Increased SARS-CoV-2 Infection, Protease, and Inflammatory Responses in Chronic Obstructive Pulmonary Disease Primary Bronchial Epithelial Cells Defined with Single-Cell RNA Sequencing.
Increased SARS-CoV-2 Infection, Protease, and Inflammatory Responses in Chronic Obstructive Pulmonary Disease Primary Bronchial Epithelial Cells Defined with Single-Cell RNA Sequencing.
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DOI：
10.1164/rccm.202108-1901oc
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发表时间：
2022-09-15
影响因子：
24.7
通讯作者：
中科院分区：
文献类型：
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作者：
慢性阻塞性肺疾病（COPD）患者发生更严重的冠状病毒病（COVID-19）;然而，尚不清楚他们是否更容易感染严重急性呼吸综合征冠状病毒2（SARS-CoV-2），以及导致严重疾病的机制。确定SARS-CoV-2接种COPD患者的原代支气管上皮细胞（pBECs）是否支持更大的感染，并阐明相关的影响和机制。我们在SARS-CoV-2接种后7天对来自健康受试者和COPD患者的分化的pBEC进行了单细胞RNA测序分析。我们将变化与病毒滴度、促炎反应和IFN产生相关联。单细胞RNA测序显示，COPD pBEC的感染比健康细胞高24倍，这得到了空斑试验的支持。俱乐部/杯状和基底细胞是主要的人口感染和表达的mRNA参与病毒复制。参与SARS-CoV-2进入/感染的蛋白酶（TMPRSS 2和CTSB）在COPD中增加，蛋白酶抑制剂（丝氨酸蛋白酶抑制剂）下调更多。与COPD急性加重和严重COVID-19相关的炎性细胞因子增加，而IFN反应减弱。共表达分析揭示了具有高1/2型IFN应答的俱乐部/杯状细胞的突出群体，其是健康和COPD pBEC中对感染的免疫应答的重要驱动因素。蛋白酶和炎症失衡的治疗性抑制降低了病毒滴度和细胞因子应答，特别是在COPD pBEC中。COPD pBEC更容易受到SARS-CoV-2感染，因为辅助受体表达增加和蛋白酶失衡，并具有更大的炎症反应。在感染过程中出现了一个突出的IFN反应性俱乐部/杯状细胞群，这可能是免疫的重要驱动因素。治疗干预抑制SARS-CoV-2复制和随后的炎症。
Patients with chronic obstructive pulmonary disease (COPD) develop more severe coronavirus disease (COVID-19); however, it is unclear whether they are more susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and what mechanisms are responsible for severe disease. To determine whether SARS-CoV-2 inoculated primary bronchial epithelial cells (pBECs) from patients with COPD support greater infection and elucidate the effects and mechanisms involved. We performed single-cell RNA sequencing analysis on differentiated pBECs from healthy subjects and patients with COPD 7 days after SARS-CoV-2 inoculation. We correlated changes with viral titers, proinflammatory responses, and IFN production. Single-cell RNA sequencing revealed that COPD pBECs had 24-fold greater infection than healthy cells, which was supported by plaque assays. Club/goblet and basal cells were the predominant populations infected and expressed mRNAs involved in viral replication. Proteases involved in SARS-CoV-2 entry/infection (TMPRSS2 and CTSB) were increased, and protease inhibitors (serpins) were downregulated more so in COPD. Inflammatory cytokines linked to COPD exacerbations and severe COVID-19 were increased, whereas IFN responses were blunted. Coexpression analysis revealed a prominent population of club/goblet cells with high type 1/2 IFN responses that were important drivers of immune responses to infection in both healthy and COPD pBECs. Therapeutic inhibition of proteases and inflammatory imbalances reduced viral titers and cytokine responses, particularly in COPD pBECs. COPD pBECs are more susceptible to SARS-CoV-2 infection because of increases in coreceptor expression and protease imbalances and have greater inflammatory responses. A prominent cluster of IFN-responsive club/goblet cells emerges during infection, which may be important drivers of immunity. Therapeutic interventions suppress SARS-CoV-2 replication and consequent inflammation.