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.

本提案旨在建立一种实验方法，以检测在模型生物中引发肢体再生的因素，