Mechanisms and Therapeutic Modulation of T Cell Autoimmune Responses in Sjogren's Syndrome

项目摘要 Sjӧ-gren综合征是一种自身免疫性疾病，影响400万人的口腔和全身健康 美国人。SS以慢性炎症和外分泌腺功能障碍为特征，会导致口干， 干眼和各种全身健康问题，没有治愈或有效的生物疗法可用。这个 本R01项目的目的是阐明IL-7和Th1细胞因子对SjS Treg功能的负面影响 并测试增强Tregs中的TCF1和TIM-3是否可以改善和增强其抑制功能， 对IL-7/Th1细胞因子的抵抗，以及改善SjS的能力。倾向于干燥综合征的非肥胖型糖尿病小鼠的Tregs 是功能受损的。促炎细胞因子，包括Th1型细胞因子干扰素γ和IL-12，可以损害 Tregs促进自身免疫性炎症的功能和稳定性。我们的初步研究取得了重要的成果 我们的中心假说IL-7和Th1细胞因子参与异常Treg形成的证据 在SjS条件下发挥作用，部分是通过下调TCF1和TIM-3，并增强 TCF1和TIM-3的表达/活性可改善和促进免疫抑制功能、稳定性和 SJS--这些树的减毒能力。在目标1中，我们将确定TCF1在Tregs和 慢病毒基因表达载体可改善/增强其免疫抑制功能，在 炎症状况，以及改善SjS的能力。体外Treg功能检测和体内Treg转移 将采用TO SjS小鼠模型。在目标2中，我们将确定是否增强TIM-3的活性或表达 Tregs可以改善和提高其免疫调节和Sjs的减毒功能。我们将增强TIM-3 使用几种方法激活或表达，包括体内注射TIM-3配体，体外刺激 携带Tim-3配体的Tregs和慢病毒载体在Tregs中强制表达TIM-3。我们将测试是否增强了 TIM-3的表达/激活可改善/增强正常及干燥综合征患者的免疫抑制功能 老鼠和人类的树。在目标3中，我们将全面定义小鼠和 IL-7和Th1细胞因子联合高通量NGS RNA诱导与SjS相关的人Treg 测序，这将识别新的分子玩家和潜在的Treg功能缺陷的途径 SJS，IL-7和Th1细胞因子诱导。该项目的完成将解决一个关键的知识差距，并 促进对Treg功能和稳定性的基本理解和翻译调节，这可能会导致 以Treg为基础的策略的发展，以改善未来的干燥综合征和其他各种自身免疫性疾病。

Project Summary Sjӧgren’s syndrome (SjS) is an autoimmune disease affecting the oral and systemic health of 4 million Americans. Characterized by chronic inflammation and dysfunction of exocrine glands, SS causes dry mouth, dry eyes and various systemic health problems, with no cure or effective biological therapy available. The objective of this R01 project is to elucidate the negative impact of IL-7 and Th1 cytokines on Treg function in SjS disease, and to test if enhancing TCF1 and Tim-3 in Tregs can improve and enhance their suppressor function, resistance to IL-7/Th1 cytokines, and the ability to ameliorate SjS. Tregs in SjS-prone non-obese diabetic mice are functionally impaired. Pro-inflammatory cytokines, including Th1 cytokines IFNγ and IL-12, can impair the function and stability of Tregs to promote autoimmune inflammation. Our preliminary studies yielded important evidence for the formation of our central hypothesis that IL-7 and Th1 cytokines contribute to aberrant Treg function in SjS conditions, in part by downregulation of TCF1 and Tim-3, and that enhancing the expression/activity of TCF1 and Tim-3 may improve and boost the immunosuppressive function, stability and SjS-attenuating ability of these Tregs. In Aim 1, we will determine if forced expression of TCF1 in Tregs with lentiviral gene expression vectors can improve/enhance their immunosuppressive function, stability under inflammatory conditions, and ability to ameliorate SjS. In vitro Treg functional assays and in vivo Treg transfer to SjS mouse model will be employed. In Aim 2, we will determine if enhancing Tim-3 activity or expression in Tregs can improve and boost their immune-regulatory and SjS-attenuating function. We will enhance Tim-3 activation or expression using several approaches, including in vivo injection of Tim-3 ligand, in vitro stimulation of Tregs with Tim-3 ligand, and force-expressing Tim-3 in Tregs using lentiviral vectors. We will test if enhancing Tim-3 expression/activation can improve/boost the immunosuppressive function of normal and SjS-affected mouse and human Tregs. In Aim 3, we will comprehensively define the transcriptomic changes in mouse and human Tregs associated with SjS and induced by IL-7 and Th1 cytokines with high throughput NGS RNA- sequencing, which will identify new molecular players and pathways underlying the defective Treg function in SjS and induced by IL-7 and Th1 cytokines. Completion of this project will address a critical knowledge gap and advance both basic understanding and translational modulation of Treg function and stability, which could lead to development of Treg-based strategies for ameliorating SjS and various other autoimmune diseases in future.