Elucidating the role of cardiac myofibroblasts on matrix and vasculature remodeling

阐明心肌成纤维细胞对基质和脉管系统重塑的作用

基本信息

批准号：
10469639
负责人：
EMILY OLSZEWSKI
金额：
$ 0.26万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-16 至 2022-06-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10469639
关键词：
3-Dimensional Affect Aging Aortic Valve Stenosis Apoptosis Area Arteries Artificial Heart Atomic Force Microscopy Automobile Driving Biochemical Biometry Blood Vessels Cardiac Cardiac Myocytes Cardiovascular Physiology Cause of Death Cells Characteristics Chemicals Chemistry Collagen Collagen Fiber Confocal Microscopy Contracts Coronary Cues Data Deposition Disease Endothelial Cells Endothelium Engineering Environment Extracellular Matrix Extracellular Space Fibroblasts Fibrosis Freeze Drying Gel Gene Activation Generations Genetic Models Goals Heart Heart Diseases Heart failure Hypertension Hypertrophy Image In Vitro Injury Lectin Left Length Liquid Chromatography Longitudinal Studies MAP2K6 gene Maps Measurement Measures Mechanics Mediating Methods Modeling Molecular Mus Myocardial Infarction Myofibroblast Nature Operative Surgical Procedures Organ Pathway interactions Perfusion Perivascular Fibrosis Phenotype Phosphotransferases Physiological Physiology Preparation Process Property Protein Kinase Proteomics Pump Regulation Role Signal Transduction Stimulus Stress Stretching Structure System Therapeutic Three-Dimensional Image Time Tissues Traction Transgenes Transgenic Mice Vascular remodeling Ventricular Work blebbistatin cell growth cell motility cell type combinatorial coronary fibrosis density experimental study extracellular heart function hypertensive image reconstruction in vitro Model in vivo in vivo Model interstitial knock-down mathematical model mechanical properties mechanical signal microscopic imaging migration overexpression preservation pressure response scaffold second harmonic tandem mass spectrometry two-photon

项目摘要

In every form of heart disease, the secretion of extracellular matrix (ECM) by activated fibroblasts, or myofibroblasts, results in cardiac fibrosis. Fibrosis impedes compliance and pumping function, ultimately leading to heart failure due to left ventricular dilation and loss of mechanical function. Little is known about endothelial cell and vessel adaptations to the environment or how local mechanics and chemistry impact vessel structure and flow in vivo. Combinatorial fibroblast and ECM mechanical and chemical crosstalk with endothelial cells are unknown. Moreover, in vitro models aiming to assess vascular adaptations to an extracellular environment lack physiologically relevant ECMs and instead provide exogenous ECM components to optimize control of variables. A system for in vivo, cell-specific phenotypic manipulation will allow for controlled perturbations at an organ level while maintaining relevant, native ECM remodeling over time. Thus, I propose to examine transgenic mice with cardiac fibroblast-specific overexpression of a constitutively active mitogen-activated protein kinase kinase 6 (MKK6) to study vascular remodeling with respect to the fibroblasts and the ECM they secrete. These mice were previously shown to develop interstitial and perivascular fibrosis after 16-20 weeks of the MKK6 gene activation without an injury stimulus, serving as an effective model of the interstitial fibrosis preserved across the results of aging, hypertension, aortic stenosis, and other diseases of the heart. Importantly, the remodeling seen in these diseases does not involve a massive loss of cardiomyocytes, as in a myocardial infarction, but rather a conserved fibroblast phenotypic change, an altered extracellular space, and/or restricted vascular flow over time. Similarly, manipulation of the MKK6 pathway allows for overexpression or knockdown of cardiac fibroblast activation, corresponding to increased ECM or the inability to secrete ECM as a response to a stimulus, respectively. First, I propose to study the biochemical, structural, and mechanical properties of the ECM as well as the macro- and microvascular responses to activated, quiescent, and control cardiac fibroblast phenotypes in vivo. Second, three-dimensional vessel-like structures with controlled fibroblasts and ECMs will be engineered as in vitro platforms to define the molecular regulators of vascular remodeling induced by microenvironmental cues. The effects of combined signaling will be resolved by global characterization along with a reductionist method. The goal of this work is to inform heart therapies by providing targets for steering cardiac vascular remodeling.

在各种形式的心脏病中，激活细胞外基质（ECM）的分泌成纤维细胞或肌纤维细胞会导致心脏纤维化。纤维化阻碍了合规性和抽水功能，最终导致左心室扩张和丧失导致心力衰竭机械功能。关于内皮细胞和容器适应对环境或局部力学和化学如何影响血管结构和体内流动。组合成纤维细胞以及ECM机械和化学串扰与内皮细胞是未知。此外，旨在评估细胞外血管适应的体外模型环境缺乏生理相关的ECM，而是提供外源性ECM 组件以优化对变量的控制。体内，细胞特异性表型的系统操纵将允许在维持相关性的同时，在器官级别受控扰动，随着时间的推移，本地ECM重塑。因此，我建议用心脏检查转基因小鼠组成型活性有丝分裂原活性蛋白激酶的成纤维细胞特异性过表达激酶6（MKK6）研究成纤维细胞和ECM的血管重塑分泌。这些小鼠先前被证明会在没有损伤刺激的MKK6基因激活16-20周，可作为有效在衰老，高血压，主动脉的结果中保存的间质纤维化模型狭窄和心脏的其他疾病。重要的是，在这些疾病中看到的重塑不像心肌梗塞一样涉及大量心肌细胞的损失，而是保守的成纤维细胞表型变化，细胞外空间改变和/或受限随时间流动的血管流动。同样，对MKK6途径的操作允许过表达或敲低心脏成纤维细胞激活，对应于ECM增加或无法分别分泌ECM作为对刺激的反应。首先，我建议研究 ECM的生化，结构和机械性能以及宏观和宏观和机械性能对激活，静止和控制心脏成纤维细胞表型的微血管反应体内。第二，具有受控成纤维细胞和ECM的三维管子结构将被设计为体外平台，以定义血管重塑的分子调节剂由微环境提示诱导。组合信号的影响将通过全球表征以及简化主义方法。这项工作的目的是告知心脏通过提供转向心脏血管重塑的靶标的疗法。