Expression of miR-17-92 enhances anti-tumor activity of T-cells transduced with the anti-EGFRvIII chimeric antigen receptor in mice bearing human GBM xenografts.
Expression of miR-17-92 enhances anti-tumor activity of T-cells transduced with the anti-EGFRvIII chimeric antigen receptor in mice bearing human GBM xenografts.
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DOI：
10.1186/2051-1426-1-21
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发表时间：
2013
影响因子：
10.9
通讯作者：
Okada H
中科院分区：
文献类型：
--
作者：
Ohno M;Ohkuri T;Kosaka A;Tanahashi K;June CH;Natsume A;Okada H
miR-17-92的表达增强t细胞存活和干扰素(IFN)-γ的产生。我们之前报道了miR-17-92在胶质母细胞瘤（GBM）患者的t细胞中下调。我们假设转基因衍生的miR17-92和嵌合抗原受体（CAR）在t细胞中的共表达将提高对GBM过继性转移治疗的疗效。我们为miR-17-92构建了新的慢病毒载体（FG12-EF1a-miR-17/92）和一种CAR，该CAR由表皮生长因子受体变体III （EGFRvIII）特异性，单链可变片段（scFv）偶联到t细胞受体CD3ζ链信号模块和CD137 （4-1BB）和CD28的共刺激基元串联（pELNS-3C10-CAR）组成。用这些慢病毒载体转染人t细胞，并在体内和体外对其抗肿瘤作用进行了评价。car转导的t细胞（car -t细胞）对稳定表达EGFRvIII （U87-EGFRvIII）的U87 GBM细胞表现出有效的、抗原特异性的细胞毒性活性，当与miR-17-92共转导时，与未与miR-17-92共转导的car -t细胞相比，在替莫莫胺（TMZ）存在下表现出更高的存活率。在接受颅内U87-EGFRvIII异种移植的小鼠中，带有或不带有转基因来源的miR-17-92表达的car - t细胞显示出相似水平的治疗效果，而没有显示出任何car - t细胞不受控制的生长。然而，当这些小鼠在其大脑中再次受到U87-EGFRvIII细胞的攻击时，接受共转导car -t细胞的小鼠与未接受miR-17-92共转导的car -t细胞处理的小鼠相比，表现出更好的保护作用。这些结果保证了新型car - t细胞策略的发展，包括miR-17-92，以提高治疗效力，特别是在GBM患者中。
Expression of miR-17-92 enhances T-cell survival and interferon (IFN)-γ production. We previously reported that miR-17-92 is down-regulated in T-cells derived from glioblastoma (GBM) patients. We hypothesized that transgene-derived co-expression of miR17-92 and chimeric antigen receptor (CAR) in T-cells would improve the efficacy of adoptive transfer therapy against GBM. We constructed novel lentiviral vectors for miR-17-92 (FG12-EF1a-miR-17/92) and a CAR consisting of an epidermal growth factor receptor variant III (EGFRvIII)-specific, single-chain variable fragment (scFv) coupled to the T-cell receptor CD3ζ chain signaling module and co-stimulatory motifs of CD137 (4-1BB) and CD28 in tandem (pELNS-3C10-CAR). Human T-cells were transduced with these lentiviral vectors, and their anti-tumor effects were evaluated both in vitro and in vivo. CAR-transduced T-cells (CAR-T-cells) exhibited potent, antigen-specific, cytotoxic activity against U87 GBM cells that stably express EGFRvIII (U87-EGFRvIII) and, when co-transduced with miR-17-92, exhibited improved survival in the presence of temozolomide (TMZ) compared with CAR-T-cells without miR-17-92 co-transduction. In mice bearing intracranial U87-EGFRvIII xenografts, CAR-T-cells with or without transgene-derived miR-17-92 expression demonstrated similar levels of therapeutic effect without demonstrating any uncontrolled growth of CAR-T-cells. However, when these mice were re-challenged with U87-EGFRvIII cells in their brains, mice receiving co-transduced CAR-T-cells exhibited improved protection compared with mice treated with CAR-T-cells without miR-17-92 co-transduction. These results warrant the development of novel CAR-T-cell strategies that incorporate miR-17-92 to improve therapeutic potency, especially in patients with GBM.