Mechanisms underlying tendon regeneration and attachment site pattern restoration

肌腱再生和附着部位模式恢复的机制

• 批准号: 10061555
• 负责人: JENNA L GALLOWAY
• 金额: $ 35.85万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10061555
• 关键词: Ablation; Adult; Affect; Biological Assay; Biology; Biomechanics; Cartilage; Cell Proliferation; Cell-Matrix Junction; Cells; Chemicals; Collagen; Congenital Abnormality; Congenital Disorders; Connective Tissue; Contracture; Defect; Deformity; Development; Disease; Elements; Embryo; Etiology; Event; Extracellular Matrix; Gene Expression; Genetic; Genetic Models; Goals; Growth; Human; Image; Individual; Injury; Knowledge; Ligaments; Location; Mammals; Mediating; Mesoderm; Methods; Modeling; Molecular; Morphogenesis; Morphology; Movement; Muscle; Musculoskeletal; Musculoskeletal System; Natural regeneration; Neural Crest; Pain; Pathway interactions; Pattern; Population; Positioning Attribute; Process; Property; Quality of life; Regenerative Medicine; Regenerative response; Regulation; Role; Signal Pathway; Signal Transduction; Site; Source; System; Tendon Injuries; Tendon structure; Testing; Tissues; Transgenic Organisms; Vertebrates; Visualization; Work; Zebrafish; age related; aging population; bone; cell behavior; cell regeneration; chemical genetics; constriction; foot; healing; histological stains; insight; joint mobilization; malformation; migration; musculoskeletal injury; novel; progenitor; recruit; regeneration potential; regenerative; regenerative biology; restoration; scleraxis; skeletal; small molecule; tendon development; tool; transcription factor

Galloway, Jenna L Project Summary Tendons make essential connections between the forming musculoskeletal tissues, enabling coordinated movement. Disruption to the development and patterning of the musculoskeletal system can result in skeletal malformations or contractures, a congenital abnormality that results in constriction in the movement of joints. In adults, musculoskeletal injuries are common among active individuals and the aging population. Tendon injuries, in particular, are complicated by a slow and limited healing, which can pose significant mobility, pain, and quality of life issues. Comprehensive knowledge of the molecular pathways that guide tendon development and regeneration would have broad impact in our understanding of the etiology of congenital defects as well as in regenerative medicine approaches to tendon injuries. This proposal aims to use the zebrafish to understand the mechanisms underlying tendon cell regeneration and the re-establishment of attachment site pattern. Our previous studies have shown that zebrafish and mammalian tendons are similar in gene expression, developmental regulation, and ultrastructural properties, making them an excellent genetic system for studying tendon biology. We also find zebrafish have robust abilities to regenerate their tendon tissue unlike adult mammals. Building from this novel work, we propose to use a genetic cell ablation model to dissect the cellular and molecular mechanisms underlying tendon regeneration and the restoration of the attachment pattern. We will identify the source of the newly regenerating tendon cells using cell proliferation assays, genetic lineage tracing, and live imaging. Examination of BMP Responsive Element transgenic zebrafish and functional analysis indicate BMP signaling in the regeneration of specific attachment sites. In addition, a high-throughput chemical screen identified compounds with tendon promoting activities and whose targets may intersect the BMP pathway. Using chemical and genetic functional assays, we will dissect the role of BMP signaling in tendon regeneration and the re-establishment of a specific attachment site. We will also test if the chemicals and the pathways they target can augment the regenerative response through their potential intersection with the BMP pathway. Our proposal combines novel tools with live imaging and functional studies and together, this will provide unprecedented visualization of tendon cell behaviors during regeneration. We believe these studies will add crucial insight into tendon development and regeneration, which could impact our understanding of congenital disorders and regenerative medicine approaches to tendon injuries.

Galloway，Jenna L 肌腱在形成的肌肉骨骼组织之间建立重要的联系， 使运动协调一致。破坏人类的发展和模式 肌肉骨骼系统可能导致骨骼畸形或挛缩，先天性 导致关节活动受限的不正常现象。在成人中，肌肉骨骼 受伤在活跃的个人和老龄化人口中是常见的。肌腱损伤， 特别是，由于缓慢和有限的愈合而变得复杂，这可能造成显著的移动性，疼痛， 和生活质量问题。全面了解引导肌腱的分子通路 发育和再生将对我们理解的病因学产生广泛的影响， 先天性缺陷以及再生医学中肌腱损伤的方法。这 这项提案的目的是利用斑马鱼来了解肌腱细胞的机制， 再生和附着位点模式的重建。我们以往的研究 显示斑马鱼和哺乳动物的肌腱在基因表达、发育 调节和超微结构特性，使其成为研究的优良遗传系统 肌腱生物学我们还发现斑马鱼有强大的能力再生肌腱组织 不像成年哺乳动物。在这项新工作的基础上，我们建议使用遗传细胞消融 模型来解剖肌腱再生的细胞和分子机制， 恢复依恋模式。我们将确定新再生肌腱的来源 使用细胞增殖测定、遗传谱系追踪和活体成像来检测细胞。BMP检查 响应元件转基因斑马鱼和功能分析表明BMP信号在 特定附着位点的再生。此外，高通量化学筛选确定了 具有腱促进活性的化合物，其靶点可能与BMP途径交叉。 利用化学和遗传功能分析，我们将剖析BMP信号在肌腱中的作用， 再生和重新建立特定的附着位点。我们还将测试 化学物质及其靶向途径可以通过它们的作用增强再生反应， 与BMP通路的潜在交叉点。我们的建议将新颖的工具与实时成像相结合 和功能研究，这将为肌腱细胞提供前所未有的可视化 再生过程中的行为。我们相信这些研究将增加对肌腱的重要了解 发育和再生，这可能会影响我们对先天性疾病的理解， 肌腱损伤的再生医学方法。