Understanding the impact of AAK1 on T cell chemokine receptor expression and chemotaxis

项目总结/摘要 T细胞直接和间接地浸润到大脑中，在各种各样的神经元损伤中促进神经元损伤。 神经炎性疾病，包括痴呆、多发性硬化（MS）和癫痫。因此，限制T细胞 浸润到中枢神经系统可能对这些患者具有治疗益处。衔接蛋白2 相关激酶1（Aak 1）最近被鉴定为T细胞趋化性进入炎症的重要调节因子， 在体内正向遗传筛选中的组织。本项目的主要目标是了解AAK 1 调节T细胞的趋化性，次要目标是建立AAK 1的翻译潜力， 神经炎性疾病的治疗靶点。这些目标将通过两个目标来实现。目标1将 使用原代T细胞确定AAK 1是否调节T细胞表面上的趋化因子受体表达。目的2 将使用体外迁移测定和体内T细胞趋化性测定来确定AAK 1调节T细胞趋化性的程度。 使用MS的Theiler小鼠脑脊髓炎病毒（TMEV）模型将细胞运输到脑中。 该提案具有几个创新方面，包括产生新的T细胞特异性Aak 1敲除小鼠， Aak 1作为T细胞浸润的遗传调节剂的验证，新型Aak 1的功能和机制测试 Aak 1作为限制T细胞趋化性进入炎症的新的治疗靶点的评价 组织.该项目的成功完成将使许多疾病环境广泛受益，因为研究结果可以很容易地 翻译为其他炎症性疾病，其中T细胞的募集驱动致病性，并可能导致 更好地治疗免疫性疾病

PROJECT SUMMARY/ABSTRACT T cells infiltrating into the brain directly and indirectly promote neuronal impairment in a wide variety of neuroinflammatory diseases, including dementia, multiple sclerosis (MS), and epilepsy. Thus, limiting T cell infiltration into the central nervous system could have therapeutic benefit for these patients. Adapter protein 2 associated kinase 1 (Aak1) was recently identified as an important regulator of T cell chemotaxis into inflamed tissues in an in vivo forward genetic screen. The primary objective of this project is to understand how AAK1 regulates T cell chemotaxis, with a secondary goal of establishing the translational potential of AAK1 as a therapeutic target in neuroinflammatory diseases. These goals will be accomplished in two aims. Aim 1 will determine if AAK1 regulates chemokine receptor expression on the T cell surface using primary T cells. Aim 2 will define the extent to which AAK1 regulates T cell chemotaxis using in vitro migration assays and in vivo T cell trafficking into the brain using the Theiler’s murine encephalomyelitis virus (TMEV) model of MS. This proposal has several innovative aspects, including generation of a novel, T cell specific Aak1 knockout mouse, validation of Aak1 as a genetic regulator of T cell infiltration, 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 broadly benefit many disease settings, as findings can easily be translated to other inflammatory conditions where recruitment of T cells drives pathogenicity and may lead to better treatments of immunologic diseases.