Dynamics and Regulations of Bone Stem Cells in Vivo

体内骨干细胞的动力学和调控

基本信息

批准号：
10477641
负责人：
Noriaki Ono
金额：
$ 36.68万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10477641
关键词：
Adipocytes Adult Alleles Behavior Biological Assay Bone Diseases Bone Growth Bone Marrow Bone Marrow Cells Cells Characteristics Child Chondrocytes Clinical Collagen Type II Data Deformity Development Epiphysial cartilage Exhibits Family Gene Expression Genes Genetic Growth High Fat Diet Hypertrophy Immunodeficient Mouse In Vitro Knowledge Label Lead Life LoxP-flanked allele Maintenance Marrow Mesenchymal Modality Modeling Molecular Natural regeneration Operative Surgical Procedures Osteoblasts PTH gene Patients Physiologic Ossification Process Property Publishing Regenerative engineering Regulation Reporter Research Rest Reticular Cell Role Signal Transduction Societies Source Stromal Cell-Derived Factor 1 Stromal Cells Testing Tibial Fractures Transgenic Organisms Transplantation Well in self adipocyte differentiation base beta catenin bone clinical application experimental study improved in vivo intramembranous bone formation novel parathyroid hormone-related protein postnatal progenitor regenerative approach response to injury retention rate rosiglitazone self-renewal smoothened signaling pathway stem stem cells stromal progenitor transcription factor transcriptome sequencing translational applications young adult

项目摘要

PROJECT SUMMARY/ABSTRACT Bone disorders and deformities are prevalent in children and young adults. Due to lack of effective modalities to regenerate growing bones, these young patients often undergo multiple surgical interventions, posing a significant burden on them, their family and society. During bone growth, chondrocytes and osteoblasts are continuously generated to make bones bigger and stronger. Endogenous bone stem cells that serve as the source of these cells have not been completely understood. Fundamental knowledge of how these bone stem cells coordinate the two processes of endochondral and intramembranous ossification is essential to develop a reliable approach to regenerate growing bones. In this project, the characteristics of distinct types of bone stem cells that actively promote bone growth will be identified. We hypothesize that a subset of resting chondrocytes in the postnatal growth plate behave as growth-associated bone stem cells, and become a source of mesenchymal stromal progenitor cells in bone marrow; these two types of bone stem/progenitor cells concertedly promote proper bone growth and maintenance. Identifying characteristics and molecular regulations of bone stem cells will facilitate our endeavor to reproduce these cells through regenerative engineering. In Aim1, we will identify molecular mechanisms regulating properties and fates of resting chondrocytes. The working hypothesis is that resting chondrocytes of the postnatal growth plate exhibit unique characteristics as growth-associated bone stem cells, whose properties and fates are regulated by Hedgehog signaling. We will first identify a self-renewing multipotent subpopulation of resting chondrocytes using in vitro colony-forming assays and in vivo transplantation of isolated growth plate cells. We will second identify the unique gene expression profiles of self-renewing colony-forming resting chondrocytes. We will further define roles of Hedgehog signaling in determining self-renewal and differentiation of resting chondrocytes by modulating its signaling components, while simultaneously tracing their behavior both in vivo and in vitro. In Aim2, we will define formation and fates of bone marrow mesenchymal stromal progenitors in growing bones. The working hypothesis is that growth plate chondrocytes undergo hypertrophy and transform into Cxcl12- abundant reticular (CAR) cells in bone marrow that behave as regional and reactive mesenchymal stromal progenitor cells. We will first define differentiation potentials of CAR cells into osteoblasts and adipocytes in vivo through intermittent PTH administration and a high-fat diet containing rosiglitazone. We will second determine CAR cells' response to injury using a semistabilized tibial fracture model. We will also identify effects of these manipulations on CAR cells' expression levels of key transcription factors that regulate cell fate choice. We will third define the properties of CAR cells as mesenchymal stromal progenitors through in vitro colony- forming assays and in vivo transplantation of isolated bone marrow cells. We will further define roles of β- catenin signaling as a cell fate determinant of osteoblast versus adipocyte differentiation using its floxed allele.

项目摘要/摘要儿童和年轻人普遍存在骨骼疾病和畸形。由于缺乏有效的方式为了再生生长的骨骼，这些年轻患者经常接受多种手术干预，提出对他们，他们的家人和社会的重要伯恩。在骨骼生长过程中，软骨细胞和成骨细胞为连续生成以使骨骼变大和结实。内源性骨干细胞，作为这些细胞的来源尚未完全理解。这些骨干的基本知识细胞协调内侧软骨和膜内骨化的两个过程对于发展A是至关重要的可靠的方法来再生生长的骨骼。在这个项目中，骨干的不同类型的特征将确定积极促进骨生长的细胞。我们假设一部分静止软骨细胞在产后生长板作为生长相关的骨干细胞的行为，并成为骨髓中的间充质基质祖细胞；这两种类型的骨干/祖细胞协同促进适当的骨骼生长和维护。识别特征和分子骨干细胞的法规将促进我们通过再生来再现这些细胞的努力工程。在AIM1中，我们将确定调节静止特性和命运的分子机制软骨细胞。工作假设是产后生长板的休息软骨细胞具有独特特征作为生长相关的骨干细胞，其特性和命运受刺猬调节信号。我们将首先确定使用体外的静息软骨细胞的自我更新多能亚群菌落形成测定和分离生长板细胞的体内移植。我们将第二确定自我更新菌落形成的静息软骨细胞的独特基因表达谱。我们将进一步定义刺猬信号传导在确定静止软骨细胞的自我更新和分化中的作用调节其信号传导成分，同时在体内和体外追踪其行为。 AIM2，我们将定义生长骨骼中骨髓间充质祖细胞的形成和命运。有效的假设是生长板软骨细胞经历肥大并转化为CXCL12- 骨髓中丰富的网状细胞，其表现为区域和反应性间质基质祖细胞。我们将首先将CAR细胞的分化潜力定义为成骨细胞和脂肪细胞通过间歇性PTH给药和含有罗格列酮的高脂饮食的体内。我们将第二使用半稳定的胫骨断裂模型来确定CAR细胞对损伤的反应。我们还将确定效果这些对CAR细胞的表达水平的操作中的关键转录因子调节细胞命运选择的水平。我们将通过体外菌落 - 将汽车细胞作为间充质基质祖细胞的特性定义为间质基质祖细胞的特性。形成分离的骨髓细胞的测定和体内移植。我们将进一步定义β-的作用 Catenin信号传导是骨细胞与脂肪细胞分化的细胞脂肪确定剂，使用其floxed等位基因。

项目成果

期刊论文数量（30）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Cranial Base Synchondrosis Lacks PTHrP-Expressing Column-Forming Chondrocytes.

DOI：
10.3390/ijms23147873
发表时间：
2022-07-17
期刊：
International journal of molecular sciences
影响因子：
5.6
作者：
通讯作者：

Flow Cytometry-Based Analysis of the Mouse Bone Marrow Stromal and Perivascular Compartment.

DOI：
10.1007/978-1-0716-1425-9_7
发表时间：
2021
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Matsushita Y;Ono W;Ono N
通讯作者：
Ono N

A three-dimensional analysis of primary failure of eruption in humans and mice.

DOI：
10.1111/odi.13249
发表时间：
2020-03
期刊：
Oral diseases
影响因子：
3.8
作者：
Tokavanich N;Gupta A;Nagata M;Takahashi A;Matsushita Y;Yatabe M;Ruellas A;Cevidanes L;Maki K;Yamaguchi T;Ono N;Ono W
通讯作者：
Ono W

The diverse origin of bone-forming osteoblasts.