Anthrax toxin receptor 1 is the cellular receptor for Seneca Valley virus
Linde A. Miles, … , John T. Poirier, Charles M. Rudin
Linde A. Miles, … , John T. Poirier, Charles M. Rudin
Published June 26, 2017
Citation Information: J Clin Invest. 2017;127(8):2957-2967. https://doi.org/10.1172/JCI93472.
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Research Article Virology

Anthrax toxin receptor 1 is the cellular receptor for Seneca Valley virus

Abstract

Seneca Valley virus (SVV) is an oncolytic picornavirus with selective tropism for neuroendocrine cancers. It has shown promise as a cancer therapeutic in preclinical studies and early-phase clinical trials. Here, we have identified anthrax toxin receptor 1 (ANTXR1) as the receptor for SVV using genome-wide loss-of-function screens. ANTXR1 is necessary for permissivity in vitro and in vivo. However, robust SVV replication requires an additional innate immune defect. We found that SVV interacts directly and specifically with ANTXR1, that this interaction is required for SVV binding to permissive cells, and that ANTXR1 expression is necessary and sufficient for infection in cell lines with decreased expression of antiviral IFN genes at baseline. Finally, we identified the region of the SVV capsid that is responsible for receptor recognition using cryoelectron microscopy of the SVV-ANTXR1-Fc complex. These studies identify ANTXR1, a class of receptor that is shared by a mammalian virus and a bacterial toxin, as the cellular receptor for SVV.

Authors

Linde A. Miles, Laura N. Burga, Eric E. Gardner, Mihnea Bostina, John T. Poirier, Charles M. Rudin

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Figure 5

SVV interacts directly with ANTXR1.

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SVV interacts directly with ANTXR1. (A) SVV was coimmunoprecipitated wit...
(A) SVV was coimmunoprecipitated with decreasing amounts of an ANTXR1-Fc chimera. Bound proteins were eluted and analyzed by Western blot using an anti-SVV antibody. Input SVV was immunoblotted as a positive control. Blot representative of 2 independent experiments. (B) SVV was coimmunoprecipitated with the ANTXR1-Fc chimera. Washes were performed with increasing concentrations of NaCl up to 2 M. Bound proteins were eluted and analyzed as described in A. Blot representative of 2 independent experiments. (C) SVV was coimmunoprecipitated with the ANTXR1-Fc chimera or decreasing amounts of ANTXR2-Fc chimera and analyzed as described in A. Blot representative of 2 independent experiments. (D) SVV-GFP was preincubated with the ANTXR1-Fc chimera, ANTXR2-Fc chimera, or IgG-Fc isotype control before an 8-hour incubation with parental H446 cells. Cell nuclei were stained with a NucBlue Live ReadyProbe. Scale bars: 100 μm. Images representative of 3 independent experiments. (E) ANTXR1-KO (blue) and TEX2-KO (red) cells were incubated with SVV-Cy5 and analyzed by flow cytometry. Parental H446 (black) and DMS114 (gray) cells were used as positive and negative controls for SVV binding, respectively. Data representative of 2 independent experiments. (F) Cryo-EM density map of SVV capsid (blue) bound to ANTXR1-Fc chimera (green).