Use of specific transcription factors to promote limb regeneration capacity

利用特定转录因子促进肢体再生能力

• 批准号: 7685935
• 负责人: JOHN W DAY
• 金额: $ 8.18万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7685935
• 关键词: Amphibia; Amputation; Animal Model; Biological Assay; Body part; Cell Differentiation process; Cells; Chromatin; Development; Developmental Biology; Disease; Fibroblasts; Flow Cytometry; Gene Targeting; Genes; Goals; Healed; Health; Human; Knowledge; Laboratories; Limb Bud; Limb structure; Methods; Muscle; Muscle Development; Muscle Fibers; Muscular Dystrophies; Musculoskeletal; Mutation; Myopathy; Natural regeneration; Performance; Phenotype; Pilot Projects; Process; Rana; Research; Skeletal Muscle; Specific qualifier value; Staging; Stem cells; System; Tadpoles; Technology; To specify; Work; Xenopus; Xenopus laevis; appendage; design; healing; induced pluripotent stem cell; limb regeneration; regenerative; stem cell technology; tissue/cell culture; transcription factor

This proposal seeks to establish an assay for factors provoking limb regeneration in a model organism, which will inform parallel efforts to design methods for enabling muscle, skeletal structures or even whole limbs of human beings to regenerate. Tadpoles of Xenopus laevis can regenerate limbs at early developmental stages but not at later stages. The rationale for the current proposal is whatever regenerative mechanism is lost is probably missing in mammalian limbs and responsible for their inability to regenerate: limb fibroblasts can be re-specified to a limb bud phenotype by specific transcription factors. A selective system will isolate cells that have been appropriately re-specified, and performance of the regenerated muscle fibers, discrete muscles, and entire regenerated appendages will be assessed quantitatively with the help of the Muscle Force Assessment Core. This project uses methods that have seldom, if ever, been used to study muscle development, differentiation and regeneration: iPS technology to re-define cells; tissue culture methods to investigate regeneration; use of flow cytometry to select the desired cells. Specific Aims of the proposal are: Specific Aim 1; To respecify froglet limb-derived fibroblasts to a. limb bud state. The working hypothesis is that this can be achieved by introduction of genes encoding the limb bud state, together with genes to unwrap the chromatin, followed by a selective step for positively responding cells. Specific Aim 2: To rescue regenerative ability in Xenopus froglet limbs by re-introduction of limb bud cells. The working hypothesis is that this can be achieved by injecting limb bud cells subcutaneously, allowing to heal, and amputation through the graft region.

该建议旨在为模型生物体中引起肢体再生的因素建立一个测定法， 这将为设计方法提供肌肉，骨骼结构甚至整个方法的平行努力 人类的肢体要再生。爪蟾laevis的t可​​以在早期再生四肢 发展阶段，但不在以后的阶段。当前建议的理由是任何再生 哺乳动物的四肢可能缺少机制，并且负责其无法再生： 肢体成纤维细胞可以通过特定的转录因子重新指定为肢体芽表型。选择性 系统将隔离已适当重新指定的细胞，并进行重生的性能 肌肉纤维，离散的肌肉和整个再生窝将被定量评估 肌肉力量评估核心的帮助。该项目使用很少使用（如果有的话）的方法 研究肌肉发育，分化和再生：IPS技术以重新定义细胞；组织培养 研究再生的方法；使用流式细胞仪选择所需的细胞。特定目标 提案是： 具体目标1；将Froglet肢体衍生的成纤维细胞重新定位为a。肢体芽状态。工作假设 是可以通过引入编码肢体芽状态的基因以及基因与 解开染色质，然后是一个选择性的步骤，用于积极响应的细胞。 特定目的2：通过重新引入肢体芽中的爪蟾果蝇中的再生能力 细胞。工作假设是可以通过皮下注射肢体芽细胞来实现， 允许通过移植区愈合和截肢。