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

项目概要 干燥综合征 (SjS) 是一种影响 400 万人口腔和全身健康的自身免疫性疾病 美国人。 SS 以慢性炎症和外分泌腺功能障碍为特征，导致口干、 干眼症和各种全身健康问题，目前尚无治愈或有效的生物疗法。的 该 R01 项目的目标是阐明 IL-7 和 Th1 细胞因子对 SjS 中 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 细胞因子的抵抗力以及改善 SjS 的能力。易发生 SjS 的非肥胖糖尿病小鼠中的 Tregs 功能受损。促炎细胞因子，包括 Th1 细胞因子 IFNγ 和 IL-12，可损害 Tregs 的功能和稳定性促进自身免疫炎症。我们的初步研究取得了重要成果 IL-7 和 Th1 细胞因子导致异常 Treg 的中心假设形成的证据 在 SjS 条件下发挥功能，部分是通过 TCF1 和 Tim-3 的下调，并且增强 TCF1 和 Tim-3 的表达/活性可以改善和增强免疫抑制功能、稳定性和 这些 Tregs 的 SjS 减弱能力。在目标 1 中，我们将确定 TCF1 在 Tregs 中是否强制表达 慢病毒基因表达载体可以改善/增强其免疫抑制功能、稳定性 炎症状况和改善 SjS 的能力。体外 Treg 功能测定和体内 Treg 转移 将采用SjS小鼠模型。在目标 2 中，我们将确定是否增强 Tim-3 活性或表达 Tregs 可以改善和增强其免疫调节和 SjS 减弱功能。我们将增强 Tim-3 使用多种方法激活或表达，包括体内注射 Tim-3 配体、体外刺激 使用 Tim-3 配体构建 Tregs，并使用慢病毒载体在 Tregs 中强制表达 Tim-3。我们将测试是否增强 Tim-3 表达/激活可以改善/增强正常和 SjS 受影响的免疫抑制功能 小鼠和人类 Tregs。在目标 3 中，我们将全面定义小鼠和小鼠中的转录组变化。 人类 Tregs 与 SjS 相关并由 IL-7 和 Th1 细胞因子诱导，具有高通量 NGS RNA- 测序，这将识别 Treg 功能缺陷背后的新分子参与者和通路 SjS 并由 IL-7 和 Th1 细胞因子诱导。该项目的完成将解决关键的知识差距 推进对 Treg 功能和稳定性的基本理解和翻译调节，这可能导致 未来开发基于 Treg 的策略来改善 SjS 和各种其他自身免疫性疾病。

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.