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

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施加的力和作用的生物物理信号特别敏感。用于血管治疗或输送的工程材料。对于细胞来说，一个关键的挑战是了解细胞用来感知生物物理线索的机制 Syndecans 是由以下成分组成的蛋白聚糖：由于硫酸乙酰肝素糖胺聚糖链存在于蛋白质核心上。细胞表面及其与细胞骨架和粘着斑相关分子、细胞的相互作用表面蛋白聚糖的位置非常理想，可用作细胞的机械传感器我们课题组最近发现syndecan-1（SDC1）调节血管平滑肌。肌细胞分化和内皮细胞对剪切应力的反应。 SDC1 是一种用于底物介导信号的机械传感器，可用于改变内皮细胞在目标 1 中，我们将通过生物材料相互作用更好地控制这些工程材料上的细胞功能。检查 SDC1 在调节底物血管机械传感中的机制作用我们将创建具有表达或纳米拓扑结构的内皮细胞系。 SDC1 突变并确定这些改变如何影响工程化机械传感具体来说，我们将研究具有不同顺应性和纳米拓扑结构的基板。 SDC1 通过 Hippo 途径等途径调节机械介导的信号传导，此外，我们将检查表型调节和内皮-GTP 酶。在生化物质存在的情况下，内皮细胞在这些基质上的间质转化在目标 2 中，我们将开发一种新型的、基于 FRET 的分子力生物传感器，该传感器将能够实现。我们检查活细胞与工程细胞相互作用期间施加到 SDC1 的力该生物传感器将使我们能够探究分子子域的重要性。机械传感中的 SDC1，包括糖基化位点和细胞质结构域。我们的研究结果将有助于我们对细胞力转导的理解，并提供所需的信息开发改进的血管装置和细胞输送支架的知识。