Modeling the efficiency of filovirus entry into cells in vitro: Effects of SNP mutations in the receptor molecule

Kwang Su Kim; Tatsunari Kondoh; Yusuke Asai; Ayato Takada; Shingo Iwami

doi:10.1371/journal.pcbi.1007612

Modeling the efficiency of filovirus entry into cells in vitro: Effects of SNP mutations in the receptor molecule

PLoS Comput Biol. 2020 Sep 28;16(9):e1007612. doi: 10.1371/journal.pcbi.1007612. eCollection 2020 Sep.

Authors

Kwang Su Kim¹, Tatsunari Kondoh², Yusuke Asai³, Ayato Takada², Shingo Iwami^{1

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Affiliations

¹ Department of Biology, Kyushu University, Fukuoka, Japan.
² Research Center for Zoonosis Control, Hokkaido University, Sapporo, Japan.
³ Disease Control and Prevention Center, National Center for Global Health and Medicine, Tokyo, Japan.
⁴ MIRAI, Japan Science and Technology Agency, Saitama, Japan.
⁵ CREST, Japan Science and Technology Agency, Saitama, Japan.
⁶ Science Groove Inc., Fukuoka, Japan.

Abstract

Interaction between filovirus glycoprotein (GP) and the Niemann-Pick C1 (NPC1) protein is essential for membrane fusion during virus entry. Some single-nucleotide polymorphism (SNPs) in two surface-exposed loops of NPC1 are known to reduce viral infectivity. However, the dependence of differences in entry efficiency on SNPs remains unclear. Using vesicular stomatitis virus pseudotyped with Ebola and Marburg virus GPs, we investigated the cell-to-cell spread of viruses in cultured cells expressing NPC1 or SNP derivatives. Eclipse and virus-producing phases were assessed by in vitro infection experiments, and we developed a mathematical model describing spatial-temporal virus spread. This mathematical model fit the plaque radius data well from day 2 to day 6. Based on the estimated parameters, we found that SNPs causing the P424A and D508N substitutions in NPC1 most effectively reduced the entry efficiency of Ebola and Marburg viruses, respectively. Our novel approach could be broadly applied to other virus plaque assays.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Ebolavirus / genetics
Ebolavirus / pathogenicity
Ebolavirus / physiology*
Humans
Marburgvirus / genetics
Marburgvirus / pathogenicity
Marburgvirus / physiology*
Models, Biological*
Mutation
Polymorphism, Single Nucleotide
Viral Plaque Assay
Virus Internalization*

Grants and funding

This study was supported in part by the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education (2019R1A6A3A12031316 to K.S.K.); JSPS Scientific Research (KAKENHI) B 18KT0018 (to S.I.), 18H01139 (to S.I.), 16H04845 (to S.I.), Scientific Research in Innovative Areas 20H05042 (to S.I.), 19H04839 (to S.I.), 18H05103 (to S.I.); AMED CREST 19gm1310002 (to S.I.); AMED J-PRIDE 18fm0208101 (to A.T.), 19fm0208006s0103, 19fm0208014h0003, and19fm0208019h0103 (to S.I.); AMED Japan Program for Infectious Diseases Research and Infrastructure, 20wm0325007h0001, 20wm0325004s0201, 20wm0325012s0301, 20wm0325015s0301 (to S.I.); AMED Research Program on HIV/AIDS 19fk0410023s0101 (to S.I.); AMED Research Program on Emerging and Re-emerging Infectious Diseases 19fk0108156h0001 and 20fk0108140s0801 (to S.I.); AMED Program for Basic and Clinical Research on Hepatitis 19fk0210036h0502 (to S.I.); AMED Program on the Innovative Development and the Application of New Drugs for Hepatitis B 19fk0310114h0103 (to S.I.); JST MIRAI (to S.I.); Mitsui Life Social Welfare Foundation (to K.W. and S.I.); Shin-Nihon of Advanced Medical Research (to S.I.); Suzuken Memorial Foundation (to S.I.); Life Science Foundation of Japan (to S.I.); SECOM Science and Technology Foundation (to S.I.); The Japan Prize Foundation (to S.I.); Fukuoka Financial Group, Inc. (to S.I.); Kyusyu Industrial Advancement Center Gapfund Program (to S.I.); Foundation for the Fusion of Science and Technology (to S.I.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.