Interaction of protein-targeted therapeutics and ciliary dynamics

项目摘要 常染色体多囊肾病(ADPKD)患者通常经历高血压和其他 20多岁开始出现心血管症状，并形成越来越多的充满液体的肾囊肿 中年，通常在终末期肾病(ESRD)和需要透析或肾脏 在以后的生活中替代。此应用程序的目标是获得将改善临床的机械性见解 ADPKD患者的预后。ADPKD是由PKD1功能减弱或消除的突变引起的 或者PKD2基因，它编码多囊蛋白：在细胞纤毛处异源二聚体的大跨膜蛋白，以及 影响多个下游信号蛋白的活性。临床前实验和临床试验 研究表明，靶向多囊蛋白依赖的信号缺陷可以减缓疾病的进展。我们的预赛 在小鼠模型中的研究表明，抑制蛋白质伴侣HSP90对 减轻ADPKD症状，而抑制Aurora-A(AURKA)是有害的，并开始定义相关 信令机制。有趣的是，最近的报告表明，ADPKD的严重表现取决于 部分在于维持完整的纤毛，而我们发现抑制AURKA可以稳定纤毛。这 提示在ADPKD患者中使用AURKA抑制剂和类似活性的蛋白质可能是有害的。 相反，我们对HSP90、其蛋白客户和多囊蛋白调节的信号效应器的研究表明 这些可能部分通过促进纤毛吸收而起作用。该提案将探讨行动机制。 这些药物在ADPKD囊变的纤毛模型的背景下，测试了它们的部分 活动产生于对纤毛动力学的控制，并进行临床前测试，旨在表明改善 ADPKD的治疗。在这项提案中，Aim 1将使用小鼠模型来评估HSP90抑制剂ganetespib 与其他有希望的治疗ADPKD与非ADPKD囊性综合征的方法相结合， 并将使用最近开发的多路复用激酶抑制物珠(MIB)技术来描述相互作用 具有ADPKD特异性信号的ganetespib。目标2将补充这一目标，评估单独使用ganetespib 联合治疗影响纤毛动力学和纤毛依赖信号，在ADPKD与 非ADPKD肾细胞和组织。最后，35-40%的ADPKD患者会发展成某种形式的 在他们的有生之年，许多人会患癌症，许多人将接受系统性癌症治疗。AURKA和AURKA的抑制剂 功能相关的蛋白质在癌症治疗中变得越来越常见。AIM 3将使用鼠标模型来测试 认为AURKA抑制剂和其他预计可以稳定纤毛的药物对ADPKD患者构成风险。

Project Summary Autosomal polycystic kidney disease (ADPKD) patients typically experience hypertension and other cardiovascular symptoms commencing in their 20s, and develop an increasing burden of fluid-filled renal cysts in middle age, culminating typically in end stage renal disease (ESRD) and the need for dialysis or kidney replacement in later life. The goal of this application is to gain mechanistic insights that will improve clinical outcomes in patients with ADPKD. ADPKD arises from mutations reducing or eliminating function of the PKD1 or PKD2 genes, which encode polycystins: large transmembrane proteins that heterodimerize at cell cilia, and influence activity of multiple downstream signaling proteins. Pre-clinical experiments and clinical trials have shown that targeting polycystin-dependent signaling defects can slow disease progression. Our preliminary studies in mouse models have shown that inhibition of the protein chaperone HSP90 is extremely beneficial in reducing ADPKD symptoms, while inhibition of Aurora-A (AURKA) is deleterious, and begun to define related signaling mechanisms. Interestingly, recent reports indicate that severe manifestation of ADPKD depends in part on the maintenance of intact cilia, while we have found that AURKA inhibition stabilizes cilia. This suggests use of inhibitors of AURKA and proteins with similar activity may be harmful in ADPKD patients. Conversely, our studies of HSP90, its protein clients, and polycystin-regulated signaling effectors indicate these may act in part by contributing to ciliary resorption. This proposal will explore the mechanisms of action of these drugs in the context of a ciliary model for ADPKD cystogenesis, testing the hypothesis that part of their activity arises from control of ciliary dynamics, and perform preclinical tests intended to suggest improved therapy for ADPKD. In this proposal, Aim 1 will use mouse models to evaluate the HSP90 inhibitor ganetespib in combination with other promising therapies for efficacy in ADPKD versus in non-ADPKD cystic syndromes, and will use recently developed multiplexed kinase inhibitor beads (MIBs) technology to profile the interaction of ganetespib with ADPKD-specific signaling. Aim 2 will complement this aim, evaluating how ganetespib alone and in therapeutic combinations influences ciliary dynamics and cilia-dependent signaling, in ADPKD versus non-ADPKD renal cells and tissue. Finally, 35-40% of individuals with ADPKD will develop some form of cancer in their lifetime, and many will be treated with systemic cancer therapies. Inhibitors of AURKA and functionally related proteins are becoming common in cancer therapy. Aim 3 will use mouse models to test the idea that AURKA inhibitors and other drugs predicted to stabilize cilia pose risks for patients with ADPKD.