Aak1 to increase infiltration of adoptively transferred cells into solid tumors

项目摘要/摘要 在像CAR-T这样的过继转移疗法中，T细胞浸润不足是一个主要挑战。因此，一个 改进治疗的策略是加强T细胞向肿瘤的运输。然而，目前针对T细胞的治疗 细胞活动很大程度上是由免疫检查点调节器组成的，而在 针对肿瘤内蓄积的T细胞内在调节因子的治疗设计。这在一定程度上是由于 对T细胞转运所涉及的调控途径的认识不全面。我们最近确定了适配器 蛋白2相关激酶1(Aak1)在体内T细胞趋化肿瘤中的重要调节作用 正向基因筛查。该项目的主要目标是测量AAK1AS的翻译潜力 癌症的治疗目标是加强过继转移疗法，另一个目标是更好地 了解Aak1作为趋化因子受体CXCR3内化调节因子的分子功能。这些 目标将分三个目标实现。目标1将量化Aak1基因修饰对肿瘤的影响 在临床前实体瘤模型中过继转移的T细胞的渗透。目标2将决定Aak1是否 趋化因子诱导原代T细胞CXCR3内化所需的激酶活性。目标3将测量 在过继转移疗法中Aak1修饰对治疗效果的影响程度。这项建议 有几个创新方面，包括表征一种新型的T细胞特异性Aak1基因敲除小鼠， 一种新的Aak1突变结构的功能和机制测试，并对Aak1作为一种新的结构进行评估 治疗的目标是限制T细胞对炎症组织的趋化作用。这一项目的顺利完成将使其受益 实体肿瘤的新治疗策略的开发，其结果可广泛应用于任何T细胞 采用转让方式，并不限于单个CAR或TCR工程平台。

PROJECT SUMMARY/ABSTRACT Insufficient T cell infiltration is a major challenge in adoptive transfer therapies like CAR-T. Therefore, one strategy to improve therapy is to enhance T cell trafficking into tumors. However, current therapies targeting T cell activities largely consist of immune checkpoint modulators, and very little innovation has occurred in therapeutic design targeting T cell intrinsic regulators of intratumoral accumulation. This is due, in part, to an incomplete understanding of the regulatory pathways involved in T cell trafficking. We recently identified Adapter protein 2 associated kinase 1 (Aak1) as an important regulator of T cell chemotaxis into tumors in an in vivo forward genetic screen. The primary objective of this project is to measure the translational potential of AAK1 as a therapeutic target in cancer to augment adoptive transfer therapies, with the additional goal of better understanding molecular functions of Aak1 as a regulator of chemokine receptor Cxcr3 internalization. These goals will be accomplished in three aims. Aim 1 will quantify the impact of genetic modification of Aak1 on tumor infiltration of adoptively transferred T cells in a preclinical solid tumor model. Aim 2 will determine whether Aak1 kinase activity is required for chemokine-induced internalization of Cxcr3 in primary T cells. Aim 3 will measure the degree to which Aak1 modification impacts therapeutic efficacy in adoptive transfer therapies. This proposal has several innovative aspects, including characterization of a novel, T cell specific Aak1 knockout mouse, functional and mechanistic testing of a novel Aak1 mutant construct, and evaluation of Aak1 as a novel therapeutic target to limit T cell chemotaxis into inflamed tissue. Successful completion of this project will benefit development of novel treatment strategies for solid tumors, and findings can broadly be applied to any T cell adoptive transfer approach and is not limited to individual CAR or TCR engineered platforms.