Investigating a novel role of DRAK2 in T cell migration and synapse formation

项目摘要 自身免疫性疾病是一种日益严重的威胁，影响着2400多万美国人。目前的许多治疗方法 80多种不同的自身免疫性疾病试图广泛抑制免疫系统。然而，这导致了 许多有害的影响，并使患者面临感染和癌症的风险。有一件急事 需要开发在不损害自身反应性免疫反应的情况下选择性抑制自身反应的治疗方法 对病原体和肿瘤免疫。然而，我们缺乏对分子途径的完整了解 不同地影响自身反应性T细胞和病原体特异性T细胞。DRAK2是一种丝氨酸/苏氨酸激酶 淋巴细胞含量高。Drak2-/-小鼠对1型糖尿病和多发性骨髓瘤小鼠模型中的疾病具有抵抗力 硬化症。对这些自身免疫性疾病的抵抗力部分是由于T细胞在靶体内积累的减少 Drak2-/-小鼠的器官与野生型小鼠的比较。然而，Drak2-/-小鼠保持了有效清除 肿瘤和病原体类似于野生型小鼠，Drak2-/-T细胞能够进入外周器官 对病原体的反应。因此，DRAK2在不同地影响自身反应的信号通路中发挥作用 和病原体特异性T细胞。然而，DRAK2在T细胞激活、迁移和功能上的不同作用 目前仍不清楚。进一步了解DRAK2是如何影响T细胞功能的，将提供对 分子通路明显影响自身反应性和病原体特异性T细胞，这对于 为自身免疫性疾病开发改进的治疗方法。为了填补这一知识空白，我将评估 DRAK2在T细胞迁移和突触形成的关键方面，这是最佳T细胞所必需的 激活。我的初步数据显示，与迁移、突触形成和肌动蛋白相关的基因 在缺乏Drak2的情况下，调节在T细胞中差异表达。此外，将显示Drak2-/-T单元格 与野生型T细胞相比，向趋化因子CCL19迁移减少，突触形态异常。 因此，我假设DRAK2调节T细胞突触的形成和迁移，这损害了最佳T细胞 激活。为了解决这个假设，在目标1中，我将通过以下方式评估在没有Drak2的情况下T细胞的迁移 评估趋化因子受体表面的表达、向趋化因子的迁移、肌动蛋白突起的形成以及 活体迁移。在目标2中，我将确定Drak2-/-T细胞和抗原提呈细胞之间是否形成突触 与野生型T细胞相比，在大小、动力学或组织结构上不同。总之，这些研究将调查如何 DRAK2改变了T细胞迁移和突触形成的关键功能。进一步深入了解 DRAK2在T细胞激活和功能中的作用将揭示影响自身反应和病原体的不同途径。 特异性T细胞，这将为开发新的治疗靶点提供重要信息 自身免疫性疾病。

Project Summary Autoimmune disease is a rising threat that affects over 24 million Americans. Many of the current therapies for the over 80 distinct autoimmune diseases seek to broadly suppress the immune system. However, this leads to many deleterious effects and leaves patients at risk for development of infection and cancer. There is an urgent need to develop therapeutics that selectively inhibit the autoreactive immune response without compromising immunity to pathogens and tumors. However, we lack a complete understanding of molecular pathways that differentially impact autoreactive and pathogen-specific T cells. DRAK2 is a serine/threonine kinase expressed highly in lymphocytes. Drak2-/- mice are resistant to disease in mouse models of type 1 diabetes and multiple sclerosis. Resistance to these autoimmune diseases is partly due to reduced accumulation of T cells in the target organs of Drak2-/- mice compared to wildtype mice. However, Drak2-/- mice maintain the ability to effectively clear tumors and pathogens similar to wildtype mice, and Drak2-/- T cells are able to enter peripheral organs in response to pathogens. Thus, DRAK2 functions in a signaling pathway that differentially impacts autoreactive and pathogen-specific T cells. However, the distinct roles of DRAK2 on T cell activation, migration, and function remain unclear. A further understanding of how DRAK2 impacts T cell function will provide essential insight into molecular pathways distinctly affecting autoreactive and pathogen-specific T cells, which is critical in order to develop improved therapies for autoimmune diseases. To fill this gap in knowledge, I will evaluate the role of DRAK2 in critical aspects of T cell migration and synapse formation, which are essential for optimal T cell activation. My preliminary data demonstrate that genes associated with migration, synapse formation, and actin regulation are differentially expressed in T cells in the absence of Drak2. Additionally, Drak2-/- T cells display decreased migration to the chemokine, CCL19 and abnormal synapse morphology compared to wildtype T cells. Thus, I hypothesize that DRAK2 regulates T cell synapse formation and migration, which impairs optimal T cell activation. To address this hypothesis, in Aim 1, I will assess T cell migration in the absence of Drak2 by evaluating chemokine receptor surface expression, migration to chemokines, actin protrusion formation, and in vivo migration. In Aim 2, I will determine if synapses formed between Drak2-/- T cells and antigen presenting cells differ in size, kinetics, or organization compared to wildtype T cells. Together, these studies will investigate how DRAK2 alters the critical T cell functions of migration and synapse formation. Further insight into the role of DRAK2 in T cell activation and function will reveal pathways that differentially impact autoreactive and pathogen- specific T cells, which will provide important information for developing novel therapeutic targets to treat autoimmune disease.