Intratumor heterogeneity of cancer/testis antigens expression in human cutaneous melanoma is methylation-regulated and functionally reverted by 5-aza-2′-deoxycytidine

被引:168
作者
Sigalotti, L
Fratta, E
Coral, S
Tanzarella, S
Danielli, R
Colizzi, F
Fonsatti, E
Traversari, C
Altomonte, M
Maio, M
机构
[1] Univ Hosp Siena, Dept Oncol, Div Med Oncol & Immunotherapy, I-53100 Siena, Italy
[2] Ist Ricovero & Cura Carattere Sci, Canc Bioimmunotherapy Unit, Dept Med Oncol, Ctr Riferimento Oncol, Aviano, Italy
[3] MolMed SpA, Milan, Italy
关键词
D O I
10.1158/0008-5472.CAN-04-1442
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Cancer/testis antigens (CTA) are suitable targets for immunotherapy of human malignancies, and clinical trials are mainly focusing on MAGE-A3. However, the heterogeneous intratumor expression of CTA may hamper the effectiveness of CTA-directed vaccination through the emergence of CTA-negative neoplastic clones. We investigated the intratumor heterogeneity of CTA in human melanoma and the underlying molecular mechanism(s) at clonal level using 14 single cell clones generated from the melanoma lesion Mel 313. Reverse transcription-PCR revealed a highly heterogeneous expression of MADE-A1, -A2, -A3, -A4, -A6, GAGE 1-6, SSX 1-5, and PRAME among melanoma clones. Only nine clones expressed MAGE-A3 and competitive reverse transcription-PCR identified relative differences in the number of mRNA molecules of up to 130-fold between clones 5 and 14. This clonal heterogeneity of MAGE-A3 expression correlated with the methylation status of specific CpG dinucleotides in MAGE-A3 promoter: i.e., hypomethylated CpG dinucleotides at positions -321, -151, -19, -16, -5, -2, +21, and +42 were round in clones expressing high but not low levels of MAGE-A3. Supporting the role of DNA methylation in generating the intratumor heterogeneity of CTA, the DNA hypomethylating agent 5-aza-2'-deoxycytidine (5-AZA-dCyd) invariably induced their expression in all CTA-negative clones. Furthermore, 5-AZA-dCyd-treatment reduced to 6 folds the differential expression of MAGE-A3 between clones 5 and 14, which became recognized to a similar extent by T cells specific for a MAGE-A-encoded peptide. These findings identify promoter methylation as directly responsible for the intratumoral heterogeneity of therapeutic CTA in melanoma and foresee the use of 5-AZA-dCyd to overcome the limitations set by their intratumor heterogeneous expression to CTA-based vaccine therapy.
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页码:9167 / 9171
页数:5
相关论文
共 28 条
[1]  
ALTOMONTE M, 1993, CANCER RES, V53, P3343
[2]   Prolonged upregulation of the expression of HLA class I antigens and costimulatory molecules on melanoma cells treated with 5-aza-2′-deoxycytidine (5-AZA-CdR) [J].
Coral, S ;
Sigalotti, L ;
Gasparollo, A ;
Cattarossi, I ;
Visintin, A ;
Cattelan, A ;
Altomonte, M ;
Maio, M .
JOURNAL OF IMMUNOTHERAPY, 1999, 22 (01) :16-24
[3]  
Coral S, 2002, CLIN CANCER RES, V8, P2690
[4]   Demethylation of a hypermethylated P15/INK4B gene in patients with myelodysplastic syndrome by 5-Aza-2′-deoxycytidine (decitabine) treatment [J].
Daskalakis, M ;
Nguyen, TT ;
Nguyen, C ;
Guldberg, P ;
Köhler, G ;
Wijermans, P ;
Jones, PA ;
Lübbert, M .
BLOOD, 2002, 100 (08) :2957-2964
[5]  
De Smet C, 1999, MOL CELL BIOL, V19, P7327
[6]   STRUCTURE, CHROMOSOMAL LOCALIZATION, AND EXPRESSION OF 12 GENES OF THE MAGE FAMILY [J].
DEPLAEN, E ;
ARDEN, K ;
TRAVERSARI, C ;
GAFORIO, JJ ;
SZIKORA, JP ;
DESMET, C ;
BRASSEUR, F ;
VANDERBRUGGEN, P ;
LETHE, B ;
LURQUIN, C ;
BRASSEUR, R ;
CHOMEZ, P ;
DEBACKER, O ;
CAVENEE, W ;
BOON, T .
IMMUNOGENETICS, 1994, 40 (05) :360-369
[7]  
dos Santos NR, 2000, CANCER RES, V60, P1654
[8]   Characterization of an antigen that is recognized on a melanoma showing partial HLA loss by CTL expressing an NK inhibitory receptor [J].
Ikeda, H ;
Lethe, B ;
Lehmann, F ;
VanBaren, N ;
Baurain, JF ;
DeSmet, C ;
Chambost, H ;
Vitale, M ;
Moretta, A ;
Boon, T ;
Coulie, PG .
IMMUNITY, 1997, 6 (02) :199-208
[9]   Simultaneous humoral and cellular immune response against cancer-testis antigen NY-ESO-1:: Definition of human histocompatibility leukocyte antigen (HLA)-A2-binding peptide epitopes [J].
Jäger, E ;
Chen, YT ;
Drijfhout, JW ;
Karbach, J ;
Ringhoffer, M ;
Jäger, D ;
Arand, M ;
Wada, H ;
Noguchi, Y ;
Stockert, E ;
Old, LJ ;
Knuth, A .
JOURNAL OF EXPERIMENTAL MEDICINE, 1998, 187 (02) :265-270
[10]  
Jager E, 1997, INT J CANCER, V71, P142, DOI 10.1002/(SICI)1097-0215(19970410)71:2<142::AID-IJC3>3.0.CO