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

基于人 iPSC 的马凡综合征模型中主动脉平滑肌细胞的力学生物学

• 批准号: 10215619
• 负责人: Robert Wiener
• 金额: $ 4.44万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-24 至 2022-07-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10215619
• 关键词: 3-Dimensional; Abdomen; Actins; Affect; Anatomy; Aneurysm; Animal Model; Animals; Antihypertensive Agents; Aorta; Aortic Aneurysm; Biochemical; Bioinformatics; Biological; Biological Markers; Biomedical Engineering; Calcium Signaling; Cardiovascular system; Cell Culture Techniques; Cell Differentiation process; Cell model; Cells; Cessation of life; Clinical; Clinical Trials; Congenital Abnormality; Connective Tissue Diseases; Core Facility; Critiques; Cytoskeleton; DNA Sequence Alteration; Data; Data Set; Descending aorta; Development; Disease; Dissection; Engineering; Exhibits; Experimental Designs; Extracellular Matrix; FBN1; Fellowship; Fluorescence; Future; Gene Expression Profile; Gene set enrichment analysis; Genes; Germ; Germ Layers; Goals; High Prevalence; Human; In Vitro; Incidence; Individual; Intervention; Investigation; Journals; Lead; Length; Location; Losartan; Marfan Syndrome; Measures; Mechanics; Medial; Mentors; Modeling; Neural Crest; Neuroectoderm; Outcome; Output; Paraxial Mesoderm; Patients; Phenotype; Physiological; Plant Roots; Population; Predisposition; Property; Proteomics; Publishing; Research; Research Personnel; Role; Rupture; Scanning Probe Microscopes; Serum; Signal Transduction; Smooth Muscle Myocytes; Stimulus; Stress; Students; Sudden Death; Technical Expertise; Techniques; Testing; Therapeutic; Therapeutic Intervention; Time; Tissue Engineering; Tissue Model; Tissue-Specific Gene Expression; Tissues; Training; Transforming Growth Factor beta; Vascular remodeling; abdominal aorta; ascending aorta; base; bioinformatics pipeline; biomarker discovery; body system; career; cell dimension; collaborative environment; confocal imaging; drug candidate; effective therapy; healthy volunteer; hemodynamics; heritable connective tissue disorder; human model; hydrogel scaffold; improved; in vitro Model; induced pluripotent stem cell; insight; lead candidate; machine learning algorithm; mechanotransduction; meetings; multidimensional data; nanoindentation; next generation; novel; prophylactic; protein biomarkers; protein expression; regional difference; response; skills; stem; stem cell model; stem cells; symposium; symptom management; targeted treatment; transcriptome sequencing; transcriptomics; two-dimensional; vascular tissue engineering

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.

项目摘要 Marfan综合征（MFS）是最常见的遗传结缔组织障碍之一，影响5,000分之一 个人并在多个器官系统中具有破坏性表现；值得注意的是心血管系统。 MFS是一种由原纤维蛋白-1基因的基因突变引起的常染色体显性疾病，导致异常 TGFβ信号传导，并经常导致主动脉瘤，解剖和死亡。有趣的是，关联 主动脉壁内的变性几乎总是发生在主动脉根或上升主动脉中，而不是在 下降或腹主动脉；虽然这预定反映了血液动力学应激的区域差异，但 仅降压治疗在MFS中管理主动脉瘤就无法有效。或者，它也是 主动脉平滑肌细胞（ASMC）占主导地位的情况下 血管壁具有基于其独特的发育生殖器的异质亚型 解剖位置； Neurocectoderm（NE）起源会引起上升的ASMC和近期中胚层（PM） 来源引起降落的ASMC。该项目将使用特定于原点的ASMC与诱导的 来自MFS和健康对照患者的多能干细胞（IPSC），以检验一个新的假设，即 发育起源引起与媒体退化相关的ASMC的位置特异性异常 MFS。此外，它将探索Marfan综合征的前先天性缺陷的生物标志物 确定预防治疗干预的新型靶标。这些研究将表征表型 人类ASMC亚型在细胞和组织水平上的差异随干细胞培养和血管的差异 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 这些细胞和组织模型上的蛋白质组学分析，以培养生物信息学丰富的生物学特征 分析。此外，我们将开发出生物信息学管道，以阐明新型预防目标 本质上负责使用我们的独特组合 表型，转录组和蛋白质组学结果作为输入。最后，根据我们的生物信息输出，我们将 测试我们对基于人类IPSC的体外模型的干预措施，并与Losartan进行比较 常用的抗高血压药物也表现出独特的抗塑料特性，并显示了 有望管理动物和患者中MF的症状。该奖学金的培训计划将 专注于技术技能，实验设计和批判性分析，批评已发表的科学数据以及 演讲技巧。这将通过常规的心理会议，期刊俱乐部，会议演讲，双 年度委员会会议和高级课程。大多数培训将发生在Costa Lab的 ISMMS的心血管研究中心，这是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。在 Ramirez Lab与Marfan综合征和生化研究技术的高级研究人员。