Extracellular vimentin is an attachment factor that facilitates SARS-CoV-2 entry into human endothelial cells.
Extracellular vimentin is an attachment factor that facilitates SARS-CoV-2 entry into human endothelial cells.
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DOI：
10.1073/pnas.2113874119
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发表时间：
2022-02-08
影响因子：
11.1
通讯作者：
Rahimi N
中科院分区：
文献类型：
--
作者：
Amraei R;Xia C;Olejnik J;White MR;Napoleon MA;Lotfollahzadeh S;Hauser BM;Schmidt AG;Chitalia V;Mühlberger E;Costello CE;Rahimi N
人血管紧张素转换酶2（ACE 2）是SARS-CoV-2最广为人知的进入受体。其他细胞成分参与病毒进入机制的可能性仍然未知。波形蛋白在人内皮细胞中表达，与SARS-CoV-2-spike结合，加速SARS-CoV-2进入。用纯化的波形蛋白处理肺ACE 2/A549癌细胞或ACE 2/A549细胞与表达波形蛋白的HEK-293细胞共培养增加ACE 2依赖性病毒进入。CR 3022抗体阻断波形蛋白与SARS-CoV-2-spike的相互作用并抑制SARS-CoV-2进入。波形蛋白可能促进SARS-CoV-2感染，并导致与COVID-19相关的血管并发症。SARS-CoV-2进入宿主细胞是病毒嗜性、传播和致病的关键步骤。血管紧张素转换酶2（ACE 2）已被确定为SARS冠状病毒的主要进入受体-2，然而，在病毒进入的其他细胞成分的可能参与尚未完全阐明。在这里，我们描述了波形蛋白（Vim），广泛表达于间充质来源的细胞中的中间丝蛋白，作为SARS冠状病毒-2对人内皮细胞的重要附着因子的鉴定。使用液相色谱-串联质谱法，我们确定Vim是一种与SARS-CoV-2刺突（S）蛋白结合的蛋白。我们发现，S-蛋白受体结合结构域（RBD）是足够的S-蛋白与Vim相互作用。进一步的分析表明，细胞外Vim结合SARS-CoV-2 S蛋白，并促进SARS-CoV-2感染，如通过用表达S的假型病毒和感染性SARS-CoV-2进行的进入试验所确定的。Vim与ACE 2的共表达增加了SARS-CoV-2在HEK-293细胞中的进入，并且shRNA介导的Vim敲低显著降低了SARS-CoV-2对人内皮细胞的感染。此外，表达ACE 2的A549细胞与纯化的Vim孵育增加了假型SARS-CoV-2-S的进入。CR 3022抗体识别SARS-CoV-2-S-RBD上的一个独特表位，但不干扰刺突与ACE 2的结合，可抑制Vim与CoV-2 S-RBD的结合，并中和病毒进入人内皮细胞，表明Vim在SARS-CoV-2感染内皮细胞中起关键作用。这项工作提供了对与血管系统相关的COVID-19发病机制的深入了解，对治疗和疫苗的开发具有重要意义。
Human angiotensin-converting enzyme 2 (ACE2) is the most widely known entry receptor for SARS-CoV-2. The possible involvement of other cellular components in viral entry mechanisms remains unknown. Vimentin is expressed in human endothelial cells, binds to SARS-CoV-2-spike, and expedites SARS-CoV-2 entry. Treatment of lung ACE2/A549 carcinoma cells with purified vimentin or coculture of ACE2/A549 cells with HEK-293 cells expressing vimentin increased ACE2-dependent viral entry. CR3022 antibody blocked vimentin interaction with SARS-CoV-2-spike and inhibited SARS-CoV-2 entry. Vimentin could facilitate SARS-CoV-2 infection and contribute to vascular complications associated with COVID-19. SARS-CoV-2 entry into host cells is a crucial step for virus tropism, transmission, and pathogenesis. Angiotensin-converting enzyme 2 (ACE2) has been identified as the primary entry receptor for SARS-CoV-2; however, the possible involvement of other cellular components in the viral entry has not yet been fully elucidated. Here we describe the identification of vimentin (VIM), an intermediate filament protein widely expressed in cells of mesenchymal origin, as an important attachment factor for SARS-CoV-2 on human endothelial cells. Using liquid chromatography–tandem mass spectrometry, we identified VIM as a protein that binds to the SARS-CoV-2 spike (S) protein. We showed that the S-protein receptor binding domain (RBD) is sufficient for S-protein interaction with VIM. Further analysis revealed that extracellular VIM binds to SARS-CoV-2 S-protein and facilitates SARS-CoV-2 infection, as determined by entry assays performed with pseudotyped viruses expressing S and with infectious SARS-CoV-2. Coexpression of VIM with ACE2 increased SARS-CoV-2 entry in HEK-293 cells, and shRNA-mediated knockdown of VIM significantly reduced SARS-CoV-2 infection of human endothelial cells. Moreover, incubation of A549 cells expressing ACE2 with purified VIM increased pseudotyped SARS-CoV-2-S entry. CR3022 antibody, which recognizes a distinct epitope on SARS-CoV-2-S-RBD without interfering with the binding of the spike with ACE2, inhibited the binding of VIM with CoV-2 S-RBD, and neutralized viral entry in human endothelial cells, suggesting a key role for VIM in SARS-CoV-2 infection of endothelial cells. This work provides insight into the pathogenesis of COVID-19 linked to the vascular system, with implications for the development of therapeutics and vaccines.
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影响因子：
64.5
作者：
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通讯作者：
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影响因子：
16.6
作者：
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作者：
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通讯作者：
Clevers, Hans