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.

Pharmacogenetic variants in TPMT alter cellular responses to cisplatin in inner ear cell lines

“…Furthermore, indirect evidence supporting a cisplatin- TPMT drug-gene interaction was recently reported where the transcriptomic and metabolomic responses of cisplatin-treated ES cells were described demonstrating that cisplatin-treatment increased TPMT expression[26]. We similarily observed a significant upregulation of TPMT expression in response to 10 μM cisplatin.…”

Pharmacogenomics of Cisplatin‐Induced Ototoxicity: Successes, Shortcomings, and Future Avenues of Research

“…These experiments showed that TPMT*3A was associated with an increase in both cisplatin biosensor response and cisplatin‐induced cytotoxicity. In addition, von Stechow et al . demonstrated that TPMT expression is induced by cisplatin in embryonic stem cells, providing further evidence for this drug–gene relationship.…”

“…These experiments showed that TPMT*3A was associated with an increase in both cisplatin biosensor response and cisplatin-induced cytotoxicity. In addition, von Stechow et al 29 demonstrated that TPMT expression is induced by cisplatin in embryonic stem cells, providing further evidence for this drug-gene relationship. The TPMT and COMT associations have been investigated in multiple cohorts, and although some studies have replicated these associations, 23,[30][31][32][33] others were unable to confirm these findings 9,13,16,30,[32][33][34][35][36] (Table 1 and Table S1).…”

Advances in applications of metabolomics in pluripotent stem cell research

“…Combining transcriptomic and metabolomic analysis, Stechow et al [40] identified cisplatin-regulated pathways in human PSCs, including nucleotide metabolism, the urea cycle, and arginine and proline metabolism. Several anti-oxidant associated metabolites and p53-regulated enzymes also showed significant enrichment due to genotoxic stress induced by cisplatin.…”