Abstract 523: miR-206 inhibits pleural mesothelioma by targeting an active KRAS/CDK4/CCND1 pathway

Abstract

Abstract Introduction: Since there remains no effective, durable chemotherapy against malignant pleural mesothelioma (MPM), we propose to leverage microRNA(miR)-based approaches that recently reached phase I trial testing. We posit there are many miR with therapeutic potential yet to be identified in MPM. Methods/ Results: Our prior miRNA expression profiling results of MPM (GSE40345) were cross-referenced with a novel p-value minimization analytic technique to identify prognostic miRNA. Both MPM cell lines and tissues (tumors and normal pleura) were assessed using quantitative and functional biologic assays. In silico algorithms identified gene targets that were verified by 3'-UTR luciferase assay.In silico analyses showed underexpression of miR-206 by 12.1-fold in MPM tumors compared to normal pleura. Using our p-value minimization technique, we assessed the clinical impact of this underexpressed miR in TCGA data (n=73 MPM) and noted a significant association to worse survival for patients with lower miR-206 expression (p=0.024). We confirmed that miR-206 was significantly underexpressed in tumor tissues by qRT-PCR analysis of a new, randomly selected MPM cohort (n=41) versus normal pleura (n=14). In a set of established MPM cell lines with low endogenous miR-206 level, ectopic re-expression of miR-206 dramatically suppressed cell proliferation, invasiveness, colony foci formation, and growth in soft-agar. Treatment of non-malignant mesothelial cells MeT-5a and LP9 with ectopic miR-206 showed no untoward effects. In silico gene target predictions for miR-206 were summated across 3 databases and screened by prognostic effect based on Kaplan-Meier results from TCGA data (n=87 MPM). Interestingly, we noted several overexpressed MPM-prognostic genes (p<0.05) regulated by miR-206: KRAS, CDK4, and CCND1. qRT-PCR analysis of tissues confirmed elevated transcripts of KRAS, CDK4, and CCND1 in MPM. This signaling axis of KRAS/CDK4/CCND1 is important in MPM as it summates well-known dysregulated tyrosine kinase receptors (EGF, IGF-1, VEGF, MET, etc) that are upstream. The KRAS/CDK4/CCND1 axis is associated with cell cycle progression and survival of cancer cells, but is not an easily druggable target. miR-206 treatment significantly downregulated KRAS, CDK4 and CCND1 genes in MPM cell lines. Conclusion: In vitro, miR-206 exerts tumor suppressive effects in MPM via inhibition of KRAS/CDK4/CCND1 signaling. miR-206 restoration in MPM mimics simultaneous blocking of multiple tyrosine kinases. Loss of miR-206 and concomitant overexpression of KRAS, CDK4 and CCND1 formed a poor prognostic signature of MPM. Our results identify miR-206 as a rational therapeutic agent to studied further in preclinical MPM models. Citation Format: Anand Singh, Roma Pahwa, Li Zhang, Nathanael Pruett, R Taylor Ripley, David S. Schrump, Chuong D. Hoang. miR-206 inhibits pleural mesothelioma by targeting an active KRAS/CDK4/CCND1 pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 523.