Abstract B58: The role of mutant P53 in repetitive element regulation and the immune response in ovarian cancer

Abstract

Abstract While therapies that activate the host immune system have shown tremendous promise for a wide variety of solid tumors, less than 10% of ovarian cancer (OC) patients have responded. Previous work from our group showed that DNA methyltransferase inhibitors (DNMTis) activate the interferon response in tumors through upregulation of repetitive elements (REs) including endogenous retroviruses (ERVs), remnants of exogenous retroviruses that integrated into the germline several million years ago. DNMTis increase ERVs in a mouse model of ovarian cancer, activating interferon signaling and recruiting CD8+ T cells to kill the tumors. Importantly, nearly all high-grade serous OCs harbor a mutation in the TP53 gene, encoding the tumor suppressor protein P53. p53 may transcriptionally repress ERVs to ensure genome stability; 30% of all p53 binding sites are in ERV-derived sequences. We treated seven human cancer cell lines, including four OC lines (3 TP53 wild-type, 4 TP53 mutant) with DNMTi and analyzed repetitive element expression (RNA-Seq), open chromatin (ATAC-Seq), and DNA methylation (MeDIP + MRE). Interestingly, RE upregulation was significantly higher in the P53 mutant cell lines. This was true for the SINE, LINE, and ERV groups of repetitive elements. The DNA methylation of these elements was not significantly different between TP53 wild-type and mutant cell lines, but the open chromatin peaks (ATAC-Seq) were significantly increased in the TP53 mutant cell lines. We thus hypothesize that wild-type TP53 may partner with DNA methylation to transcriptionally repress RE transcription. Treating TP53 wild-type cells with Nutlin-3A, which activates P53, represses transcription of some ERVs. When we overexpressed R175H or R273H mutant P53 constructs in the SKOV3 P53-null cell line and treated with DNMTi, we observed much higher upregulation of ERVs after DNMTi treatment than when we overexpressed the control wild type P53 vector. We predicted that higher ERVs would induce a higher interferon response and perhaps change the microenvironment in P53 mutant tumors. We used the ID8 MOSE mouse ovarian cancer model modified by the MacNeish laboratory to compare p53+/+ and p53-/- tumor response to DNMTi treatment. Characterization of the tumor microenvironment indicated a significantly higher frequency of lymphocytes, Treg cells, MDSCs, and myeloid cells in the p53-/- model in comparison to the p53 wild-type model. In current work, we are creating isogenic cell lines of human and mouse wild-type, mutant, and null p53 and comparing the response to DNMTi in vitro and in vivo. We will then determine whether P53 binding directly regulates repetitive elements transcriptionally, and more broadly, whether mutant P53 cells are more sensitive to DNMTi upregulation of repetitive elements and induction of an immune response. Results of this work will be particularly relevant for high-grade serous OC, which is driven by mutant P53 and is generally unresponsive to immunotherapies. Citation Format: Stephanie Gomez, James I. McDonald, Elisa Arthofer, Noor Diab, Aneil Srivastava, Paul Austin, Katherine B. Chiappinelli. The role of mutant P53 in repetitive element regulation and the immune response in ovarian cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research; 2019 Sep 13-16, 2019; Atlanta, GA. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(13_Suppl):Abstract nr B58.