Impacts of mechanosensation and matrix architecture on cell fate specification in traumatic heterotopic ossification

机械感觉和基质结构对创伤性异位骨化细胞命运规范的影响

• 批准号: 10613582
• 负责人: Benjamin Levi
• 金额: $ 43.34万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10613582
• 关键词: Anisotropy; Architecture; Automobile Driving; Burn injury; Cell Lineage; Cells; Chemicals; Clinical; Collagen; Collagen Receptors; Collagen Type I; Communication; Cre lox recombination system; Data; Data Set; Devices; Disease; Elements; Exposure to; Extracellular Matrix; Focal Adhesion Kinase 1; Gene Expression Profile; Genetic; Genetic Transcription; Heterotopic Ossification; Human; Immobilization; In Vitro; Injury; Intervention; Joints; Label; Lesion; Ligands; Limb structure; Link; Mechanical Stress; Mesenchymal Stem Cells; Modeling; Molecular; Mus; Osteogenesis; PTK2 gene; Pathologic; Pathway interactions; Patients; Periodicity; Phase; Physical therapy; Physiologic Ossification; Process; Protein Tyrosine Kinase; Protocols documentation; Radial; Regulation; Role; Series; Signal Transduction; Site; Skeletal bone; Stretching; System; Therapeutic; Therapeutic Intervention; Traumatic injury; Work; cell fate specification; clinical translation; clinically relevant; combat; discoidin domain receptor 2; discoidin receptor; fibula; hip replacement arthroplasty; in vivo; in vivo Model; inducible Cre; inhibitor therapy; insight; joint mobilization; lipid biosynthesis; mechanotransduction; mouse model; musculoskeletal injury; novel; osteochondral tissue; prevent; progenitor; programs; single-cell RNA sequencing; targeted treatment; tibia; ulna

Project Summary Heterotopic ossification (HO) is the pathologic formation of extra-skeletal bone forming almost exclusively at sites of mechanical stress, that occurs in ~20% of patients after hip arthroplasty, burns or musculoskeletal injury. Currently, no therapeutics or physical therapy-based protocols exist to prevent HO. In this regard, there is a void in our understanding of the causative mechanotransductive pathways behind this debilitating process. Our unbiased transcription profiles in mouse HO-mesenchymal progenitor cells (MPCs) recovered from HO sites in combination with immunostaining of mouse and human HO revealed that a series of mechanotranduction-linked pathways, including discoidin receptor 2 (DDR2), FAK and the Hippo effectors, YAP and TAZ, are highly upregulated in tandem with observed changes in extracellular matrix (ECM) alignment. Using a novel, regional MPC-specific inducible Cre system (Hoxa11-CreERT2), we have compiled preliminary data that support critical roles for DDR2 signaling and stage-specific immobilization in both triggering FAK/YAP/TAZ signaling and MPC lineage commitment, but also an unexpected function in controlling ECM alignment. Together, these observations have led to our central hypothesis that MPC DDR2 signaling is necessary for mobility-induced changes in ECM alignment that trigger aberrant osteochondral differentiation at HO sites and can be blocked by DDR2 inhibition or injury stage-specific immobilization. Aim 1: Define the role of DDR2 as a critical upstream regulator of FAK/YAP/TAZ signaling in controlling the induction and progression of HO. We hypothesize that DDR2-mechanotransductive signaling alters osteochondral differentiation and HO in vivo and can be targeted with cell specific deletion models or translatable clinical therapies. Aim 2: Determine the optimal post-injury timing during which MPCs can be redirected away from aberrant osteochondral fate and pathologic ECM alignment through immobilization-based intervention. We hypothesize that immobilization during the early proliferative phase after injury will block pathologic changes in ECM alignment with disease-ameliorating effects on MPC fate determination and aberrant ossification. Aim 3: Characterize the role of mobilization-induced DDR2 activation on collagen alignment/anisotropy and mechanotransductive signaling. We hypothesize that DDR2 activity drives ECM alignment independently of limb mobility in vivo or cyclic stretch in vitro. Impact: The proposed studies will provide a comprehensive and mechanistic understanding of how DDR2 and joint mobility regulate ECM alignment, cell fate and HO using conditional deletion models and clinical therapies.

项目概要 异位骨化 (HO) 是骨骼外骨形成的病理性形成，几乎完全在 约 20% 的患者在髋关节置换术、烧伤或肌肉骨骼损伤后发生机械应力。 目前，尚无基于治疗或物理治疗的方案可预防 HO。在这方面，存在空白 帮助我们理解这一令人衰弱的过程背后的致病机制。我们的 从 HO 位点恢复的小鼠 HO 间充质祖细胞 (MPC) 中的无偏转录谱 与小鼠和人类 HO 的免疫染色相结合，揭示了一系列机械转导相关的 通路，包括盘状蛋白受体 2 (DDR2)、FAK 和 Hippo 效应器、YAP 和 TAZ，高度 与观察到的细胞外基质（ECM）排列变化同时上调。使用新颖的、区域性的 MPC 特异性诱导 Cre 系统 (Hoxa11-CreERT2)，我们已经编制了支持关键的初步数据 DDR2 信号传导和特定阶段固定在触发 FAK/YAP/TAZ 信号传导和 MPC 中的作用 谱系承诺，而且在控制 ECM 对齐方面也是一个意想不到的功能。在一起，这些 观察结果得出我们的中心假设：MPC DDR2 信号对于移动引起的 ECM 排列的变化会引发 HO 位点异常骨软骨分化，并且可以通过以下方法阻断： DDR2 抑制或损伤阶段特异性固定。 目标 1：定义 DDR2 作为 FAK/YAP/TAZ 信号控制的关键上游调节器的作用 HO 的诱导和进展。我们假设 DDR2 机械传导信号改变 骨软骨分化和 H2O 体内，可以通过细胞特异性缺失模型或可翻译来靶向 临床治疗。 目标 2：确定受伤后的最佳时机，在此期间 MPC 可以被重新引导离开 通过基于固定的干预来实现异常的骨软骨命运和病理 ECM 排列。 我们假设损伤后早期增殖期的固定将阻止病理变化 ECM 与 MPC 命运决定和异常骨化的疾病改善作用相一致。 目标 3：表征动员诱导的 DDR2 激活对胶原排列/各向异性的作用 和机械传导信号。我们假设 DDR2 活动独立驱动 ECM 对齐 体内肢体活动性或体外循环拉伸。 影响：拟议的研究将为 DDR2 和 使用条件缺失模型和临床疗法，关节活动性调节 ECM 排列、细胞命运和 HO。