Q493K and Q498H substitutions in Spike promote adaptation of SARS-CoV-2 in mice

被引：93

作者：

Huang, Kun ^{[1
,2
,3
]}

Zhang, Yufei ^{[1
,2
,3
]}

Hui, Xianfeng ^{[1
,2
,3
]}

Zhao, Ya ^{[1
,2
,3
]}

Gong, Wenxiao ^{[1
,2
,3
]}

Wang, Ting ^{[1
,2
,3
]}

Zhang, Shaoran ^{[1
]}

Yang, Yong ^{[1
,2
,3
]}

Deng, Fei ^{[4
,5
]}

Zhang, Qiang ^{[1
,2
,3
]}

Chen, Xi ^{[2
]}

Yang, Ying ^{[1
,2
,3
]}

Sun, Xiaomei ^{[1
,2
,3
]}

Chen, Huanchun ^{[1
,2
]}

Tao, Yizhi J. ^{[6
]}

Zou, Zhong ^{[1
,2
,3
]}

Jin, Meilin ^{[1
,2
,3
]}

机构：

[1] Huazhong Agr Univ, State Key Lab Agr Microbiol, Wuhan 430070, Peoples R China

[2] Huazhong Agr Univ, Coll Vet Med, Wuhan 430070, Peoples R China

[3] Minist Agr, Key Lab Dev Vet Diagnost Prod, Wuhan 430070, Peoples R China

[4] Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China

[5] Chinese Acad Sci, Wuhan Inst Virol, Natl Virus Resource Ctr, Wuhan 430071, Peoples R China

[6] Rice Univ, Dept BioSci, Houston, TX 77005 USA

来源：

EBIOMEDICINE | 2021年 / 67卷

关键词：

SARS-CoV-2; Angiotensin-converting enzyme 2; Mouse-adapted strain; Adaptive mutations; TLR7/8; agonist; SARS-CORONAVIRUS; PROTEIN; INFECTION; PATHOGENESIS; REPLICATION; MORTALITY; COVID-19; DELETION; DISEASE; VARIANT;

D O I：

10.1016/j.ebiom.2021.103381

中图分类号：

R5 [内科学];

学科分类号：

1002 ; 100201 ;

摘要：

Background: An ideal animal model to study SARS-coronavirus 2 (SARS-CoV-2) pathogenesis and evaluate therapies and vaccines should reproduce SARS-CoV-2 infection and recapitulate lung disease like those seen in humans. The angiotensin-converting enzyme 2 (ACE2) is a functional receptor for SARS-CoV-2, but mice are resistant to the infection because their ACE2 is incompatible with the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein . Methods: SARS-CoV-2 was passaged in BALB/c mice to obtain mouse-adapted virus strain. Complete genome deep sequencing of different generations of viruses was performed to characterize the dynamics of the adaptive mutations in SARS-CoV-2. Indirect immunofluorescence analysis and Biolayer interferometry experiments determined the binding affinity of mouse-adapted SARS-CoV-2 WBP-1 RBD to mouse ACE2 and human ACE2. Finally, we tested whether TLR7/8 agonist Resiquimod (R848) could also inhibit the replication of WBP-1 in the mouse model. Findings: The mouse-adapted strain WBP-1 showed increased infectivity in BALB/c mice and led to severe interstitial pneumonia. We characterized the dynamics of the adaptive mutations in SARS-CoV-2 and demonstrated that Q493K and Q498H in RBD significantly increased its binding affinity towards mouse ACE2. Additionally, the study tentatively found that the TLR7/8 agonist Resiquimod was able to protect mice against WBP-1 challenge. Therefore, this mouse-adapted strain is a useful tool to investigate COVID-19 and develop new therapies. Interpretation: We found for the first time that the Q493K and Q498H mutations in the RBD of WBP-1 enhanced its interactive affinities with mACE2. The mouse-adapted SARS-CoV-2 provides a valuable tool for the evaluation of novel antiviral and vaccine strategies. This study also tentatively verified the antiviral activity of TLR7/8 agonist Resiquimod against SARS-CoV-2 in vitro and in vivo. (C) 2021 The Author(s). Published by Elsevier B.V.

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