Institut National de la Santé et de la Recherche Médicale, UMR 892, Nantes, France ; University of Nantes, Nantes, France ; Centre national de la recherche scientifique, UMR 6299, Nantes, France.
PLoS One. 2013 Sep 25;8(9):e75421. doi: 10.1371/journal.pone.0075421. eCollection 2013.
The melanoma antigens MELOE-1 and MELOE-2 are encoded by a messenger, called meloe, overexpressed in melanomas compared with other tumour cell types and healthy tissues. They are both able to elicit melanoma-specific T cell responses in melanoma patients, and MELOE-1-specific CD8 T cells have been involved in melanoma immunosurveillance. With the aim to develop immunotherapies targeting this antigen, we investigated the transcriptional mechanisms leading to the preferential expression of meloe messenger in the melanocytic lineage. We defined the minimal promoter region of meloe gene and identified binding motifs for a set of transcription factors. Using mutagenesis, co-transfection experiments and chromatin immunoprecipitation, we showed that transcription factors involved in meloe promoter activity in melanomas were the melanocytic specific SOX9 and SOX10 proteins together with the activated P-CREB protein. Furthermore, we showed that meloe promoter was hypomethylated in melanomas and melanocytes, and hypermethylated in colon cancer cell lines and mesotheliomas, thus explaining the absence of P-CREB binding in these cell lines. This was a second key to explain the overerexpression of meloe messenger in the melanocytic lineage. To our knowledge, such a dual transcriptional control conferring tissue-specificity has never been described for the expression of tumour antigens.
黑色素瘤抗原 MELOE-1 和 MELOE-2 由一种称为 meloe 的信使编码,与其他肿瘤细胞类型和健康组织相比,在黑色素瘤中过度表达。它们都能够在黑色素瘤患者中引发黑色素瘤特异性 T 细胞反应,并且 MELOE-1 特异性 CD8 T 细胞已参与黑色素瘤免疫监视。为了开发针对该抗原的免疫疗法,我们研究了导致 meloe 信使在黑素细胞谱系中优先表达的转录机制。我们定义了 meloe 基因的最小启动子区域,并确定了一组转录因子的结合基序。通过突变、共转染实验和染色质免疫沉淀,我们表明参与黑色素瘤中 meloe 启动子活性的转录因子是黑素细胞特异性的 SOX9 和 SOX10 蛋白以及激活的 P-CREB 蛋白。此外,我们表明 meloe 启动子在黑色素瘤和黑素细胞中呈低甲基化,在结肠癌细胞系和间皮瘤中呈高甲基化,从而解释了 P-CREB 结合在这些细胞系中缺失的原因。这是解释 meloe 信使在黑素细胞谱系中过度表达的第二个关键因素。据我们所知,这种赋予组织特异性的双重转录控制从未在肿瘤抗原的表达中描述过。