，并证明了TMPRSS 2蛋白酶抑制的功效，验证了用于病毒进入人肺泡细胞的假定机制。我们的模型系统揭示了一个关键的肺靶细胞对感染的细胞内在反应，为进一步的药物开发提供了一个生理相关的平台，并促进了对COVID-19发病机制的更深入理解。严重COVID-19肺炎的一个标志是SARS-CoV-2感染肺泡的兼性祖细胞，肺泡上皮2型细胞（AT 2 s）。然而，无法从患者体内获得这些细胞，特别是在疾病的早期阶段，限制了对疾病开始的理解。在这里，我们提出了一个体外人体模型，通过使用已适应气液界面培养的诱导多能干细胞衍生的AT 2来模拟SARS-CoV-2对肺泡上皮的初始顶端感染。我们发现感染细胞中的快速转录组学变化，其特征在于向炎症表型的转变，伴有NF-κ B信号的上调和成熟肺泡程序的丧失。药物测试证实了remdesivir以及TMPRSS 2蛋白酶抑制的功效，验证了用于病毒进入肺泡细胞的假定机制。我们的模型系统揭示了一个关键的肺靶细胞对SARS-CoV-2感染的细胞内在反应，并应有助于药物开发。
The most severe and fatal infections with SARS-CoV-2 result in the acute respiratory distress syndrome, a clinical phenotype of coronavirus disease 2019 (COVID-19) that is associated with virions targeting the epithelium of the distal lung, particularly the facultative progenitors of this tissue, alveolar epithelial type 2 cells (AT2s). Little is known about the initial responses of human lung alveoli to SARS-CoV-2 infection due in part to inability to access these cells from patients, particularly at early stages of disease. Here we present an in vitro human model that simulates the initial apical infection of the distal lung epithelium with SARS-CoV-2, using AT2s that have been adapted to air-liquid interface culture after their derivation from induced pluripotent stem cells (iAT2s). We find that SARS-CoV-2 induces a rapid global transcriptomic change in infected iAT2s characterized by a shift to an inflammatory phenotype predominated by the secretion of cytokines encoded by NF-kB target genes, delayed epithelial interferon responses, and rapid loss of the mature lung alveolar epithelial program. Over time, infected iAT2s exhibit cellular toxicity that can result in the death of these key alveolar facultative progenitors, as is observed in vivo in COVID-19 lung autopsies. Importantly, drug testing using iAT2s confirmed an antiviral dose-response to remdesivir and demonstrated the efficacy of TMPRSS2 protease inhibition, validating a putative mechanism used for viral entry in human alveolar cells. Our model system reveals the cell-intrinsic responses of a key lung target cell to infection, providing a physiologically relevant platform for further drug development and facilitating a deeper understanding of COVID-19 pathogenesis.A hallmark of severe COVID-19 pneumonia is SARS-CoV-2 infection of the facultative progenitors of lung alveoli, the alveolar epithelial type 2 cells (AT2s). However, inability to access these cells from patients, particularly at early stages of disease, limits an understanding of disease inception. Here, we present an invitro human model that simulates the initial apical infection of alveolar epithelium with SARS-CoV-2 by using induced pluripotent stem cell-derived AT2s that have been adapted to air-liquid interface culture. We find a rapid transcriptomic change in infected cells, characterized by a shift to an inflammatory phenotype with upregulation of NF-kappaB signaling and loss of the mature alveolar program. Drug testing confirms the efficacy of remdesivir as well as TMPRSS2 protease inhibition, validating a putative mechanism used for viral entry in alveolar cells. Our model system reveals cell-intrinsic responses of a key lung target cell to SARS-CoV-2 infection and should facilitate drug development.