Lack of specific γ-retroviral vector long terminal repeat promoter silencing in patients receiving genetically engineered lymphocytes and activation upon lymphocyte restimulation
Lack of specific γ-retroviral vector long terminal repeat promoter silencing in patients receiving genetically engineered lymphocytes and activation upon lymphocyte restimulation
复制标题
DOI：
10.1182/blood-2009-01-199216
复制
发表时间：
2009-10-01
期刊：
影响因子：
20.3
通讯作者：
Morgan, Richard A.
中科院分区：
文献类型：
--
作者：
Burns, William R.;Zheng, Zhili;Morgan, Richard A.
肿瘤抗原特异性T细胞受体（TCR）基因的逆转录病毒转导进入淋巴细胞重定向T细胞以裂解肿瘤。此外，这些淋巴细胞的过继转移在转移性癌症患者中介导了客观应答。从2004年到2006年，超过40名患者在国家癌症研究所接受了表达黑色素瘤抗原特异性TCR的自体基因修饰淋巴细胞治疗。对18名这样的患者进行了体内持久性和基因表达分析。此外，研究了表观遗传沉默和淋巴细胞再刺激的影响。虽然基因修饰的淋巴细胞在体内持续存在，但观察到TCR转基因表达的关闭。亚硫酸氢盐测序分析和离体DNA甲基转移酶抑制表明，基因表达的减少并不导致DNA甲基化。令人惊讶的是，载体驱动的转基因转录活性的下调不是载体特异性的，而是模仿内源基因的。然而，TCR转基因表达的减少在淋巴细胞刺激后被逆转。这些数据表明，在过继转移的人淋巴细胞中缺乏γ-逆转录病毒启动子特异性基因沉默，并支持转基因表达在很大程度上受全球细胞机制的影响。使用免疫调节佐剂，例如疫苗接种或细胞因子治疗，用于体内T细胞活化可能有助于克服这种代谢静止，从而增强基于细胞免疫治疗的癌症治疗。（血。2009; 114：2888-2899）
Retroviral transduction of tumor antigen-specific T-cell receptor (TCR) genes into lymphocytes redirects T cells to lyse tumors. Furthermore, adoptive transfer of these lymphocytes has mediated objective responses in patients with metastatic cancer. From 2004 to 2006, more than 40 patients were treated with autologous gene-modified lymphocytes expressing a melanoma antigen-specific TCR at the National Cancer Institute. Eighteen such patients were analyzed for persistence and gene expression in vivo. In addition, the impact of epigenetic silencing and of lymphocyte restimulation was studied. Although gene-modified lymphocytes persisted in vivo, the shutdown of TCR transgene expression was observed. Bisulfite sequencing analysis and ex vivo DNA methyltransferase inhibition demonstrated that the decrease in gene expression did not result from DNA methylation. Surprisingly, down-regulation of vector-driven transgene transcriptional activity was not vector specific but mimicked that of endogenous genes. The decrease in TCR transgene expression, however, was reversed upon lymphocyte stimulation. These data demonstrate a lack of gamma-retroviral promoter-specific gene silencing in adoptively transferred human lymphocytes and support that transgene expression is largely affected by global cellular mechanisms. The use of immunomodulatory adjuvants, eg, vaccination or cytokine therapy, for in vivo T-cell activation may help overcome this metabolic quiescence and thus augment cellular immunotherapy-based cancer therapy. (Blood. 2009; 114: 2888-2899)