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: 9805445
• 负责人: Olena Zhulyn
• 金额: $ 11.19万
• 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 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9805445
• 关键词: Acute; Address; Advisory Committees; Affect; Ambystoma; Area; Cartilage; Cell Differentiation process; Cell Survival; Cells; Chondrocytes; Code; Complement; Complex; Coupled; Development; Digit structure; Disease; Embryonic Development; Ensure; FRAP1 gene; Gatekeeping; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Models; Genetic Transcription; Glycobiology; Goals; Health; Heterogeneity; Homeostasis; Human; Immunoprecipitation; In Vitro; Injury; Investigation; Label; Limb structure; Mammals; Mass Spectrum Analysis; Measurement; Mediating; Messenger RNA; Methods; Modeling; Molecular; Mus; Natural regeneration; Organoids; Output; Pathway interactions; Phase; Play; Post-Translational Protein Processing; Process; Production; Protein Biosynthesis; Proteins; Proteome; Regenerative Medicine; Regulation; Research; Research Personnel; Ribosomal Proteins; Ribosomes; Role; Salamander; Signal Pathway; Signal Transduction; Site; Spinal Cord; Stress; System; Testing; Therapeutic; Therapeutic Intervention; Time; Tissue-Specific Gene Expression; Tissues; Trans-Activators; Transcript; Translating; Translational Regulation; Translations; Universities; Up-Regulation; Variant; Vertebrates; Work; Wound Healing; articular cartilage; base; cell type; collaborative environment; gene product; in vivo; in vivo Model; knock-down; limb amputation; limb regeneration; mTOR Signaling Pathway; multidisciplinary; novel; polysome profiling; regenerative; repaired; response; small molecule inhibitor; stem cell differentiation; tissue degeneration; tissue regeneration; tissue repair; transcriptome sequencing; 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.

项目概要/摘要 在从基因到基因的旅程的最后一步，翻译已经成为一个新的、重要的监管层。 这种调节部分是通过特化核糖体的活性来实现的，这些核糖体的活性各不相同。 核糖体蛋白的补充，并具有选择性翻译特定 mRNA 子集的能力。 虽然这种机制可能为细胞提供无与伦比的快速调整蛋白质输出的能力，但其程度和影响 核糖体介导的翻译调控在终末分化细胞中尚未得到解决 我的初步研究表明，翻译调控可能与组织水平有关。 对于促进蝾螈（一种高度再生的物种）的组织再生特别重要 我以蝾螈截肢作为模型，证明与哺乳动物不同的是，蝾螈的严重损伤 该物种触发快速翻译反应，选择性地靶向核糖体蛋白的翻译。 我在这项提案中研究了蝾螈和小鼠中受调节的蛋白质合成的作用； 蝾螈肢体再生过程中的特殊核糖体，我将这项研究的范围扩展到 鼠软骨细胞和新型人类软骨类器官系统，以确定专门的作用 核糖体在脊椎动物组织的发育、再生和修复中的作用 这项工作的长期目标是。 利用体外和体内模型，包括微团鼠软骨细胞培养物和人类软骨 类器官、小鼠遗传模型和蝾螈再生模型，加上最先进的质量 光谱测定法和核糖体/多核糖体分析方法来定义核糖体介导的翻译作用 组织发育和疾病的调节，并识别具有治疗潜力的新基因和因子 本申请中提出的研究将促进组织再生，特别是软骨健康。 在斯坦福大学高度协作的环境中进行，并得到了 多学科咨询委员会在翻译控制、糖生物学、蝾螈再生和 干细胞分化完成后，候选人的目标是继续这项工作。 学术研究环境中的独立调查员。