Elucidating the Endothelial-Smooth Muscle Cell Interactions in Marfan Syndrome Using iPSCs

使用 iPSC 阐明马凡氏综合症中内皮-平滑肌细胞的相互作用

基本信息

批准号：
10156365
负责人：
Franklyn Hall
金额：
$ 4.6万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-31 至 2025-12-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10156365
关键词：
3-Dimensional Abnormal Cell Affect Animal Model Apoptosis Architecture Behavior Binding Biocompatible Materials Biological Assay Blood Vessels Caliber Cell Communication Cells Coculture Techniques Collagen Confocal Microscopy Connective Tissue Connective Tissue Diseases Contractile Proteins Cues Data Deposition Development Disease Disease Progression Early Diagnosis Elasticity Elastin Endothelial Cells Endothelium Enzyme-Linked Immunosorbent Assay Etiology Event Exposure to Extracellular Matrix Extracellular Matrix Proteins FBN1 Fibrin Fibronectins Genes Genetic Diseases Genetic study Human Impairment Lead MMP2 gene MMP9 gene Marfan Syndrome Matrix Metalloproteinases Mechanics Mediator of activation protein Methods Microfibrils Modeling Mutation NADPH Oxidase 1 Paracrine Communication Pathogenesis Pathology Patients Perfusion Phenotype Physiological Production Property Reactive Oxygen Species Regulation Signal Transduction Small Interfering RNA Smooth Muscle Myocytes Source Stimulus TGFB1 gene Tensile Strength Testing Therapeutic Therapeutic Intervention Time Tissues Transforming Growth Factor beta Transgenic Mice Tubular formation Vascular Graft Vascular Smooth Muscle acetovanillone base cell behavior diagnostic biomarker falls fastins fibrillin fibulin-4 hemodynamics in vitro Model in vivo induced pluripotent stem cell inhibitor/antagonist insight interdisciplinary approach mechanical force mechanical properties mouse model new therapeutic target novel overexpression pressure protein expression response scaffold shear stress therapeutic development therapeutic target

项目摘要

Project Summary Marfan Syndrome (MFS) is a genetic disorder of the connective tissue caused by mutations in the fibrillin- 1(FBN1) gene. These mutations cause increased vascular smooth muscle cell (vSMC) apoptosis, increased matrix metalloproteinase (MMP) expression, excessive elastin degeneration and collagen overexpression; weakening the vascular architecture, impair hemodynamic regulation, and ultimately leading to severe vascular complications. Primary cells, tissue explants, and transgenic mouse models suffer from insufficiencies that inhibit development of novel insights into disease pathogenesis or potential therapeutics; warranting new modeling platforms. Recent advances in biomaterials have generated vascular grafts that mimic properties of the extracellular matrix (ECM) in the natural vasculature. Human induced pluripotent stem cells (hiPSC) are an ideal, patient specific renewable cell source that provides an avenue to study how genetic diseases effect cell mechanobiology, signaling and function to better development therapeutics. Here, using a natural fibrin-based vascular graft and hiPSCs from patients with Marfan Syndrome, we aim to: (1) fully characterized both ECs and contractile vSMCs from hiPSCs derived from Marfan patients, (2) study the responses of hiPSC-ECs to shear force and hiPSC-vSMCs to circumferential strain, (3) study the impacts of mechanical forces (i.e. shear force and circumferential strain) and paracrine signaling in a co-culture model, specifically reactive oxygen species(ROS), on vSMC phenotype and rate of graft degradation using the fibrin-based vascular graft. Our multidisciplinary approach underpinning this project will elucidate key mediators of MFS disease progression towards early detection and therapeutic targets.

项目概要马凡氏综合症 (MFS) 是一种由原纤维蛋白突变引起的结缔组织遗传性疾病。 1（FBN1）基因。这些突变导致血管平滑肌细胞 (vSMC) 凋亡增加、基质金属蛋白酶（MMP）表达、弹性蛋白过度变性和胶原蛋白过度表达；削弱血管结构，损害血流动力学调节，最终导致严重的血管损伤并发症。原代细胞、组织外植体和转基因小鼠模型存在缺陷，抑制对疾病发病机制或潜在治疗方法的新见解的发展；保证新的建模平台。生物材料的最新进展已经产生了模仿血管特性的血管移植物自然脉管系统中的细胞外基质（ECM）。人类诱导多能干细胞 (hiPSC) 是一种理想的、患者特异性的可再生细胞源，为研究遗传疾病如何影响细胞提供了途径机械生物学、信号传导和功能，以更好地开发治疗方法。在这里，使用基于天然纤维蛋白的对于来自马凡氏综合症患者的血管移植物和 hiPSC，我们的目标是：(1) 充分表征这两种 EC 和来自马凡患者 hiPSC 的收缩性 vSMC，(2) 研究 hiPSC-EC 对剪切力和 hiPSC-vSMCs 对周向应变的影响，(3) 研究机械力（即剪切力）的影响力和周向应变）和共培养模型中的旁分泌信号传导，特别是活性氧种（ROS），对 vSMC 表型和使用基于纤维蛋白的血管移植物的移植物降解率的影响。我们的支持该项目的多学科方法将阐明 MFS 疾病进展的关键介质实现早期检测和治疗目标。