工程化表达 ACE2 的 MRC5 细胞可作为针对 SARS-CoV-2 的抗病毒药物发现的模型系统。

MRC5 cells engineered to express ACE2 serve as a model system for the discovery of antivirals targeting SARS-CoV-2.

机构信息

Drug Discovery and Disease Research Laboratory, Shionogi & Co., Ltd, Osaka, Japan.

Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.

出版信息

Sci Rep. 2021 Mar 8;11(1):5376. doi: 10.1038/s41598-021-84882-7.

Abstract

Although the spread of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) has resulted in a worldwide pandemic, there are currently no virus-specific drugs that are fully effective against SARS-CoV-2. Only a limited number of human-derived cells are capable of supporting SARS-CoV-2 replication and the infectivity of SARS-CoV-2 in these cells remains poor. In contrast, monkey-derived Vero cells are highly susceptibility to infection with SARS-CoV-2, although they are not suitable for the study of antiviral effects by small molecules due to their limited capacity to metabolize drugs compared to human-derived cells. In this study, our goal was to generate a virus-susceptible human cell line that would be useful for the identification and testing of candidate drugs. Towards this end, we stably transfected human lung-derived MRC5 cells with a lentiviral vector encoding angiotensin-converting enzyme 2 (ACE2), the cellular receptor for SARS-CoV-2. Our results revealed that SARS-CoV-2 replicates efficiently in MRC5/ACE2 cells. Furthermore, viral RNA replication and progeny virus production were significantly reduced in response to administration of the replication inhibitor, remdesivir, in MRC5/ACE2 cells compared with Vero cells. We conclude that the MRC5/ACE2 cells will be important in developing specific anti-viral therapeutics and will assist in vaccine development to combat SARS-CoV-2 infections.

摘要

虽然严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)的传播导致了全球大流行,但目前还没有针对 SARS-CoV-2 的完全有效的病毒特异性药物。只有有限数量的人源细胞能够支持 SARS-CoV-2 的复制,而这些细胞中的 SARS-CoV-2 感染性仍然很差。相比之下,猴源 Vero 细胞极易感染 SARS-CoV-2,但由于与人类来源的细胞相比,它们对药物的代谢能力有限,因此不适合用于研究小分子的抗病毒作用。在这项研究中,我们的目标是生成一种对病毒敏感的人源细胞系,用于鉴定和测试候选药物。为此,我们使用慢病毒载体将血管紧张素转换酶 2(ACE2),即 SARS-CoV-2 的细胞受体,稳定转染到人肺衍生的 MRC5 细胞中。我们的结果表明,SARS-CoV-2 在 MRC5/ACE2 细胞中高效复制。此外,与 Vero 细胞相比,在 MRC5/ACE2 细胞中,病毒 RNA 复制和子代病毒产生对复制抑制剂瑞德西韦的处理明显减少。我们得出结论,MRC5/ACE2 细胞将在开发特异性抗病毒治疗药物方面发挥重要作用,并有助于开发针对 SARS-CoV-2 感染的疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ad45/7940632/9ea1bc4a6efc/41598_2021_84882_Fig1_HTML.jpg

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