Regulation of Skeletal progenitor cells in Osteogenesis Imperfecta

成骨不全中骨骼祖细胞的调节

• 批准号: 10528208
• 负责人: Brendan Lee
• 金额: $ 66.24万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-13 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10528208
• 关键词: Affect; Animal Model; Biological; Bone Diseases; Bone Injury; Bone Marrow; Bone Regeneration; Breathing; CCR5 gene; Cell Count; Cell Proliferation; Cell model; Cell physiology; Cells; Characteristics; Clinical; Clinical Trials; Collagen; Craniofacial Abnormalities; Defect; Deformity; Disease; Environment; Extracellular Matrix; Face; Fracture; Gene Expression Profile; Genetic Diseases; Goals; Heterogeneity; Human; Impaired healing; Injury; KDR gene; Label; Ligands; Location; Maintenance; Mastication; Methods; Modeling; Modification; Molecular; Mus; Mutation; Natural regeneration; Oral; Osteoblasts; Osteogenesis Imperfecta; Patients; Periosteal Cell; Periosteum; Population; Procollagen; RANTES; Regulation; Reporter; Series; Severities; Signal Transduction; Site; Source; Surface; Tail; Technology; Testing; Transforming Growth Factor beta; Transplantation; Vascular Endothelial Growth Factors; Wild Type Mouse; Work; base; bone; bone healing; bone repair; bone strength; cell motility; clinically relevant; combinatorial; cortical bone; craniofacial; craniofacial bone; craniofacial disorder; fracture risk; improved; in vivo; insight; intravital imaging; long bone; migration; mouse model; neutralizing antibody; novel; osteoprogenitor cell; postnatal; progenitor; receptor; restoration; selective expression; single cell analysis; single-cell RNA sequencing; skeletal; skeletal disorder; skeletal stem cell; stem; stem cell function; stem cells; success

PROJECT SUMMARY Osteogenesis Imperfecta (OI) is a congenital bone and oral-facial disorder that mainly affects bone with less well characterized alterations in bone healing and repair. Like other skeletal diseases, at least some of the progressive bone and craniofacial defects in OI patients have been attributed to changes in the populations and functions of their stem/progenitor cells. In general, stem cells require a specialized environment for their maintenance and function. Nevertheless, how normal or defective structural components of collagen regulate skeletal stem/progenitor cells (SSPCs) is essentially unknown. Therefore, the goal of this proposal is to define the in vivo characteristics and function of skeletal stem/progenitor cells in OI and to understand how an abnormal OI extracellular matrix alters these stem/progenitor cells in the context of bone regeneration and repair. We previously showed that the Mx1Cre and αSMAGFP combination can selectively label skeletal stem cells in the periosteum and that these Mx1ÈαSMAGFPÈ periosteal cells are long-term repopulating stem cell subsets responsible for lifelong regeneration of periosteal osteoblasts and bone repair. Moreover, our preliminary study revealed that these periosteal SSPCs are Prx1GFP positive and selectively express KDR (VEGFR2). Human primary periosteal cells also express KDR with multi-lineage differentiation potentials. Notably, we found that these KDRÈ periosteal progenitor cells are significantly decreased in OI bones. Hence, we hypothesize that the abnormal OI matrix deregulates the number and function of periosteal and bone marrow SSPCs and the OI- associated molecular changes in stem/progenitor cells are critical for the progressive deformity and delayed or defective healing of bones. By using a series of OI animal models and SSPC reporter mice, in which we can differentially label periosteal and bone marrow SSPC subsets, in combination with intravital imaging and the latest single-cell RNA-sequencing technology, we plan to pursue the following specific aims. In aim 1, we will define the in vivo characteristics and function of periosteal and bone marrow SSPCs in clinically relevant OI mouse models. In aim 2, we will define key OI matrix factors that regulate SSPC function and improve both craniofacial and long bone healing. Upon completion of this work, we will achieve new biological insights into a better understanding of the molecular and cellular mechanisms that differentially regulate SSPCs under OI pathophysiological conditions.

项目摘要 成骨不完美（OI）是一种先天性骨和口腔疾病，主要影响骨骼的良好良好 表征了骨骼愈合和修复的改变。像其他骨骼疾病一样，至少有一些 OI患者的进行性骨骼和颅面缺陷归因于种群的变化和 其茎/祖细胞的功能。通常，干细胞需要专门的环境 维护和功能。然而，胶原蛋白调节的正常或有缺陷的结构成分 骨骼干/祖细胞（SSPC）本质上是未知的。因此，该提议的目的是定义 OI中骨骼干/祖细胞的体内特征和功能，并了解 异常的OI细胞外基质在骨再生的背景下改变了这些茎/祖细胞 和维修。我们先前表明MX1CRE和αSmAGFP组合可以选择性地标记骨骼茎 骨膜中的细胞和这些MX1èαSmagfpè骨膜细胞是长期重脑细胞 负责骨膜成骨细胞和骨修复的终身再生的子集。而且，我们的初步 研究表明，这些骨膜SSPC为PRX1GFP阳性，有选择地表达KDR（VEGFR2）。人类 原发性骨膜细胞还具有多条纹分化电位的KDR。值得注意的是，我们发现 OI骨骼中这些KDRè骨膜祖细胞显着降低。因此，我们假设 异常的OI基质消除了骨髓和骨髓SSPC的数量和功能以及OI- 茎/祖细胞中相关的分子变化对于进行性畸形和延迟或 骨骼的缺陷愈合。通过使用一系列OI动物模型和SSPC报告基因小鼠，我们可以在其中 差异标记骨髓和骨髓SSPC子集，并结合浸润成像和 最新的单细胞RNA测序技术，我们计划追求以下特定目标。在AIM 1中，我们将 定义骨膜和骨髓SSPC在临床相关的OI中的体内特征和功能 鼠标模型。在AIM 2中，我们将定义调节SSPC功能并提高两者的关键OI矩阵因子 颅面和长骨愈合。完成这项工作后，我们将获得新的生物学见解 更好地了解在OI下对SSPC不同调节的分子和细胞机制 病理生理状况。