Signal Transduction by Oxysterols

氧甾醇的信号转导

基本信息

批准号：
8920626
负责人：
DOUGLAS F COVEY
金额：
$ 40.2万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-05 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8920626
关键词：
Accounting Adherence Administrative Personnel Affinity Labels Agonist Alanine Amino Acids Arbitration Atherosclerosis Authorship Binding Binding Sites Biochemical Biological Biological Assay Biology Biotin Boxing Budgets Cell Communication Cell Extracts Cell physiology Cells Cellular biology Chemicals Chemistry Cholesterol Clinic Collaborations Committee Members Communication Conflict (Psychology)Congenital Abnormality Coupled Data Data Analyses Development Digestion Disease Drug Targeting Electronic Mail Ensure Erinaceidae Ethics Experimental Designs Functional disorder Funding Generations Goals Grant Human Human Resources Hydroxycholesterols Image Inborn Errors of Metabolism Inflammation Institution Integral Membrane Protein Intellectual Property Laboratories Leadership Legal patent Ligands Lipids Location Malignant Neoplasms Maps Mass Spectrum Analysis Mediating Metabolic Diseases Methods Molecular Mutagenesis Natural regeneration Nature Nerve Degeneration Oncogene Proteins Organic Chemistry Organic Synthesis Outcome Paper Patch Tests Pathway interactions Peptides Pharmaceutical Preparations Phase Photoaffinity Labels Play Portraits Preparation Process Progress Reports Protein Biochemistry Proteins Publications Published Comment Publishing Receptor Signaling Recruitment Activity Regulation Relative (related person)Research Research Personnel Research Project Grants Resistance Resolution Resources Role Safety Sampling Scanning Second Messenger Systems Signal Pathway Signal Transduction Signaling Molecule Site Steroids Sterols Streptavidin Structure Suggestion Synthesis Chemistry System Techniques Technology Transfer Telephone Testing Therapeutic Time TimeLine United States National Institutes of Health Universities Washington Work affinity labeling analog base biochemical tools conflict resolution cyclopamine data sharing design experience gene replacement hedgehog signal transduction human SMO protein immune function inhibitor/antagonist interdisciplinary collaboration lipid metabolism meetings mutant novel oncology organizational structure oxidation programs receptor research study response scaffold second messenger small molecule smoothened signaling pathway symposium

项目摘要

DESCRIPTION (provided by applicant): Oxysterols are a class of endogenous cellular lipids derived from cholesterol that have been implicated in the pathophysiology of atherosclerosis, inborn errors of metabolism, inflammation and cancer. In many cases, the proteins and molecular pathways through which these enigmatic molecules exert their powerful biological effects remain unknown. We established collaboration between a synthetic organic chemist and a cell biologist to understand how oxysterols activate the Hedgehog (Hh) signaling system, a pathway that plays important roles in development, regeneration and cancer. We discovered that a specific oxysterol, 20(S)-OHC, is an allosteric activator of the 7-pass transmembrane protein Smoothened (Smo), a human oncoprotein and key drug target in oncology. This finding significantly expands the regulatory scope of oxysterols as signaling molecules, demonstrating their capacity to function as direct agonists for both a human on co-protein and a signaling receptor. Based on preliminary work, we hypothesize that endogenous 20(S)-OHC functions as a second-messenger in Hh signaling. Using a combination of mutagenesis, photo affinity labeling, and mass spectrometry, we will map the region of Smo that interacts with 20(S)-OHC to provide a biochemical portrait of this novel class of receptor-ligand interaction (Aim 1). Using quantitative mass spectrometry and a click chemistry-based imaging assay, we will ask if Hedgehog signaling can alter cellular levels or distribution of 20(S)-OHC (Aim 2). Finally, we will develop and characterize novel Hh pathway inhibitors that are inspired by oxysterol scaffolds (Aim 3). We expect three major outcomes to emerge from the successful completion of this project: (1) An answer to the question of how Smo is regulated in cells, perhaps the longest-standing mystery in the Hh pathway, (2) a biochemical understanding of how oxysterols engage and regulate 7-pass signaling receptors, and (3) the development of an integrative toolkit that can be deployed to dissect any other oxysterol- regulated cellular process. To accomplish these goals, we have recruited a team of investigators with complementary expertise in cell biology, protein biochemistry, synthetic chemistry, and mass spectrometry.

描述（由申请人提供）：氧甾醇是一类源自胆固醇的内源性细胞脂质，其与动脉粥样硬化、先天性代谢缺陷、炎症和癌症的病理生理学有关。在许多情况下，这些神秘分子发挥其强大生物效应的蛋白质和分子途径仍然未知。我们与合成有机化学家和细胞生物学家建立了合作，以了解氧甾醇如何激活 Hedgehog (Hh) 信号系统，这是一种在发育、再生和癌症中发挥重要作用的途径。我们发现一种特定的氧甾醇 20(S)-OHC 是 7 次跨膜蛋白 Smoothened (Smo) 的变构激活剂，Smo 是一种人类癌蛋白，也是肿瘤学中的关键药物靶点。这一发现显着扩大了氧甾醇作为信号分子的调节范围，证明了它们作为人类辅助蛋白和信号受体的直接激动剂的能力。基于前期工作，我们假设内源性 20(S)-OHC 在 Hh 信号传导中充当第二信使。通过结合诱变、光亲和标记和质谱分析，我们将绘制 Smo 与 20(S)-OHC 相互作用的区域图，以提供此类新型受体-配体相互作用的生化肖像（目标 1）。使用通过定量质谱分析和基于点击化学的成像分析，我们将询问 Hedgehog 信号传导是否可以改变 20(S)-OHC 的细胞水平或分布（目标 2）。最后，我们将受氧甾醇支架启发，开发并表征新型 Hh 通路抑制剂（目标 3）。我们预计该项目的成功完成将产生三个主要成果：(1) 回答 Smo 在细胞中如何调节的问题，这可能是 Hh 途径中最古老的谜团，(2) 对 Smo 如何在细胞中调节的生化理解氧甾醇参与并调节7次传递信号受体，以及(3)开发可用于剖析任何其他氧甾醇调节的细胞过程的综合工具包。为了实现这些目标，我们招募了一支在细胞生物学、蛋白质生物化学、合成化学和质谱方面具有互补专业知识的研究人员团队。