Abstract 3089: YAP and TAZ induce MCM protein expression to facilitate tumor-supporting properties in liver cancer

Abstract

Abstract Overexpression and nuclear enrichment of the Hippo pathway-regulated transcriptional co-activator Yes-Associated Protein (YAP) is frequently detected in human hepatocellular carcinoma (HCC) and correlates with poor clinical outcome of cancer patients1. In mice, inducible overexpression of the constitutively active isoform YAPS127A induces hepatocellular proliferation leading to hepatomegaly followed by tumor formation within 12-15 weeks. How oncogenic YAP and its paralogue TAZ (syn. WWTR1, WW Domain Containing Transcription Regulator 1) induce uncontrolled proliferation in liver cells is not known, yet. Analysis of expression profiling data, derived from human liver cancer cells after siRNA-mediated inhibition of endogenous YAP, revealed that the Minichromosome Maintenance (MCMs) family members MCM2-7 were positively regulated. MCMs are key components of the pre-replication complex and involved in the formation of the replication fork, which is essential for efficient DNA duplication followed by mitosis2. We confirmed that silencing of YAP and TAZ by independent siRNAs reduced the mRNA and protein expression of MCM2-7 in liver cancer cell lines. In vitro analyses revealed that transcription factors of the TEAD family were the most relevant YAP binding partners needed for the transcriptional regulation of MCMs. Using chromatin immunoprecipitation and Dual Luciferase Reporter Assay, we showed that YAP, TAZ and TEAD4 directly bound the MCM promoter. First in vitro data suggested that viability of liver cancer cells, subjected to drug induced replication stress, was exclusively reduced in cells with reduced YAP, TAZ or MCM protein expression. Administration of the YAP/TAZ/TEAD inhibitor Verteporfin reduced the expression of all MCMs in liver cancer cells. Vice versa, the overexpression of YAPS127A in transgenic mice showed significantly increased levels of the proliferation marker Ki67 and MCM2-7. Expression data from 242 HCC patients illustrated an association between YAP and MCM mRNA levels, with elevated MCMs significantly correlating with worse overall survival and early cancer recurrence3. Immunohistochemical stains of HCC tissue micro-arrays revealed a highly significant correlation between Ki67 expression, nuclear YAP overexpression and nuclear MCM2-7 enrichment. In summary, our results strongly suggest that YAP and TAZ facilitate their tumor-supporting properties through the regulation of MCM2-6, a complete protein complex. We conclude that combined inhibition of YAP and TAZ or perturbation of MCM activity might represent an efficient therapeutic approach for the treatment of a subgroup of HCC patients. 1Tschaharganeh D, et al., Gastroenterol. 2013; 2Deegan TD, et al., Curr Opin Struct Biol. 2016; 3Roessler S, et al., Cancer Res. 2010. Citation Format: Maria Knaub, Sofia Weiler, Stefan Thomann, Peter Schirmacher, Kai Breuhahn. YAP and TAZ induce MCM protein expression to facilitate tumor-supporting properties in liver cancer [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 3089.