5-aza-2'-deoxycytidine-induced expression of functional cancer testis antigens in human renal cell carcinoma: Immunotherapeutic implications

• Published in: Clinical cancer research 2002-08, Vol.8 (8), p.2690-2695
• Main Authors: CORAL, Sandra, SIGALOTTI, Luca, ALTOMONTE, Maresa, ENGELSBERG, Arne, COLIZZI, Francesca, CATTAROSSI, Ilaria, MARASKOVSKY, Eugene, JAGER, Elke, SELIGER, Barbara, MAIO, Michele
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal; Antigens, Neoplasm - biosynthesis; Antimetabolites, Antineoplastic - pharmacology; Antineoplastic agents; Azacitidine - analogs & derivatives; Azacitidine - pharmacology; Biological and medical sciences; Blotting, Western; Carcinoma, Renal Cell - drug therapy; Carcinoma, Renal Cell - therapy; Chemotherapy; Electrophoresis, Polyacrylamide Gel; Eye Proteins - biosynthesis; Humans; Immunotherapy - methods; Kidney Neoplasms - drug therapy; Kidney Neoplasms - therapy; Medical sciences; Melanoma-Specific Antigens; Membrane Proteins; Mitogen-Activated Protein Kinases; Neoplasm Proteins - biosynthesis; Pharmacology. Drug treatments; Precipitin Tests; Protein Biosynthesis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Time Factors; Tumor Cells, Cultured
• ISSN: 1078-0432

Limited therapeutic options are presently available for advanced renal cell carcinoma (RCC). This study was designed to define the clinical potential of the DNA hypomethylating agent 5-aza-2'-deoxycytidine (5-AZA-CdR) in human RCC, through its control of the expression of "therapeutic targets" of the cancer testis antigen (CTA) family, and of the tumor-associated antigen RAGE-1, in RCC cells. Reverse transcription (RT)-PCR assays of a panel of RCC cells treated with 5-AZA-CdR, investigated the induction of the expression of several CTAs and of RAGE-1. Immunoprecipitation and Western blotting assessed whether the expression of CTA-specific mRNA induced by 5-AZA-CdR resulted in a translated protein of appropriate molecular weight. The functional activity of de novo expressed CTA was evaluated using (51)Cr release cytotoxicity assays of 5-AZA-CdR-treated HLA-A2-positive RCC cells using HLA-A2-restricted NY-ESO-1-specific CTLs. Exposure to 5-AZA-CdR invariably induced the expression of the CTA MAGE-1, -2, -3, and -4, GAGE 1-6, and NY-ESO-1 in all of the RCC cells investigated. De novo expression of NY-ESO-1 was persistent, being still detectable 60 days after the end of treatment, and generated a functional protein efficiently recognized by HLA-A2-restricted NY-ESO-1-specific CTLs. 5-AZA-CdR also induced RAGE-1 expression in RAGE-1-negative RCC and sarcoma cells but not in neoplastic cells of different histology. This study provides the scientific rationale to establish new strategies of chemoimmunotherapy in RCC patients. The well-defined immunogenicity of the investigated CTAs and of RAGE-1 suggest that systemic administration of 5-AZA-CdR represents a promising strategy to enhance the constitutively poor immunogenic potential of RCC cells, and to propose that virtually all RCC patients receive active and/or adoptive CTA- or RAGE-1-based immunotherapy.