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的进步使我们更接近能够产生具有生理功能的组织（包括软骨、骨骼、肌肉和肌腱）的目标，但肌肉骨骼系统的功能要求这些组织与一个组织进行组织和整合。另一个与神经系统的成功运作有关。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，并且未来可能会继续加速，但在更多灾难性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。这一关键领域进展甚少的一个主要原因是缺乏可处理的动物模型来解决这个问题。虽然斑马鱼可以再生它们的鳍并且非常适合分子和遗传分析，但鳍能够再生的部分（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、蝾螈及其近亲。然而，由于缺乏实验工具，两栖动物肢体再生的研究仍然相对停滞。该提案利用蝾螈（一种非常适合在实验室环境中使用的蝾螈）开发方法和转基因动物，从而在现代基因水平上解决肢体再生问题。这将包括创建一个用于组织特异性操纵基因活性的系统（这样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（这样肢体的发育可以正常发生，并且基因激活或敲除） -down只能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明确的基因操作之外，还将建立利用病毒载体高效快速改变基因表达的方法。建立这样的遗传系统将为未来在高等生物体中再生综合肌肉骨骼系统的工作带来变革。 公共健康相关性：该项目的目标是开发一个遗传系统，用于研究诸如肢体中的综合肌肉骨骼系统的再生。蝾螈的转基因方法将用于开发一种通过组织特异性和时间控制来激活和敲低基因功能的方法。