Abstract
Abstract Introduction: The purpose of this study was to develop a Sleeping Beauty (SB)-induced mouse model of metastatic vulvar carcinoma with which we could identify novel tumor-driving mutations. Experimental Procedures: An existing transgenic mouse model known to induce development of benign vaginal papillomas, (PrCre/+;KrasG12D), was bred to mice containing the components of the SB system to render (PrCre/+;KrasG12D;SB) mice. The SB system consists of a transposable element (transposon) and an activating enzyme (transposase) that induces excision of the transposon from one site to another in the host cell genome. Insertional mutations that enhance cell survival and proliferation are selected for, resulting in the development of cancer. We utilize the SB system to identify candidate driver mutations since the transposon is not only the mutagen but also serves as a sequence tag for localization of mutations. Mice were aged and monitored for phenotypic signs of disease. Upon necropsy, gynecologic and other grossly abnormal organs were harvested. Genomic DNA was extracted from each sample, prepared for genetic analysis, and sequenced using the Illumina platform. Candidate genes were identified using an unbiased Monte-Carlo approach in conjunction with a gene-centric Chi-Squared analysis. Paraffin-embedded human tumor samples were obtained from the University of Iowa Hospitals and Clinics for immunohistochemical labeling of protein expression for selected genes. Results: Vulvar carcinoma developed in 29 of 29 PrCre/+;KrasG12D;SB female mice with metastases by seven months of age confirmed in 13 animals. Organs within the abdominal cavity including the uterus and ovaries contained metastatic lesions in some mice. In one mouse, a distant metastasis within the subcutaneous tissue of the thoracic back was observed. Illumina sequencing of primary lesions from all mice revealed 77 candidate tumor drivers. Conclusions: To our knowledge, the PrCre/+;KrasG12D;SB mouse is the first reported metastatic model of vulvar carcinoma. Protein expression of candidate genes identified by our SB screen are being assessed in human tissue samples. Patient clinical data will be incorporated into analyses to determine whether candidate genes are correlated to patient outcomes. This work will permit insights into the pathophysiology and future treatment of vulvar cancer. Note: This abstract was not presented at the meeting. Citation Format: Amy Guimaraes-Young, Traci Neff, David K. Meyerholz, Adam J. Dupuy, Michael J. Goodheart. Identification of novel genetic drivers of vulvar carcinoma in a Sleeping Beauty model of spontaneous cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4192. doi:10.1158/1538-7445.AM2015-4192
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