Prostate cancer (PCa) is one of the leading causes of death from cancer in men. Several prognostic factors allow differentiating low-grade from high-grade PCa that are often refractory to chemical castration but are still treated with hormone therapy to which docetaxel or cabazitaxel are added when they become resistant to the anti-androgen. Despite many clinical trials with other chemotherapeutic agents, response rates remain low, pointing towards the need for new alternative therapies to treat these aggressive tumors. Using a rational in silico approach based on the NCI60-cell line panel, we recently identified a signature of 6 genes, the expression of which could predict at the functional level, sensitivity to oxaliplatin but not to cisplatin in DU145, LNCaP and C42B prostate cancer cell lines (Puyo et al. Mol. Pharmacol, 2012). Among them, we focused on SHMT2, the mitochondrial isoform of serine hydroxymethyl transferase involved in the biosynthesis of purines. Downregulation of SHMT2 or absence of SHMT2 catalytic activity was associated with a resistance to oxaliplatin, whereas it had no effect on cisplatin sensitivity, a selectivity that was attributed to the DACH moiety of platinum derivatives. Here, we investigated the role of DNA methylation in this selective response to platinum compounds since SHMTs can indirectly regulate the methylation status of DNA. Variations in global level of methylation as measured by the methylation status of LINE-1 retrotransposon, was associated with differences in sensitivity of PCa cells to platinum compounds. Using our in silico approach, we found significant correlations between SHMT2 expression and cell sensitivity to both demethylating agents azacytidine and decitabine. We also found that treatment of DU145 cells, with high level of global DNA methylation, sensitized cells to platinum compounds. In order to evaluate whether methylation could impair the formation of Pt-adducts in vitro, we used purified oligonucleotides containing a unique site of platination. We show that methylation at specific CpG in the vicinity of the platination site could reduce the kinetics of DNA adducts formation. This inhibition was more pronounced for DACH platin than for cisplatin. We also assessed the effect of transient repression of SHMT2 in LNCaP cells on the methylation status of ∼450,000 CpG sites using the Infinium Meth450K beadchip from Illumina. Preliminary results show that significant changes in methylation status was only observed in a reduced number of CpGs which were not located in genes that are known to be involved in cell response to Pt adducts formation. Together our results demonstrate that SHMT2-mediated specific response to oxaliplatin can be due to both methylation in the vicinity of the platination site and the modulation of the expression of genes that are not directly linked to the processing of DNA-adducts.

Citation Format: Stéphane Puyo, Nadine Houédé, Marina Hamant, Pierre Richaud, Jacques Robert, Philippe Pourquier. SHMT2 modulates DNA methylation and differentially affects prostate cancer cell response to platinum derivatives. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4478. doi:10.1158/1538-7445.AM2013-4478