Abstract 4356: Identification of the anthrax toxin receptor (ANTXR1) as the high affinity cellular receptor for Seneca Valley Virus (SVV)
Authors: Miles Linde A. , Poirier J.T. , Rudin Charles M. 
Published: July 2016

Small cell lung cancer (SCLC) is an extremely aggressive and almost universally lethal form of lung cancer, which accounts for approximately 15% of all lung cancer diagnoses annually. Novel and more effective therapies are desperately needed for SCLC, as the standard treatment has not improved in almost three decades. Recently, oncolytic viruses have shown strong anti-cancer efficacies in tumors with high levels of selectivity for cancer cells.

The oncolytic picornavirus, Seneca Valley Virus (SVV), has previously been shown to infect and lyse cancer cells with neuroendocrine features, notably SCLC and pediatric brain tumors. Pre-clinical mouse models and early phase clinical trials have confirmed the ability of SVV to home directly to the tumor through the vasculature, specifically infect tumor cells without affecting normal cells, and replicate to high viral titers intratumorally even after the production of neutralizing antibodies. However, the clinical development of SVV has been hindered by the inability to identify patients who would benefit from SVV virotherapy because essential host proteins for viral entry, including the cellular receptor, have not been characterized.

Using genome wide loss-of-function CRISPR screens in the SCLC cell line H446 and haploid cell line HAP1, we identified the anthrax toxin receptor 1 (ANTXR1) as an essential host protein for SVV. Interestingly, ANTXR1 is also one of the receptors for the toxin of Bacillus anthracis and has been previously described as being upregulated in tumor endothelial cells. Secondary screens performed in both cell lines confirmed gene knockout of ANTXR1 conferred resistance to SVV. We established that in multiple permissive SCLC and pediatric cancer cell lines that loss of ANTXR1 protein expression using CRISPR gene knockout leads to a loss of SVV permissivity. Re-expression of ANTXR1 in ANTXR1 knockout cell lines was sufficient to rescue SVV permissivity. Furthermore, the expression of the ANTXR1 protein in two non-permissive SCLC cell lines led to a conversion in SVV permissivity. A direct and high affinity interaction was confirmed between SVV and ANTXR1 using co-immunoprecipitation assays. Additionally, we discovered that ANTXR1 is the major binding determinant for SVV on cells using a binding assay with fluorescently labeled SVV. Lastly we determined using previously published gene expression data from the Cancer Cell Line Encyclopedia (CCLE) that non-permissive lines which do express ANTXR1 can be explained by the presence of an intact and active innate immune response pathway. These findings identify ANTXR1 as the high-affinity cellular receptor for SVV. Furthermore, these data identify a predictive biomarker for SVV permissivity that could lead to defined selection criteria for subsequent clinical trials testing SVV in both SCLC and pediatric brain cancer patients.

Citation Format: Linde A. Miles, J.T. Poirier, Charles M. Rudin. Identification of the anthrax toxin receptor (ANTXR1) as the high affinity cellular receptor for Seneca Valley Virus (SVV). [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4356.