In vivo two-photon imaging of vascular invasion and stem cell translocation in calvarial bone

颅骨血管侵袭和干细胞易位的体内双光子成像

基本信息

批准号：
10603163
负责人：
Andy Y Shih
金额：
$ 29.54万
依托单位：
SEATTLE CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10603163
关键词：
Address Adult Blood Vessels Bone Development Bone Marrow Bone Regeneration Bone Tissue Bone remodeling Calvaria Cartilage Cell Lineage Cells Defect Development Developmental Process Distant Embryonic Development Endothelial Cells Esthetics Fluorescent Dyes Future GLI gene Goals Histologic Homeostasis Iatrogenesis Image Injury Intravenous Invaded Knowledge Label Maintenance Mesenchyme Molecular Mus Natural regeneration Osteoblasts Osteoclasts Osteogenesis Pericytes Periosteum Physiologic Ossification Population Regulation Reporter Research Research Project Grants Research Proposals Site Skeleton Surgical sutures Tamoxifen Techniques Testing Time Transgenic Mice Trauma Vascularization Visualization analog bone bone healing bone repair cell behavior congenital anomaly cortical bone craniofacial development healing in vivo imaging in vivo two-photon imaging injured injury and repair intramembranous bone formation long bone novel therapeutic intervention osteoclastogenesis postnatal postnatal development regenerative serial imaging skeletal stem cell stem cell niche stem cells substantia spongiosa two-photon

项目摘要

PROJECT SUMMARY Calvarial bone defects commonly occur as a result of trauma, congenital anomalies and iatrogenic conditions. The healing of bone defects relies on the regenerative capability of calvarial bones to replace damaged bone tissues and the availability of skeletal stem cells (SSCs) is one of the key factors for generating new bones to restore both structural and functional integrity. Calvarial SSCs that express marker gene Gli1, Axin2 or Prx1 reside in the sutural stem cell niche and contribute to calvarial bone homeostasis and healing after injury. It remains unclear how calvarial SSCs translocate from the sutural niche to distant regions of the calvarial bones for bone repair after injury of sites remote to the suture. Our preliminary analysis through 2-photon imaging indicates that Gli1-expressing calvarial SSCs not only reside in the sutural niche but also line the walls of blood vessels distributed widely throughout the postnatal calvarial bones. In this project, we will test the hypothesis that calvarial SSCs in the sutural niche could translocate with vascular invasion that initiates at the postnatal suture, and could reside within calvarial bones to contribute to bone regeneration after injury. We propose two specific aims for this project: 1) Determine if vascular invasion from the suture results in bone marrow cavity formation in postnatal calvarial bones, and 2) Determine if calvarial SSCs in the sutural niche translocate with vascular invasion to establish the SSC niche in bone marrow cavities of postnatal calvarial bones. This research proposal leverages the unique expertise of two investigative labs to address a key gap in our knowledge of calvarial healing. Successful completion of this exploratory research will set the stage for further mechanistic studies such as molecular regulation of calvarial SSC translocation, which could facilitate the development of new therapeutic approaches for rapid bone regeneration after injury.

项目概要颅骨骨缺损通常是由外伤、先天异常和医源性疾病引起的。骨缺损的愈合依赖于颅骨的再生能力来替代受损的骨组织和骨骼干细胞（SSC）的可用性是生成新骨骼的关键因素之一恢复结构和功能的完整性。表达标记基因 Gli1、Axin2 或 Prx1 的颅骨 SSC 存在于骨缝干细胞生态位中，有助于颅骨骨稳态和损伤后的愈合。它目前尚不清楚颅骨 SSC 如何从骨缝微环境转移到颅骨的远处区域用于远离缝合部位损伤后的骨修复。我们通过 2 光子成像进行初步分析表明表达 Gli1 的颅骨 SSC 不仅存在于骨缝微环境中，而且还排列在血壁上血管广泛分布在出生后的颅骨中。在这个项目中，我们将测试假设骨缝微环境中的颅骨 SSC 可能会随着出生后开始的血管侵袭而易位缝合线，并且可以驻留在颅骨内，有助于受伤后的骨再生。我们建议两个该项目的具体目标：1) 确定缝线的血管侵入是否会导致骨髓腔出生后颅骨的形成，以及 2) 确定颅缝微环境中的颅骨 SSC 是否易位血管侵袭以在出生后颅骨的骨髓腔中建立 SSC 生态位。这研究提案利用两个研究实验室的独特专业知识来解决我们的关键差距颅骨愈合的知识。这项探索性研究的成功完成将为进一步的研究奠定基础机制研究，例如颅骨 SSC 易位的分子调控，这可以促进开发损伤后快速骨再生的新治疗方法。