Regulation of the Hedgehog Pathway by new cilium proteins in Development and Disease

新纤毛蛋白在发育和疾病中对刺猬通路的调节

• 批准号: 10517927
• 负责人: Xuecai Ge
• 金额: $ 32.35万
• 依托单位: UNIVERSITY OF CALIFORNIA, MERCED
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10517927
• 关键词: Adaptor Signaling Protein; Biotinylation; Brain; Cell Line; Cell division; Cell physiology; Cell surface; Cells; Cilia; Congenital Abnormality; Cultured Cells; Data; Defect; Development; Disease; Drosophila genus; Embryo; Endocytosis; Erinaceidae; Future; G-Protein-Coupled Receptors; Goals; In Vitro; Intervention; Knockout Mice; Label; Ligands; Light; Malignant Neoplasms; Mediating; Methods; Molecular; Numbness; Organelles; Pathway interactions; Phase; Phenocopy; Process; Proteins; Proteomics; Regulation; Research; Rest; Role; Signal Transduction; Signaling Protein; Testing; Time; Transducers; Transportation; Work; base; developmental disease; effective therapy; hedgehog signal transduction; in vivo; innovation; insight; mouse model; nerve stem cell; neurodevelopment; neurogenesis; neuroregulation; novel; premature; progenitor; protein transport; receptor; receptor internalization; smoothened signaling pathway; stable cell line; success; therapeutic development; tool; trafficking

PROJECT SUMMARY Defects in Hedgehog (Hh) signaling is widely implicated in various birth defects and cancers. The transduction of Hh signaling relies on the primary cilium, a miniature cell surface organelle known as the antenna of the cell. To activate Hh signaling, almost all protein transducers need to transit through the cilium. However, the molecular mechanisms of these protein transportation are not completely understood; and the signaling cascade within the cilium also remains unclear in the field. Our goal is to fill this gap with mechanistic studies of novel cilium proteins in the regulation of Hh signaling in vitro and in vivo. In preliminary studies, we have leveraged a new proximity-labeling tool and built an experimental platform to discovery new signaling proteins in the cilium with quantitative proteomics. We discovered surprising cilium localization of a molecule involved in receptor internalization during neural development. In this proposal, we aim to 1) decipher how the new cilium molecule control protein transport in the cilium during Hh signal activation; 2) determine its role in the Hh-controlled neural progenitor proliferation in the developing brain; and 3) apply the proximity biotinylation approach to identify new proteins involved in Smo trafficking and signaling during the time course of Hh activation. Our approach is innovative because it will reveal a new function of an old protein in the cilium during Hh transduction, and it employs a newly developed biotinylation tool to gain the systematic view of signaling proteins during Hh signaling activation. Upon completion, our study will shed light on long standing questions in the Hh pathway and may highlight new methods for the intervention of Hh related developmental disorders.

项目摘要 Hedgehog（Hh）信号传导缺陷广泛涉及各种出生缺陷和癌症。的 Hh信号转导依赖于初级纤毛，这是一种称为纤毛的微型细胞表面细胞器。 手机的天线。为了激活Hh信号传导，几乎所有的蛋白质转换器都需要通过 纤毛然而，这些蛋白质转运的分子机制并不完全 纤毛内的信号级联在该领域也仍然不清楚。我们的目标是 填补这一空白的机制研究的新纤毛蛋白在调节Hh信号在体外， in vivo.在初步研究中，我们利用了一种新的邻近标记工具，并建立了一个实验性的 通过定量蛋白质组学发现纤毛中新的信号蛋白的平台。我们发现 令人惊讶的纤毛定位的分子参与受体内化在神经 发展在这项提议中，我们的目标是1）破译新的纤毛分子如何控制蛋白质 Hh信号激活过程中纤毛的转运; 2）确定其在Hh控制的神经细胞中的作用 在发育中的脑中的祖细胞增殖;和3）将邻近生物素化方法应用于 鉴定在Hh活化的时间过程中参与Smo运输和信号传导的新蛋白质。 我们的方法是创新的，因为它将揭示纤毛中一种旧蛋白质的新功能， 它采用了一种新开发的生物素化工具，以获得系统的观点， 在Hh信号激活过程中的信号蛋白。完成后，我们的研究将揭示长期 Hh通路中的长期问题，并可能突出Hh相关干预的新方法 发育障碍