Investigating the role of matrix metalloproteinases in SARS-CoV-2 infection

Professor Chu, Hin   (Principal Investigator (PI))

24

2022-06-30

82000

Investigating the role of matrix metalloproteinases in SARS-CoV-2 infection

matrix metalloproteinase, SARS-CoV-2, virus entry

VirologyMicrobiology

202111159140

Seed Fund for PI Research – Basic Research

2021

Completed

The causative agent of Coronavirus Disease 2019 (COVID-19), severe acute respiratory coronavirus 2 (SARS-CoV-2), was emerged in late 2019 and has resulted in an unprecedented pandemic, causing severe mortality and morbidity among humans. As of December 2021, the virus has infected over 282 million people with more than 5.4 million associated deaths. The treatment options of SARS-CoV-2 infection remain limited and the protection efficiency of vaccines is still suboptimal, particularly against the emerging SARS-CoV-2 variants including Delta and Omicron. In this regard, understanding on the fundamental biology of SARS-CoV-2 infection will not only provide basic knowledge on the virus but will also reveal novel targets for potential intervention strategies against the pandemic. Coronavirus entry requires virus attachment, receptor-binding, follows by membrane fusion (1). Virus attachment of SARS-CoV-2 on the cell surface can be mediated by heparan sulphate (2), follows by receptor recognition through angiotensin-converting enzyme 2 (ACE2) and the viral spike protein (3). Simultaneously, SARS-CoV-2 spike is cleavage activated by the transmembrane serine protease, transmembrane protease, serine 2 (TMPRSS2), which triggers fusion of the viral spike with the host cell membrane (3). Despite utilizing the same host receptor, ACE2, for virus entry, SARS-CoV-2 and SARS-CoV-1 appear to differ dramatically in transmissibility and infectability. SARS-CoV-1 infected approximately 8000 patients and quickly disappeared in about one year. In contrast, SARS-CoV-2 has infected over 200 million people and the pandemic is still ongoing. In line with these observations, SARS-CoV-2 replicates significantly more efficiently in the human respiratory tract than that of SARS-CoV-1 (4, 5), and appears to have a boarder tissue tropism than SARS-CoV-1. Notably, SARS-CoV-2 spike contains a four amino acid insertion at the S1/S2 junction that results in a ""RRAR"" sequence, which corresponded to a canonical furin-like cleavage site (6, 7). These residues are absent in the spike protein of SARS-CoV-1, SARS-CoV-related coronavirus (SARSr-CoV), and the closely related bat coronavirus RaTG13 (8). In addition to furin, a recent study demonstrated that the inserted furin-like cleavage site could similarly promote SARS-CoV-2 spike cleavage by other host proteases (9). Collectively, these findings suggest that the high transmissibility and infectability of SARS-CoV-2 may be contributed by its broad protease usage. Extending from the existing evidence, in the pilot study of this proposal we investigated the role of a comprehensive list of transmembrane protease in facilitating SARS-CoV-2 entry. Interestingly, our results suggested that multiple members of the transmembrane serine protease family in addition to TMPRSS2, including TMPRSS11D, TMPRSS11E, TMPRSS11F, TMPRSS13, and TMPRSS14, could facilitate SARS-CoV-2 entry. Moreover, members of other transmembrane protease family including that from the transmembrane matrix metalloproteinases (MMP) family were found to similarly facilitate SARS-CoV-2 entry. Thus, these preliminary findings revealed that in addition to TMPRSS2, a repertoire of additional transmembrane proteases can also facilitate SARS-CoV-2 entry, which may contribute to its efficient infection and transmission among humans. In the proposed study, we aim to evaluate the physiological relevance of the identified matrix metalloproteinases in SARS-CoV-2 entry and replication. Overall, the proposed study will provide novel knowledge on SARS-CoV-2 entry and reveal new targets of intervention against COVID-19. Reference: 1. Li F. Receptor recognition mechanisms of coronaviruses: a decade of structural studies. J Virol 2015; 89: 1954-1964. 2. Chu H, Hu B, Huang X, Chai Y, Zhou D, Wang Y, Shuai H, Yang D, Hou Y, Zhang X, Yuen TT, Cai JP, Zhang AJ, Zhou J, Yuan S, To KK, Chan IH, Sit KY, Foo DC, Wong IY, Ng AT, Cheung TT, Law SY, Au WK, Brindley MA, Chen Z, Kok KH, Chan JF, Yuen KY. Host and viral determinants for efficient SARS-CoV-2 infection of the human lung. Nat Commun 2021; 12: 134. 3. Hoffmann M, Kleine-Weber H, Schroeder S, Kruger N, Herrler T, Erichsen S, Schiergens TS, Herrler G, Wu NH, Nitsche A, Muller MA, Drosten C, Pohlmann S. SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. Cell 2020; 181: 271-280 e278. 4. Chu H, Chan JF, Wang Y, Yuen TT, Chai Y, Hou Y, Shuai H, Yang D, Hu B, Huang X, Zhang X, Cai JP, Zhou J, Yuan S, Kok KH, To KK, Chan IH, Zhang AJ, Sit KY, Au WK, Yuen KY. Comparative replication and immune activation profiles of SARS-CoV-2 and SARS-CoV in human lungs: an ex vivo study with implications for the pathogenesis of COVID-19. Clin Infect Dis 2020. 5. Hui KPY, Cheung MC, Perera R, Ng KC, Bui CHT, Ho JCW, Ng MMT, Kuok DIT, Shih KC, Tsao SW, Poon LLM, Peiris M, Nicholls JM, Chan MCW. Tropism, replication competence, and innate immune responses of the coronavirus SARS-CoV-2 in human respiratory tract and conjunctiva: an analysis in ex-vivo and in-vitro cultures. Lancet Respir Med 2020; 8: 687-695. 6. Coutard B, Valle C, de Lamballerie X, Canard B, Seidah NG, Decroly E. The spike glycoprotein of the new coronavirus 2019-nCoV contains a furin-like cleavage site absent in CoV of the same clade. Antiviral Res 2020; 176: 104742. 7. Walls AC, Park YJ, Tortorici MA, Wall A, McGuire AT, Veesler D. Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein. Cell 2020; 181: 281-292 e286. 8. Andersen KG, Rambaut A, Lipkin WI, Holmes EC, Garry RF. The proximal origin of SARS-CoV-2. Nat Med 2020; 26: 450-452. 9. Jaimes JA, Millet JK, Whittaker GR. Proteolytic Cleavage of the SARS-CoV-2 Spike Protein and the Role of the Novel S1/S2 Site. iScience 2020; 23: 101212.