Signaling at the primary cilium in development and disease

发育和疾病中初级纤毛的信号传导

• 批准号: 10330492
• 负责人: Saikat Mukhopadhyay
• 金额: $ 32.8万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330492
• 关键词: Address; Adenylate Cyclase; Affect; Cilia; Complex; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cyst; Development; Disease; Embryology; Erinaceidae; Functional disorder; Homeostasis; Human; Human Genetics; Kidney; Microtubules; Morphogenesis; Morphology; Nephrology; Neural Tube Closure; Neural tube; Outcome; Pathogenesis; Pathology; Pathway interactions; Pattern; Phenotype; Process; Property; Proteomics; Regulation; Repression; Research; Role; Signal Transduction; Signaling Molecule; Testing; Tissues; Tubular formation; Vertebrates; Work; appendage; base; cell type; ciliopathy; extracellular; innovation; insight; morphogens; mouse model; neuropathology; preservation; prevent; public health relevance; response; smoothened signaling pathway; trafficking

Abstract The primary cilium is a microtubule-based dynamic cellular appendage that is found in many cell types. Cilia transduce cellular responses to extracellular signals, particularly to the morphogen hedgehog in vertebrates during differentiation and proliferation, regulating morphogenesis in multiple tissues. However, the mechanisms by which cilia-specific signals are maintained and propagated to direct downstream pathways during morphogenesis is not well understood. Understanding signaling at cilia requires mechanistic understanding of trafficking to cilia, isolating ciliary from extraciliary functions of signaling molecules, and studying functional consequences directly in tissues without disrupting cilia. My group is one of the foremost in studying cilia-specific signaling from subcellular to organismal scales, while preserving ciliary morphology. We identified the ciliary trafficking adapter Tulp3 and key repressors of hedgehog pathway, Gpr161 and Ankmy2, both of which function via cAMP signaling regulated by cilia. We postulate that the inherent complexity of ciliary signaling can be understood by examining how signals are maintained in and propagated uniquely by cilia (compartmentalization) and how cilia direct positive and negative regulation in downstream pathways (counterregulatory signaling). Over the next five years, we will directly study how compartmentalization and counterregulatory signaling at cilia regulates morphogenesis in different tissues. By leveraging our expertise in ciliary trafficking and hedgehog pathway repression, and by using innovative mouse models, we will study the effect of ciliary signaling in the following contexts. First, we will determine how counterregulatory signaling in cilia regulates renal tubular homeostasis. We hypothesize that Tulp3 cargoes function as cystogenic ciliary signals that are normally inhibited by polycystins. We propose to identify cystogenic drivers in cilia by identifying and perturbing cargoes of Tulp3 in preventing cysts. Second, we will determine the role of ciliary cAMP signaling in neural tube patterning and closure. We hypothesize that hedgehog pathway repression by cAMP-protein kinase A signaling at cilia regulates neural tube closure. We will determine role of adenylyl cyclase and protein kinase A compartmentalization in the cilium-centrosomal complex in regulating hedgehog signaling strength, neural tube patterning, and closure. Third, we will determine how cilia regulated repression of hedgehog pathway affects tissue morphogenesis. We will test ciliary contributions to repression thresholds required for specific morpho-phenotypic outcomes by perturbing ciliary compartmentalization of Gpr161 and adenylyl cyclases. Through this research we will identify the features and consequences distinctive to signaling by cilia in directing tissue emergent properties. Our work is cross-disciplinary and is supported by collaborators with expertise in proteomics, nephrology, neuropathology, human genetics and embryology. Our research will expose new entry points for understanding complex ciliopathy phenotypes and define translational opportunities for treating diseases caused by ciliary dysfunction.

抽象的 初级纤毛是一种基于微管的动态细胞附属物，存在于许多细胞类型中。纤毛 转导细胞对细胞外信号的反应，特别是脊椎动物中的形态发生素刺猬 在分化和增殖过程中，调节多个组织的形态发生。然而，这些机制 通过它维持纤毛特异性信号并将其传播到直接下游途径 形态发生尚不清楚。了解纤毛的信号传导需要机械地了解 运输到纤毛，将纤毛与信号分子的纤毛功能分离，并研究功能 直接影响组织而不破坏纤毛。我的团队是研究纤毛特异性的最重要的团队之一 从亚细胞到生物体尺度的信号传导，同时保留纤毛形态。我们确定了睫状体 转运接头 Tulp3 和刺猬通路的关键抑制因子 Gpr161 和 Ankmy2，两者均发挥作用 通过纤毛调节的 cAMP 信号传导。我们假设纤毛信号传导的固有复杂性可以是 通过检查信号如何在纤毛中维持和传播来理解（区室化） 以及纤毛如何指导下游途径的正向和负向调节（反调节信号传导）。超过 未来五年，我们将直接研究纤毛的区隔化和反调节信号传导 调节不同组织的形态发生。利用我们在纤毛贩运和刺猬方面的专业知识 途径抑制，并通过使用创新的小鼠模型，我们将研究纤毛信号传导在 以下上下文。首先，我们将确定纤毛中的反调节信号如何调节肾小管 体内平衡。我们假设 Tulp3 货物充当通常被抑制的囊原性纤毛信号 通过多囊蛋白。我们建议通过识别和扰动 Tulp3 的货物来识别纤毛中的囊原性驱动因素 以预防囊肿。其次，我们将确定睫状 cAMP 信号在神经管模式形成中的作用和 关闭。我们假设纤毛上的 cAMP-蛋白激酶 A 信号传导抑制刺猬通路，从而调节 神经管闭合。我们将确定腺苷酸环化酶和蛋白激酶 A 区室化在 纤毛中心体复合物调节刺猬信号强度、神经管模式和闭合。 第三，我们将确定纤毛如何调节刺猬通路的抑制影响组织形态发生。我们 将测试纤毛对特定形态表型结果所需的抑制阈值的贡献 干扰 Gpr161 和腺苷酸环化酶的纤毛区室化。通过这项研究我们将确定 纤毛信号在指导组织突现特性方面的独特特征和后果。我们的工作 是跨学科的，并得到具有蛋白质组学、肾病学、神经病理学专业知识的合作者的支持， 人类遗传学和胚胎学。我们的研究将揭示理解复杂性的新切入点 纤毛病表型并定义治疗纤毛功能障碍引起的疾病的转化机会。