Temporal roles of Sonic and Indian Hedgehog in tunnel integration following anterior cruciate ligament reconstruction

Sonic 和 Indian Hedgehog 在前十字韧带重建后隧道整合中的时间作用

• 批准号: 10684062
• 负责人: Timur Kamalitdinov
• 金额: $ 4.77万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10684062
• 关键词: Ablation; Adult; Anterior Cruciate Ligament; Area; Biological Assay; Biological Models; Biomechanics; Bone Marrow; Bypass; Cell Proliferation; Cells; Cicatrix; Clinical; Clinical Treatment; Collagen Fiber; Data; Development; Differentiated Gene; Embryo; Erinaceidae; Fellowship; Fibrocartilages; Growth; Growth and Development function; Healthcare; Histologic; Infiltration; Injections; Injury; Knowledge; Lasers; Ligaments; Ligand Binding; Ligands; Mechanics; Mentors; Microfluidics; Modeling; Monitor; Mus; Musculoskeletal; Normal tissue morphology; Orthopedics; Outcome; Pathway interactions; Pennsylvania; Phase; Physiologic calcification; Play; Positioning Attribute; Procedures; Process; Productivity; Proliferating; Proteoglycan; Qualifying; Quality of life; Research; Role; Rotator Cuff; SHH gene; Scientist; Series; Signal Pathway; Signal Transduction; Site; Specific qualifier value; Stains; Stromal Cells; Tamoxifen; Tendon Injuries; Tendon structure; Testing; Therapeutic; Tissues; Training; Transducers; Transgenic Mice; Transgenic Organisms; United States; Universities; Work; aging population; anterior cruciate ligament reconstruction; bone; bone repair; career; experience; healing; improved; innovation; insight; laser capture microdissection; ligament injury; loss of function; mineralization; mouse model; musculoskeletal injury; new therapeutic target; novel; novel strategies; novel therapeutics; postnatal; progenitor; reconstruction; repair model; repaired; smoothened signaling pathway; spatiotemporal; stem cell proliferation; stem cells; tendon graft

Project Summary This F31 fellowship application provides a training plan with a collaborative team in the McKay Orthopaedic Research Labs at the University of Pennsylvania and an innovative research plan to prepare Mr. Timur Kamalitdinov to be an independent scientist in the musculoskeletal field. Approximately 30% of adults in the United States suffer from a musculoskeletal condition, with almost half of those conditions being tendon and ligament injuries. These injuries often require repair (e.g., rotator cuff tendons) or reconstruction (e.g., anterior cruciate ligament (ACL)) to re-integrate the tendon midsubstance with the underlying mineralized bone. Recreating the zonal tendon-to-bone insertion site (i.e., enthesis) is critical for restoring normal tissue function. Zonal enthesis formation involves anchoring collagen fibers, synthesizing proteoglycan-rich fibrocartilage, and mineralizing this fibrocartilage. The Hedgehog (Hh) signaling pathway is a potent regulator of zonal tendon-to- bone insertion formation during growth and development as it promotes the formation of the unmineralized and mineralized fibrocartilage zones. Unfortunately, studying this pathway in traditional tendon-to-bone repair has been challenging because these repair models do not sufficiently anchor collagen fibers to bone, much less produce zones of fibrocartilage. Conversely, ligament reconstructions where a tendon graft is passed through bone tunnels can produce zonal attachments in the tunnels. Therefore, we utilize innovative transgenic murine ACL reconstruction models to elucidate the mechanisms of zonal tendon-to-bone attachment formation in order to develop novel therapies to improve traditional tendon repair outcomes. Our preliminary data demonstrate that Hh signaling promotes zonal tendon-to-bone integration following ACL reconstruction. The objective of this proposal is to define the spatiotemporal roles of Sonic hedgehog (Shh) and Indian hedgehog (Ihh) in promoting adult tendon-to-bone repair. Our hypothesis is that the Hh pathway plays a biphasic role during zonal tendon-to- bone attachment formation following ACL reconstruction, with Shh acting in the early stages of repair to promote expansion of the progenitor pool and Ihh facilitating fibrocartilage differentiation and maturation. We will use multiplexed mineralized cryohistology and laser capture microdissection with microfluidic qPCR arrays to monitor the zonal attachment formation and novel mechanical testing to assess tunnel integration strength. Targeting specific ligands of the Hh pathway will provide key mechanistic insights into whether certain ligands are more important during specific stages of the tunnel integration process, opening new avenues of research into novel therapeutic targets to improve adult tendon-to-bone repair.

项目摘要 该F31奖学金申请提供了与McKay骨科的协作团队的培训计划 宾夕法尼亚大学的研究实验室和一项创新的研究计划，以准备Timur先生 kamalitdinov是肌肉骨骼领域的独立科学家。大约30％的成年人 美国患有肌肉骨骼状况，其中几乎一半是肌腱和 韧带受伤。这些伤害通常需要修复（例如，肩袖肌腱）或重建（例如， 交叉韧带（ACL））将肌腱中缩质与潜在的矿化骨重新整合。 重新创建区域肌腱到骨插入位点（即源）对于恢复正常组织功能至关重要。 区域埃文的形成涉及锚定胶原纤维，合成富含蛋白聚糖的纤维球纤维和 矿物质矿物质。刺猬（HH）信号通路是区域肌腱的有效调节剂 骨插入在生长和发育过程中的形成，因为它促进了非矿化和 矿化纤维球区。不幸的是，在传统的肌腱到骨修复中研究这种途径 之所以具有挑战性，是因为这些维修模型不足以将胶原蛋白纤维固定在骨头上，更不用 产生纤维球卫生区域。相反，通过肌腱移植的韧带重建 骨隧道可以在隧道中产生纬向附件。因此，我们利用创新的转基因鼠 ACL重建模型以阐明纬向肌腱对骨附着的机理按顺序形成 开发新的疗法以改善传统的肌腱修复结果。我们的初步数据表明 HH信号传导在ACL重建后促进了区域肌腱到骨的整合。这个目的 提案是定义Sonic刺猬（SHH）和印度刺猬（IHH）的时空角色 成人肌腱对骨修复。我们的假设是，HH途径在区域肌腱之间起双相作用 ACL重建后的骨附着形成，SHH在修复的早期阶段起作用以促进 祖细胞池的扩展和IHH的促进纤维球药物分化和成熟。我们将使用 用微流体QPCR阵列进行多重矿化的冷冻组织学和激光捕获显微解剖以监测 区域附着形成和新型的机械测试，以评估隧道整合强度。定位 HH途径的特定配体将为某些配体是否更多的配体提供关键的机械见解 在隧道整合过程的特定阶段中很重要，开辟了新的研究途径 改善成年肌腱对骨修复的治疗靶标。