The Cellular Basis of Heterotopic Ossification

异位骨化的细胞基础

基本信息

批准号：
8269566
负责人：
DAVID J GOLDHAMER
金额：
$ 32.79万
依托单位：
UNIVERSITY OF CONNECTICUT STORRS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-17 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8269566
关键词：
ACVR1 gene Affect Amputation BMP2 gene Biological Assay Bone Matrix Bone Morphogenetic Proteins Cartilage Cell Culture Techniques Cell Transplantation Cell surface Cells Development Dyes Electron Microscopy Endothelial Cells Ensure Exclusion Exclusion Criteria Fluorescence Fluorescence-Activated Cell Sorting Functional disorder Gene Expression Genetic Genetic Recombination Goals Health Hematopoietic Hereditary Disease Heterotopic Ossification Histology Immunofluorescence Immunologic Immunohistochemistry Individual Inflammation Inflammatory Injury Intramuscular Injections Joints Label Lead Lesion Life Life Expectancy Mediating Methodology Methods Modality Modeling Molecular Mus Muscle Mutation Operative Surgical Procedures Osteogenesis Pain Pericytes Peripheral Phosphorylation Physiological Population Prevalence Prevention Prevention strategy Property Quality of life Reporter Reporter Genes Research Role Rotation SCID Mice Severities Signal Transduction Skeletal Muscle Soft Tissue Injuries Sorting - Cell Movement Source Specificity Stem cells Swelling Tamoxifen Technology Testing Therapeutic Time Tissues Total Hip Replacement Transgenic Mice Transplantation Trauma Vascular Smooth Muscle Western Blotting base bone bone morphogenetic protein receptor type I cadherin 5 cell type cellular targeting combat common treatment disability implantation in vivo inclusion criteria matrigel mouse model mutant novel osteogenic progenitor progressive myositis ossificans public health relevance receptor recombinase research study response small molecule soft tissue tool translational study whole body imaging

项目摘要

DESCRIPTION (provided by applicant): Heterotopic ossification (HO), defined as the inappropriate formation of bone in skeletal muscles and associated soft tissues, causes pain, swelling, decreased rotation of affected joints, and other health consequences. HO is a common consequence of certain surgeries and other physical traumas, and also is the defining feature of the severely debilitating condition, fibrodysplasia ossificans progressiva (FOP), a genetic disorder of dysregulated bone morphogenetic protein (BMP) signaling. Despite its prevalence and health consequences, the pathophysiology of HO is poorly understood and the progenitor cells responsible for HO have not been identified. The overarching objective of the proposed research is to identify and characterize the cellular precursors that are responsible for HO. The following hypothesis will be tested: cells of the peripheral vasculature function as osteogenic progenitor cells in response to excessive or dysregulated BMP signaling. In Aims 1 and 2, Cre/lox lineage-tracing methods will be used to determine the contribution of endothelial cells, hematopoietic cells, vascular smooth muscle, pericytes and muscle SP stem cells to HO. HO will be induced by intramuscular injection of BMP2, and the contribution of labeled cell populations to heterotopic cartilage and bone determined by immunohistochemistry using cell-specific markers. In a complementary, independent approach, a cell transplantation bioassay will assess the osteogenic potential of cellular subfractions sorted on the basis of lineage-restricted reporter gene expression, cell surface marker expression or dye exclusion properties (muscle SP). In Aim 3, a genetic mouse model for HO will be developed and characterized. The model is based on Cre recombinase-based conditional expression of the mutant Type I BMP receptor, ACVR1(R206H), which was recently found to cause FOP. In Aim 4, HO in ACVR1(R206H)-expressing mice will be analyzed by whole-body imaging and immunohistochemistry to determine the consequence of cell-specific and temporally controlled expression of the mutant receptor. Effects of injury on the occurrence and severity of ACVR1(R206H)-mediated HO also will be evaluated. These studies will define the cellular progenitors and developmental window in which dysregulated intracellular BMP signaling causes HO. Understanding the cellular basis of HO will identify specific cellular targets and suggest therapeutic strategies for HO. Additionally, the mouse models developed herein will be valuable tools for translational studies aimed at developing and testing small molecule therapies and other modalities for the treatment or prevention of HO. PUBLIC HEALTH RELEVANCE: Heterotopic ossification is a painful and debilitating condition characterized by the inappropriate formation of bone in skeletal muscles and associated soft tissues. The proposed studies will identify and characterize the progenitor cell populations that are responsible for heterotopic ossification, thereby defining cellular targets and suggesting therapeutic strategies for the prevention and treatment of this common condition.

描述（由申请人提供）：异位骨化（HO），定义为骨骼肌和相关软组织中骨骼不适当形成，会导致疼痛，肿胀，受影响关节的旋转减少以及其他健康后果。 HO是某些手术和其他身体创伤的普遍结果，也是严重使人衰弱的疾病的定义特征，即纤维化肿瘤骨化剂（FOP），这是一种失调的骨形态发生蛋白（BMP）信号的遗传疾病。尽管其患病率和健康后果，但HO的病理生理学知之甚少，尚未鉴定出负责HO的祖细胞。拟议研究的总体目标是识别和表征负责HO的细胞前体。将测试以下假设：外周血管的细胞对过度或失调的BMP信号传导起着成骨祖细胞的作用。在目标1和2中，将使用CRE/LOX谱系方法来确定内皮细胞，造血细胞，血管平滑肌，周细胞和肌肉SP干细胞对HO的贡献。 HO将通过肌内注射BMP2的诱导，并使用细胞特异性标记物通过免疫组织化学确定标记的细胞群对异位软骨和骨的贡献。在互补的独立方法中，细胞移植生物测定将根据谱系限制的报告基因表达，细胞表面标记表达或染料排除特性（Muscle SP）评估细胞分支的成骨潜力。在AIM 3中，将开发和表征用于HO的遗传小鼠模型。该模型基于基于CRE重组酶I型突变体BMP受体ACVR1（R206H）的条件表达，最近发现它会引起FOP。在AIM 4中，将通过全身成像和免疫组织化学分析ACVR1（R206H）中表达小鼠的HO，以确定突变受体的细胞特异性和时间控制表达的结果。还将评估损伤对ACVR1（R206H）介导的HO的发生和严重程度的影响。这些研究将定义细胞祖细胞和发育窗口，其中不调节细胞内BMP信号导致HO。了解HO的细胞基础将确定特定的细胞靶标，并提出HO的治疗策略。此外，本文开发的小鼠模型将是用于转化研究的有价值的工具，旨在开发和测试小分子疗法和其他用于治疗或预防HO的方式。公共卫生相关性：异位骨化是一种痛苦且令人衰弱的状况，其特征是骨骼肌和相关软组织中骨骼不适当形成。拟议的研究将确定并表征负责异位骨化的祖细胞群体，从而定义细胞靶标，并提出预防和治疗这种常见状况的治疗策略。