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.

项目摘要异位骨化（HO）是几乎仅在髋关节置换术，烧伤或肌肉骨骼损伤后约20％的患者发生机械应激部位。目前，尚无治疗或基于物理治疗的方案以防止HO。在这方面，有一个空白在理解这一使人衰弱过程背后的致病机械转移途径。我们的小鼠HO-间充质祖细胞（MPC）中的无偏转录曲线从HO地点回收与小鼠和人HO的免疫染色结合表明，一系列机械界线连接包括盘状受体2（DDR2），FAK和河马效应器，YAP和TAZ的途径高度高度与观察到的细胞外基质（ECM）比对的变化相连。使用小说，区域性 MPC特异性诱导CRE系统（HOXA11-CREERT2），我们编制了支持关键的初步数据 DDR2信号传导和阶段特异性固定的作用在触发FAK/YAP/TAZ信号传导和MPC中的作用血统承诺，但也是控制ECM对齐的意外功能。在一起，这些观察结果导致了我们的中心假设，即MPC DDR2信号对于迁移率诱导是必需的 ECM对齐的变化在HO站点触发异常的骨软骨分化，并且可以阻止 DDR2抑制或损伤阶段特异性固定化。 AIM 1：将DDR2的作用定义为FAK/YAP/TAZ信号的关键上游调节器 HO的诱导和进展。我们假设DDR2-Mechanotransductive信号改变了骨软骨分化和体内HO，可以用特异性缺失模型或可翻译的靶向临床疗法。目标2：确定可以将MPC重定向到的最佳伤害后时间安排通过基于固定的干预措施，异常的骨软骨命运和病理ECM对齐。我们假设损伤后早期增生阶段的固定将阻止病理变化在ECM与疾病的影响对MPC命运确定和异常骨化的影响中。 AIM 3：表征动员诱导的DDR2激活在胶原蛋白排列/各向异性上的作用和机械转移信号传导。我们假设DDR2活动可以独立地驱动ECM对齐体内的肢体迁移率或体外环状拉伸。影响：拟议的研究将对DDR2和联合迁移率使用条件缺失模型和临床疗法调节ECM比对，细胞命运和HO。