RSUME AND VHL ASSOCIATION IN VHL RENAL CELL CARCINOMA TUMORS PHENOTYPE

GONILSKI PACIN, DAVID; FUERTES, MARIANA; SENIN, SERGIO A; POLLAK, CORA; GARCIA MARCHIÑENA, PATRICIO A.; JURADO, ALBERTO M; ISOLA, MARIANA; LABANCA, MARÍA J; PALAZZO, MARTÍN; GUREVICH MESSINA, JUAN M; KNOTT, MARÍA E; MARTINEFSKI, MANUELA; YANKILEVICH, PATRICIO; MONGE, MARÍA EUGENIA; ELGUERO, MARÍA BELÉN; ARZT, EDUARDO

Mar del Plata

Congreso; Reunión Anual de Biociencia 2019 - SAIC-SAFE-SAB-SAP-NANOMEDar-AACyTAL-HSC; 2019

RSUME or RWDD3 is a hypoxia inducible protein isolated from highly tumorigenic and angiogenic cells, which targets Von Hippel-Lindau (VHL) protein. Renal Cell Carcinoma (RCC) tumors, are the main common cause of death in VHL patients. Survival analysis available at The Human Protein Atlas with a recent dataset of Cancer Genome Atlas Research Network (TCGA) shows that 20.07 % of the 528 RCC tumors samples studied express elevated levels of RSUME, which correlate with a 23 % decrease of patients? survival rate. Using the same bioinformatics? platform, we observed that patients carrying VHL mutations have increased levels of RSUME compared with those without VHL mutations (p= 0.048). RSUME expression levels were significantly higher in stage IV tumors than those in stages I-II and III (p= 0.036 and 0.021). Analyzing in more detail the levels of RSUME in stage IV, we observed increased levels of RSUME in patients carrying VHL mutations and specifically in those carrying VHL missense mutations compared with those patients without VHL mutations (p= 0.012 and p= 0.027, respectively). We validated these data in a RCC cell line in which we observed by a Luciferase assay of the RSUME promoter and by WB that, while VHL wild type transfection decreases RSUME expression levels, VHL missense mutants transfection do not. Moreover, in cells cultured under HPX (1 % O2) and next exposed to NMX, we observed by WB that when VHL wild type was transfected RSUME protein levels were lower at 0, 5 and 30 minutes compared with VHL missense mutants transfection. Data about RSUME expression were validated in paraffin samples of RCC patients by immunohistochemical staining. A first metabolomics analysis in the RCC cell line in which RSUME is silenced show a differential pattern in acetylcarnitine levels. These in silico and experimental results strongly support the association of VHL mutants and RSUME in the phenotype of RCC tumors. Supported by ANPCyT, CONICET, UBA and FOCEM (COF 03/11). Keywords: VHL, RSUME, RCC.