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，肌肉骨骼再生带来了特定的挑战。尽管干细胞生物学和组织工程的进步使我们更接近能够生产生理能力的组织，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的功能彼此组织和集成，并与神经系统相互融合。因此，尽管增强局部肌肉或骨骼损害的维修能力一直在不断提高，并且将来可能会以加速的速度继续这样做，但是在灾难性损失更多后，再生功能性肌肉骨骼系统的能力仍然是一个遥远的梦想。在这个关键领域的进展很少的主要原因是缺乏解决该动物的动物模型。尽管斑马鱼可以再生其鳍片，并且非常适合分子和遗传分析，但鳍的一部分能够再生（鳍片）既不包含耳软骨骨，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Urodele两栖动物，Salamanders及其亲戚。但是，由于缺乏实验工具，对两栖动物肢体再生的研究仍然是相对的回水。该建议利用Axolotl是一种非常适合在实验室环境中使用的sal，是为了开发方法和转基因动物，这将使肢体再生问题能够在现代的遗传水平上解决。这将包括创建一个用于组织基因活性的组织特异性操纵系统（可以评估基因在特定再生组织中的作用），以调节基因表达（这样的肢体的发展，并且只能在再生过程的特定阶段启动肢体激活或敲门，并且可以启动肢体的发展）。此外，除了转基因动物的确定基因操纵之外，还将建立用于高效率上高效率下基因表达的方法。建立这种遗传体系将是对未来在较高生物体中再生综合肌肉骨骼系统的转变。 公共卫生相关性：该项目的目的是开发一种遗传系统，用于研究肢体中发现的综合肌肉骨骼系统的再生。 Axolotls中的转基因方法将用于开发一种激活和击倒组织特异性和时间对照的基因功能的方法。