Unveiling the Role of RSUME in Renal Cell Carcinoma Cell Metabolism by means of a Mass Spectrometry-Based Metabolic Fingerprinting Strategy

MONGE, MARIA EUGENIA; MANUELA MARTINEFSKI; BELEN ELGUERO; MARÍA ELENA KNOTT; DAVID GOLNISKI; LUCAS TEDESCO; JUAN MANUEL GUREVICH MESSINA; CORA POLLAK; EDUARDO ARZT

Conferencia Virtual

Conferencia; 17th Annual Conference of the Metabolomics Society Metabolomics2020 Online; 2021

The Metabolomics Society

Clear cell renal cell carcinoma (ccRCC) is a heterogeneous disease with 50−80% patients exhibiting mutations in the von Hippel−Lindau (VHL) gene, which is involved in the expression of hypoxia-inducible factors 1α and 2α (HIF-1α and -2α). High RSUME expression has been associated with poor prognosis in ccRCC, and it acts as a negative regulator of VHL function in normoxia. Considering that the HIF is a key regulator of metabolic reprogramming in this cancer type, we hypothesized that RSUME participates in metabolic changes in ccRCC tumor progression. Thus, we optimized a protocol for harvesting, quenching, extracting, and fingerprinting the endometabolome derived from a human ccRCC cell line 786-O and three other transformed cell systems (n = 102) with different expressions of RSUME and VHL by means of UHPLC-QTOF-MS. Cross-validated orthogonal projection to latent structures discriminant analysis models were built on positive, negative, and a combination of positive- and negative-ion mode MS data sets. Discriminant feature panels selected by an iterative multivariate classification allowed differentiating cells with different expressions of RSUME and VHL. Fifteen identified discriminant metabolites with level 1, including glutathione, butyrylcarnitine, and acetylcarnitine, contributed to understand the role of RSUME in ccRCC. Altered pathways associated with the RSUME expression were validated by biological and bioinformatics analyses. Combined results showed that in the absence of VHL, RSUME is involved in the downregulation of the antioxidant defense system, whereas in the presence of VHL, it acts in rerouting energy-related pathways, negatively modulating the lipid utilization, and positively modulating the fatty acid synthesis.