Interaction of protein-targeted therapeutics and ciliary dynamics

 描述(申请人提供)：常染色体多囊肾病(ADPKD)患者通常从20多岁开始出现高血压和其他心血管症状，中年时出现越来越多的充满液体的肾囊肿，最终通常出现终末期肾脏疾病(ESRD)，并在以后的生活中需要透析或肾脏置换。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是由于编码多囊蛋白的PKD1或PKD2基因的功能突变而引起的，多囊蛋白是一种大的跨膜蛋白，在细胞纤毛上异源二聚体，并影响多个下游信号蛋白的活性。临床前实验和临床试验表明，针对依赖多囊蛋白的信号缺陷可以减缓疾病的进展。我们在小鼠模型上的初步研究表明，抑制蛋白伴侣HSP90在缓解ADPKD症状方面非常有益，而抑制Aurora-A(AURKA)则是有害的，并开始确定相关的信号机制。有趣的是，最近的报道表明，ADPKD的严重表现部分取决于维持完整的纤毛，而我们发现抑制AURKA可以稳定纤毛。这表明使用AURKA抑制剂和类似活性的蛋白质对ADPKD患者可能是有害的。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。这项建议试图在ADPKD囊变的纤毛模型的背景下探索这些药物的作用机制，并进一步改进对该疾病的治疗。在这项建议中，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方法在ADPKD与非ADPKD囊性综合征中的疗效，并将使用最近开发的多路蛋白激酶抑制物珠(MIBs)技术来分析ganetespib与ADPKD特异性信号的相互作用。目标2将补充这一目标，评估在ADPKD和非ADPKD肾细胞和组织中，Ganetespib单独和联合治疗如何影响纤毛动力学和纤毛依赖信号。最后，35-40%的ADPKD患者会在他们的一生中患上某种形式的癌症，其中许多人将接受全身癌症治疗。AURKA和功能相关蛋白的抑制剂在癌症治疗中变得越来越常见。Aim 3将使用小鼠模型来测试AURKA抑制剂和其他预计用于稳定纤毛的药物对ADPKD患者构成风险的想法。

 DESCRIPTION (provided by applicant): 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 seeks to explore the mechanisms of action of these drugs in the context of a ciliary model for ADPKD cystogenesis, and to further improve therapy for this disease. 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.