Biochemical mechanisms of Hedgehog signal transduction through primary cilia

Hedgehog通过初级纤毛信号转导的生化机制

基本信息

批准号：
10447446
负责人：
David R Raleigh
金额：
$ 20.19万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10447446
关键词：
Address Adult Animals Antibodies Applications Grants Architecture Ascorbic Acid Binding Biochemical Biological Assay Biophysical Process CRISPR interference CRISPR/Cas technology Cell Proliferation Cells Cilia Congenital Disorders Critical Pathways Data Development Disease Erinaceidae Event Family Future Gene Expression Genes Genetic Genetic Transcription Goals Health Homeostasis Human Human Biology Immunofluorescence Immunologic Investigation Kinetics LLC-PK1 Cells Label Ligands Light Lipid Binding Lipids Luciferases Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Membrane Transport Proteins Molds Molecular Normal Cell Oncogenic Output Pathway interactions Patients Peroxidases Phenotype Physiological Proteins Proteomics Reaction Regulation Reporter Research Project Grants Role SHH gene Sampling Signal Transduction Signaling Molecule Specific qualifier value Surface Testing Tissues Tumor Cell Line Vertebrates cell type experimental study functional genomics hedgehog signal transduction high dimensionality improved in vivo innovation lipidomics migration neoplastic cell programs response smoothened signaling pathway spatiotemporal trafficking transcription factor

项目摘要

Project summary Spatiotemporal regulation of signal molecules is critical for development and adult tissue homeostasis. Indeed, misactivation or mislocalization of signaling events can cause congenital disorders and cancers, and understanding how information transfer is regulated in cells is essential for understanding and improving human health. The Hedgehog pathway is conserved across metazoan animals where it controls cell proliferation, differentiation, migration, and homeostasis. In vertebrates, Hedgehog signals are transduced through primary cilia that project from the surface of most cells, including cells in cancers driven by misactivation of the Hedgehog pathway. Cilia are required for vertebrate Hedgehog signaling for reasons that remain unknown. Our central hypothesis is that the ciliary microenvironment enables protein and lipid interactions that are necessary for Hedgehog signal transduction. Our lab has identified an important role for cilia-associated lipids that bind to Smoothened, accumulate Smoothened to cilia, and activate the Hedgehog pathway. When the Hedgehog pathway is off, Smoothened accumulation and activity in cilia is inhibited by Patched1. How Patched1 and Smoothened accumulate and function in cilia, and how Patched1 inhibits Smoothened, are poorly understood despite increasing recognition that these activities of the Hedgehog pathway are critical in health and disease. To address these key gaps in our understanding of human biology, the objectives of this proposal are to define the protein interactions that are necessary for Patched1 and Smoothened accumulation and activity in primary cilia, and to determine if Patched1 inhibited Smoothened through compartmentalization of ciliary lipids. Recently, we have begun to appreciate that the Hedgehog pathway can gather information directly from the ciliary microenvironment. Our discovery of ciliary lipids that specify Hedgehog pathway output establishes a new paradigm for information input onto the Hedgehog transcriptional program, one in which changes in the ciliary microenvironment reprogram the Hedgehog response. In this proposal, we will systematically examine the dynamic repertoire of protein and lipid interactions in cilia that specify the Hedgehog pathway across normal and tumor cells. We will leverage recent technical advances in proteomic proximity-labeling mass spectrometry, lipidomic mass spectrometry, and functional genomics to interpret high-dimensional landscapes of cell states induced by Hedgehog pathway activation. The long-term goal of this proposal is to understanding mechanisms underlying Hedgehog signal transduction well enough to develop new treatments for Hedgehog-associated cancers. To do so, the data generated by this proposal will establish a rationale to interrogate these mechanisms in vivo and in samples from human patients with Hedgehog-associated cancers. More broadly, this proposal will elucidate why cilia are required for Hedgehog signaling and how information transfer is regulated in cells, an elusive and fundamental question of human biology.

项目摘要信号分子的时空调节对于发育和成人组织稳态至关重要。实际上，信号事件的误导或错误定位会导致先天性疾病和癌症，以及了解在细胞中如何调节信息传递对于理解和改善人类至关重要健康。刺猬途径是在控制细胞增殖的，跨多宗动物中保守的，分化，迁移和体内平衡。在脊椎动物中，刺猬信号通过初级传递从大多数细胞表面投射的纤毛，包括疾病犯罪驱动的癌中细胞路径。纤毛是脊椎动物刺猬信号所必需的，其原因尚不清楚。我们的中心假设是睫状微环境实现蛋白质和脂质相互作用刺猬信号转导。我们的实验室已经确定了与纤毛相关的脂质结合的重要作用平滑，积累平滑至纤毛，并激活刺猬途径。当刺猬途径关闭，平滑的积累和纤毛的活性受到补丁1的抑制。如何修补1和在纤毛中平滑积累和功能，以及如何抑制平滑的修补方法，对尽管人们越来越认识到刺猬途径的这些活动在健康和疾病中至关重要。为了解决我们对人类生物学的理解时的这些关键差距，该提议的目标是定义修补1所需的蛋白质相互作用，并在原发性纤毛，并确定修补了1是否通过纤毛脂质的分室化而抑制了平滑。最近，我们开始欣赏刺猬途径可以直接从睫状微环境。我们发现指定刺猬途径输出的睫状脂质建立了一个新的范式输入刺猬转录程序的信息，其中一个纤毛变化微环境重新编程了刺猬响应。在此提案中，我们将系统地检查纤毛中蛋白质和脂质相互作用的动态曲目指定正常和肿瘤细胞。我们将利用蛋白质组学接近标记的质谱法的最新技术进步，脂质组质谱和功能基因组学解释细胞态的高维景观由刺猬途径激活诱导。该建议的长期目标是了解刺猬信号的机制转导足够好，可以为刺猬相关的癌症开发新的治疗方法。为此，数据该提案产生的将建立一个理由，以在体内询问这些机制与刺猬相关的癌症患者。更广泛地，该提议将阐明纤毛为什么刺猬信号传导所必需的以及如何在单元格中调节信息传递的方式，这是一种难以捉摸和基本的人类生物学问题。