Identification and characterization of influenza variants resistant to a viral endonuclease inhibitor

Song, Min-Suk; Kumar, Gyanendra; Shadrick, William R.; Zhou, Wei; Jeevan, Trushar; Li, Zhenmei; Slavish, P. Jake; Fabrizio, Thomas P.; Yoon, Sun-Woo; Webb, Thomas R.; Webby, Richard J.; White, Stephen W.

doi:10.1073/pnas.1519772113

Title: Identification and characterization of influenza variants resistant to a viral endonuclease inhibitor

Journal Article · Mon Mar 14 00:00:00 EDT 2016 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1519772113· OSTI ID:1247357

Song, Min-Suk ^[1]; Kumar, Gyanendra ^[2]; Shadrick, William R. ^[3]; Zhou, Wei ^[4]; Jeevan, Trushar ^[1]; Li, Zhenmei ^[2]; Slavish, P. Jake ^[3];

^[1]; Yoon, Sun-Woo ^[1]; Webb, Thomas R. ^[4]; Webby, Richard J. ^[1]; White, Stephen W. ^[2]

St. Jude Children's Research Hospital, Memphis, TN (United States). Dept. of Infectious Diseases
St. Jude Children's Research Hospital, Memphis, TN (United States). Dept. of Structural Biology
St. Jude Children's Research Hospital, Memphis, TN (United States). Dept. of Chemical Biology and Therapeutics
SRI International, Menlo Park, CA (United States). Bioscience Division

The influenza endonuclease is an essential subdomain of the viral RNA polymerase. It processes host pre-mRNAs to serve as primers for viral mRNA and is an attractive target for antiinfluenza drug discovery. Compound L-742,001 is a prototypical endonuclease inhibitor, and we found that repeated passaging of influenza virus in the presence of this drug did not lead to the development of resistant mutant strains. Reduced sensitivity to L-742,001 could only be induced by creating point mutations via a random mutagenesis strategy. Furthermore, these mutations mapped to the endonuclease active site where they can directly impact inhibitor binding. Engineered viruses containing the mutations showed resistance to L-742,001 both in vitro and in vivo, with only a modest reduction in fitness. Introduction of the mutations into a second virus also increased its resistance to the inhibitor. When using the isolated wild-type and mutant endonuclease domains, we used kinetics, inhibitor binding and crystallography to characterize how the two most significant mutations elicit resistance to L-742,001. These studies lay the foundation for the development of a new class of influenza therapeutics with reduced potential for the development of clinical endonuclease inhibitor-resistant influenza strains.

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Research Organization:: St. Jude Children's Research Hospital, Memphis, TN (United States); SRI International, Menlo Park, CA (United States)

Sponsoring Organization:: USDOE; National Inst. of Health (NIH) (United States); American Lebanese Syrian Associated Charities (ALSAC) (United States)

Grant/Contract Number:: W‐31‐109‐Eng‐38; AI098757; CA21765; HHSN266200700005C; HHSN272201400006C

OSTI ID:: 1247357

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Vol. 113, Issue 13; ISSN 0027-8424

Publisher:: National Academy of Sciences, Washington, DC (United States)Copyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 44 works

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