Modulation of cystogenesis

PKD 1突变导致超过85%的常染色体显性多囊肾病病例 （ADPKD）。我们和其他人的一些研究表明，ADPKD蛋白多囊蛋白-1和-2 (PC1和PC 2）调节许多细胞事件和信号传导途径，如Ca 2+信号传导、JAK- STAT、mTOR、环AMP（cAMP）和平面细胞极性（PCP）。多囊蛋白如何调节这些途径， 然而，仍然难以捉摸。这些信号事件的顺序是未知的。一个主要的挑战是， 已经使用不同的模型系统、不同的细胞类型和在不同的环境下进行了实验。 条件更好地了解ADPKD的囊形成机制和疾病进展是一个高 临床护理优先。我们这项建议的长期目标是确定和调整控制因素， 使用多学科方法研究ADPKD囊肿形成和扩大。我们之前报道了一个 神经元中PC 1与蛋白激酶C和酪蛋白激酶底物的生化相互作用2（Pacsin 2），一种与细胞骨架组织和囊泡运输有关的细胞质磷蛋白。 通过体外研究，我们发现PC 1、Pacsin 2和N-Wasp处于同一蛋白复合体中，且存在缺陷 PC 1或Pacsin 2的缺失导致肌动蛋白细胞骨架和细胞迁移的缺陷。这里我们 我建议将我们对Pacsin 2的研究扩展到多个ADPKD的正向小鼠模型， 旨在了解抑制囊肿形成的机制。我们的目标是开发一种体内成像 细胞迁移的协议，并阐明Pascin 2在囊肿发生中的作用，使用多学科 方法包括人和小鼠的成囊蛋白质组和磷酸化蛋白质组的分析 ADPKD模型使用最新的定量蛋白质组学技术，再加上创新的生物信息学 工具. 拟议的研究可能会发现新的治疗靶点，这些靶点对膀胱发生的早期事件至关重要。

Mutations in PKD1 are responsible for over 85% of cases in autosomal dominant polycystic kidney disease (ADPKD). A number of studies by us and others have implicated that the ADPKD proteins polycystin-1 and -2 (PC1 and PC2) modulate a number of cellular events and signaling pathways such as Ca2+ signaling, JAK- STAT, mTOR, cyclic AMP (cAMP), and planar cell polarity (PCP). How polycystins modulate these pathways, however, remains elusive. The sequence of these signaling events is unknown. A major challenge is that many experiments have been performed using different model systems, different cell types, and under different conditions. Better understanding of the cystogenic mechanisms and disease progression of ADPKD is a high priority for clinical care. Our long-term goal of this proposal is to identify and modulate the factors controlling cyst formation and enlargement in ADPKD using multidisciplinary approaches. We previously reported a biochemical interaction between PC1 and protein kinase C and casein kinase substrate in neurons 2 (Pacsin 2), a cytoplasmic phosphoprotein that has been implicated in cytoskeletal organization and vesicle trafficking. By in vitro studies, we found that PC1, Pacsin 2 and N-Wasp are in the same protein complex and deficiency of either PC1 or Pacsin 2 leads to defects in actin cytoskeleton and cell migration in cultured cells. Here we propose to extend our study of Pacsin 2 to multiple orthologous mouse models of ADPKD that we have developed to understand mechanisms suppressing cystogenesis. We aim to develop an in vivo imaging protocol of cell migration and to elucidate the role of Pascin 2 in cystogenesis using multidisciplinary approaches including the analysis the cystogenic proteome and phosphoproteome in human and mouse ADPKD models using the latest quantitative proteomics technology, coupled with innovative bioinformatics tools. The proposed studies will likely discover novel therapeutic targets central to the early event(s) in cystogenesis.