Identifying host and viral correlates for coronavirus pathogenesis

识别冠状病毒发病机制的宿主和病毒相关性

基本信息

批准号：
10642898
负责人：
VINEET D MENACHERY
金额：
$ 49.79万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-15 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10642898
关键词：
Attenuated Automobile Driving Binding Body Weight decreased CRISPR/Cas technology Cells Chimera organism Chiroptera Conserved Sequence Coronavirus Coronavirus Infections Coronavirus spike protein Data Disease Drug usage Epidemic Family Goals Human Immune response Impairment In Vitro Infection Integration Host Factors Lung Mediating Middle East Respiratory Syndrome Coronavirus Monitor Mus Mutation Pathogenesis Peptide Hydrolases Pharmacotherapy Proteins Respiratory Tract Infections SARS coronavirus Serine Protease Severe Acute Respiratory Syndrome Shapes Structural Models System Testing Tropism Viral Virulence Virus Work Zoonoses attenuation cross-species transmission experimental study in vivo insight knockout gene mutant novel novel coronavirus receptor receptor binding restoration reverse genetics transmission process zoonotic coronavirus

项目摘要

Abstract The coronavirus (CoV) spike protein is a key viral determinant responsible for receptor binding and fusion/entry. The spike protein has also been predicted to be the major factor driving cross-species transmission, allowing the emergence of epidemic strains like SARS- and MERS-CoV. In the first decade after SARS-CoV emergence, changes to the epidemic spike that allowed binding to a new host receptor were thought to underlie this zoonotic emergence. However, our work has shown that bat species already harbor SARS-like CoVs with spike proteins capable of infecting human cells. These results argue that for a subset of bat CoVs, receptor binding and infection of human cells is not the major barrier for emergence. We found that despite equivalent replication in vitro, chimeric viruses containing bat CoV spikes have reduced virulence in vivo. Mice infected with a chimeric SARS-CoV expressing the bat derived SHC014-CoV spike had reduced weight loss and lethality compared to SARS-CoV controls. Importantly, this attenuation occurs despite equivalent replication to SARS-CoV in the lung. The results indicate that virulence is dictated by more than just the ability to infect host cells in vitro. Notably, we also found that the SHC014 spike chimera has reduced infection of the large airways of the lung. These preliminary data shaped our central hypothesis that SARS- CoV virulence is predicated on both host interactions with and viral motifs in the CoV spike protein. Understanding the host and viral mechanisms that drive reduced airway infection may predict in vivo pathogenesis and have critical implications for zoonotic emergence. In this proposal, we explore the host factors and CoV spike changes that attenuate the zoonotic SHC014 spike in vivo. In part one, we examine tropism changes finding that the zoonotic SHC014 spike has impaired upper airway infection. We predict that this incompatibility relates to differences in host protease activity. We subsequently define the specific host proteases that mediate this attenuation using both in vitro and in vivo approaches. In part two, we use mouse-adaptation and structural analysis to predict spike changes responsible for attenuation of the SHC014 spike. We subsequently generate mutant viruses and restore the SHC014 spike or attenuate the SARS spike in vivo. Finally, we evaluate the mechanism of attenuation focusing on spike interactions with host proteases. Together, the proposal identifies host proteases and spike interactions that alter airway infection and dictate virulence following coronavirus infection. These findings provide critical insights for understanding virulence as well as have important implications for emergence and transmission of coronaviruses.

抽象的冠状病毒（COV）尖峰蛋白是负责受体结合和融合/输入。尖峰蛋白也被预测是驱动跨物种的主要因素传播，允许SARS-和MERS-COV等流行病的出现。在之后的头十年 SARS-COV的出现，允许与新宿主受体结合的流行病的变化是被认为是这种人畜共努性出现的基础。但是，我们的工作表明蝙蝠物种已经藏有具有尖峰蛋白的SARS样COV能够感染人类细胞。这些结果表明，对于人类细胞的BAT COV，受体结合和感染并不是出现的主要障碍。我们发现，尽管体外复制等效，但含有BAT COV尖峰的嵌合病毒却降低了体内毒力。用嵌合SARS-COV感染的小鼠表达蝙蝠衍生的SHC014-cov尖峰有与SARS-COV对照相比，体重减轻和致死性降低。重要的是，尽管等效复制与肺中的SARS-COV。结果表明毒力由超过只是在体外感染宿主细胞的能力。值得注意的是，我们还发现SHC014尖峰嵌合体减少了肺部大气道的感染。这些初步数据塑造了我们的中心假设，即 COV毒力是在COV尖峰蛋白中与宿主相互作用和病毒基序的鉴定的。了解驱动降低气道感染的宿主和病毒机制可能预测体内发病机理，对人畜共患病具有关键意义。在此提案中，我们探讨了宿主因素和COV尖峰变化，从而减弱了人畜共动性SHC014 Spike 体内。在第一部分中，我们检查了潮流的变化，发现人畜共动性SHC014尖峰已损害上部气道感染。我们预测这种不兼容性与宿主蛋白酶活性的差异有关。我们随后，定义了使用体外和体内介导这种衰减的特定宿主蛋白酶方法。在第二部分中，我们使用鼠标适应和结构分析来预测峰值变化负责SHC014 SPIKE的衰减。随后，我们生成突变病毒并恢复 SHC014尖峰或衰减体内Sars Spike。最后，我们评估了衰减的机制专注于与宿主蛋白酶的尖峰相互作用。该提案一起确定宿主蛋白酶和尖峰改变气道感染并决定冠状病毒感染后的毒力的相互作用。这些发现提供理解毒力的关键见解，并对出现和冠状病毒的传播。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Binding and entering: COVID finds a new home.

DOI：
10.1371/journal.ppat.1009857
发表时间：
2021-08
期刊：
PLoS pathogens
影响因子：
6.7
作者：
Vu MN;Menachery VD
通讯作者：
Menachery VD

Immune Profiling to Determine Early Disease Trajectories Associated With Coronavirus Disease 2019 Mortality Rate: A Substudy from the ACTT-1 Trial.

DOI：
10.1093/infdis/jiab035
发表时间：
2021-04-23
期刊：
The Journal of infectious diseases
影响因子：
0
作者：
Thiede JM;Gress AR;Libby SD;Ronayne CE;Matchett WE;Noren B;Billings JL;Menachery VD;Langlois RA;Kline S;Bold TD
通讯作者：
Bold TD

Nucleocapsid mutations in SARS-CoV-2 augment replication and pathogenesis.

DOI：
10.1101/2021.10.14.464390
发表时间：
2022-03-28
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Johnson, Bryan A;Zhou, Yiyang;Menachery, Vineet D
通讯作者：
Menachery, Vineet D

The variant gambit: COVID-19's next move.

DOI：
10.1016/j.chom.2021.02.020
发表时间：
2021-04-14
期刊：
Cell host & microbe
影响因子：
30.3
作者：
Plante JA;Mitchell BM;Plante KS;Debbink K;Weaver SC;Menachery VD
通讯作者：
Menachery VD

The N501Y spike substitution enhances SARS-CoV-2 infection and transmission.

DOI：
10.1038/s41586-021-04245-0
发表时间：
2022-03
期刊：
Nature
影响因子：
64.8
作者：
Liu Y;Liu J;Plante KS;Plante JA;Xie X;Zhang X;Ku Z;An Z;Scharton D;Schindewolf C;Widen SG;Menachery VD;Shi PY;Weaver SC
通讯作者：
Weaver SC