Emergence and Magnitude of ML336 Resistance in Venezuelan Equine Encephalitis Virus Depend on the Microenvironment

被引:9
作者
Lee, Jasper [1 ]
Parvathareddy, Jyothi [2 ]
Yang, Dong [2 ]
Bansal, Shruti [2 ]
O'Connell, Kathryn [3 ]
Golden, Jennifer E. [4 ]
Jonsson, Colleen B. [1 ,2 ]
机构
[1] Univ Tennessee, Hlth Sci Ctr, Dept Microbiol Immunol & Biochem, Memphis, TN 38112 USA
[2] Univ Tennessee, Hlth Sci Ctr, Reg Biocontainment Lab, Memphis, TN 38112 USA
[3] Univ Tennessee, Hlth Sci Ctr, Lab Anim Care Unit, Memphis, TN USA
[4] Univ Wisconsin, Sch Pharm, Pharmaceut Sci Div, 425 N Charter St, Madison, WI 53706 USA
关键词
ML336; Venezuelan encephalitis virus; Alphavirus; antiviral agents; next-generation sequencing; INFECTION;
D O I
10.1128/JVI.00317-20
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Venezuelan equine encephalitis virus (VEEV) is a New World Alphavirus that can cause neurological disease and death in humans and equines following transmission from infected mosquitoes. Despite the continued epidemic threat of VEEV, and its potential use as a bioterrorism agent, there are no FDA-approved antivirals or vaccines for treatment or prevention. Previously, we reported the discovery of a small molecule, ML336, with potent antiviral activity against VEEV. To further explore the population-level resistance profiles of ML336, we developed a wholegenome next-generation sequencing (NGS) approach to examine single nucleotide polymorphisms (SNPs) from virus passaged in dose escalation studies in a nonhuman primate kidney epithelial and a human astrocyte cell line, Vero 76 and SVGA, respectively. We passaged VEEV TC-83 in these two cell lines over seven concentrations of ML336, starting at 50 nM. NGS revealed several prominent mutations in the nonstructural protein (nsP) 3 and nsP4 genes that emerged consistently in these two distinct in vitro environments-notably, a mutation at Q210 in nsP4. Several of these mutations were stable following passaging in the absence of ML336 in Vero 76 cells. Network analyses showed that the trajectory of resistance differed between Vero and SVGA. Moreover, the penetration of SNPs was lower in SVGA. In conclusion, we show that the microenvironment influenced the SNP profile of VEEV TC-83. Understanding the dynamics of resistance in VEEV against newly developed antiviral compounds will guide the design of optimal drug candidates and dosing regimens for minimizing the emergence of resistant viruses. IMPORTANCE RNA viruses, including Venezuelan equine encephalitis virus (VEEV), have high mutation rates that allow for rapid adaptation to selective pressures in their environment. Antiviral compounds exert one such pressure on virus populations during infections. Next-generation sequencing allows for examination of viruses at the population level, which enables tracking of low levels of single-nucleotide polymorphisms in the population over time. Therefore, the timing and extent of the emergence of resistance to antivirals can be tracked and assessed. We show here that in VEEV, the trajectory and penetration of antiviral resistance reflected the microenvironment in which the virus population replicates. In summary, we show the diversity of VEEV within a single population under antiviral pressure and two distinct cell types, and we show that population dynamics in these viruses can be examined to better understand how they evolve over time.
引用
收藏
页数:17
相关论文
共 27 条
[1]   Endemic Venezuelan equine encephalitis in the Americas: hidden under the dengue umbrella [J].
Aguilar, Patricia V. ;
Estrada-Franco, Jose G. ;
Navarro-Lopez, Roberto ;
Ferro, Cristina ;
Haddow, Andrew D. ;
Weaver, Scott C. .
FUTURE VIROLOGY, 2011, 6 (06) :721-740
[2]   Discovery of a Novel Compound with Anti-Venezuelan Equine Encephalitis Virus Activity That Targets the Nonstructural Protein 2 [J].
Chung, Dong-Hoon ;
Jonsson, Colleen B. ;
Tower, Nichole A. ;
Chu, Yong-Kyu ;
Sahin, Ergin ;
Golden, Jennifer E. ;
Noah, James W. ;
Schroeder, Chad E. ;
Sotsky, Julie B. ;
Sosa, Melinda I. ;
Cramer, Daniel E. ;
McKellip, Sara N. ;
Rasmussen, Lynn ;
White, E. Lucile ;
Schmaljohn, Connie S. ;
Julander, Justin G. ;
Smith, Jeffrey M. ;
Filone, Claire Marie ;
Connor, John H. ;
Sakurai, Yasuteru ;
Davey, Robert A. .
PLOS PATHOGENS, 2014, 10 (06)
[3]   Approved Antiviral Drugs over the Past 50 Years [J].
De Clercq, Erik ;
Li, Guangdi .
CLINICAL MICROBIOLOGY REVIEWS, 2016, 29 (03) :695-747
[4]   Clinical Evidence and Bioinformatics Characterization of Potential Hepatitis C Virus Resistance Pathways for Sofosbuvir [J].
Donaldson, Eric F. ;
Harrington, Patrick R. ;
O'Rear, Julian J. ;
Naeger, Lisa K. .
HEPATOLOGY, 2015, 61 (01) :56-65
[5]  
Drake JWH, 1999, P NATL ACAD SCI USA, V96, P4
[6]   REGULATION OF THE INTERFERON SYSTEM - EVIDENCE THAT VERO CELLS HAVE A GENETIC DEFECT IN INTERFERON-PRODUCTION [J].
EMENY, JM ;
MORGAN, MJ .
JOURNAL OF GENERAL VIROLOGY, 1979, 43 (APR) :247-252
[7]   Characterization of substitutions in the neuraminidase of A(H7N9) influenza viruses selected following serial passage in the presence of different neuraminidase inhibitors [J].
Farrukee, R. ;
Butler, J. ;
Reading, P. C. ;
Hurt, A. C. .
ANTIVIRAL RESEARCH, 2019, 168 :68-75
[8]   Influenza Virus Drug Resistance: A Time-Sampled Population Genetics Perspective [J].
Foll, Matthieu ;
Poh, Yu-Ping ;
Renzette, Nicholas ;
Ferrer-Admetlla, Anna ;
Bank, Claudia ;
Shim, Hyunjin ;
Malaspinas, Anna-Sapfo ;
Ewing, Gregory ;
Liu, Ping ;
Wegmann, Daniel ;
Caffrey, Daniel R. ;
Zeldovich, Konstantin B. ;
Bolon, Daniel N. ;
Wang, Jennifer P. ;
Kowalik, Timothy F. ;
Schiffer, Celia A. ;
Finberg, Robert W. ;
Jensen, Jeffrey D. .
PLOS GENETICS, 2014, 10 (02)
[9]   Antiviral drug resistance as an adaptive process [J].
Irwin, Kristen K. ;
Renzette, Nicholas ;
Kowalik, Timothy F. ;
Jensen, Jeffrey D. .
VIRUS EVOLUTION, 2016, 2 (01)
[10]   Apoptotic cell death is an important cause of neuronal injury in experimental Venezuelan equine encephalitis virus infection of mice [J].
Jackson, AC ;
Rossiter, JP .
ACTA NEUROPATHOLOGICA, 1997, 93 (04) :349-353