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.

项目概要/摘要由于缺乏有效的治疗方法，骨骼疾病和畸形在儿童和年轻人中普遍存在。为了再生正在生长的骨骼，这些年轻患者经常接受多次手术干预，造成在骨骼生长过程中，软骨细胞和成骨细胞是他们、他们的家庭和社会的重大负担。不断生成使骨骼更大更强的内源性骨干细胞。这些细胞的来源尚未完全了解这些骨干的基本知识。细胞协调软骨内骨化和膜内骨化的两个过程对于发展在这个项目中，研究了不同类型骨干的特征。我们将鉴定出积极促进骨骼生长的细胞。在出生后生长板中表现为与生长相关的骨干细胞，并成为骨干细胞的来源骨髓中的间充质基质祖细胞；这两种类型的骨干/祖细胞共同促进适当的骨骼生长和维护。识别特征和分子。骨干细胞的调控将有助于我们通过再生来繁殖这些细胞在 Aim1 中，我们将确定调节静息特性和命运的分子机制。工作假设是出生后生长板的静息软骨细胞表现出独特的特性。具有与生长相关的骨干细胞的特征，其特性和命运由 Hedgehog 调节我们将首先使用体外鉴定静息软骨细胞的自我更新多能亚群。我们将其次鉴定分离的生长板细胞的集落形成测定和体内移植。我们将进一步定义自我更新集落形成静息软骨细胞的独特基因表达谱。 Hedgehog 信号在决定静息软骨细胞自我更新和分化中的作用调节其信号成分，同时追踪它们在体内和体外的行为。目标2，我们将定义骨髓间充质基质祖细胞在骨骼生长中的形成和命运。工作假设是生长板软骨细胞肥大并转化为 Cxcl12- 骨髓中丰富的网状（CAR）细胞，充当区域性和反应性间充质基质我们首先定义 CAR 细胞向成骨细胞和脂肪细胞的分化潜力。体内通过间歇性 PTH 给药和含有罗格列酮的高脂肪饮食我们将其次。使用半稳定胫骨骨折模型确定 CAR 细胞对损伤的反应我们还将确定效果。这些操作对 CAR 细胞调节细胞命运选择的关键转录因子的表达水平进行了影响。第三，我们将通过体外集落来定义 CAR 细胞作为间充质基质祖细胞的特性 - 我们将进一步确定 β- 的作用。使用其 floxed 等位基因，连环蛋白信号传导作为成骨细胞与脂肪细胞分化的细胞命运决定因素。

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.