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

项目摘要 马凡综合征(MFS)是一种由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(MMPs)表达，弹性蛋白过度变性，胶原过度表达； 削弱血管结构，损害血流动力学调节，最终导致严重的血管 并发症。原代细胞、组织外植体和转基因小鼠模型存在不足 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产生了模仿血管特性的血管移植物 天然血管中的细胞外基质(ECM)。人诱导多能干细胞是一种 理想的、患者特定的可再生细胞来源，为研究遗传病如何影响细胞提供了一条途径 机制生物学，信号和功能，以更好地开发治疗学。在这里，使用一种基于天然纤维蛋白的 来自马凡综合征患者的血管移植物和HiPSCs，我们的目标是：(1)充分表征这两种内皮细胞 和来自Marfan患者的HiPSC的收缩VSMCs，(2)研究HiPSC-ECs对 剪切力和HiPSC-vSMCs对周向应变的影响；(3)研究机械力(即剪切)对周向应变的影响 力和周向应变)和旁分泌信号在共培养模型中，特别是活性氧 使用基于纤维蛋白的血管移植物的vSMC表型和移植物降解率。我们的 支持该项目的多学科方法将阐明MFS疾病进展的关键中介因素 向早期发现和治疗靶点迈进。

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.