Interaction of protein-targeted therapeutics and ciliary dynamics

项目摘要 常染色体多囊肾病（ADPKD）患者通常患有高血压和其他疾病。 20多岁时开始出现心血管症状，并逐渐增加充满液体的肾囊肿的负担 在中年时，通常在终末期肾病（ESRD）中达到顶峰，需要透析或肾脏移植。 生活中的替代品。此应用程序的目标是获得机制见解，这将改善临床 ADPKD患者的结局。ADPKD是由PKD 1基因突变引起的， 或PKD 2基因，其编码多囊蛋白：在细胞纤毛处异二聚化的大跨膜蛋白，和 影响多种下游信号蛋白的活性。临床前实验和临床试验 表明靶向多囊蛋白依赖性信号传导缺陷可以减缓疾病进展。我们的初步 在小鼠模型中的研究表明，抑制蛋白质伴侣HSP 90在 减少ADPKD症状，而Aurora-A（AURKA）的抑制是有害的，并开始定义相关的 信号机制。有趣的是，最近的报告表明，ADPKD的严重表现取决于 部分维持完整的纤毛，而我们已经发现，AURKA抑制稳定纤毛。这 提示使用AURKA抑制剂和具有类似活性的蛋白质可能对ADPKD患者有害。 相反，我们对HSP 90、其蛋白质客户和多囊蛋白调节的信号效应物的研究表明， 这些可能部分地通过促进睫状体再吸收而起作用。本提案将探讨行动机制 这些药物的背景下，睫状体模型的ADPKD囊肿，测试的假设，他们的一部分， 活动产生于纤毛动力学的控制，并进行临床前试验，旨在表明改善 治疗ADPKD。在该提案中，Aim 1将使用小鼠模型来评估HSP 90抑制剂ganetespib 与其他有希望的治疗方法联合用于ADPKD与非ADPKD囊性综合征的疗效， 并将使用最近开发的多重激酶抑制剂珠（MIBs）技术来分析相互作用 与ADPKD特异性信号传导有关。目标2将补充这一目标，评估单用ganetespib 在治疗组合中影响纤毛动力学和纤毛依赖性信号传导，在ADPKD与 非ADPKD肾细胞和组织。最后，35-40%的ADPKD患者会出现某种形式的 在他们的一生中，许多人将接受全身癌症治疗。AURKA抑制剂和 功能相关的蛋白质在癌症治疗中变得普遍。Aim 3将使用小鼠模型来测试 认为AURKA抑制剂和其他预计可稳定纤毛的药物对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.