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

基本信息

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
项目类别：
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

项目摘要

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）具有众所周知，SSCs 在骨修复中发挥着重要作用。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以及它们在骨骼愈合中的作用尚不清楚，因此，我们建议的目标是定义损伤。 P-SSC 的体内特性和功能以及调节 P-SSC 控制骨的机制我们还将探讨内源性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。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) 可以选择性标记仅存在于骨膜形成层的内源性 P-SSC 此外，连续体内成像显示 Mx1 αSMAGFP P-SSC，而非 BM-SSC，能够快速响应损伤并持续提供新的成骨细胞用于损伤修复这些 P-SSC 表现出较高的 CCR3/CCR5 表达，即趋化因子（C-C 基序）配体 5 的受体。 (CCL5)，比BM-SSCs和其他骨细胞更重要，而且，我们首次发现骨膜。 CCL5 的施用刺激这些 P-SSC 向人体内损伤部位实时迁移。原代骨膜细胞也表达 CCR5 并进行 CCL5 介导的迁移。 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。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因此，我们计划在活体动物中研究骨髓及其在单细胞水平上的分化。在目标 1 中，我们将确定 CCL5 等炎症刺激是否是必要的。我们将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。还检查 CCL5 是否针对 P-SSC 迁移。在目标 2 中，我们将确定 P-SSC 及其是否具有特异性。通过对 P-SSC 进行局部消融并生成条件性损伤修复需要迁移 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性 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骼愈合。完成这项工作后，我们将获得新的生物学见解。进入骨膜 SSC 并将确定逆转骨疾病和缺陷的新治疗靶点。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Targeting periosteal SSCs for aged bone defects.

针对老年骨缺损的骨膜 SSC。

DOI：
发表时间：
2020
期刊：
影响因子：
0
作者：
Jeong, Youngjae;Park, Dongsu
通讯作者：
Park, Dongsu

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

通过四种转录因子建立稳定可扩增的诱导生肌干细胞。

DOI：
发表时间：
2018
期刊：
影响因子：
9
作者：
Lee, Eun;Kim, Minhyung;Kim, Yong Deuk;Chung, Myung;Elfadl, Ahmed;Ulah, H M Arif;Park, Dongsu;Lee, Sunray;Park, Hyun;Kim, Tae;Hwang, Daehee;Jeong, Kyu
通讯作者：
Jeong, Kyu

Prognostic Role of TMED3 in Clear Cell Renal Cell Carcinoma: A Retrospective Multi-Cohort Analysis.

TMED3 在透明细胞肾细胞癌中的预后作用：回顾性多队列分析。

DOI：
发表时间：
2019
期刊：
影响因子：
0
作者：
Ha, Mihyang;Moon, Hwan;Choi, Dongwook;Kang, Wonmo;Kim, Ji;Lee, Keon Jin;Park, Dongsu;Kang, Chi;Oh, Sae;Han, Myoung;Kim, Yun Hak;Lee, Dongjun
通讯作者：
Lee, Dongjun

GABRQ expression is a potential prognostic marker for patients with clear cell renal cell carcinoma.

GABRQ 表达是透明细胞肾细胞癌患者的潜在预后标志物。

DOI：
发表时间：
2019-12
期刊：
影响因子：
2.9
作者：
Lee, Dongjun;Ha, Mihyang;Hong, Chae Mi;Kim, Jayoung;Park, Su Min;Park, Dongsu;Sohn, Dong Hyun;Shin, Ho Jin;Yu, Hak;Kim, Chi Dae;Kang, Chi;Han, Myoung;Oh, Sae;Kim, Yun Hak
通讯作者：
Kim, Yun Hak

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

不同骨骼区室中骨骼干/祖细胞的差异调节。