Syndecan-1 in Mechanosensing of Engineered Microenvironments

Syndecan-1 在工程微环境机械传感中的应用

基本信息

批准号：
9387690
负责人：
Aaron Blair Baker
金额：
$ 25.17万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-15 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9387690
关键词：
Adhesions Affect Atherosclerosis Biochemical Biocompatible Materials Biological Assay Biophysics Biosensor Blood Vessels Blood capillaries Cell Differentiation process Cell Line Cell Surface Receptors Cell physiology Cell surface Cells Cellular Mechanotransduction Collagen Fiber Core Protein Cues Cytoplasmic Tail Cytoskeleton Development Devices Disease Endothelial Cells Engineering Environment Fluorescence Resonance Energy Transfer Focal Adhesions Generations Genetic Goals Grant Heparitin Sulfate Homeostasis Human Cell Line Imagery Implant Inflammatory Ion Channel Knock-out Knowledge Lentivirus Vector Leukocytes Measures Mechanics Mediating Medical Mesenchymal Microfabrication Molecular Mutate Mutation Nanotopography Pathogenesis Pathway interactions Phenotype Process Proteoglycan Regulation Role Signal Pathway Signal Transduction Site Site-Directed Mutagenesis Stretching Surface Techniques Testing Therapeutic Tissue Engineering Tissues Transforming Growth Factor beta Vascular Diseases Vascular Graft Vascular Smooth Muscle Vascular System base capillary cell behavior design effective therapy genetically modified cells glycosylation implant material improved inflammatory marker insight knock-down link protein lithography macrophage mechanotransduction nanopattern nanoscale novel polysulfated glycosaminoglycan response restenosis rho GTP-Binding Proteins scaffold sensor shear stress small hairpin RNA syndecan tool

Adhesions Affect Atherosclerosis Biochemical Biocompatible Materials Biological Assay Biophysics Biosensor Blood Vessels Blood capillaries Cell Differentiation process Cell Line Cell Surface Receptors Cell physiology Cell surface Cells Cellular Mechanotransduction Collagen Fiber Core Protein Cues Cytoplasmic Tail Cytoskeleton Development Devices Disease Endothelial Cells Engineering Environment Fluorescence Resonance Energy Transfer Focal Adhesions Generations Genetic Goals Grant Heparitin Sulfate Homeostasis Human Cell Line Imagery Implant Inflammatory Ion Channel Knock-out Knowledge Lentivirus Vector Leukocytes Measures Mechanics Mediating Medical Mesenchymal Microfabrication Molecular Mutate Mutation Nanotopography Pathogenesis Pathway interactions Phenotype Process Proteoglycan Regulation Role Signal Pathway Signal Transduction Site Site-Directed Mutagenesis Stretching Surface Techniques Testing Therapeutic Tissue Engineering Tissues Transforming Growth Factor beta Vascular Diseases Vascular Graft Vascular Smooth Muscle Vascular System base capillary cell behavior design effective therapy genetically modified cells glycosylation implant material improved inflammatory marker insight knock-down link protein lithography macrophage mechanotransduction nanopattern nanoscale novel polysulfated glycosaminoglycan response restenosis rho GTP-Binding Proteins scaffold sensor shear stress small hairpin RNA syndecan tool

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项目摘要

The cells of the vascular system are uniquely sensitive to biophysical cues from applied forces and their local cellular microenvironment. In engineering materials for vascular therapies or for delivery of cells, a key challenge is to understand the mechanisms that cells use to sense biophysical cues from their environment to enable more effective therapies. Syndecans are proteoglycans that consist of a protein core modified with heparan sulfate glycosaminoglycan chains. Due to their presence on the cell surface and their interaction with cytoskeletal and focal adhesion associated molecules, cell surface proteoglycans are ideally located to serve as mechanosensors of the cellular microenvironment. Our group recently found that syndecan-1 (SDC1) regulates vascular smooth muscle cell differentiation and the response to shear stress in endothelial cells. We hypothesize that SDC1 is a mechanosensor for substrate-mediated cues and can be targeted to alter endothelial cell- biomaterial interactions to better control cell function on these engineered materials. In Aim 1, we will examine the mechanistic role of SDC1 in regulating vascular mechanosensing of substrate compliance and nanotopology. We will create endothelial cell lines with altered expression or mutation in SDC1 and determine how these alterations affect mechanosensing on engineered substrates with varying compliance and nanotopology. Specifically, we will investigate the ability of SDC1 to regulate mechanically mediated signaling through pathways including the Hippo pathway, TGF-β and Rho-GTPases. In addition, we will examine phenotypic regulation and endothelial-to- mesenchymal transition of endothelial cells on these substrates in the presence of biochemical treatments. In Aim 2, we will develop a novel, FRET-based molecular force biosensor that will enable us to examine the forces that are applied to SDC1 during interactions of live cells with the engineered substrates. This biosensor will enable us to probe the importance of the molecular subdomains of SDC1 in mechanosensing, including the glycosylation sites and the cytoplasmic domains. Together, our findings will contribute to our understanding of cellular mechanotransduction and provide needed knowledge for the development of improved vascular devices and cell delivery scaffolds.

血管系统的细胞对来自应用力和他们当地的细胞微环境。用于血管疗法的工程材料或提供细胞，关键挑战是了解细胞用来感知生物物理提示的机制他们的环境使得更有效的疗法。 syndecans是由蛋白质核心用硫酸乙酰肝素糖胺聚糖链修饰。由于他们在细胞表面及其与细胞骨架和局灶性粘合剂相关分子的相互作用，细胞表面蛋白聚糖是理想位置的，可作为细胞的机械学微环境。我们的小组最近发现Syndecan-1（SDC1）调节血管平滑肌肉细胞分化以及对内皮细胞剪切应力的反应。我们假设这一点 SDC1是底物介导的提示的机械力学方法，可以针对改变内皮细胞 - 生物材料相互作用以更好地控制这些工程材料的细胞功能。在AIM 1中，我们将检查SDC1在控制底物的血管机械传感中的机械作用合规性和纳米学。我们将创建具有更改表达或更改的内皮细胞系或 SDC1中的突变并确定这些变化如何影响工程的机制求解具有不同依从性和纳米学的底物。具体来说，我们将研究 SDC1通过包括河马途径的途径来调节机械介导的信号传导， TGF-β和Rho-GTPases。此外，我们将检查表型调节和内皮调节在存在生化的情况下，内皮细胞在这些底物上的间充质转变治疗。在AIM 2中，我们将开发一种基于FRET的新型分子力生物传感器，将启用我们检查活细胞与工程的相互作用期间应用于SDC1的力基材。该生物传感器将使我们能够探究分子亚域的重要性机械感应中的SDC1，包括糖基化位点和细胞质结构域。一起，我们的发现将有助于我们对蜂窝机械转移的理解，并提供了所需的开发改进的血管设备和细胞输送支架的知识。