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

项目总结/摘要 翻译已经成为一个新的和重要的调节层，在从基因到基因的旅程的最后一步。 基因产物这种调节部分是通过特殊核糖体的活性来实现的，这些核糖体在其功能上各不相同。 核糖体蛋白的互补，并有能力选择性地翻译特定的mRNA子集。 虽然这种机制可能为细胞提供了无与伦比的快速调节蛋白质输出的能力，但其程度和影响仍然是未知的。 核糖体介导的翻译调控尚未在终末分化细胞中得到解决， 组织或在生物体水平上。我的初步研究表明，翻译调控可能是 在促进美西螈的组织再生方面特别重要--美西螈是一种高度再生的 火蜥蜴使用蝾螈肢体截肢作为一个模型，我证明，与哺乳动物不同， 该物种触发选择性靶向核糖体蛋白翻译的快速翻译反应。在 这个建议我检查的作用，调节蛋白质合成的蝾螈和小鼠;我建立的作用， 在蝾螈肢体再生的过程中，专门的核糖体，我扩大了这项研究的范围， 小鼠软骨细胞和一种新的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软骨 类器官，小鼠遗传建模和蝾螈再生建模，加上最先进的质量 光谱和核糖体/多聚核糖体分析方法来确定核糖体介导的翻译的作用 调节组织发育和疾病，并确定新的基因和因子与治疗潜力， 组织再生，特别强调软骨健康。本申请中提出的研究将 在斯坦福大学的高度协作环境中进行，并得到 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K99阶段完成后，候选人的目标是继续这项工作， 在学术研究环境中的独立调查员。

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.