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

描述（由申请人提供）：使用嵌合抗原受体（CAR）转导的T细胞（CAR-T细胞）在实体瘤（如GBM）中成功应用过继性细胞转移（ACT）将需要T细胞提高肿瘤归巢能力。最近对中枢神经系统（CNS）自身免疫性疾病的研究表明，与CNS自身抗原反应的产生白细胞介素-17（IL-17）的17型T细胞比其他T细胞亚群具有更高的向CNS实质迁移的能力，这是因为它们能够通过表达IL-17/IL-22穿透血脑屏障（BBB）。此外，17型T细胞上的趋化因子受体CCR 6允许它们通过与其配体CCL 20相互作用来渗透CNS。尽管应避免诱导CNS自身免疫，但这些数据使我们假设，与1型T细胞相比，经工程改造以定向胶质母细胞瘤（GBM）抗原的17型T细胞将安全地显示出上级抗胶质瘤活性。为了在临床前研究中评估我们的假设，我们将采用同基因小鼠胶质瘤模型（Aim 1）和人异种移植模型以及人GBM患者来源的CAR-T细胞的ACT（Aim 2）。目标1：与1型T细胞相比，小鼠17型T细胞将在同基因胶质瘤模型中表现出上级的体内持久性、肿瘤归巢和抗肿瘤作用，而没有CNS自身免疫。在我们的初步数据中，17型T细胞在体外表现出比1型对应物更持久的抗肿瘤免疫应答。此外，我们的17型T细胞表达CXCR 3和CCR 6，并诱导神经胶质瘤细胞分别产生其同源配体CXCL 10和CCL 20。基于这些数据，我们假设，与1型细胞相比，17型T细胞将表现出对脑内胶质瘤的上级治疗功效，这是由于其更大的体内持久性和运输至CNS肿瘤的能力。使用C57 BL/6小鼠同基因模型，我们将确定CXCL 10-CXCR 3和CCL 20-CCR 6趋化因子轴是否促进CNS胶质瘤的17型细胞浸润。我们还将确保不存在CNS自身免疫。目的2：用CAR转导的人GBM患者来源的17型T细胞将在体内安全地介导针对人GBM异种移植物的抗肿瘤作用。我们将评估我们是否可以使用GBM患者来源的T细胞增殖17型CAR-T细胞，以及关键表型（例如，在Aim 1中的同基因模型中观察到的趋化因子受体表达）可以在用CAR转导的人GBM患者来源的17型T细胞中重现。最后，使用在脑中携带人GBM患者来源的异种移植物的免疫受损的NOD/scid/γ c（-/-）（NSG）小鼠，我们将确定静脉内（即，与单一抗原靶向方法相比，施用人17型CAR-T细胞，特别是EGFRvIII-和EphA 2-汽车两者，实现了针对人GBM异种移植物的有效和持久的治疗应答，而不引起显著的CNS毒性。

DESCRIPTION (provided by applicant): 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/gamma c (-/-) (NSG) mice bearing human GBM patient-derived xenografts in the brain, we will determine whether intravenous (i.e.) 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 responses against the human GBM xenografts without causing significant CNS toxicity.