Targeting Microvesicles in Kidney Disease

项目摘要 细胞外囊泡（EV）将信号分子运输并递送到远处的细胞以支持代谢 过程、肾脏发育和体内平衡。EV信号的失调与代谢相关， 慢性肾病EV货物抑制葡萄糖转运蛋白，导致胰岛素抵抗。此外，EV 刺激促纤维化因子以驱动糖尿病患者的肾纤维化。此外，EV还涉及 多囊肾的发病机制尽管越来越多的人认识到EV在肾脏中起着至关重要的作用， 疾病，我们对控制EV形成和货物装载的分子原理的基本知识仍然存在 从根本上限制。 在预测性动物模型中识别和靶向控制EV生物发生的分子具有潜力， 从而产生有益于肾病患者的新的和协同的治疗剂。 EV释放是一个进化保守的过程，从藻类到苍蝇再到人类，这表明广泛地说， 对于该方法存在可推广的分子要求。黑腹果蝇在人类进化史上 发现与人类疾病相关的保守信号动力学，包括肾病。开始 为了探索EV释放和负载的分子机制，我们开发了一种体内EV标记方法， 果蝇上皮组织中的免疫调节系统。该系统允许在实时成像中可视化EV分泌步骤 接近。重要的是，我们已经将该系统用于EV蛋白质组学研究，旨在鉴定新的 EV生物发生和货物装载的调节剂。此外，该系统适用于靶向遗传突变体， 分析，允许稳健的功能验证推定的EV监管机构。在这个探索性的R21赠款中，我们 建议使用我们的系统来识别新的EV调节器。经验证的监管机构将构成以下内容的基础- up R 01基金专注于使用组织培养和小鼠模型描绘潜在的机制， 慢性肾病

PROJECT SUMMARY Extracellular vesicles (EV) transport and deliver signaling molecules to distant cells to support metabolic processes, kidney development and homeostasis. Deregulation of EV signals are associated with metabolic and chronic kidney disease. EV cargo inhibit glucose transporters, leading to insulin resistance. In addition, EV stimulate profibrotic factors to drive kidney fibrosis in diabetic patients. Furthermore, EV are implicated in the pathogenesis of polycystic kidney disease. Despite the growing realization that EV play a vital role in kidney disease, our basic knowledge of the molecular principles that control EV formation and cargo loading remains fundamentally limited. Identifying and targeting molecules that control EV biogenesis in predictive animal models has the potential to lead to novel and synergizing therapeutics for the benefit of kidney disease patients. EV release is an evolutionary conserved process from algae to flies to humans, suggesting that broadly generalizable molecular requirements for this process exist. Drosophila melanogaster has been invaluable in the discovery of conserved signaling dynamics relevant to human diseases, including nephropathies. To begin to explore the molecular mechanism of EV release and cargo loading, we have developed an in vivo EV labeling system in Drosophila epithelial tissues. This system permits the visualization of EV secretion steps in live imaging approaches. Importantly, we have adapted this system for EV proteomic studies aimed at identifying novel regulators of EV biogenesis and cargo loading. Further, the system is amenable to targeted genetic mutant analyses, permitting robust functional validation of putative EV regulators. In this exploratory R21 grant, we are proposing to use our systems to identify novel EV regulators. Validated regulators will form the basis of a follow- up R01 grant focused on delineating the underlying mechanisms using tissue culture and murine models of chronic kidney disease.