Three-Dimensional Human Alveolar Stem Cell Culture Models Reveal Infection Response to SARS-CoV-2.
Three-Dimensional Human Alveolar Stem Cell Culture Models Reveal Infection Response to SARS-CoV-2.
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DOI：
10.1016/j.stem.2020.10.004
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发表时间：
2020-12-03
期刊：
影响因子：
23.9
通讯作者：
Lee JH
中科院分区：
文献类型：
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作者：
Youk J;Kim T;Evans KV;Jeong YI;Hur Y;Hong SP;Kim JH;Yi K;Kim SY;Na KJ;Bleazard T;Kim HM;Fellows M;Mahbubani KT;Saeb-Parsy K;Kim SY;Kim YT;Koh GY;Choi BS;Ju YS;Lee JH
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是目前流行病的原因，其感染人肺泡2型（hAT 2）细胞。表征发病机制对于开发疫苗和治疗剂至关重要。然而，缺乏反映hAT 2细胞的细胞生理学和病理学的模型限制了这项研究。在这里，我们开发了一种无饲养层的，长期的，三维（3D）培养技术的hAT 2细胞来源于原代人肺组织和调查感染反应的SARS冠状病毒2。通过基于成像的分析和单细胞转录组分析，我们揭示了快速的病毒复制和感染的hAT 2细胞中干扰素相关基因和促炎基因的表达增加，表明了强大的内源性先天免疫反应。进一步追踪在传播过程中获得的病毒突变，可以有效地从单个病毒进入中识别单个细胞的完全感染。我们的研究为SARS-CoV-2的发病机制和定义的3D hAT 2培养物作为呼吸系统疾病模型的应用提供了深入的见解。SARS-CoV-2在化学限定条件下长期3D培养原代hAT 2细胞感染体外hAT 2模型，并发生实质性超微结构变化单细胞RNA-seq发现两种不同先天免疫表型的细胞状态突变分析显示SARS-CoV-2进入hAT 2细胞进行完全感染为了研究对SARS-CoV-2的感染反应，Youk et al.在化学限定的条件下开发了人肺泡干细胞的3D培养模型。他们揭示了SARS-CoV-2有效地感染肺泡2型细胞，导致先天性免疫反应。单个病毒进入显示出个体细胞的完全感染。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is the cause of a present pandemic, infects human lung alveolar type 2 (hAT2) cells. Characterizing pathogenesis is crucial for developing vaccines and therapeutics. However, the lack of models mirroring the cellular physiology and pathology of hAT2 cells limits the study. Here, we develop a feeder-free, long-term, three-dimensional (3D) culture technique for hAT2 cells derived from primary human lung tissue and investigate infection response to SARS-CoV-2. By imaging-based analysis and single-cell transcriptome profiling, we reveal rapid viral replication and the increased expression of interferon-associated genes and proinflammatory genes in infected hAT2 cells, indicating a robust endogenous innate immune response. Further tracing of viral mutations acquired during transmission identifies full infection of individual cells effectively from a single viral entry. Our study provides deep insights into the pathogenesis of SARS-CoV-2 and the application of defined 3D hAT2 cultures as models for respiratory diseases. Long-term 3D culture of primary hAT2 cells in chemically defined conditions SARS-CoV-2 infects in vitro hAT2 models with substantial ultrastructural changes Single-cell RNA-seq finds two cellular states of distinct innate immune phenotypes Mutation analysis reveals single SARS-CoV-2 entry to a hAT2 cell for full infection To study infection response to SARS-CoV-2, Youk et al. developed a 3D culture model of human alveolar stem cells in chemically defined conditions. They reveal that SARS-CoV-2 productively infects alveolar type 2 cells, resulting in an innate immune response. Full infection of individual cells was exhibited from a single viral entry.