Ribosomes and Regeneration: Defining the Role of Protein Synthesis in Tissue Development, Homeostasis and Repair.

核糖体和再生：定义蛋白质合成在组织发育、稳态和修复中的作用。

• 批准号: 10407363
• 负责人: Olena Zhulyn
• 金额: $ 4.75万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10407363
• 关键词: Address; Advisory Committees; Ambystoma; Area; Biological; Cartilage; Cell Differentiation process; Cells; Chondrocytes; Code; Complement; Coupled; Development; Disease; Embryo; Enzymes; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Gatekeeping; Gene Expression Regulation; Gene Proteins; Genes; Genetic Models; Glycobiology; Glycoproteins; Goals; Grant; Guided Tissue Regeneration; Health; Homeostasis; Human; Immunoprecipitation; In Vitro; Label; Limb structure; Mammalian Cell; Mammals; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Methods; Modeling; Modification; Molecular; Morphology; Mus; Natural regeneration; Organoids; Output; Pathway interactions; Phase; Play; Polyribosomes; Post-Translational Protein Processing; Process; Production; Protein Biosynthesis; Proteins; Puromycin; Regulation; Research; Research Personnel; Ribosomal Proteins; Ribosomes; Role; Salamander; Sedimentation process; Source; Spinal Cord; Sucrose; System; Testing; Therapeutic; Therapeutic Intervention; Time; Tissue-Specific Gene Expression; Tissues; Trans-Activators; Transcript; Translating; Translational Regulation; Translations; Universities; Up-Regulation; Work; articular cartilage; cartilage cell; cell type; collaborative environment; cost; efficacy validation; embryonic stem cell; gene product; glycosyltransferase; in vivo Model; inhibitor/antagonist; interest; limb amputation; limb regeneration; mouse model; multidisciplinary; novel; polysome profiling; regeneration model; regenerative; repaired; response; ribosome profiling; severe injury; stem cell differentiation; stressor; tissue degeneration; tissue regeneration; wound closure

PROJECT SUMMARY/ABSTRACT Translation has emerged as a new and important layer of regulation at the last step in the journey from gene to gene product. This regulation is conferred in part through the activity of specialized ribosomes which vary in their complement of ribosomal proteins and have the capacity to selectively translate specific subsets of mRNAs. While this mechanism may offer cells an unparalleled ability to rapidly adjust protein output, the extent and impact of ribosome-mediated translational regulation has not yet been addressed in terminally differentiated cells and tissues or at the organismal level. My preliminary studies demonstrate that translational regulation may be of particular importance in promoting tissue regeneration in the axolotl - a species of highly regenerative salamander. Using axolotl limb amputation as a model, I demonstrate that unlike in mammals, severe injury in this species triggers a rapid translational response which selectively targets translation of ribosomal proteins. In this proposal I examine the role of regulated protein synthesis in the axolotl and mouse; I establish the role of specialized ribosomes in the process of axolotl limb regeneration and I extend the scope of this research to murine chondrocytes and a novel human cartilage organoid system, in order to define the role of specialized ribosomes in the development, regeneration and repair of vertebrate tissues. The long-term goal of this work is to utilize in vitro and in vivo models, including micromass murine chondrocyte culture and human cartilage organoids, genetic modeling in mice and regeneration modeling in axolotls, coupled with state-of-the-art mass spectrometry and ribosome/polysome profiling methods to define the role of ribosome-mediated translational regulation in tissue development and disease and to identify new genes and factors with therapeutic potential in tissue regeneration, with a particular emphasis on cartilage health. The research proposed in this application will be carried out within the highly collaborative environment of Stanford University and supported by a multidisciplinary advisory committee with expertise in translation control, glycobiology, axolotl regeneration and stem cell differentiation. Upon completion of the K99 phase, the candidate’s goals are to continue this work as an independent investigator in an academic research setting.

项目摘要/摘要 在从基因到基因的旅程的最后一步中，翻译已成为一个新的重要监管层 基因产物。该调节是通过专门的核糖体的活动来赋予的 核糖体蛋白质的补充，并具有选择性地翻译MRNA的特定子集的能力。 尽管该机制可能为细胞提供无与伦比的能力，以快速调节蛋白质输出，但程度和影响 核糖体介导的翻译调节尚未在终末分化的细胞和 组织或有机水平。我的初步研究表明，翻译调节可能是 在促进Axolotl的组织再生方面特别重要 - 高度再生的一种 使用Axolotl肢体截肢为模型，我证明了与哺乳动物不同，在 该物种触发了快速的翻译反应，该反应有选择地靶向核糖体蛋白的翻译。在 该建议我研究了调节蛋白质合成在Axolotl和小鼠中的作用。我确定了 在Axolotl肢体再生过程中的专门核糖体，我将这项研究的范围扩展到 鼠软骨细胞和一种新型的人体软骨器官系统，以定义专业的作用 核糖体的发育，再生和修复脊椎动物组织。这项工作的长期目标是 利用体外和体内模型，包括微瘤鼠软骨细胞培养和人体软骨 类器官，小鼠中的遗传建模和轴突的再生建模，再加上最先进的质量 光谱法和核糖体/多层分析方法来定义核糖体介导的翻译的作用 组织发育和疾病的调节，并确定具有治疗潜力的新基因和因素 组织再生，特别强调软骨健康。本应用程序中提出的研究将 在斯坦福大学高度协作环境中进行，并在 多学科咨询委员会具有翻译控制，糖生物学，Axolotl再生和 干细胞分化。 K99阶段完成后，候选人的目标是继续这项工作 学术研究环境中的独立研究者。