Targeting the host factor HGS–viral membrane protein interaction in coronavirus infection
Xubing Long, Rongrong Chen, Rong Bai, Buyun Tian, Yu Cao, Kangying Chen, Fuyu Li, Yiliang Wang, Yongjie Tang, Qi Yang, Liping Ma, Fan Wang, Maoge Zhou, Xianjie Qiu, Yongzhi Lu, Jie Zheng, Peng Zhou, Xinwen Chen, Qian Liu, Xuepeng Wei, Yongxia Shi, Yanhong Xue, Jincun Zhao, Wei Ji, Liqiao Hu, Jinsai Shang, Tao Xu, Zonghong Li
Xubing Long, Rongrong Chen, Rong Bai, Buyun Tian, Yu Cao, Kangying Chen, Fuyu Li, Yiliang Wang, Yongjie Tang, Qi Yang, Liping Ma, Fan Wang, Maoge Zhou, Xianjie Qiu, Yongzhi Lu, Jie Zheng, Peng Zhou, Xinwen Chen, Qian Liu, Xuepeng Wei, Yongxia Shi, Yanhong Xue, Jincun Zhao, Wei Ji, Liqiao Hu, Jinsai Shang, Tao Xu, Zonghong Li
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Research Article Cell biology Microbiology Virology

Targeting the host factor HGS–viral membrane protein interaction in coronavirus infection

Abstract

While current antivirals primarily target viral proteins, host-directed strategies remain underexplored. Here, we performed a genome-wide CRISPR inhibition (CRISPRi) screening to identify the host protein, hepatocyte growth factor-regulated tyrosine kinase substrate (HGS), facilitating the pan-coronavirus infection both in vitro and in vivo. Mechanistically, HGS interacts with the viral membrane (M) protein, facilitating its trafficking to the ER-Golgi intermediate compartment for virion assembly. Conversely, HGS deficiency caused M retention in the ER, blocking assembly. Leveraging this interaction, we designed M-derived peptides and screened over 5,000 FDA-approved or commonly used drugs, identifying riboflavin tetrabutyrate (RTB). Both the peptides and RTB bind HGS and disrupt its interaction with the M protein, leading to M retention in the ER and subsequent blockade of virion assembly. These agents demonstrated broad anti-pan-coronavirus activity in vitro and in vivo. Collectively, our findings establish HGS as a druggable host target and identify RTB as a promising broad-spectrum antiviral candidate.

Authors

Xubing Long, Rongrong Chen, Rong Bai, Buyun Tian, Yu Cao, Kangying Chen, Fuyu Li, Yiliang Wang, Yongjie Tang, Qi Yang, Liping Ma, Fan Wang, Maoge Zhou, Xianjie Qiu, Yongzhi Lu, Jie Zheng, Peng Zhou, Xinwen Chen, Qian Liu, Xuepeng Wei, Yongxia Shi, Yanhong Xue, Jincun Zhao, Wei Ji, Liqiao Hu, Jinsai Shang, Tao Xu, Zonghong Li

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Figure 1

Genome-wide CRISPRi screens identify HGS as a host target for pan-coronavirus therapy.

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Genome-wide CRISPRi screens identify HGS as a host target for pan-corona...
(A) Schematic illustrating genome-wide CRISPRi screens based on surface membrane LAMP1 for the identification of host factors for coronavirus assembly and egress. dCas9-KRAB–expressing 17Cl-1 cells transduced with a genome-wide sgRNA library were infected with MHV for 7 h (MOI = 1). The 15% of cells with low ratio of cell surface LAMP1/total LAMP1 were enriched, and sgRNA abundance was determined by next-generation sequencing. (B) Gene enrichment for CRISPRi screen. Enrichment scores were determined by MaGECK analysis, and genes were colored by biological function. Dotted line indicates log10 (enrichment score) = 1. All genes and enrichment scores can be found in Supplemental Table 1. (C–G) RT-qPCR analysis of extracellular SARS-CoV-2 (C), HCoV-OC43 (D), WIV1 (E), and MHV (F) viral gRNA and HSV-1 (G) viral gDNA levels (hpi = 24 h, 48 h, 96 h, respectively; MOI = 1). N = 3 independent biological replications. (H) Schematic illustrating conditional Hgs-KO K18-hACE2 mice infected with SARS-CoV-2. (I) Immunohistochemical staining of HGS in lung of ctr or Hgs-KO-K18-hACE2 mice. Scale bar = 300 μm. (J, K, and M) Survival curves (J), body weight changes (K), and histopathology of hematoxylin and eosin–stained (HE-stained) lung tissues on day 7 (M). Quantitative analysis of pathological severity scores based on the percentage of affected area in lung tissues. (N = 9 for K18-hACE2 group and N = 8 for Hgs-KO group.) (L) Viral titration by focus-forming assay (FFA) with the supernatant of homogenized lung tissues on day 2 (N = 9 for each group). FFU, focus-forming units. Data are the mean ± SD. Significance testing for C–G was performed with 1-way ANOVA and Tukey’s multiple-comparison test. Significance testing for K–M was performed with a 2-tailed t test. *P ≤ 0.05, **P ≤ 0.005, ***P ≤ 0.0005, ****P ≤ 0.0001.