Identification of markers for the selection of patients undergoing renal cell carcinoma‐specific immunotherapy

Abstract

Renal cell carcinoma (RCC) represents the most common malignant tumor in the kidney and is resistant to conventional therapies. The diagnosis of RCC is often delayed leading to progression and metastatic spread of the disease. Thus, validated markers for the early detection of the disease as well as selection of patients undergoing specific therapy is urgently needed. Using treatment with the monoclonal antibody (mAb) G250 as a model, proteome-based strategies were implemented for the identification of markers which may allow the discrimination between responders and nonresponders prior to application of G250-mediated immunotherapy. Flow cytometry revealed G250 surface expression in approximately 40% of RCC cell lines, but not in the normal kidney epithelium cell lines. G250 expression levels significantly varied thereby distinguishing between low, medium and high G250 expressing cell lines. Comparisons of two-dimensional gel electrophoresis expression profiles of untreated RCC cell lines versus RCC cell lines treated with a mAb directed against G250 and the characterization of differentially expressed proteins by mass spectrometry and/or Edman sequencing led to the identification of proteins such as chaperones, antigen processing components, transporters, metabolic enzymes, cytoskeletal proteins and unknown proteins. Moreover, some of these differentially expressed proteins matched with immunoreactive proteins previously identified by proteome analysis combined with immunoblotting using sera from healthy donors and RCC patients, a technique called PROTEOMEX. Immunohistochemical analysis of a panel of surgically removed RCC lesions and corresponding normal kidney epithelium confirmed the heterogeneous expression pattern found by proteome-based technologies. In conclusion, conventional proteome analysis as well as PROTEOMEX could be successfully employed for the identification of markers which may allow the selection of patients prior to specific immunotherapy.

Ubiquitin COOH-Terminal Hydrolase 1: A Biomarker of Renal Cell Carcinoma Associated with Enhanced Tumor Cell Proliferation and Migration[?Q1: Running head: UCHL1, a Biomarker of RCC. Short title OK?Q1]

“…Distinct UCHL1 expression pattern in RCC subtypes and normal kidney epithelium. Using RCC tissue microarrays, the cellular distribution of the UCHL1 protein was monitored by immunohistochemistry in 87 primary lesions of different RCC subtypes, 9 oncocytomas, and autologous normal kidney epithelia as described previously (36). Abundant cytoplasmic staining of UCHL1 was observed in normal kidney epithelium, such as the distal and proximal tubule system, the mesangium of the glomerula, and the epithelium of the Bowman's capsule.…”

“…Negative controls were done by omitting the primary antibody. The design of the microtissue arrays used for the immunohistochemical evaluations has been described previously (36).…”

Application of immunoproteomics to leptospirosis: towards clinical diagnostics and vaccine discovery

“…renal cell carcinoma. These include the 'Serological and proteomic evaluation of antibody responses' (SPEAR) in the study by Unwin et al [17], 'Serological proteomic analysis' (SERPA) in the study by Klade et al [18] and 'Proteomex' in reports by Seliger and colleagues [19][20][21]. These techniques were named differently but all have similar analytical strategy that combines 2-D PAGE, MS identification, and immunoblotting, using cancer tissues as the antigens and sera from patients as the antibodies.…”

“…These techniques were named differently but all have similar analytical strategy that combines 2-D PAGE, MS identification, and immunoblotting, using cancer tissues as the antigens and sera from patients as the antibodies. With this approach to compare patients' sera with healthy normal sera, numerous candidate immunogens have been identified [17][18][19][20][21]. These candidate immunogens may potentially be useful for clinical diagnostics and therapeutics in renal cell carcinoma.…”

Rapid and Sensitive Identification of Major Histocompatibility Complex Class I-associated Tumor Peptides by Nano-LC MALDI MS/MS

“…The lysates were subjected to sonification (2 ϫ 5 cycles, 0.5 s at 100% power, UW 2070 sonicator, MS 73 needle; Bandelin, Berlin, Germany). Isoelectric focusing, second dimension SDS-PAGE separation, gel staining with colloidal Coomassie Blue, silver staining, and gel documentation were performed as described previously (26,27). For gel analyses, the ProteomWeaver software package (Version 2.1; Definiens AG, Munich, Germany) was used.…”