Signal Transduction by Oxysterols

氧甾醇的信号转导

• 批准号: 8694262
• 负责人: DOUGLAS F COVEY
• 金额: $ 41.8万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 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/8694262
• 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; 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; tool

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）：氧固醇是一类来源于胆固醇的内源性细胞脂质，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。我们在合成有机化学家和细胞生物学家之间建立了合作，以了解氧固醇如何激活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，一种特定的氧固醇，20（S）-OHC，是一种变构激活剂的7-通过跨膜蛋白Smoothened（Smo），人类癌蛋白和关键药物靶点在肿瘤学。这一发现显着扩大了氧固醇作为信号分子的调节范围，证明它们有能力作为人类辅助蛋白和信号受体的直接激动剂。基于前期工作，我们假设内源性20（S）-OHC在Hh信号转导中作为第二信使发挥作用。使用诱变，光亲和标记和质谱的组合，我们将映射与20（S）-OHC相互作用的Smo区域，以提供这类新型受体-配体相互作用的生化特征（目的1）。使用 通过定量质谱和基于点击化学的成像测定，我们将询问Hedgehog信号传导是否可以改变20（S）-OHC（Aim 2）的细胞水平或分布。最后我们将 开发和表征受氧固醇支架启发的新型Hh途径抑制剂（目标3）。我们预计，成功完成这一项目将产生三大成果：（1）Smo在细胞中如何调节的问题的答案，也许是Hh通路中存在时间最长的谜团，（2）对氧固醇如何参与和调节7-pass信号受体的生物化学理解，以及（3）开发一个综合工具包，该工具包可以用于剖析任何其他氧固醇调节的细胞过程。为了实现这些目标，我们招募了一组在细胞生物学，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。