Hox Gene Regulation of Skeletal Repair

Hox 基因对骨骼修复的调控

• 批准号: 10550118
• 负责人: Katharine A. Hubert
• 金额: $ 3.49万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10550118
• 关键词: Acute; Adipocytes; Adult; Alleles; Animals; Anterior; Behavior; Biological; Biological Assay; Bone Marrow; Bone Matrix; Bone callus; Bone remodeling; Cartilage; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cells; Chondrocytes; Chondrogenesis; Collagen; Collection; Control Animal; Data; Defect; Development; Developmental Process; Embryonic Development; Event; Exhibits; Fracture; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Recombination; Goals; Growth; Histocytochemistry; Histology; Homeobox Genes; Homeostasis; Injury; Knowledge; Label; Laboratories; Life; Maintenance; Measures; Mediating; Mesenchymal; Mesenchymal Stem Cells; Modeling; Molecular; Mus; Natural regeneration; Organ; Osteoblasts; Osteocytes; Osteogenesis; Pathway interactions; Pattern; Play; Population; Process; Radial; Reporter; Reporting; Roentgen Rays; Role; Skeleton; Small Interfering RNA; Stromal Cells; Testing; Time; Tomatoes; Work; base; bone; bone cell; bone fracture repair; cartilage cell; conditional mutant; differential expression; fibula; gene function; healing; injury and repair; insight; loss of function; microCT; mouse model; mutant; osteogenic; paralogous gene; postnatal; progenitor; repaired; response; response to injury; self-renewal; single cell analysis; single cell sequencing; skeletal; skeletal stem cell; stem; stem cell population; stem cells; tibia; tool; transcription factor; transcriptome sequencing; transcriptomics; ulna

PROJECT SUMMARY/ABSTRACT Hox genes are a group of evolutionarily conserved transcription factors important for several developmental processes, including patterning of the anterior-posterior axis of the skeleton. The Hox11 paralogous gene group, which is expressed in the zeugopod region (radius/ulna and fibula/tibia), are necessary for proper patterning of the zeugopod. In the past few years, work from the Wellik laboratory has shown that these developmentally important Hox transcription factors remain expressed in the skeleton throughout life, specifically in progenitor-enriched mesenchymal stem cells (MSCs). Rigorous genetic lineage labeling from the lab demonstrated that these cells give rise to all three mesenchymal lineages, osteoblasts, chondrocytes and adipocytes, and exhibit life-long self-renewal, providing strong evidence that this population of cells are skeletal stem cells. A key question based on this information is whether Hox gene function is important in these stem cells throughout life. We recently reported that temporal deletion of Hox11 at adult stages results in defects in osteoblastogenesis, wherein differentiation is initiated, but osteoblasts and osteocytes fail to mature. Adult conditional loss of Hox11 function results in a progressively weakened bone matrix where collagen does not properly assemble in remodeling bone. In this study, I will use a temporally-controlled, conditional loss-of- function model to assess defects in response to fracture repair (Aim 1). Preliminary data shows that temporally-deleted, ROSACreERT2/+;Hoxa11eGFP/-;Hoxd11LoxP/LoxP mice are unable to repair after fracture. Additionally, preliminary data suggest that the populations of osteoblasts and chondrocytes appear to be in abnormal in mutants. Using Hoxa11CreERT2 to enact both deletion and lineage labeling, I can mark the cells that have undergone recombination for isolation and transcriptomic analyses (Hoxa11eGFP/CreERT2;Hoxd11LoxP/LoxP; ROSAtd-Tomato/+, Aim 2). Fracture injury induces an acute response in which stem/progenitor expansion and differentiation to both skeletal lineages is occurring simultaneously, providing an excellent model to isolate single cells and identify the pathways and targets Hox genes regulate in these processes. Preliminary data shows that a large proportion of GFP+ cells are available for collection from the fracture callus, making single cell sequencing not only possible, but a highly effective tool to investigate transcriptomic change in Hox- expressing and Hox-lineage cells. The overall goal of this project is to define Hox genetic function in fracture repair and to identify the molecular mechanisms by which Hox genes regulate skeletal behavior in this process.

项目总结/文摘