Abstract 6185: <i>In vivo</i> screening in HPV positive and HPV negative head and neck squamous cell carcinoma
Authors: Mohamed K. Abdelhakiem, Liangpeng Yang, Tongxin Xie, Manish Kumar, David Molkentine, Aakash Sheth, Mitchell Frederick, Jeffrey Myers, Curtis R. Pickering, Heath D. Skinner
Journal: Cancer Research (2020)

Abstract Background: The pathogenesis of head and neck squamous cell carcinoma (HNSCC) largely occurs through two pathways, human papillomavirus (HPV) mediated or chemical carcinogenesis secondary to alcohol and tobacco use. HPV status is an important prognostic factor and marker for sensitivity to radiation therapy in HNSCC patients. HPV negative HNSCC has been shown to have increased radioresistance compared to HPV positive HNSCC leading to poorer prognosis. The goal of this work is to utilize in vivo screening technology to identify key differential targets between HPV positive and negative tumors for both therapeutic purposes as well as to identify differential pathways between these two malignancies. Methods: Pooled, barcoded shRNA libraries focused on DNA damage repair (DDR) and currently targetable genes were transduced into three HPV positive (UM-SCC-147, UD-SCC-2, UPCI:SCC152) and three HPV negative (UM-SCC-22A, Cal-27, HN31) cell lines. Each cell line was injected into the mouse flank and allowed to grow to ~100mm3. Tumors were divided into control and radiation groups; the latter being treated with 6-10 Gy at 2 Gy/treatment. For each tumor type radiation was titrated to generate an approximate 20% reduction in tumor size. Tumors were then allowed to grow for an additional 2 weeks and then harvested. Barcodes from each tumor were sequenced and compared to both the reference input cells as well as untreated control tumors. Barcodes observed at decreased frequency in treated tumors indicated a potential target for therapeutic sensitization. Both p-values as well as percentile median fold change for each gene were calculated. These values were averaged for the three HPV negative and three HPV positive cell lines and used for analysis. Results: Multiple genes in the DDR and FDA libraries were identified from the HPV negative and HPV positive groups that may be associated with radioresistance in vivo. Example genes that were identified primarily in the HPV negative group include BTK, TXN, TTK, while those in the HPV positive group include RNF168, RAD50, ESR1. Inhibition of these genes may lead to increased cell death following radiation therapy through diminished DNA repair of radiation-induced damage. Conclusion: Our study identifies targetable genes that may contribute to radioresistance in both HPV negative and HPV positive head and neck squamous cell carcinoma and warrants further analysis. Citation Format: Mohamed K. Abdelhakiem, Liangpeng Yang, Tongxin Xie, Manish Kumar, David Molkentine, Aakash Sheth, Mitchell Frederick, Jeffrey Myers, Curtis R. Pickering, Heath D. Skinner. In vivo screening in HPV positive and HPV negative head and neck squamous cell carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6185.