A genetic system for the study of vertebrate limb regeneration

用于研究脊椎动物肢体再生的遗传系统

• 批准号: 8121590
• 负责人: CLIFFORD J. TABIN
• 金额: $ 18.22万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-05 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8121590
• 关键词: Address; Ambystoma; Amphibia; Animal Model; Area; Birds; Cartilage; Cell Lineage; Cells; Dissection; Distant; Dreams; Engineering; Future; Gene Activation; Gene Expression; Genes; Genetic; Goals; Laboratories; Life Cycle Stages; Limb Development; Limb structure; Mediating; Methods; Molecular Genetics; Muscle; Musculoskeletal; Musculoskeletal System; Natural regeneration; Nervous system structure; Organism; Pathway interactions; Pattern; Process; Regenerative Medicine; Relative (related person); Research; Role; Salamander; Signal Transduction; Specificity; Staging; Structure; System; Tendon structure; Testing; Time; Tissue Engineering; Tissue-Specific Gene Expression; Tissues; Transgenic Animals; Transgenic Organisms; Vertebrates; Viral; Viral Vector; Work; Zebrafish; blastema; bone; cell type; gene function; genetic analysis; improved; knock-down; limb regeneration; public health relevance; receptor; recombinase; regenerative; repaired; skeletal; stem cell biology; tool; transgene expression

DESCRIPTION (provided by applicant): Within the broad framework of regenerative medicine, musculoskeletal regeneration poses particular challenges. While advances in stem cell biology and tissue engineering have brought us closer to the goal of being able to produce physiologically competent tissues, including cartilage, bone, muscle and tendons, the function of the musculoskeletal system requires that these tissues be organized and integrated with one another and with the nervous system to function successfully. Thus, while the ability to enhance repair of local muscle or skeletal damage is improving all the time and will likely continue to do so at an accelerating rate in the future, the capacity to regenerate a functional musculoskeletal system after more catastrophic loss remains a distant dream. A major reason that there has been so little progress in this critical area has been the lack of a tractable animal model for addressing it. While zebrafish can regenerate their fins and are well suited to molecular and genetic analyses, the portion of the fin capable of regenerating (the fin rays) contains neither endochondral bone nor muscle and tendons. Hence, the only class of vertebrates capable of regenerating entire musculoskeletal structures is the urodele amphibians, the salamanders and their relatives. However, research on amphibian limb regeneration has remained a relative backwater due to the lack of experimental tools. Utilizing the axolotl, a salamander that is highly suited for use in a laboratory setting, this proposal is to develop methods and transgenic animals that will allow the problem of limb regeneration to be addressed on a modern genetic level. This will include creating a system for tissue-specific manipulation of gene activity (such that the roles of genes in specific regenerating tissues can be assessed), for conditionally regulating gene expression (such that development of the limb can occur normally and gene activation or knock-down can be initiated only during specific stages of the regenerative process). Moreover, methods will be established for rapidly altering gene expression at a high efficiency with viral vectors in addition to definitive gene manipulation in transgenic animals. Establishing such a genetic system will be transformative for future work towards regenerating an integrative musculoskeletal system in higher organisms. PUBLIC HEALTH RELEVANCE: The goal of this project is to develop a genetic system for studying the regeneration of integrated musculoskeletal systems such as is found in the limb. Transgenic approaches in axolotls will be utilized to develop a means of activating and knocking down gene function with tissue-specificity and temporal control.

描述（由申请人提供）：在再生医学的广泛框架内，肌肉骨骼再生提出了特殊的挑战。虽然干细胞生物学和组织工程学的进步使我们更接近能够产生生理活性组织的目标，包括软骨，骨骼，肌肉和肌腱，但肌肉骨骼系统的功能要求这些组织相互组织和整合，并与神经系统成功地发挥作用。因此，虽然增强局部肌肉或骨骼损伤修复的能力一直在提高，并且将来可能会继续以加速的速度这样做，但在更灾难性的损失后再生功能性肌肉骨骼系统的能力仍然是一个遥远的梦想。在这个关键领域进展如此之少的一个主要原因是缺乏一种易于处理的动物模型来解决这个问题。虽然斑马鱼可以再生鳍，并且非常适合分子和遗传分析，但鳍的能够再生的部分（鳍条）既不包含软骨内骨，也不包含肌肉和肌腱。因此，唯一能够再生整个肌肉骨骼结构的脊椎动物是有尾目两栖动物，蝾螈及其亲属。然而，由于缺乏实验工具，两栖动物肢体再生的研究一直处于相对的停滞状态。利用蝾螈，一种非常适合在实验室环境中使用的蝾螈，这项建议是开发方法和转基因动物，使肢体再生的问题能够在现代遗传水平上得到解决。这将包括创建一个系统，用于组织特异性操纵基因活性（以便可以评估基因在特定再生组织中的作用），用于有条件地调节基因表达（以便肢体的发育可以正常发生，并且基因激活或敲低可以仅在再生过程的特定阶段启动）。此外，除了在转基因动物中进行确定性基因操作之外，还将建立用病毒载体以高效率快速改变基因表达的方法。建立这样一个遗传系统将是未来工作的变革，在高等生物体中再生一个综合的肌肉骨骼系统。 公共卫生相关性：该项目的目标是开发一个遗传系统，用于研究肢体等综合肌肉骨骼系统的再生。转基因方法在蝾螈将被用来开发一种手段，激活和敲低基因功能的组织特异性和时间控制。