Novel adoptive transfer therapy for glioma using CAR-transduced Type17 T-cells

摘要 嵌合抗原受体(CAR)转导T细胞过继细胞转移(ACT)的成功应用 实体肿瘤中的细胞(CAR-T细胞)，如GBM，将需要T细胞提高肿瘤归巢能力。 最近对中枢神经系统(CNS)自身免疫状况的研究表明，白细胞介素17(IL-17)- 产生对中枢神经系统自身抗原有反应的17型T细胞具有更高的迁移能力 中枢神经系统实质比其他T细胞亚群具有穿透血脑屏障的能力 (BBB)通过IL-17/IL-22的表达。此外，17型T细胞上的趋化因子受体CCR6使它们能够 通过与其配体CCL20相互作用来渗透CNS。尽管中枢神经系统自身免疫的诱导应该 避免这些数据，这些数据使我们假设，17型T细胞被改造成导向胶质母细胞瘤(GBM) 与1型T细胞相比，抗原将安全地显示出更好的抗胶质瘤活性。评估 我们的假设在临床前研究中，我们将使用同基因小鼠胶质瘤模型(Aim 1)和 人基底膜患者来源CAR-T细胞的异种移植模型和ACT(目标2)。 目的1.小鼠17型T细胞将在体内表现出优越的持久性、肿瘤归巢和抗肿瘤活性。 同基因脑胶质瘤模型中无中枢神经系统自身免疫的肿瘤效应与1型T细胞的比较。在……里面 我们的初步数据显示，17型T细胞在体外表现出比过去更持久的抗肿瘤免疫反应 类型1对应对象。此外，我们的17型T细胞同时表达CXCR3和CCR6并诱导胶质瘤细胞 以产生它们的同源配体，分别为CXCL10和CCL20。根据这些数据，我们假设 与其他T细胞相比，17型T细胞对脑内胶质瘤的治疗效果更好 类型1细胞，因为它们在体内的持久性更强，并有能力运输到中枢神经系统肿瘤。使用C57BL/6 小鼠同基因模型，我们将确定CXCL10-CXCR3和CCL20-CCR6趋化素轴 促进中枢神经系统胶质瘤中17型细胞的侵袭。我们还将确保中枢神经系统自身免疫的缺失。 目的2.CAR转导的人GBM患者来源的17型T细胞将安全地介导抗肿瘤 体内抗人GBM异种移植的实验研究我们将评估我们是否可以传播类型17 CAR-T 细胞使用GBM患者来源的T细胞，以及临界表型(如趋化因子受体) 在AIM 1中的同基因模型中观察到的)表达)可以在人类GBM患者来源的情况下概括 17型T细胞是与汽车一起转换的。最后，使用免疫低下的NOD/SCID/？c(-/-)(NSG) 小鼠脑内携带人GBM患者来源的异种移植物，我们将确定静脉注射(i.v.) 人17型CAR-T细胞的管理，特别是EGFRvIII-和EphA2-CARS与 单一抗原靶向方法，实现对人类的有效和持久的治疗反应 GBM异种移植不会引起明显的CNS毒性。

ABSTRACT The successful application of adoptive cell transfer (ACT) using chimeric antigen receptor (CAR)-transduced T- cells (CAR-T-cells) in solid tumors, such as GBM, will require improved tumor-homing ability by the T-cells. Recent studies of autoimmune conditions in the central nervous system (CNS) show that interleukin-17 (IL-17)- producing Type17 T-cells, which are reactive to CNS autoantigens, have a higher migratory capability to the CNS parenchyma than other T-cell subpopulations due to their ability to penetrate the blood brain barrier (BBB) via expression of IL-17/IL-22. Moreover, a chemokine receptor, CCR6, on Type17 T-cells allows them to infiltrate the CNS by interacting with its ligand, CCL20. Although the induction of CNS autoimmunity should be avoided, these data led us to hypothesize that Type17 T-cells engineered to direct glioblastoma (GBM) antigens will safely demonstrate superior anti-glioma activity compared with Type1 T-cells. To evaluate our hypothesis in preclinical studies, we will employ both a syngeneic mouse glioma model (Aim 1) and a human xenograft model and ACT with human GBM patient-derived CAR-T-cells (Aim 2). Aim 1. Mouse Type17 T-cells will demonstrate superior in vivo persistence, tumor-homing, and anti- tumor effects without CNS autoimmunity in syngeneic glioma models compared with Type1 T-cells. In our preliminary data, Type17 T-cells showed a more sustained anti-tumor immune response in vitro than did Type1 counterparts. Furthermore, our Type17 T-cells express both CXCR3 and CCR6 and induce glioma cells to produce their cognate ligands, CXCL10 and CCL20, respectively. Based on these data, we hypothesize that Type17 T-cells will demonstrate superior therapeutic efficacy against intra-cerebral gliomas compared with Type1 cells due to their greater in vivo persistence and their ability to traffic to CNS tumors. Using a C57BL/6 mouse syngeneic model, we will determine whether the CXCL10-CXCR3 and CCL20-CCR6 chemokine axes promote Type17 cell infiltration of CNS gliomas. We will also ensure the absence of CNS autoimmunity. Aim 2. Human GBM patient-derived Type17 T-cells transduced with CAR will safely mediate anti-tumor effects against human GBM xenograft in vivo. We will evaluate whether we can propagate Type17 CAR-T cells using GBM patient-derived T-cells, and whether the critical phenotype (e.g., chemokine receptor expression) observed in the syngeneic model in Aim 1 can be recapitulated in human GBM patient-derived Type17 T-cells that are transduced with the CAR. Finally, using immunocompromised NOD/scid/¿c(-/-) (NSG) mice bearing human GBM patient-derived xenografts in the brain, we will determine whether intravenous (i.v.) administration of human Type17 CAR-T-cells, especially both EGFRvIII- and EphA2-CARs compared with the single antigen-targeting approach, achieves effective and long-lasting therapeutic response against the human GBM xenografts without causing significant CNS toxicity.