Genetic Modification to Harness the Regenerative Power of the African Spiny Mouse

利用非洲刺鼠再生能力的基因改造

• 批准号: 10015365
• 负责人: Malcolm Maden
• 金额: $ 22.88万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10015365
• 关键词: Acomys; Adipose tissue; Adult; Affect; African; Animals; Biological Assay; Biological Models; Breeding; Case Study; Cells; Cicatrix; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen Type I; Collagen Type III; Communities; Complementary DNA; Complex; Cues; Cytoskeletal Proteins; Dermis; Disease; Ear; Embryo; Epidermis; Excision; Exhibits; Failure; Fibroblasts; Fibrosis; Gene Targeting; Gene-Modified; Generations; Genes; Genetic; Genome; Genomics; Goals; Growth Factor; Guide RNA; Hair; Hair follicle structure; Histology; In Vitro; Knock-out; Mammals; Mechanics; Mediating; Methods; Modeling; Modification; Molecular; Mus; Muscle; Myocardial Infarction; Myofibroblast; Natural regeneration; Normal tissue morphology; Operative Surgical Procedures; Pilum; Plasmids; Play; Procedures; Property; Proteins; Protocols documentation; Recombinants; Regenerative Medicine; Reproductive Biology; Reproductive system; Research; Ribonucleoproteins; Role; Sebaceous Glands; Skeletal Muscle; Skeletal muscle injury; Skin; Smooth Muscle; Superovulation; Surrogate Mothers; System; Technical Expertise; Testing; Thick; Time; Tissues; Traction; Transfection; Transgenic Organisms; United States National Institutes of Health; base; burn wound; cell growth; egg; experience; experimental study; gene transplantation for gene therapy; in vivo; interest; knockout gene; new therapeutic target; pup; regenerative; response; scaffold; skin regeneration; skin wound; stem cells; success; tissue regeneration; tissue repair; transcriptome; wound; wound healing; wound response; zygote

PROJECT ABSTRACT A fundamental goal of regenerative medicine is to replace fibrosis and scarring in damaged mammalian tissues with functional tissue. Thus far there has been very little progress towards this goal because of a lack of an adult mammalian model system that demonstrates functional regeneration, and instead the typical response of damaged mammalian skin is to form a hairless, collagenous scar. However, recent studies revealed that African spiny mouse (Acomys), following full thickness skin removal, can regenerate all the tissues removed: the epidermis, hair, sebaceous glands, erector pili muscles, dermis and skeletal muscle of the panniculus carnosus, all without scarring. In our experiments to date characterizing the differences between regenerating Acomys skin wounds and scarring Mus skin wounds, we have identified many molecular differences but we have no real handle on what exactly are responsible for regeneration vs scarring. To reveal molecular mechanisms underlying this remarkable capacity of Acomys to regenerate normal tissue, the scientific community needs a way to modify genes in the animals. For instance, we previously identified that collagen type III is particularly abundant in Acomys skin regeneration, whereas Mus exhibits a much higher ratio of collagen type I to type III during wound scarring1. However, we do not know that how this abundant collagen type III plays a role in a higher regenerative capacity of Acomys. In the present proposal, we will knockout the Col3a1 gene within Acomys fibroblasts as well as the whole animals as an exemplar with which to study the role of specific genes in higher wound repair capacity of the animal. The establishment of an efficient gene editing method with Acomys will open a revolutionary new avenue for mammalian regeneration research.

项目摘要 再生医学的一个基本目标是取代受损组织中的纤维化和瘢痕形成。 哺乳动物组织与功能组织。到目前为止，在这方面进展甚微。 这一目标是因为缺乏成年哺乳动物模型系统， 再生，相反，受损哺乳动物皮肤的典型反应是形成无毛， 胶原性疤痕。然而，最近的研究表明，非洲刺鼠（Acomys）， 全层皮肤切除，可以再生所有被切除的组织：表皮，毛发， 皮脂腺、竖皮利肌、真皮和肉脂膜的骨骼肌， 都没有疤痕在我们迄今为止的实验中， 再生Acomys皮肤伤口和瘢痕小鼠皮肤伤口，我们已经确定了许多 但是我们没有真实的方法来确定到底是什么导致了再生 vs疤痕。 为了揭示Acomys这种非凡的再生能力的分子机制， 科学界需要一种方法来修改动物的基因。比如说， 我们先前发现III型胶原蛋白在Acomys皮肤再生中特别丰富， 而小鼠在伤口结疤期间表现出高得多的I型胶原与III型胶原的比率1。 然而，我们不知道这种丰富的III型胶原蛋白是如何在更高的 Acomys的再生能力在本发明的提议中，我们将敲除Col3a1基因， 以Acomys成纤维细胞以及整个动物为例，研究 特定基因在动物的伤口修复能力较高。建立高效 Acomys的基因编辑方法将为哺乳动物基因编辑开辟革命性的新途径 再生研究