NF-κB 抑制剂 α 控制 ACE2 过表达的人气道类器官中的 SARS-CoV-2 感染。
NF-κB inhibitor alpha controls SARS-CoV-2 infection in ACE2-overexpressing human airway organoids.
发表日期：2024 Jul 04
作者：
Camille R Simoneau, Pei-Yi Chen, Galen K Xing, Jennifer M Hayashi, Irene P Chen, Mir M Khalid, Nathan L Meyers, Taha Y Taha, Kristoffer E Leon, Rahul K Suryawanshi, Maria McCavitt-Malvido, Tal Ashuach, Krystal A Fontaine, Lauren Rodriguez, Bastian Joehnk, Keith Walcott, Sreelakshmi Vasudevan, Xiaohui Fang, Mazharul Maishan, Shawn Schultz, Jeroen P Roose, Michael A Matthay, Anita Sil, Mehrdad Arjomandi, Nir Yosef, Melanie Ott
来源：
Stem Cell Research & Therapy
摘要：
随着 SARS-CoV-2 继续在世界范围内传播，需要易于处理的初级气道细胞模型来概括细胞对出现的病毒变异的内在反应。在这里，我们描述了一种过表达 ACE2 受体 (ACE2-OE) 的成体干细胞衍生的人气道类器官模型，该模型支持强大的病毒复制，同时保持气道上皮的 3D 结构和细胞多样性。 ACE2-OE 类器官感染 SARS-CoV-2 变体并进行单细胞 RNA 测序。干扰素 lambda 在低水平感染的细胞中上调，而 NF-kB 抑制剂 α 基因（编码 IkBa）在受感染的细胞中持续上调，其表达与感染水平呈正相关。共聚焦显微镜显示，感染细胞中的 IkBa 表达量高于旁观者细胞，但发现 IkBa 通常会阻止的同时发生的 NF-kB 核转位。过度表达不可降解的 IkBa 突变体会减少 NF-kB 易位并增加病毒感染。这些数据证明了 ACE2-OE 类器官在 SARS-CoV-2 研究中的功能，并强调受感染细胞中 NF-kB 反馈环路的强度控制着病毒复制。© 2024。作者。
As SARS-CoV-2 continues to spread worldwide, tractable primary airway cell models that recapitulate the cell-intrinsic response to arising viral variants are needed. Here we describe an adult stem cell-derived human airway organoid model overexpressing the ACE2 receptor (ACE2-OE) that supports robust viral replication while maintaining 3D architecture and cellular diversity of the airway epithelium. ACE2-OE organoids were infected with SARS-CoV-2 variants and subjected to single-cell RNA-sequencing. Interferon-lambda was upregulated in cells with low-level infection while the NF-kB inhibitor alpha gene (encoding IkBa) was consistently upregulated in infected cells, and its expression positively correlated with infection levels. Confocal microscopy showed more IkBa expression in infected than bystander cells, but found concurrent nuclear translocation of NF-kB that IkBa usually prevents. Overexpressing a nondegradable IkBa mutant reduced NF-kB translocation and increased viral infection. These data demonstrate the functionality of ACE2-OE organoids in SARS-CoV-2 research and underscore that the strength of the NF-kB feedback loop in infected cells controls viral replication.© 2024. The Author(s).