Mechanisms of Primary Cilia Regulating Tendon Enthesis Development and Regeneration

初级纤毛调节肌腱附着点发育和再生的机制

• 批准号: 10707864
• 负责人: Fei Fang
• 金额: $ 58.44万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707864
• 关键词: Ablation; Acute; Adult; Affect; Age; Architecture; Biochemical; Biological Assay; Biology; Biomechanics; Cartilage; Cell Transplantation; Cell surface; Cells; Chondrocytes; Chondrogenesis; Cicatrix; Cilia; Clinical; Data; Deterioration; Development; Disease; Environment; Epiphysial cartilage; Erinaceidae; Event; Failure; Fibrocartilages; General Population; Genetic Transcription; Hypertrophy; Immunohistochemistry; Impairment; In Situ Hybridization; Injury; Knockout Mice; Knowledge; Label; Length; Maintenance; Mammalian Cell; Mechanical Stimulation; Mechanics; Mediating; Medical; Modeling; Molecular; Mus; Muscle; Natural regeneration; Neonatal; Older Population; Operative Surgical Procedures; Organelles; Outcome; Pain; Pathogenesis; Pathology; Pathway interactions; Patients; Pilot Projects; Population; Process; Protocols documentation; Regenerative capacity; Regulation; Reporter; Reporting; Role; Rotator Cuff; Shoulder; Shoulder Pain; Signal Transduction; Sports; Stimulus; Tendon structure; Testing; Therapeutic; Tissues; Transgenic Mice; United States; Work; biomechanical test; bone; bone fracture repair; cost; disability; experience; healing; improved; in vivo; injured; mechanical force; mechanical load; mechanical properties; mechanical signal; mechanotransduction; mineralization; mouse model; muscle regeneration; novel; novel therapeutic intervention; pharmacologic; postnatal; programs; repaired; response; rotator cuff tear; single molecule; single-cell RNA sequencing; smoothened signaling pathway; soft tissue; stem cell function; stem cell population; stem cells; stemness; therapeutically effective; transcriptomics

PROJECT SUMMARY 40% of the population over the age of 60 experiences a rotator cuff tear. The high failure rates of rotator cuff tear after surgical repair or non-surgical treatment make it a major clinical challenge. The outcome failures accompany the formation of scar tissues at the tendon-to-bone insertion (tendon enthesis) with disorganized architecture and deteriorated function. Development of effective therapeutics has been hampered by limited knowledge of enthesis development biology and mechanobiology and an incomplete understanding of endogenous mechanisms governing enthesis pathogenesis and healing. To bridge this knowledge gap, the current proposal seeks to elucidate how tendon enthesis responds to its mechanical and biochemical environment during development and healing processes. It is known that a combination of mechanical force and distinct pathways, including hedgehog (Hh) signaling, drive enthesis formation, promote remodeling of mature enthesis, and affect enthesis healing. Recently, our studies have indicated that the primary cilium, a solitary antenna protruding from mammalian cell surface, potentially functions as a hub for mechanotransduction and Hh signaling. Building on our previous work, the objective of this proposal is to gain a mechanistic understanding of the role of primary cilia in concentrating and synchronizing mechanical and Hh signals during enthesis development and healing. To achieve this objective, we will determine identities and activities of ciliated enthesis cells during enthesis development and mechanical adaptation (Specific Aim 1) and evaluate the regenerative capacity of ciliated enthesis cells for improving enthesis healing (Specific Aim 2). The approaches we will use include cilia-labeled and cilia-deleted transgenic mouse models, different established loading models, cell transplantation, and transcriptomics analysis, combined with structural, compositional, and biomechanical evaluation assays. At the conclusion of this project, we expect to identify new cilia-regulated mechano- transduction pathways during in vivo enthesis mechanical adaptation and suggest novel mechanisms by which cilia convert mechanical cues to cellular signaling events. The new findings of the role of primary cilia in enthesis healing will guide the development of novel pharmacological and mechanobiology therapeutics for treating rotator cuff tears.

项目摘要 60岁以上的40％的人口经历了肩袖的撕裂。肩袖撕裂的高故障率 手术修复或非手术治疗后，它成为了主要的临床挑战。结果失败 伴随着肌腱到骨插入（肌腱插入）的形成疤痕组织的形成 架构和功能恶化。有限的有效疗法的开发受到了限制 对ENTEMES发展生物学和机械生物学的了解以及对 内源性机制管理发病机理和愈合。为了弥合这个知识差距， 当前的提案旨在阐明肌腱衰减如何对其机械和生化的反应 开发和康复过程中的环境。众所周知，机械力和 不同的途径，包括刺猬（HH）信号传导，驱动灌注形成，促进成熟的重塑 灌感并影响灌注愈合。最近，我们的研究表明，主要的纤毛是孤立的 天线从哺乳动物细胞表面突出，可能充当机械转导的枢纽和 HH信号传导。在我们以前的工作的基础上，该提案的目的是获得机械理解 纤毛在灌感过程中原发性纤毛在浓缩和同步的机械和HH信号中的作用 发展和治愈。为了实现这一目标，我们将确定纤毛分节的身份和活动 灌感开发和机械适应过程中的细胞（特定目标1）并评估再生 纤毛灌肠细胞改善埃文愈合的能力（特定目的2）。我们将使用的方法 包括纤毛标记的和纤毛的转基因小鼠模型，不同的已建立的加载模型，细胞 移植和转录组学分析与结构，组成和生物力学结合 评估测定法。在该项目的结尾，我们希望确定新的纤毛调节的机械 体内插入机械适应过程中的转导途径，并提出新的机制 纤毛将机械提示转换为细胞信号事件。原发性纤毛在埃文中的作用的新发现 康复将指导新型药理学和机械生物学治疗剂用于治疗 肩袖撕裂。