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.

摘要 初级纤毛是一种以微管为基础的动态细胞附件，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，特别是对脊椎动物中的形态发生因子刺猬的反应 在分化和增殖过程中，调节多种组织的形态发生。然而，这些机制 纤毛特异性信号通过其维持并传播到直接下游通路 形态发生还不是很清楚。理解纤毛上的信号需要机械性地理解 纤毛运输，分离纤毛和信号分子的胞外功能，并研究功能 在不破坏纤毛的情况下直接在组织中产生后果。我的团队是最早研究纤毛专一性的团队之一。 从亚细胞到有机体鳞片的信号，同时保持纤毛的形态。我们确认了纤毛 运输适配子Tulp3和Hedgehog途径的关键抑制因子GPR161和Ankmy2，两者都发挥作用 通过纤毛调控的cAMP信号。我们假设纤毛信号的内在复杂性可以是 通过检查纤毛中信号是如何保持和唯一传播的(区隔)来理解 纤毛如何在下游通路中引导正向和负向调节(反调节信号)。完毕 在接下来的五年里，我们将直接研究纤毛的区划和反调节信号 调节不同组织中的形态发生。通过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在 以下是上下文。首先，我们将确定纤毛中的反向调节信号如何调节肾小管 动态平衡。我们假设Tulp3作为通常被抑制的生囊纤毛信号起作用。 被多囊藻毒素感染。我们建议通过识别和干扰Tulp3的货物来确定纤毛中的致囊驱动因素 在预防囊肿病方面。第二，我们将确定纤毛cAMP信号在神经管模式形成和 结案了。我们假设纤毛上cAMP-蛋白激酶A信号对Hedgehog通路的抑制起调节作用。 神经管关闭。我们将确定腺酰环化酶和蛋白激酶A区隔化在 纤毛-中心体复合体在调节刺猬信号强度、神经管构型和闭合中的作用。 第三，我们将确定纤毛调节的Hedgehog途径的抑制如何影响组织形态发生。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和腺苷环化酶的纤毛区划。通过这项研究，我们将确定 纤毛在引导组织突现特性中所特有的信号特征和后果。我们的工作 是跨学科的，并得到了在蛋白质组学、肾脏病、神经病理学、 人类遗传学和胚胎学。我们的研究将为理解复杂性提供新的切入点 纤毛疾病表型和定义治疗由睫毛功能障碍引起的疾病的翻译机会。