Potassium Molybdate Blocks APN-dependent Coronavirus Entry by Degrading Receptor Via PIK3C3-mediated Autophagy

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2024 Dec 6

PMID 39641621

Authors

Yunhang Zhang

Na Zhang

Yue Zhang

Yang Li

Ning Yang

Yifei Cai

Chen Tan

Jing Zhao

Wenjie Li

Yuanyuan Liu

Xue Rui

Junfei Wu

Yuguang Fu

Guangliang Liu

Affiliations

Soon will be listed here.

Abstract

Swine enteric coronaviruses pose a significant challenge to the global pig industry, inflicting severe diarrhea and high mortality rates among piglets, and resulting in substantial economic losses. In our clinical practice, we observed that the addition of potassium molybdate (PM) to the feed could dramatically reduce diarrhea and diarrhea-related mortality in piglets. However, the underlying mechanisms remain elusive and merit further investigation. In this study, we revealed that PM effectively inhibited the infection of both aminopeptidase N (APN)-dependent coronaviruses, transmissible gastroenteritis virus (TGEV), and porcine respiratory coronavirus (PRCV), both and . Specifically, PM was found to block TGEV and PRCV penetration by degrading the cell receptor APN through the upregulation of phosphatidylinositol 3-kinase catalytic subunit type 3 (PIK3C3) expression. In addition, knockdown and knockout of PIK3C3 resulted in the attenuation of PM-induced autophagy, thereby rescuing APN expression and viral infection. Correspondingly, replenishment of PIK3C3 in PIK3C3-null ST cells restored PM-mediated APN degradation and successfully blocked viral entry. Furthermore, our findings demonstrated that PM promoted the assembly of the PIK3C3-BECN1-ATG14 complex, leading to induced autophagic degradation by upregulating PIK3C3 Ser249 phosphorylation. experiments further confirmed that PM-induced PIK3C3-mediated autophagic degradation of APN, thereby limiting the pathogenicity of TGEV. In summary, our study for the first time identified the mechanism by which PM blocked TGEV and PRCV internalization by degrading the cell receptor APN via PIK3C3-mediated autophagy. This study provides valuable insights and potential strategies for preventing APN-restricted coronavirus infection.IMPORTANCEAminopeptidase N (APN) is one of the most important host receptors of coronavirus. Modulating APN expression can represent a novel approach for controlling APN-dependent coronaviruses and their variants infection. Here we found that a chemical compound potassium molybdate (PM) negatively regulates APN expression by inducing phosphatidylinositol 3-kinase catalytic subunit type 3 (PIK3C3)-mediated autophagy against APN-dependent coronavirus internalization, including transmissible gastroenteritis virus (TGEV) and porcine respiratory coronavirus (PRCV). Furthermore, PM can promote PIK3C3-BECN1-ATG14 complex assembly to induce autophagic degradation of APN by upregulating PIK3C3 Ser249 phosphorylation. Lastly, results from pig experiments also confirmed that PM can trigger PIK3C3-mediated autophagic degradation of APN to restrict TGEV pathogenicity without toxicity. Our findings underscore the promising potential of PM as an effective agent against APN-dependent coronavirus and potentially emerging viral disease entry.

References

Zhang Y, Chen H, Zou M, Oerlemans R, Shao C, Ren Y . Hypericin Inhibit Alpha-Coronavirus Replication by Targeting 3CL Protease. Viruses. 2021; 13(9). PMC: 8473218. DOI: 10.3390/v13091825. View

Zhang Y, Yang N, Li Y, Tan C, Cai Y, Rui X . Transmissible gastroenteritis virus induces inflammatory responses via RIG-I/NF-κB/HIF-1α/glycolysis axis in intestinal organoids and . J Virol. 2024; 98(6):e0046124. PMC: 11237398. DOI: 10.1128/jvi.00461-24. View

Wang P, Bai J, Liu X, Wang M, Wang X, Jiang P . Tomatidine inhibits porcine epidemic diarrhea virus replication by targeting 3CL protease. Vet Res. 2020; 51(1):136. PMC: 7656508. DOI: 10.1186/s13567-020-00865-y. View

Li L, Yu X, Zhang H, Cheng H, Hou L, Zheng Q . In vitro antiviral activity of Griffithsin against porcine epidemic diarrhea virus. Virus Genes. 2019; 55(2):174-181. PMC: 7089098. DOI: 10.1007/s11262-019-01633-7. View

Ito T, Sunada K, Nagai T, Ishiguro H, Nakano R, Suzuki Y . Preparation of cerium molybdates and their antiviral activity against bacteriophage Φ6 and SARS-CoV-2. Mater Lett. 2021; 290:129510. PMC: 7876479. DOI: 10.1016/j.matlet.2021.129510. View

Delmas B, Gelfi J, Sjostrom H, Noren O, Laude H . Further characterization of aminopeptidase-N as a receptor for coronaviruses. Adv Exp Med Biol. 1993; 342:293-8. DOI: 10.1007/978-1-4615-2996-5_45. View

Stucki J, Simon H . Mathematical modeling of the regulation of caspase-3 activation and degradation. J Theor Biol. 2005; 234(1):123-31. DOI: 10.1016/j.jtbi.2004.11.011. View

Wang C, Guan Y, Lv M, Zhang R, Guo Z, Wei X . Manganese Increases the Sensitivity of the cGAS-STING Pathway for Double-Stranded DNA and Is Required for the Host Defense against DNA Viruses. Immunity. 2018; 48(4):675-687.e7. DOI: 10.1016/j.immuni.2018.03.017. View

Li W, Luo R, He Q, van Kuppeveld F, Rottier P, Bosch B . Aminopeptidase N is not required for porcine epidemic diarrhea virus cell entry. Virus Res. 2017; 235:6-13. PMC: 7114539. DOI: 10.1016/j.virusres.2017.03.018. View

10.

Millet J, Jaimes J, Whittaker G . Molecular diversity of coronavirus host cell entry receptors. FEMS Microbiol Rev. 2020; 45(3). PMC: 7665467. DOI: 10.1093/femsre/fuaa057. View

11.

Klionsky D, Codogno P . The mechanism and physiological function of macroautophagy. J Innate Immun. 2013; 5(5):427-33. PMC: 6741458. DOI: 10.1159/000351979. View

12.

Jin S, He X, Ma L, Zhuang Z, Wang Y, Lin M . Suppression of ACE2 SUMOylation protects against SARS-CoV-2 infection through TOLLIP-mediated selective autophagy. Nat Commun. 2022; 13(1):5204. PMC: 9440653. DOI: 10.1038/s41467-022-32957-y. View

13.

Luo L, Wang S, Zhu L, Fan B, Liu T, Wang L . Aminopeptidase N-null neonatal piglets are protected from transmissible gastroenteritis virus but not porcine epidemic diarrhea virus. Sci Rep. 2019; 9(1):13186. PMC: 6742759. DOI: 10.1038/s41598-019-49838-y. View

14.

Paudel R, Lu D, Roy Chowdhury S, Monroy E, Wang J . Targeted Protein Degradation via Lysosomes. Biochemistry. 2022; 62(3):564-579. PMC: 10245383. DOI: 10.1021/acs.biochem.2c00310. View

15.

Ji C, Wang B, Zhou J, Huang Y . Aminopeptidase-N-independent entry of porcine epidemic diarrhea virus into Vero or porcine small intestine epithelial cells. Virology. 2018; 517:16-23. PMC: 7127557. DOI: 10.1016/j.virol.2018.02.019. View

16.

Hood M, Skaar E . Nutritional immunity: transition metals at the pathogen-host interface. Nat Rev Microbiol. 2012; 10(8):525-37. PMC: 3875331. DOI: 10.1038/nrmicro2836. View

17.

Galluzzi L, Pietrocola F, Levine B, Kroemer G . Metabolic control of autophagy. Cell. 2014; 159(6):1263-76. PMC: 4500936. DOI: 10.1016/j.cell.2014.11.006. View

18.

Nimitvilai S, Suputtamongkol Y, Poolvivatchaikarn U, Rassamekulthana D, Rongkiettechakorn N, Mungaomklang A . A Randomized Controlled Trial of Combined Ivermectin and Zinc Sulfate versus Combined Hydroxychloroquine, Darunavir/Ritonavir, and Zinc Sulfate among Adult Patients with Asymptomatic or Mild Coronavirus-19 Infection. J Glob Infect Dis. 2022; 14(2):69-74. PMC: 9336605. DOI: 10.4103/jgid.jgid_281_21. View

19.

Korolchuk V, Menzies F, Rubinsztein D . Mechanisms of cross-talk between the ubiquitin-proteasome and autophagy-lysosome systems. FEBS Lett. 2009; 584(7):1393-8. DOI: 10.1016/j.febslet.2009.12.047. View

20.

Sarli G, DAnnunzio G, Gobbo F, Benazzi C, Ostanello F . The Role of Pathology in the Diagnosis of Swine Respiratory Disease. Vet Sci. 2021; 8(11). PMC: 8618091. DOI: 10.3390/vetsci8110256. View