Anthrax toxin receptor 1 is the cellular receptor for Seneca Valley virus
Linde A. Miles, Laura N. Burga, Eric E. Gardner, Mihnea Bostina, John T. Poirier, Charles M. Rudin
Linde A. Miles, Laura N. Burga, Eric E. Gardner, Mihnea Bostina, John T. Poirier, Charles M. Rudin
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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 2

Knockout of ANTXR1 leads to the loss of SVV permissivity.

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Knockout of ANTXR1 leads to the loss of SVV permissivity. (A) Three of t...
(A) Three of the H446 ANTXR1-KO cell lines were challenged with increasing MOIs of SVV for 72 hours. Cell viability was determined via AlamarBlue. Parental H446 cells and the nonpermissive NSCLC cell line A549 were used as positive and negative controls, respectively. Each data point represents the average of n = 6 replicates with error bars representing SD. (B) Permissive SCLC and pediatric cancer cell lines were transduced with an sgRNA targeting ANTXR1. Parental (light gray) and ANTXR1-KO (black) cells were challenged with SVV-GFP and analyzed by flow cytometry. Each bar represents the average of n = 3 replicates with error bars representing SD. Unpaired 2-sided t tests were used to determine statistical significance. (C) Parental H446 cells (top), H446 ANTXR1-KO mCherry cells (bottom), or 1:1 mixture of parental/ANTXR1-KO mCherry cells (middle) were challenged with SVV-GFP. White arrows indicate adjacent SVV-GFP–infected parental H446 and uninfected ANTXR1-KO mCherry cells. Scale bars: 100 μm. Images representative of 3 independent experiments. (D) Parental H446 tumors (light green/dark green), H446 ANTXR1-KO mCherry tumors (pink/red), and 1:1 mixture of parental/ANTXR1-KO mCherry tumors (light purple/dark purple) were challenged with WT SVV-001 or PBS vehicle. Tumor volumes were measured every other day. Each data point corresponds to the average of n = 4–5 tumors with error bars representing SD. (E) Tumors were excised at the experiment endpoint and analyzed by flow cytometry. Each bar represents the average of n = 4–5 tumors with error bars representing SD. A 2-way ANOVA test with multiple comparisons was used to determine statistical significance. ****P ≤ 0.0001.