Profiling extracellular vesicle cargo in obesity and type 2 diabetes

项目摘要/摘要 肥胖 糖尿病 脂肪 光谱 新兴 与胰岛素抵抗和2型的高风险有关 (T2D)肥胖也主要归因于内脏 组织(VAT)，这增加了T2D风险；然而，潜在的确切分子机制 肥胖症患者血糖紊乱的炎症和代谢介质仍然很少 研究支持细胞外小泡(EVS)在T2D和肥胖中的作用。 新陈代谢 反常现象， 包括 。慢性低度炎症， 我明白了。 电动汽车是一种 不同种类的膜小泡，通过交换来参与细胞间的通讯 蛋白质、类脂和核酸。循环和脂肪细胞来源的EV已被证明在 T2D患者的肥胖，体重减轻后减少，并与血糖控制的恢复相关 在减肥手术后。脂肪来源的EVS也可能参与T2D的发病。我们假设 循环中EV的蛋白质和RNA货物有助于肥胖的代谢紊乱，以及 相应地，与减肥手术相关的葡萄糖代谢的改善。要解决这个问题 假设，我们提出了一种策略来识别EV衍生的与肥胖相关的蛋白质和RNA图谱- 相关的T2D。在目标1中，我们将 派生的 文字记录 评估血浆中蛋白质和RNA(lncRNA、miRNA和mRNA)的含量- EVS来自T2D(N=60)和血糖正常的肥胖者(N=60)。蛋白质和 显示T2D相关模式将在独立研究样本(N=120)中进行分析。 完成这一目标将导致识别和验证与以下相关的蛋白质和RNA图谱 T2D在极端肥胖中的作用。在目标2中，我们建议确定T2D相关蛋白和RNA 来自增值税的签名是通过评估从免疫选择的组份中分离出来的货物(蛋白质和RNA)而产生的 电动汽车和从增值税调节媒体获得的电动汽车。确定 涉及到的我们将建立 减肥后T2D缓解患者的T2D相关签名是否丢失 手术，但保留在糖尿病患者身上，即使体重显著下降。具体地说， 我们将测量在AIM 1中识别和验证的T2D相关蛋白质和RNA图谱 增值税是不是 在T2D的发病机制中将为进一步研究提供细胞靶点。在《目标3》中， 来源 的 电动汽车 体验T2D缓解和那些仍然患有糖尿病的人。完成这项工作 了解如何将 减肥手术患者的血浆和VAT样本及其最新分子特征 提供了一个独特的机会来确定哪些患者可能从减肥中获得最大好处 手术，提高我们对T2D发病机制的理解，并可能导致药物的开发 模仿手术的效果。 的 这个 机制 潜在的 T2D 减刑 以下是 AIM预计将加强我们的 减肥手术。

PROJECT SUMMARY/ABSTRACT Obesity diabetes adipose spectrum Emerging is linked with heightened risk of insulin resistance and type 2 (T2D) Obesity is also associated attributed primarily to visceral tissue (VAT), which increases T2D risk; however, the precise molecular mechanisms underlying the inflammatory and metabolic mediators of dysglycemia in obesity remain poorly studies support a role for extracellular vesicles (EVs) in both T2D and obesity. metabolic abnormalities, including . with chronic low-grade inflammation, of understood. EVs are a heterogeneous class of membrane vesicles that participate in cell-cell communication through exchange of proteins, lipids, and nucleic acids. Circulating and adipocyte-derived EVs have been shown to increase in obesity, decrease following weight reduction, and correlate with restoration of glycemic control in T2D patients following bariatric surgery. Adipose-derived EVs may also mediate T2D pathogenesis. We hypothesize that protein and RNA cargo of circulating EVs contribute to metabolic derangements in obesity, and correspondingly, to improvements in glucose metabolism associated with bariatric surgery. To address this hypothesis, we propose a strategy to identify EV-derived protein and RNA profiles associated with obesity- related T2D. In Aim 1, we will derived transcripts assess protein and RNA (lncRNA, miRNA, and mRNA) content in plasma- EVs obtained from T2D (N=60) and normoglycemic (N=60) individuals with obesity. Proteins and showing T2D-associated patterns will be analyzed in an independent study sample (N=120). Completion of this aim will result in the identification and validation of protein and RNA profiles associated with T2D in extreme obesity. In Aim 2, we propose to determine whether T2D-associated protein and RNA signatures emanate from VAT by evaluating cargo (protein and RNA) isolated from immuno-selected fractions of EVs and from EVs obtained from VAT-conditioned media. Determining involved we will establish whether T2D-associated signatures are lost in patients who experience T2D remission following bariatric surgery, but retained in those who remain diabetic, even in the presence of significant weight loss. In specific, we will measure T2D-associated protein and RNA profiles identified and validated in Aim 1 in individuals who whether VAT is the in the pathogenesis of T2D will provide a cellular target for further studies. In Aim 3, source of EVs experience T2D remission and those who remain diabetic. Completion of this understanding The combination of plasma and VAT samples from bariatric surgery patients and state-of-the-art molecular characterization provides a unique opportunity to identify those patients likely to receive the greatest benefit from bariatric surgery, improve our understanding of T2D pathogenesis, and perhaps lead to the development of drugs that mimic effects of surgery. of the mechanisms underlying T2D remission following aim is expected to enhance our bariatric surgery.