title: Differential proteomic analysis of renal cell carcinoma tissue interstitial fluid.
abstract: Renal cell carcinoma (RCC), the most common type of kidney cancer, currently has no biomarker of clinical utility. The present study utilized a mass spectrometry-based proteomics workflow for identifying differentially abundant proteins in RCC by harvesting shed and secreted proteins from the tumor microenvironment through sampling tissue interstitial fluid (TIF) from radical nephrectomies. Matched tumor and adjacent normal kidney (ANK) tissues were collected from 10 patients diagnosed with clear cell RCC. One-hundred thirty-eight proteins were identified with statistically significant differential abundances derived by spectral counting in tumor TIF when compared to ANK TIF. Among those proteins with elevated abundance in tumor TIF, nicotinamide n-methyltransferase (NNMT) and enolase 2 (ENO2) were verified by Western blot and selected reaction monitoring (SRM). The presence of ENO2 and thrombospondin-1 (TSP1) were verified as present and at elevated abundance in RCC patient serum samples as compared to a pooled standard control by enzyme-linked immunosorbent assay (ELISA), recapitulating the relative abundance increase in RCC as compared with ANK TIF.
authors.affiliation: Department of Pharmacology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Pang-ning
authors.initials: PN
authors.lastname: Teng
authors.forename: Brian L
authors.initials: BL
authors.lastname: Hood
authors.forename: Mai
authors.initials: M
authors.lastname: Sun
authors.forename: Rajiv
authors.initials: R
authors.lastname: Dhir
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Conrads
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis; D002292:Carcinoma, Renal Cell / Q000737:chemistry* / Q000473:pathology*; D002853:Chromatography, Liquid / Q000379:methods; D016000:Cluster Analysis; D019295:Computational Biology / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D045604:Extracellular Fluid / Q000737:chemistry*; D006801:Humans; D009363:Neoplasm Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D020543:Proteome