A yeast-based system to study SARS-CoV-2 Mpro structure and to identify nirmatrelvir resistant mutations.

Autor: Ou, Jin, Lewandowski, Eric M., Hu, Yanmei, Lipinski, Austin A., Aljasser, Ali, Colon-Ascanio, Mariliz, Morgan, Ryan T., Jacobs, Lian M. C., Zhang, Xiujun, Bikowitz, Melissa J., Langlais, Paul R., Tan, Haozhou, Wang, Jun, Chen, Yu, Choy, John S.
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Zdroj: PLoS Pathogens; 8/31/2023, Vol. 19 Issue 8, p1-19, 19p
Abstrakt: The SARS-CoV-2 main protease (Mpro) is a major therapeutic target. The Mpro inhibitor, nirmatrelvir, is the antiviral component of Paxlovid, an orally available treatment for COVID-19. As Mpro inhibitor use increases, drug resistant mutations will likely emerge. We have established a non-pathogenic system, in which yeast growth serves as an approximation for Mpro activity, enabling rapid identification of mutants with altered enzymatic activity and drug sensitivity. The E166 residue is known to be a potential hot spot for drug resistance and yeast assays identified substitutions which conferred strong nirmatrelvir resistance and others that compromised activity. On the other hand, N142A and the P132H mutation, carried by the Omicron variant, caused little to no change in drug response and activity. Standard enzymatic assays confirmed the yeast results. In turn, we solved the structures of Mpro E166R, and Mpro E166N, providing insights into how arginine may drive drug resistance while asparagine leads to reduced activity. The work presented here will help characterize novel resistant variants of Mpro that may arise as Mpro antivirals become more widely used. Author summary: The SARS-CoV-2 virus has proven to be highly adept at evading the newly developed vaccines. Antiviral drugs provide an important alternative that can reduce disease severity. Nirmatrelvir, an orally available drug approved by the FDA to treat COVID-19, inhibits the SARS-CoV-2 main or 3C-like protease (Mpro or 3CLpro) and reduces the severity of COVID-19 infections. However, continued use of nirmatrelvir will likely drive the emergence of drug resistant Mpro mutations. Our study reports a rapid, inexpensive, and non-pathogenic system using baker's yeast that can determine if potential Mpro mutations may confer drug resistance. Furthermore, we solve the structures of two Mpro mutants first characterized in yeast that provide insights into how E166 can change to drive resistance versus leading to inactivity. Our results can aid in advancing our understanding of resistance mechanisms that will be important as nirmatrelvir and other protease inhibitors become more widely used. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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