Developmental regulation of cranial tendon fibroblast diversity and ECM interactions

颅腱成纤维细胞多样性和 ECM 相互作用的发育调节

基本信息

批准号：
10583541
负责人：
Thomas F Schilling
金额：
$ 37.77万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2026-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10583541
关键词：
Address Aging Atrophic Binding Proteins Cells Cephalic Contracts Data Development Disease Embryo Extracellular Matrix Feedback Fibroblasts Gene Expression Gene Expression Profile Gene Expression Regulation Genetic Genetic Diseases Genetic Transcription Genetic study Goals Head Heterogeneity Human Image In Situ Knowledge Ligaments Limb structure Location Maintenance Mesoderm Methods Molecular Morphogenesis Mus Muscle Muscle Contraction Musculoskeletal Development Musculoskeletal System Neural Crest Cell Paralysed Pharmacological Treatment Physiological Play Population Production Property Regulation Research Role Scaffolding Protein Shock Signal Transduction Site Sorting Specific qualifier value Tendon Injuries Tendon structure Testing Thrombospondins Tissues Transforming Growth Factor beta Tretinoin Variant Weight-Bearing state Zebrafish absorption bone cell type craniofacial developmental genetics effective therapy experimental study genetic manipulation in vivo in vivo imaging insight ligament injury mechanical force mechanical load mutant novel pharmacologic progenitor repaired response scaffold scleraxis single-cell RNA sequencing skeletal stem cells tendon development thrombospondin 4 transcription factor transcriptomics

项目摘要

Project Summary/Abstract Tendons and ligaments are fundamental components of a functional musculoskeletal system. Tendons attach muscles to bone and interact with these tissues at distinct attachment sites called entheses (bone) and myotendinous junctions (MTJs, muscle). These interactions cause unique changes in gene expression and extracellular matrix (ECM) production that allow tendons to bear the forces exerted by muscle contraction. Transcription factors such as Scleraxis (Scx) specify early tendon progenitor cells (TPCs) and regulate ECM production. We previously discovered a critical ECM scaffolding protein called Thrombospondin-4b (Tsp4b) in zebrafish that is regulated by Scx, required for tendon maintenance and conserved in human tendons. How different types of tendon fibroblasts (tenocytes) are specified and influence ECM assembly at entheses or MTJs remains unclear. The current proposal addresses these issues using the advantages of the zebrafish for single cell RNA sequencing, in vivo imaging and genetic manipulation. The long-term goal of the proposed research is to understand the spatial dynamics of gene regulation and ECM assembly in tendons/ligaments and their regulation by mechanical force. Three primary hypotheses guide the research: 1) distinct subtypes of TPCs develop at entheses and MTJs in response to force, 2) ECM secreted by tenocytes regulates force-dependent signals that alter these distinct modes of gene expression in tenocytes and 3) retinoic acid is a novel force-dependent signal controlling tendon development. Aim 1 will perform scRNA-seq in tenocytes and see how force alters gene expression profiles. Aim 2 will study roles for Tsp4b, ECM, and TGF-beta signaling in force-dependent gene expression in tendons. Aim 3 will study the roles of RA signaling in tendons, and its responses to mechanical load. Each aim combines novel single cell approaches, genetic manipulation, live imaging and quantitative methods for physiological stimulation of muscles to get at mechanisms of tendon cell specification and ECM assembly in response to force.

项目摘要/摘要肌腱和韧带是功能性肌肉骨骼系统的基本成分。肌腱将肌肉固定在骨头上，并在称为恩斯的不同附着位置与这些组织相互作用（骨头）和肌无力的连接（MTJS，肌肉）。这些相互作用导致基因独特的变化表达和细胞外基质（ECM）产生，允许肌腱承受施加的力肌肉收缩。诸如硬化（SCX）之类的转录因子指定早期肌腱祖细胞（TPC）并调节ECM生产。我们以前发现了关键的ECM脚手架蛋白在斑马鱼中称为斑马鱼中的Thrombospondin-4b（TSP4B），由SCX调节，肌腱需要维护和在人肌腱中保守。不同类型的肌腱成纤维细胞（替核细胞）如何指定并影响ENHES或MTJS处的ECM组装尚不清楚。当前的建议使用斑马鱼对单细胞RNA测序的优势解决这些问题，体内成像和遗传操作。拟议研究的长期目标是了解肌腱/韧带中基因调节和ECM组装的空间动力学及其调节机械力。三个主要假设指导研究：1）TPC的不同亚型发展在响应力时，在恩特斯和mtjs上，2）tenocytes分泌的ECM调节力依赖性改变了养育细胞中基因表达的这些不同模式的信号和3）视黄酸是一种新颖的力依赖性信号控制肌腱发育。 AIM 1将在tenocytes中执行SCRNA-SEQ，并且查看力如何改变基因表达谱。 AIM 2将研究TSP4B，ECM和TGF-BETA的角色在力依赖性基因表达中的信号传导。 AIM 3将研究RA信号在中的作用肌腱及其对机械负荷的反应。每个目标结合了新型的单细胞方法，遗传操纵，实时成像和定量方法，用于肌肉的生理刺激响应力响应肌腱细胞规范和ECM组装的机理。