Laboratory of Retrovirology, The Rockefeller Universitygrid.134907.8, New York, New York, USA.
Weill Cornell/Rockefeller/Sloan-Kettering Tri-Institutional MD-PhD Program, New York, New York, USA.
mBio. 2022 Apr 26;13(2):e0300221. doi: 10.1128/mbio.03002-21. Epub 2022 Mar 1.
Emerging zoonotic viral pathogens threaten global health, and there is an urgent need to discover host and viral determinants influencing infection. We performed a loss-of-function genome-wide CRISPR screen in a human lung cell line using HCoV-OC43, a human betacoronavirus. One candidate gene, VPS29, a component of the retromer complex, was required for infection by HCoV-OC43, SARS-CoV-2, other endemic- and pandemic-threat coronaviruses, as well as ebolavirus. Notably, we observed a heightened requirement for VPS29 by the recently described Omicron variant of SARS-CoV-2 compared to the ancestral variant. However, VPS29 deficiency had no effect on certain other viruses that enter cells via endosomes and had an opposing, enhancing effect on influenza A virus infection. Deficiency in VPS29 or other retromer components caused changes in endosome morphology and acidity and attenuated the activity of endosomal proteases. These changes in endosome properties caused incoming coronavirus, but not influenza virus particles, to become entrapped therein. Overall, these data show how host regulation of endosome characteristics can influence cellular susceptibility to viral infection and identify a host pathway that could serve as a pharmaceutical target for intervention in zoonotic viral diseases. These data identify a host pathway by which VPS29 and associated factors control the endosomal environment in a manner that influences susceptibility to viral infection. This pathway could serve as a pharmaceutical target for intervention in zoonotic viral diseases, including those caused by coronaviruses, influenza viruses, and filoviruses, all of which are pandemic threats. Our findings show how host regulation of endosome characteristics can influence viral susceptibility in both a positive and negative manner.
新兴的人畜共患病毒病原体威胁着全球健康，因此迫切需要发现影响感染的宿主和病毒决定因素。我们在人肺细胞系中使用 HCoV-OC43（一种人类β冠状病毒）进行了全基因组 CRISPR 功能丧失筛选。候选基因之一 VPS29 是逆行体复合物的一个组成部分，是 HCoV-OC43、SARS-CoV-2、其他地方性和大流行威胁冠状病毒以及埃博拉病毒感染所必需的。值得注意的是，与 SARS-CoV-2 的原始变体相比，我们观察到最近描述的 SARS-CoV-2 的奥密克戎变体对 VPS29 的需求更高。然而，VPS29 缺陷对某些其他通过内体进入细胞的病毒没有影响，并且对流感 A 病毒感染具有相反的增强作用。VPS29 或其他逆行体成分的缺乏会导致内体形态和酸度发生变化，并减弱内体蛋白酶的活性。这些内体特性的变化导致进入的冠状病毒而不是流感病毒颗粒被捕获在内体中。总的来说，这些数据表明宿主如何调节内体特征可以影响细胞对病毒感染的易感性，并确定宿主途径可以作为干预人畜共患病病毒的药物靶点。这些数据确定了一种宿主途径，VPS29 和相关因子通过该途径以影响病毒感染易感性的方式控制内体环境。该途径可以作为干预人畜共患病病毒的药物靶点，包括冠状病毒、流感病毒和丝状病毒引起的人畜共患病病毒，这些病毒均构成大流行威胁。我们的研究结果表明，宿主对内体特征的调节如何以积极和消极的方式影响病毒易感性。
