Role of Small Extracellular Vesicles in Beta Cell Failure and Autoimmunity in Type 1 Diabetes Mellitus

项目摘要/摘要 1型糖尿病(T1D)的特征是由于以下原因导致产生胰岛素的β细胞选择性破坏 自身侵袭性T细胞的渗入和免疫耐受性的破坏。因此，病人会遇到 β细胞功能和质量的严重丧失，因此需要外源性胰岛素。预防性的 改善疾病结局的免疫调节治疗策略仅取得极小的成功。 因此，恢复β细胞整体功能和健康的新策略是必要的。最近的证据 提示β细胞本身促进了它们自身在T1D进展过程中的同谋死亡，因此 可作为潜在的干预治疗靶点。长期接触促炎介质会 已被证明可导致β细胞功能障碍；然而，调节这一过程的机制尚未完全阐明 已澄清。在这一应用中，我们建议研究促炎介质对β细胞的影响 通过交换循环中的纳米囊泡进行对话，称为胞外囊泡(EVS)。我们和其他人 注意到T1D Pro-EV货物在蛋白质和miRNA水平上的显著变化 炎性细胞因子诱导。然而，改变的亲和力的直接生理和机制意义-- 炎性β细胞EV转运到受体β细胞的机制尚未完全阐明。中国的长期目标是 申请者将建立一个独立的实验室，探索电动汽车的生理和机械作用 T1D的发病机制。这项应用的中心假设是β细胞对它们自己的 通过糖尿病致β细胞EV货物交换致死诱导β细胞功能障碍和增强 抗原处理和呈递。为了验证这一假设，已经提出了两个具体目标，即 有大量的初步数据支持。在目标1中，我们将检验以下假设：促炎β细胞EV 激活CXCL10：CXCR3轴诱导β细胞衰竭。在目标2中，我们将检验以下假设： 炎性β细胞EV在T1D发病过程中参与β细胞免疫调节。这两个目标都将 使用体外和体内的T1D小鼠模型和人类胰岛和细胞系来解决这些假说。这个 申请者是高度合作的胰岛再生中心(再生医学中心)的一部分，地址为 梅奥诊所，明尼苏达州罗切斯特这种出色的研究环境除了精心挑选 由NIH资助的资深调查人员组成的指导委员会将为申请者提供 必要的工具和支持，使自己成为一名独立的研究调查员。

PROJECT SUMMARY/ABSTRACT Type 1 diabetes (T1D) is characterized by the selective destruction of insulin producing β-cells due to infiltrating autoaggressive T-cells and a break in immune tolerance. As a consequence, the patient is met with a severe loss of β-cell function and mass thus requiring the need for exogenous insulin. Preventative immunomodulatory therapeutic strategies to improve disease outcome have been met with minimal success. Therefore, novel strategies to restore overall β-cell function and health are warranted. Recent evidence suggests that β-cells themselves facilitate their own complicit demise during the progression of T1D and thus may serve as potential therapeutic targets of intervention. Chronic exposure to pro-inflammatory mediators has been shown to induce β-cell dysfunction; however, the mechanisms mediating this process have yet to be fully elucidated. In this application, we propose to study the impact of pro-inflammatory mediators on the β-cell dialogue through exchange of circulating nanovesicles termed extracellular vesicles (EVs). We and others have noted significant content alterations in EV cargoes at the protein and miRNA level upon T1D pro- inflammatory cytokine induction. However, the direct physiological and mechanistic implications of altered pro- inflammatory β-cell EV cargo to recipient β-cells have yet to be fully elucidated. The long-term goal of the applicant is to establish an independent laboratory exploring the physiological and mechanistic role of EVs in the pathogenesis of T1D. The central hypothesis of the application is that β-cells contribute to their own demise through diabetogenic β-cell EV cargo exchange to induce β-cell dysfunction and enhancement of antigen processing and presentation. To test this hypothesis, two specific aims have been proposed which are backed by extensive preliminary data. In Aim 1, we will test the hypothesis that pro-inflammatory β-cell EVs activate the CXCL10:CXCR3 axis to induce β-cell failure. In Aim 2, we will test the hypothesis that pro- inflammatory β-cell EVs contribute to β-cell immunomodulation during the pathogenesis of T1D. Both Aims will use ex vivo and in vivo mouse models of T1D and human islets and cell lines to address the hypotheses. The applicant is part of the highly collaborative Islet Regeneration Center (Center for Regenerative Medicine) at Mayo Clinic, Rochester, MN. This outstanding research environment in addition to a carefully selected mentoring committee comprised of established NIH-funded investigators will provide the applicant with the necessary tools and support to establish herself as an independent research investigator.