Epigenetic modulation of MAGE-A3 antigen expression in multiple myeloma following treatment with the demethylation agent 5-azacitidine and the histone deacetlyase inhibitor MGCD0103

• Published in: Cytotherapy (Oxford, England) England), 2011-05, Vol.13 (5), p.618-628
• Main Authors: Moreno-Bost, Amberly, Szmania, Susann, Stone, Katie, Garg, Tarun, Hoerring, Antje, Szymonifka, Jackie, Shaughnessy, John, Barlogie, Bart, Grant Prentice, H, van Rhee, Frits
• Format: Article
• Language: English
• Subjects: 5-azacitidine; Advanced Basic Science; Antigens, Neoplasm - genetics; Antigens, Neoplasm - immunology; Antineoplastic Combined Chemotherapy Protocols; Azacitidine - therapeutic use; Benzamides - therapeutic use; cancer-testis antigen; Cell Line, Tumor; demethylation; DNA Methylation - drug effects; Epigenesis, Genetic - drug effects; epigenetics; Female; Gene Expression Regulation, Neoplastic - drug effects; Histone Deacetylase Inhibitors - therapeutic use; histone deactylase inhibitor; Humans; hypomethylation; MAGE-A3; Male; MGCD0103; multiple myeloma; Multiple Myeloma - genetics; Multiple Myeloma - immunology; Multiple Myeloma - therapy; Neoplasm Proteins - genetics; Neoplasm Proteins - immunology; Other; Pyrimidines - therapeutic use; Secondary Prevention; T-Lymphocytes, Cytotoxic - immunology; T-Lymphocytes, Cytotoxic - transplantation
• ISSN: 1465-3249; 1477-2566
• DOI: 10.3109/14653249.2010.529893

Abstract Background aims Immunotherapy targeting MAGE-A3 in multiple myeloma (MM) could eradicate highly aggressive and proliferative clonal cell populations responsible for relapse. However, expression of many cancer-testis antigens, including MAGE-A3, can be heterogeneous, leading to the potential for tumor escape despite MAGE-A3-induced immunity. We hypothesized that a combination of the hypomethylating agent 5-azacitidine (5AC) and the histone deacetylase inhibitor (HDACi) MGCD0103 (MGC) could induce MAGE-A3 expression in MAGE-A3-negative MM, resulting in recognition and killing of MM cells by MAGE-A3-specific cytotoxic T lymphocytes (CTL). Methods Gene expression analyses of MAGE-A3 expression in primary MM patient samples at diagnosis and relapse were completed to identify populations that would benefit from MAGE-A3 immunotherapy. MM cell lines were treated with 5AC and MGC. Real-time polymerase chain reaction (PCR) and Western blotting were performed to assess MAGE-A3 RNA and protein levels, respectively. Chromium-release assays and interferon (IFN) secretion assays were employed to ascertain MAGE-A3 CTL specificity against treated targets. Results Gene expression analysis revealed that MAGE-A3 is expressed in MM patients at diagnosis (25%) and at relapse (49%). We observed de novo expression of MAGE-A3 RNA and protein in MAGE-A3-negative cell lines treated with 5AC. MGC treatment alone did not induce expression but sequential 5AC/MGC treatment led to enhanced expression and augmented recognition by MAGE-A3-specific CTL, as assessed by51 Cr-release assays ( P = 0.047) and enzyme-linked immunosorbent assay (ELISA) for IFN-γ secretion ( P = 0.004). Conclusions MAGE-A3 is an attractive target for immunotherapy of MM and epigenetic modulation by 5AC, and MGC can induce MAGE-A3 expression and facilitate killing by MAGE-A3-specific CTL.