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bioRxiv · 10.1101/2019.12.19.882167

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

Kim, K. S.·Kondoh, T.·Asai, Y.·Takada, A.·Iwami, S.
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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. Author SummaryEbola virus belongs to the family Filoviridae, together with Marburg virus and Cueva virus. In 2015, the World Health Organization included Ebola and Marburg viruses among the infectious diseases that should be globally prioritized because these filoviruses can cause severe hemorrhagic fever in humans and nonhuman primates, and antiviral agents to these viruses are very limited. Filovirus particles bear the envelope glycoprotein (GP), which is the only viral surface protein and thus responsible for receptor binding and membrane fusion. Interaction between filovirus 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. In this study, combining in vitro experiments and mathematical models, we evaluated on the interaction between GP and wildtype and mutant NPC1, enabling us to estimate the cellular entry efficiency during plaque formation.

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Kim, K. S., Kondoh, T., Asai, Y., Takada, A., Iwami, S.. 2019-12-19. Modeling the efficiency of filovirus entry into cells in vitro: Effects of SNP mutations in the receptor molecule. https://doi.org/10.1101/2019.12.19.882167

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