Identification and characterization of a novel broad spectrum virus entry inhibitor.

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Identification and characterization of a novel broad spectrum virus entry inhibitor.

J Virol. 2016 Feb 24;

Authors: Chou YY, Cuevas C, Carocci M, Stubbs SH, Ma M, Cureton DK, Chao L, Evesson F, He K, Yang PL, Whelan SP, Ross SR, Kirchhausen T, Gaudin R

Abstract
Virus entry into cells is a multistep process that often requires the subversion of subcellular machineries. A more complete understanding of these steps is necessary to develop new antiviral strategies. While studying the potential role of the actin network and one of its master regulators, the small GTPase Cdc42, during Junin virus (JUNV) entry, we serendipitously uncovered the small molecule ZCL278, reported to inhibit Cdc42 function as an entry inhibitor for JUNV, and for vesicular stomatitis, lymphocytic choriomeningitis, and dengue virus but not for the non-enveloped Poliovirus. Although ZCL278 did not interfere with JUNV attachment to the cell surface or virus particle internalization into host cells, it prevented release of JUNV ribonucleoprotein cores into the cytosol and decreased pH-mediated viral fusion with host membranes. We also identified SVG-A astroglial-derived cells as highly permissive for JUNV infection and generated new cell lines expressing fluorescently tagged rab5 or rab7 or lacking Cdc42 using CRISPR-Cas9 gene-editing strategies. Aided by these tools, we uncovered that perturbations in the actin cytoskeleton or Cdc42 activity minimally affect JUNV entry, suggesting that the inhibitory effect of ZCL278 is not mediated by interfering with the activity of Cdc42. Instead, ZCL278 appears to redistribute the location of viral particles from endosomal to lysosomal compartments. ZCL278 also inhibited JUNV replication in a mouse model, and no toxicity was detected. Together, our data suggest unexpected antiviral activity of ZCL278 and highlight its potential for developing valuable new tools to study intracellular trafficking of pathogens.
IMPORTANCE: The Junin virus is responsible for outbreaks of Argentine hemorrhagic fever in South America, where 5 million people are at risk. Limited options are currently available to treat infection by Junin virus or other Arenaviridae, making the identification of additional tools, including small molecule inhibitors, of great importance. How Junin virus enters cells is not yet fully understood. Here we describe new cell culture models susceptible to Junin virus infection onto which we applied CRISPR-Cas9 genome engineering strategies to help characterize early steps during virus entry. We also uncovered ZCL278 as a new antiviral small molecule that potently inhibits cellular entry of the Junin virus and other enveloped viruses. Moreover, we show that ZCL278 also functions in vivo thereby preventing Junin virus replication in a mouse model, opening the possibility for the discovery of ZCL278 derivatives of therapeutic potential.

PMID: 26912630 [PubMed - as supplied by publisher]

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