Memory T Cells Expressing an NKG2D-CAR Efficiently Target Osteosarcoma Cells
Memory T Cells Expressing an NKG2D-CAR Efficiently Target Osteosarcoma Cells
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DOI：
10.1158/1078-0432.ccr-17-0075
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发表时间：
2017-10-01
影响因子：
11.5
通讯作者：
Perez-Martinez, Antonio
中科院分区：
文献类型：
--
作者：
Fernandez, Lucia;Metais, Jean-Yves;Perez-Martinez, Antonio
目的：NKG 2D配体（NKG 2DL）在肿瘤微环境中的各种肿瘤类型和免疫抑制细胞上表达，为癌症治疗提供合适的靶点。多种免疫细胞表达NKG 2D受体，包括自然杀伤（NK）细胞和CD 8 + T细胞。NKG 2DL和NKG 2D受体之间的相互作用对于NK细胞消除骨肉瘤肿瘤起始细胞至关重要。在这份报告中，我们使用NKG 2D-NKG 2DL相互作用来优化针对骨肉瘤的免疫策略。我们在体外和体内评估了表达NKG 2D-4- 1BB-CD 3 z嵌合抗原受体（CAR）的CD 45 RA（-）记忆T细胞对骨肉瘤细胞的安全性和细胞毒性能力。实验设计：用含有4-1BB和CD 3 z信号结构域的NKG 2D汽车转导来自健康供体的CD 45 RA（-）细胞。通过流式细胞术分析NKG 2D CAR表达。通过进行常规的4小时铕-TDA释放试验，评价了NKG 2D-CAR（+）CD 45 RA（-）T细胞对骨肉瘤的体外细胞毒性。结果：慢病毒转导NKG 2D-4- 1BB-CD 3 z可显著增加CD 45 RA（-）细胞表面NKG 2D的表达。在转导的细胞中保持遗传稳定性。在体外，NKG 2D-CAR+记忆T细胞对骨肉瘤细胞系的细胞溶解活性比未转导细胞显著增加，同时保持健康细胞的完整性。NKG 2DCAR（+）记忆T细胞在骨肉瘤小鼠模型中具有相当大的抗肿瘤活性，而未转导的T细胞则无效。结论：我们的结果表明NKG 2D-4- 1BB-CD 3 z CAR重定向记忆T细胞在体内和体外靶向表达NKG 2DL的骨肉瘤细胞，可能是骨肉瘤患者有希望的免疫治疗方法。(C)2017年AACR。
Purpose: NKG2D ligands (NKG2DL) are expressed on various tumor types and immunosuppressive cells within tumor microenvironments, providing suitable targets for cancer therapy. Various immune cells express NKG2D receptors, including natural killer (NK) cells and CD8+ T cells. Interactions between NKG2DL and NKG2D receptors are essential for NK-cell elimination of osteosarcoma tumor-initiating cells. In this report, we used NKG2D-NKG2DL interactions to optimize an immunotherapeutic strategy against osteosarcoma. We evaluated in vitro and in vivo the safety and cytotoxic capacity against osteosarcoma cells of CD45RA(-) memory T cells expressing an NKG2D-4-1BB-CD3z chimeric antigen receptor (CAR).Experimental Design: CD45RA(-) cells from healthy donors were transduced with NKG2D CARs containing 4-1BB and CD3z signaling domains. NKG2D CAR expression was analyzed by flow cytometry. In vitro cytotoxicity of NKG2D-CAR(+) CD45RA(-) T cells against osteosarcoma was evaluated by performing conventional 4-hour europium-TDA release assays. For the in vivo orthotopic model, 531MII YFP-luc osteosarcoma cells were used as targets in NOD-scid IL2Rg(null) mice.Results: Lentiviral transduction of NKG2D-4-1BB-CD3z markedly increased NKG2D surface expression in CD45RA(-) cells. Genetic stability was preserved in transduced cells. In vitro, NKG2D-CAR+ memory T cells showed significantly increased cytolytic activity than untransduced cells against osteosarcoma cell lines, while preserving the integrity of healthy cells. NKG2DCAR(+) memory T cells had considerable antitumor activity in a mouse model of osteosarcoma, whereas untransduced T cells were ineffective.Conclusions: Our results demonstrate NKG2D-4-1BB-CD3z CAR-redirected memory T cells target NKG2DL-expressing osteosarcoma cells in vivo and in vitro and could be a promising immunotherapeutic approach for patients with osteosarcoma. (C) 2017 AACR.