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

项目摘要/摘要 翻译作为一种新的重要的调节层出现在从基因到 基因产物。这种调节在一定程度上是通过特殊核糖体的活动来实现的，核糖体的活性在它们的 作为核糖体蛋白的补充，具有选择性地翻译特定的mRNAs亚群的能力。 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产量的能力，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 组织或在生物层面上。我的初步研究表明，翻译调控可能是 在促进组织再生方面尤为重要的是紫杉醇--一种高度再生的物种 火蜥蜴。使用Axolotl肢体截肢作为模型，我证明了与哺乳动物不同，严重损伤在 这一物种触发了快速的翻译反应，选择性地以核糖体蛋白的翻译为目标。在……里面 这项建议我研究了调节蛋白质合成在紫杉醇和小鼠中的作用；我建立了 在Axolotl肢体再生过程中的特殊核糖体，我将研究范围扩大到 小鼠软骨细胞与一种新的人软骨器官系统，以明确特化软骨的作用 核糖体与脊椎动物组织的发育、再生和修复。这项工作的长期目标是 利用体外和体内模型，包括微质量小鼠软骨细胞培养和人软骨 有机化合物，小鼠的遗传建模和Axolotls的再生建模，加上最先进的质量 光谱学和核糖体/多聚体图谱方法确定核糖体介导的翻译的作用 在组织发育和疾病中的调节以及寻找具有治疗潜力的新基因和因子 组织再生，特别强调软骨健康。本申请中提出的研究将 在斯坦福大学高度协作的环境中进行，并由 多学科咨询委员会，专长于翻译控制、糖生物学、紫杉醇再生和 干细胞分化。在完成K99阶段后，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者。

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.