Decitabine enhances tumor recognition by T cells through upregulating the MAGE-A3 expression in esophageal carcinoma
Decitabine enhances tumor recognition by T cells through upregulating the MAGE-A3 expression in esophageal carcinoma
复制标题
地西他滨通过上调食管癌中 MAGE-A3 的表达来增强 T 细胞对肿瘤的识别
DOI：
10.1016/j.biopha.2019.108632
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发表时间：
2019-04-01
影响因子：
7.5
通讯作者：
Zhang, Yi
中科院分区：
文献类型：
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作者：
Shi, Xiaojuan;Chen, Xinfeng;Zhang, Yi
睾丸癌（CT）抗原在各种类型的肿瘤中表达，代表了基于T细胞的免疫治疗的潜在靶点。CT基因表达和DNA甲基化的分析表明，某些CT基因是表观遗传调控的，研究证实，某些CT抗原是由DNA甲基化调控的。为了探讨MAGE-A3的表观遗传调控机制，提高MAGE-A3特异性T细胞治疗食管鳞癌的临床疗效，我们利用癌症基因组图谱中的分子生物学数据分析了CT基因在食管鳞癌中的表达及其与DNA甲基化的关系。我们在ESCC细胞系和ESCC组织中进行了定量逆转录PCR（qRT-PCR）、免疫组织化学和亚硫酸氢盐测序。进行功能测定，如流式细胞术、细胞毒性测定和ELISA，以确定去甲基化剂地西他滨（5-氮杂-2 '-脱氧胞苷，DAC）处理的癌细胞改善的抗原特异性T细胞应答。ESCC肿瘤细胞异种移植小鼠模型和酶联免疫斑点（ELISPOT）测定用于确定DAC治疗在ESCC中增强抗MAGE-3 T细胞应答的功能。此外，我们还对骨髓增生异常综合征（MDS）患者外周血单个核细胞（PBMC）进行了qRT-PCR和流式细胞术检测，结果显示，肿瘤抗原MAGE-A3在食管鳞癌中有不同水平的表达，并受到DNA甲基化的干扰。我们观察到地西他滨治疗后肿瘤细胞和组织中MAGE-A3表达的有效增加，并且MAGE-A3的表达受到DNA甲基化的影响。功能测定显示，DAC处理的靶细胞增强了IFN-γ的分泌和MAGE-A3抗原特异性T细胞的细胞溶解。在肿瘤细胞异种移植小鼠模型和ELISPOT测定中，DAC也增加了ESCC中MAGE-A3的表达和T细胞介导的肿瘤清除。DAC治疗MDS患者后，MAGE-A3应答性T细胞比例明显升高，提示DAC可能通过增强靶基因表达，改善MAGE-A3特异性T细胞治疗的临床疗效。
Cancer testis (CT) antigens are expressed in various types of tumors and represent the potential targets for T cell-based immunotherapy. Analysis of CT gene expression and DNA methylation have indicated that certain CT genes are epigenetically regulated and studies have confirmed that certain CT antigens are regulated by DNA methylation. In this study, we explored the epigenetic regulation of MAGE-A3 and improved the clinical outcome of MAGE-A3-specific T cell therapy in esophageal squamous cell carcinoma (ESCC).We used molecular profiling datasets in The Cancer Genome Atlas to analyze CT gene expression in ESCC and its regulation by DNA methylation. We performed quantitative reverse transcription PCR (qRT-PCR), immunohistochemistry and bisulfite sequencing in ESCC cell lines and ESCC tissues. Functional assays, such as flow cytometry, cytotoxicity assays and ELISA, were performed to determine the demethylation agent, decitabine (5-aza-2'-deoxycytidine, DAC)-treated cancer cell improved antigen specific T cells response. ESCC tumor cell-xenograft mouse model and enzyme-linked immunospot (ELISPOT) assays were used to determine the function of DAC treatment in enhancing anti-MAGE-3 T cell responses in ESCC. Furthermore, we performed qRT-PCR and flow cytometry in the peripheral blood mononuclear cells (PBMC) of myelodysplastic syndromes (MDS) patients.MAGE-A3, one of the CT antigens, expressed at various levels in ESCC and was interfered by DNA methylation. We observed an efficient increase in MAGE-A3 expression in tumor cells and tissues after the treatment of decitabine and the expression of MAGE-A3 was affected by DNA methylation. Functional assays showed enhanced secretion of IFN-gamma and cytolysis of MAGE-A3 antigen- specific T cells by DAC-treated target cells. In the tumor cell-xenograft mouse model and ELISPOT assays, DAC increased the expression of MAGE-A3 and T cell mediated tumor clearance in ESCC as well. Notably, the proportions of MAGE-A3-responsive T cells were elevated in DAC-treated patients with MDS, indicating DAC dismissed the epigenetic inhibition of MAGE-A3.DAC would probably improve the clinical outcome of MAGE-A3-specific T cell therapy by augmenting the expression of target gene.