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

基本信息

批准号：
10297550
负责人：
Benjamin Levi
金额：
$ 42.84万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
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/10297550
关键词：
Anisotropy Architecture Automobile Driving Burn injury Cell Lineage Cells Chemicals Clinical Collagen Collagen Receptors Collagen Type I Communication 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 Stretching System Therapeutic Therapeutic Intervention Traumatic injury Work base bone cell fate specification clinically relevant clinically translatable combat discoidin domain receptor 2 discoidin receptor fibula hip replacement arthroplasty in vivo in vivo Model inhibitor/antagonist insight joint mobilization lipid biosynthesis mechanotransduction mouse model musculoskeletal injury novel osteochondral tissue prevent progenitor programs single-cell RNA sequencing targeted treatment tibia ulna

项目摘要

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来预防 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 机械传导信号改变体内骨软骨分化和 H2O ，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。