Abstract B28: PTENpg1 antisense RNA mediates PTEN suppression in vemurafenib resistance and predicts clinical outcome in melanoma patients

Abstract

Abstract Approximately 50% of cutaneous melanomas carry activating mutations in the serine/threonine protein kinase BRAF. Vemurafenib is a highly potent inhibitor of BRAFV600E and prolongs survival in ~80% of melanoma patients carrying the mutation. Unfortunately, almost all patients develop resistance within 6-8 months of starting treatment. The molecular mechanism explaining this scenario is largely unclear, but loss of PTEN expression, has been suggested. In this study we reveal a role for the PTEN pseudogene encoded antisense RNA (PTENpg1 asRNA) in epigenetic suppression of PTEN in melanoma cell lines resistant to vemurafenib. Resistant cell lines showed increased PTENpg1 asRNA expression while PTEN expression was suppressed. ChIP analysis showed enrichment of the histone modification enzyme EZH2 and subsequent enrichment of H3K27me3 at the PTEN promoter in the resistant cell line. Total cellular levels of EZH2, DNMT3a and H3k27me3 were, however, unaffected thus indicating specific recruitment of these factors to the PTEN promoter in the resistant cells by a mechanism that is dictated by the increased expression of PTENpg1 asRNA. In addition, we show that PTEN is reactivated by the depletion of EZH2 and DNMT3a and that this reactivation of PTEN re-sensitizes the cells to vemurafenib treatment. Finally, we show that PTENpg1 asRNA could be a promising prognostic marker of clinical outcome of melanoma patients. PTENpg1 asRNA expression levels were measured in samples from patients that had not received any oncological treatment and were classified with stage III melanomas. We found that high PTENpg1 asRNA expression correlates with short-term survival in melanoma patients. In conclusions, the PTENpg1 asRNA appears to be an important player in vemurafenib resistance and could be an attractive therapeutic target. In addition, PTENpg1 asRNA is potentially a promising prognostic marker for clinical outcome for melanoma patients. Citation Format: Linda Vidarsdottir, Alireza Azimi, Jason Serviss, Christian Ingvar, Göran Jönsson, Håkan Olsson, Marianne Frostvik Stolt, Johan Hansson, Suzanne Egyházi Brage, Dan Grandér, Per Johnsson. PTENpg1 antisense RNA mediates PTEN suppression in vemurafenib resistance and predicts clinical outcome in melanoma patients. [abstract]. In: Proceedings of the AACR Special Conference on Noncoding RNAs and Cancer: Mechanisms to Medicines ; 2015 Dec 4-7; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2016;76(6 Suppl):Abstract nr B28.