Absolute quantitative proteomics using the total protein approach to identify novel clinical immunohistochemical markers in renal neoplasms

BMC Medicine

José L. Capelo¹  Carlos Lodeiro¹  Susana Jorge¹  Hugo M. Santos²  Jacek R. Wiśniewski³  William LaFramboise⁴  Dimitrios Korentzelos⁴  Sheldon Bastacky⁴  Gabriela Quiroga-Garza⁴  Rajiv Dhir⁴  Swati Satturwar⁴ 

[1] BIOSCOPE Group, LAQV-REQUIMTE, Chemistry Department, NOVA School of Science and Technology, FCT NOVA, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal;PROTEOMASS Scientific Society, Madan Park, 2829-516, Caparica, Portugal;BIOSCOPE Group, LAQV-REQUIMTE, Chemistry Department, NOVA School of Science and Technology, FCT NOVA, Universidade NOVA de Lisboa, 2829-516, Caparica, Portugal;PROTEOMASS Scientific Society, Madan Park, 2829-516, Caparica, Portugal;Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA, USA;Biochemical Proteomics Group, Department of Proteomics and Signal Transduction, Max-Planck-Institute of Biochemistry, Martinsried, Germany;Department of Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA, USA;

关键词: Renal neoplasms, Immunohistochemistry;    Mass spectrometry-based proteomics;    Total protein approach (TPA);    Tissue micro-array (TMA);   

DOI  :  10.1186/s12916-021-02071-9

来源: Springer

【 摘 要 】

BackgroundRenal neoplasms encompass a variety of malignant and benign tumors, including many with shared characteristics. The diagnosis of these renal neoplasms remains challenging with currently available tools. In this work, we demonstrate the total protein approach (TPA) based on high-resolution mass spectrometry (MS) as a tool to improve the accuracy of renal neoplasm diagnosis.MethodsFrozen tissue biopsies of human renal tissues [clear cell renal cell carcinoma (n = 7), papillary renal cell carcinoma (n = 5), chromophobe renal cell carcinoma (n = 5), and renal oncocytoma (n = 5)] were collected for proteome analysis. Normal adjacent renal tissue (NAT, n = 5) was used as a control. Proteins were extracted and digested using trypsin, and the digested proteomes were analyzed by label-free high-resolution MS (nanoLC-ESI-HR-MS/MS). Quantitative analysis was performed by comparison between protein abundances of tumors and NAT specimens, and the label-free and standard-free TPA was used to obtain absolute protein concentrations.ResultsA total of 205 differentially expressed proteins with the potential to distinguish the renal neoplasms were found. Of these proteins, a TPA-based panel of 24, including known and new biomarkers, was selected as the best candidates to differentiate the neoplasms. As proof of concept, the diagnostic potential of PLIN2, TUBB3, LAMP1, and HK1 was validated using semi-quantitative immunohistochemistry with a total of 128 samples assessed on tissue micro-arrays.ConclusionsWe demonstrate the utility of combining high-resolution MS and the TPA as potential new diagnostic tool in the pathology of renal neoplasms. A similar TPA approach may be implemented in any cancer study with solid biopsies.

【 授权许可】

CC BY