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 4

Re-expression of ANTXR1 reconstitutes SVV permissivity.

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Re-expression of ANTXR1 reconstitutes SVV permissivity. Cells were cotra...
Cells were cotransfected with the ANTXR1-HA and mCherry expression plasmids (A, B, and D). (A) An H446 ANTXR1-KO cell line was transfected, then challenged with SVV-GFP. Scale bars: 100 μm. Images representative of 3 independent experiments. (B) H446 and LX22cl ANTXR1-KO cell lines were transfected, challenged with SVV-GFP, and analyzed by flow cytometry. mCherry-transfected parental (P) and ANTXR1-KO cells were used as positive and negative controls, respectively. Each bar represents the average of n = 3 replicates with error bars representing SD. A 2-way ANOVA test with multiple comparisons was used to determine statistical significance. (C) ANTXR1-HA protein expression was confirmed by Western blot. Blot representative of 2 independent Westerns. (D) H69 and H146 cells were transduced with a Dox-inducible ANTXR1-HA expression lentivirus. Parental and ANTXR1-expressing cells were incubated in absence or presence of 1 μg/ml doxycycline for 72 hours, challenged with SVV-GFP, and analyzed by flow cytometry. Each bar represents the average of n = 3 replicates with error bars representing SD. A 2-way ANOVA test with multiple comparisons was used to determine statistical significance. (E) ANTXR1-HA protein expression in the presence of doxycycline was confirmed by Western blot. Blot representative of 2 independent Westerns. ****P ≤ 0.0001