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）在实体肿瘤（如GBM）中成功应用过继细胞转移（ACT），将需要t细胞提高肿瘤归家能力。最近对中枢神经系统（CNS）自身免疫性疾病的研究表明，产生白细胞介素17 （IL-17）的17型t细胞对中枢神经系统自身抗原有反应，由于它们能够通过表达IL-17/IL-22穿透血脑屏障（BBB），因此比其他t细胞亚群具有更高的向中枢神经系统实质迁移的能力。此外，17型t细胞上的趋化因子受体CCR6允许它们通过与其配体CCL20的相互作用渗透到中枢神经系统。虽然中枢神经系统自身免疫的诱导应该避免，但这些数据使我们假设，与1型t细胞相比，17型t细胞工程化引导胶质母细胞瘤（GBM）抗原将安全地表现出更强的抗胶质瘤活性。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）和人类异种移植物模型，并使用人类GBM患者来源的car - t细胞（Aim 2）进行ACT。目的1：在同基因胶质瘤模型中，与1型t细胞相比，小鼠17型t细胞在没有中枢神经系统自身免疫的情况下，将表现出更优越的体内持久性、肿瘤归巢和抗肿瘤作用。在我们的初步数据中，17型t细胞在体外比1型t细胞表现出更持久的抗肿瘤免疫反应。此外，我们的17型t细胞同时表达CXCR3和CCR6，并诱导胶质瘤细胞分别产生其同源配体CXCL10和CCL20。基于这些数据，我们假设17型t细胞对脑内胶质瘤的治疗效果优于1型t细胞，因为它们具有更强的体内持久性和转运到中枢神经系统肿瘤的能力。通过C57BL/6小鼠同基因模型，我们将确定CXCL10-CXCR3和CCL20-CCR6趋化因子轴是否促进CNS胶质瘤的17型细胞浸润。我们还将确保没有中枢神经系统自身免疫。目的2: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对人GBM异种移植物的抗肿瘤作用。我们将评估是否可以使用GBM患者衍生的t细胞繁殖17型CAR- t细胞，以及在Aim 1的同基因模型中观察到的关键表型（例如，趋化因子受体表达）是否可以在用CAR转导的GBM患者衍生的17型t细胞中重现。最后，我们将使用免疫功能低下的NOD/scid/ γ c (-/-) （NSG）小鼠，在大脑中植入人GBM患者来源的异种移植物，我们将确定静脉注射（即）人17型car -t细胞，特别是EGFRvIII-和EphA2-CARs，与单一抗原靶向方法相比，是否能在不引起显著中枢神经系统毒性的情况下，对人GBM异种移植物获得有效和持久的治疗反应。

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.