Mechanobiology of aortic smooth muscle cells in human iPSC-based models of Marfan Syndrome

项目摘要 马凡氏综合征（MFS）是最常见的遗传性结缔组织疾病之一，发病率为1/5000 个体和具有破坏性的表现在多个器官系统;特别是心血管系统。 MFS是一种常染色体显性遗传疾病，由原纤维蛋白-1基因突变引起，导致异常的免疫反应。 TGFβ信号传导，并且经常导致主动脉瘤、夹层和死亡。有趣的是，相关的 主动脉血管壁内的变性几乎总是发生在主动脉根或升主动脉中，而不是发生在主动脉根部或升主动脉中。 降主动脉或腹主动脉;虽然该脓液反映了血液动力学应激的区域差异， 单纯抗高血压治疗对MFS患者的主动脉瘤无效。或者，它也是 主动脉平滑肌细胞（ASMCs），主要是血管活性中层的情况下， 血管壁，具有源于不同发育胚层的异质亚型， 解剖位置;神经外胚层（NE）起源产生上行ASMC和轴旁中胚层（PM） 起源引起ASMC下降。本项目将使用来源特异性ASMC， 来自MFS患者和健康对照的多能干细胞（iPSC），以测试一种新的假设， 发育起源导致ASMCs中与中膜变性相关的位置特异性异常， MFS。此外，它将探索马凡氏综合征症状前先天性缺陷的生物标志物， 确定预防性治疗干预的新靶点。这些研究将描述表型 在细胞和组织水平上，人ASMC亚型与干细胞培养和血管内皮细胞的差异 组织工程技术使用最先进的核心设施，我们还将进行转录组和 对这些细胞和组织模型进行蛋白质组学分析，以培养丰富的生物学特征， 分析.此外，我们将开发一个生物信息学管道，以阐明新的预防目标 固有地负责升主动脉MFS诱导的中膜变性，使用我们独特的组合 表型、转录组和蛋白质组结果作为输入。最后，根据我们的生物信息学输出，我们将 在我们的基于人iPSC的体外模型上测试我们的干预，并与氯沙坦治疗进行比较， 一种常用的抗高血压药物，也表现出独特的抗重塑特性， 在动物和患者中管理MFS症状的承诺。该研究金的培训计划将 专注于技术技能，实验设计和批判性分析，对已发表的科学数据的批判，以及 演讲技巧它将通过定期的导师会议，期刊俱乐部，会议演示，双- 年度委员会会议和高级课程大部分培训将在Costa实验室进行， ISMMS的心血管研究中心，一个高度活跃和协作的环境， 导师和学生与我的研究主题和职业目标保持一致。其他培训将在 拉米雷斯实验室与马凡氏综合征和生化调查技术的高级研究人员。

PROJECT SUMMARY Marfan Syndrome (MFS), one of the most common heritable connective tissue disorders, affects 1 in 5,000 individuals and has destructive manifestations in multiple organ systems; notably the cardiovascular system. MFS is an autosomal dominant disease caused by a genetic mutation in the Fibrillin-1 gene leading to aberrant TGFβ signaling, and frequently results in aortic aneurysm, dissection, and death. Interestingly, the associated degeneration within the aortic vessel wall almost always occurs in the aortic root or ascending aorta and not in the descending or abdominal aorta; while this putatively reflects regional differences in hemodynamic stress, antihypertensive treatment alone is not effective in managing aortic aneurysm in MFS. Alternatively, it is also the case that aortic smooth muscle cells (ASMCs), which predominate the vasoactive medial layer of the vessel wall, have heterogeneous subtypes stemming from distinct developmental germ-layers based on their anatomical location; Neuroectoderm (NE) origin gives rise to ascending ASMCs and Paraxial mesoderm (PM) origin gives rise to descending ASMCs. This project will use origin-specific ASMCs differentiated from induced pluripotent stem cells (iPSCs) from patients with MFS and healthy controls to test a novel hypothesis that developmental origin causes location-specific abnormalities in ASMCs associated with medial degeneration in MFS. Additionally, it will explore for biomarkers of presymptomatic congenital defects in Marfan Syndrome to identify novel targets for prophylactic therapeutic intervention. These studies will characterize phenotypic differences in human ASMC subtypes at the cellular and tissue level with stem-cell culturing and vascular tissue engineering techniques. Using state-of-the-art core facilities we will also conduct transcriptomic and proteomic analysis on these cellular and tissue models to cultivate a rich biological profile for bioinformatic analysis. Furthermore, we will develop a bioinformatics pipeline to elucidate novel prophylactic targets inherently responsible for ascending aortic MFS-induced medial degeneration, using our uniquely combined phenotypic, transcriptomic, and proteomic results as input. Lastly, based on our bioinformatic outputs we will test our intervention on our human iPSC-based in vitro models and compare to treatment with Losartan, a commonly used anti-hypertensive drug that also exhibits unique anti-remodeling properties and has shown promise for managing the symptoms of MFS in animals and in patients. The training plan for this fellowship will focus on technical skills, experimental design and critical analysis, critique of published scientific data, and presentation skills. It will be achieved by regular mentor meetings, journal clubs, conference presentations, bi- annual committee meetings, and advanced coursework. The majority of training will occur in the Costa Lab at the Cardiovascular Research Center at ISMMS, a highly active and collaborative environment with available mentors and students aligned with my research topics and career goals. Additional training will occur in the Ramirez Lab with senior researchers in Marfan Syndrome and biochemical investigation techniques.