Defining periosteal skeletal stem cells and novel migration mechanisms in bone regeneration and repair in vivo

定义骨膜骨骼干细胞和体内骨再生和修复的新迁移机制

• 批准号: 10442372
• 负责人: Dongsu Park
• 金额: $ 41.53万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-09 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10442372
• 关键词: Ablation; Acceleration; Actins; Adult; Affect; Aging; Animal Model; Animals; Biological; Bone Diseases; Bone Growth; Bone Injury; Bone Marrow; Bone Regeneration; Bone Resorption; Bone Surface; Bone callus; Bone remodeling; CCR5 gene; Calvaria; Cartilage; Cell Proliferation; Cells; Chondrocytes; Clinical; Defect; Diphtheria Toxin; Drug or chemical Tissue Distribution; Fracture; Goals; Health; Human; Imaging technology; Immune; Impaired healing; In Vitro; Inflammatory; Injury; Label; Ligands; Location; Longevity; Marrow; Mediating; Modeling; Morbidity - disease rate; Mus; Myxovirus; Natural regeneration; Osteoblasts; Osteoclasts; Pain; Periosteal Cell; Periosteum; Physiological; Play; Population; Process; Proliferating; RANTES; Recovery; Regenerative capacity; Regulation; Reporter; Research Proposals; Resistance; Role; Series; Signal Transduction; Site; Skeleton; Smooth Muscle; Source; Stimulus; Stromal Cell-Derived Factor 1; Testing; Time; Tissues; Work; age related; aged; bone; bone aging; bone cell; bone fracture repair; bone fragility; bone healing; bone loss; bone repair; cell injury; chemokine receptor; clinically relevant; healing; improved; in vivo; in vivo imaging; injury and repair; insight; intravital imaging; long bone; migration; mortality; mouse model; new therapeutic target; novel; novel therapeutic intervention; preservation; prevent; progenitor; receptor; receptor expression; reconstitution; recruit; repaired; skeletal regeneration; skeletal stem cell; stem cell biomarkers; stem cell function; stem cell migration; stem cells; success

PROJECT SUMMARY/ABSTRACT Significant bone loss and defective bone healing in age-related bone diseases cause severe pain and high morbidity and mortality, and are a major health issue in the U.S. Lifelong regeneration of bone and cartilage requires skeletal stem cells (SSCs), and a skeletal stem/progenitor cell subset in periosteum (P-SSCs) has been known to play an important role for bone repair. However, due to the wide tissue distribution of SSCs and the lack of specific markers to distinguish rare P-SSCs in vivo, how endogenous P-SSCs respond to bone injury and how they function in bone healing are unknown. Therefore, the goal of our proposal is to define the in vivo identity and function of P-SSCs and the mechanisms that regulate P-SSCs in order to control bone regeneration and repair under physiologic circumstances. We will also explore the induction of endogenous P- SSC migration and proliferation with the goal of improving recovery from age-related bone injuries. Using newly generated reporter mouse models in combination with intravital imaging technology, we recently determined that a combination of SSC markers, myxovirus resistance-1 (Mx1) and alpha smooth muscle actin (αSMAGFP) can selectively label endogenous P-SSCs that are exclusively present in the periosteal cambial layer with Prx1GFP expression. In addition, sequential in vivo imaging revealed that Mx1 αSMAGFP P-SSCs, rather than BM-SSCs, rapidly respond to the injury and continually supply new osteoblasts for injury repair in vivo. These P-SSCs show higher expression of CCR3/CCR5, the receptors for chemokine (C-C motif) ligand 5 (CCL5), than BM-SSCs and other bone cells. Moreover, we found for the first time that periosteal administration of CCL5 stimulates the real-time migration of these P-SSCs toward injury sites in vivo. Human primary periosteal cells also express CCR5 and conduct CCL5-mediated migration. We thus hypothesize that P-SSCs and their migration, upon CCL5 signaling, are necessary for bone repair, and that exogenous CCL5 provision improves the healing of age-related bone defects. Our novel intravital imaging technology and the various animal models enable us to track endogenous stem cells present in the periosteum and bone marrow as well as their differentiation at the single-cell level in living animals. We thus plan to pursue the following specific aims. In aim 1, we will determine whether inflammatory stimuli such as CCL5 are necessary for P-SSC migration and bone healing by generating a conditional ablation of CCL5 in immune cells. We will also examine if CCL5 is specific for P-SSC migration. In aim 2, we will determine whether P-SSCs and their migration are required for injury repair by performing local ablation of P-SSCs and by generating a conditional deletion of the CCR5 in P-SSCs. In aim 3, we will determine whether the local provision of exogenous CCL5 induces the migration and activation of endogenous P-SSCs early in the repair process, leading to accelerated and improved bone healing in aged mice. Upon completion of this work, we will achieve new biological insights into periosteal SSCs and will define new therapeutic targets for reversing bone diseases and defects.

项目概要/摘要 与年龄相关的骨病中严重的骨质流失和骨愈合缺陷会导致严重的疼痛和高烧 发病率和死亡率，是美国的一个主要健康问题 骨和软骨的终身再生 需要骨骼干细胞（SSC），并且骨膜中的骨骼干/祖细胞亚群（P-SSC）具有 已知在骨修复中发挥重要作用。然而，由于SSC在组织中分布广泛， 缺乏特异性标记物来区分体内罕见的 P-SSC，内源性 P-SSC 如何对骨做出反应 损伤及其在骨愈合中的作用尚不清楚。因此，我们提案的目标是定义 P-SSC 的体内特性和功能以及调节 P-SSC 控制骨的机制 生理条件下的再生和修复。我们还将探索内源性 P- 的诱导 SSC 迁移和增殖，目的是改善与年龄相关的骨损伤的恢复。使用 新生成的报告小鼠模型结合活体成像技术，我们最近 确定 SSC 标记、粘病毒抗性 1 (Mx1) 和 α 平滑肌肌动蛋白的组合 (αSMAGFP) 可以选择性地标记仅存在于骨膜形成层中的内源性 P-SSC 具有 Prx1GFP 表达的层。此外，连续体内成像显示 Mx1 αSMAGFP P-SSC， 而不是 BM-SSC，快速响应损伤并持续提供新的成骨细胞用于损伤修复 体内。这些 P-SSC 显示出较高的 CCR3/CCR5 表达，即趋化因子（C-C 基序）配体 5 的受体 (CCL5)，优于 BM-SSC 和其他骨细胞。此外，我们还首次发现骨膜 CCL5 的施用刺激这些 P-SSC 向体内损伤部位实时迁移。人类 原代骨膜细胞也表达 CCR5 并进行 CCL5 介导的迁移。因此我们假设 P-SSC 及其在 CCL5 信号传导下的迁移对于骨修复是必需的，并且外源性 CCL5 的提供可改善与年龄相关的骨缺损的愈合。我们新颖的活体成像技术 各种动物模型使我们能够追踪骨膜和骨骼中存在的内源干细胞 骨髓及其在活体动物单细胞水平上的分化。因此，我们计划追求 遵循特定目标。在目标 1 中，我们将确定 CCL5 等炎症刺激是否必要 通过在免疫细胞中产生 CCL5 的条件消融来实现 P-SSC 迁移和骨愈合。我们将 还检查 CCL5 是否特定于 P-SSC 迁移。在目标 2 中，我们将确定 P-SSC 及其 通过对 P-SSC 进行局部消融并生成有条件的损伤修复需要迁移 P-SSC 中 CCR5 的缺失。在目标 3 中，我们将确定是否本地提供外源 CCL5 在修复过程的早期诱导内源性 P-SSC 的迁移和激活，从而加速修复过程 并改善老年小鼠的骨愈合。完成这项工作后，我们将获得新的生物学见解 骨膜 SSC 并将确定逆转骨疾病和缺陷的新治疗靶点。

项目成果

Differential regulation of skeletal stem/progenitor cells in distinct skeletal compartments.

• DOI: 10.3389/fphys.2023.1137063
• 发表时间: 2023
• 期刊: FRONTIERS IN PHYSIOLOGY
• 影响因子: 4
• 作者: Solidum, Jea Giezl Niedo;Jeong, Youngjae;Heralde III, Francisco;Park, Dongsu
• 通讯作者: Park, Dongsu

Establishment of stably expandable induced myogenic stem cells by four transcription factors.

• DOI: 10.1038/s41419-018-1114-8
• 发表时间: 2018-10-25
• 期刊: Cell death & disease
• 影响因子: 9
• 作者: Lee EJ;Kim M;Kim YD;Chung MJ;Elfadl A;Ulah HMA;Park D;Lee S;Park HS;Kim TH;Hwang D;Jeong KS
• 通讯作者: Jeong KS

Targeting periosteal SSCs for aged bone defects.

针对老年骨缺损的骨膜 SSC。

• DOI: 10.18632/aging.102869
• 发表时间: 2020
• 期刊: Aging
• 作者: Jeong,Youngjae;Park,Dongsu
• 通讯作者: Park,Dongsu