title: Identification of cisplatin-regulated metabolic pathways in pluripotent stem cells.
abstract: The chemotherapeutic compound, cisplatin causes various kinds of DNA lesions but also triggers other pertubations, such as ER and oxidative stress. We and others have shown that treatment of pluripotent stem cells with cisplatin causes a plethora of transcriptional and post-translational alterations that, to a major extent, point to DNA damage response (DDR) signaling. The orchestrated DDR signaling network is important to arrest the cell cycle and repair the lesions or, in case of damage beyond repair, eliminate affected cells. Failure to properly balance the various aspects of the DDR in stem cells contributes to ageing and cancer. Here, we performed metabolic profiling by mass spectrometry of embryonic stem (ES) cells treated for different time periods with cisplatin. We then integrated metabolomics with transcriptomics analyses and connected cisplatin-regulated metabolites with regulated metabolic enzymes to identify enriched metabolic pathways. These included nucleotide metabolism, urea cycle and arginine and proline metabolism. Silencing of identified proline metabolic and catabolic enzymes indicated that altered proline metabolism serves as an adaptive, rather than a toxic response. A group of enriched metabolic pathways clustered around the metabolite S-adenosylmethionine, which is a hub for methylation and transsulfuration reactions and polyamine metabolism. Enzymes and metabolites with pro- or anti-oxidant functions were also enriched but enhanced levels of reactive oxygen species were not measured in cisplatin-treated ES cells. Lastly, a number of the differentially regulated metabolic enzymes were identified as target genes of the transcription factor p53, pointing to p53-mediated alterations in metabolism in response to genotoxic stress. Altogether, our findings reveal interconnecting metabolic pathways that are responsive to cisplatin and may serve as signaling modules in the DDR in pluripotent stem cells.
authors.affiliation: Department of Toxicology, LACDR, Leiden University, Leiden, The Netherlands ; Netherlands Toxicogenomics Center, Maastricht, The Netherlands.
authors.forename: Louise
authors.initials: L
authors.lastname: von Stechow
authors.forename: Ainhoa
authors.initials: A
authors.lastname: Ruiz-Aracama
authors.forename: Bob
authors.initials: B
authors.lastname: van de Water
authors.forename: Ad
authors.initials: A
authors.lastname: Peijnenburg
authors.forename: Erik
authors.initials: E
authors.lastname: Danen
authors.forename: Arjen
authors.initials: A
authors.lastname: Lommen
mesh_terms: D000818:Animals; D001120:Arginine / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000187:drug effects*; D055442:Metabolome / Q000187:drug effects; D055432:Metabolomics; D051379:Mice; D039904:Pluripotent Stem Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D011392:Proline / Q000378:metabolism; D011687:Purines / Q000378:metabolism; D011743:Pyrimidines / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D059467:Transcriptome / Q000187:drug effects / Q000235:genetics
chemical_list: D011687:Purines; D011743:Pyrimidines; D001120:Arginine; D011392:Proline; D002945:Cisplatin; C030985:purine