Circulating Extracellular Vesicles in the Pathogenesis of Type 1 Diabetes

循环细胞外小泡在1型糖尿病发病机制中的作用 1型糖尿病(T1D)是一种自身免疫性疾病，其特征是死亡、去分化或功能障碍。 功能正常的β细胞。然而，启动β细胞丢失和功能障碍的精确分子事件或调节 自身免疫是知识上的一个重大缺口，至今仍未解决。新出现的证据表明，循环中 细胞外小泡(EVS)是已知的细胞间微通讯的媒介，可能在 T1D的发病机制。然而，它们的确切功能和分子含量，特别是在启动β 人类的细胞丢失和功能障碍在很大程度上是未知的。 在这里，我们假设T1D中循环的EV，通过它们不同的分子特征，是 对β细胞健康有细胞毒性，从而促进了T1D的发病，有可能作为 疾病早期诊断的生物标志物。我们的初步数据表明，电动汽车，但不是电动汽车耗尽的部分 在体液因子中，T1D具有细胞毒性，特别是对人β细胞，但对阿尔法细胞没有影响。具体来说， 来自T1D受试者的EV，而不是来自对照受试者的EV，诱导人类β细胞死亡，表明循环 T1D-EVS是导致体液细胞毒性的关键成分。因此，在不同的情况下表征T1D电动汽车 疾病和对照的不同阶段可以确定重要的生物标志物，用于早期发现疾病 与β细胞丢失和功能障碍有关。我们的研究计划有以下具体目标： 目的1：确定从疾病前期到晚期T1D患者循环EVS的功能效应 对人类β细胞和阿尔法细胞健康的影响。我们将在T1D受试者的血浆中表征循环EVS I)早期(1-5年)、ii)晚期(&gt；10年)和iii)儿科(&lt；18年)阶段，并与i)自身抗体进行比较 非T1D阳性，II)T2D(控制高血糖等副作用)和III)适当的年龄、种族、 以及性别匹配的健康对照非糖尿病受试者对人类β细胞和阿尔法细胞健康的影响。 目的2：在啮齿动物体内建立循环EVS在T1D发病机制中的功能作用 模特。为了提供概念验证，我们将从糖尿病雌性NOD小鼠给非糖尿病小鼠注射循环EV 糖尿病控制和受者T1D疾病进展测试(Aim2A)；我们将抑制年轻人EV的分泌 T1D前雌性NOD小鼠，并测试T1D疾病进展和量化β细胞死亡和功能(Aim2B)。 目的3：研究人T1D-的分子机制、货物组成和货物功能。 电动汽车。我们将通过研究它们的摄取来解决T1D EV在β细胞细胞毒性中的不同功能。 机制，(Aim3A)；比较T1D和对照对象的电动汽车货物(Aim3B)；检查 EV-RNAs差异表达(Aim3C)。 我们提出的研究具有临床意义，并将深刻改变我们对糖尿病进展的理解。 T1D。如果成功，我们的研究有可能确定新的生物标记物和治疗T1D的方法。

SUMMARY: Circulating Extracellular Vesicles in the Pathogenesis of Type 1 Diabetes Type 1 diabetes (T1D) is an autoimmune disease, characterized by death, dedifferentiation or dysfunction of functional beta cells. However, precise molecular events that initiate beta cell loss and dysfunction or mediate autoimmunity is a major gap in knowledge that remains unaddressed. Emerging evidence suggests that circulating extracellular vesicles (EVs), known mediators of intercellular microcommunication, may play important roles in the pathogenesis of T1D. However, their precise function and molecular contents, especially in the initiation of beta cell loss and dysfunction in humans are largely undefined. Here, we hypothesize that circulating EVs in T1D, through their distinct molecular characteristics, are cytotoxic to beta cell health, thus contributing to the pathogenesis of T1D, with the potential to serve as biomarkers for early disease diagnosis. Our preliminary data suggests that EVs, but not EV-depleted fraction in the humoral factors in T1D are cytotoxic, particularly to human beta cells, but not to alpha cells. Specifically, EVs from T1D subjects, but not from control subjects, induce human beta cell death suggesting that circulating T1D-EVs are key components contributing to humoral cytotoxicity. Therefore, characterizing T1D EVs at various stages of the disease and control could identify important biomarkers for early detection of the disease that correlate with beta cell loss and dysfunction. Our research plan has the following Specific Aims: AIM 1: To determine the functional effects of circulating EVs from pre-disease to late stage T1D subjects on human beta and alpha cell health. We will characterize circulating EVs from the plasma of T1D subjects at i) early (1-5 years), ii) late (>10 year) and iii) pediatric (<18 year) stages and compare them with i) autoantibody positive non-T1D, ii) T2D (control for secondary effects such as hyperglycemia) and iii) appropriate age, ethnicity, and sex-matched healthy control non-diabetic subjects for their effect on human beta and alpha cell health. AIM 2: To establish the functional role of circulating EVs in the pathogenesis of T1D in vivo using a rodent model. To provide proof-of-concept, we will administer circulating EVs from diabetic female NOD mice to non- diabetic controls and test T1D disease progression in the recipient (Aim2A); we will inhibit EV secretion in young pre-T1D female NOD mice and test T1D disease progression and quantify beta cell death and function (Aim2B). AIM 3: Investigate the molecular mechanisms, cargo composition and cargo function of human T1D- EVs. We will address the differential function of T1D EVs in beta cell cytotoxicity by investigating their uptake mechanisms, (Aim3A); compare EV cargo in T1D and control subjects (Aim3B); examine the function of differentially expressed EV-RNAs (Aim3C). Our proposed research is clinically relevant and will profoundly change our understanding of the progression of T1D. If successful, our study has the potential to identify new biomarkers and therapeutics for T1D.