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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要马凡氏综合症 (MFS) 是最常见的遗传性结缔组织疾病之一，影响五千分之一的人个体并在多个器官系统中具有破坏性表现；特别是心血管系统。 MFS 是一种常染色体显性遗传疾病，由 Fibrillin-1 基因的基因突变导致异常 TGFβ 信号传导，经常导致主动脉瘤、夹层和死亡。主动脉血管壁内的变性几乎总是发生在主动脉根部或升主动脉，而不是在降主动脉或腹主动脉；虽然这可能反映了血流动力学压力的区域差异，单独的抗高血压治疗对于治疗 MFS 的主动脉瘤是无效的。主动脉平滑肌细胞（ASMC）在主动脉内层的血管活性中占主导地位。血管壁，具有异质亚型，源于不同的发育胚层，基于它们的解剖位置；神经外胚层 (NE) 起源产生上行 ASMC 和轴旁中胚层 (PM) 起源产生下降的 ASMC 该项目将使用与诱导的起源特定的 ASMC 不同的。来自 MFS 患者和健康对照的多能干细胞 (iPSC) 来测试一个新假设：发育起源导致与内侧变性相关的 ASMC 的位置特异性异常此外，它将探索马凡氏综合症症状前先天性缺陷的生物标志物，以确定预防性治疗干预的新靶点这些研究将表征表型。人类 ASMC 亚型在细胞和组织水平上与干细胞培养和血管的差异我们还将使用最先进的核心设施进行转录组学和组织工程技术。对这些细胞和组织模型进行蛋白质组学分析，为生物信息学培养丰富的生物学特征此外，我们将开发一个生物信息学管道来阐明新的预防目标。使用我们独特的组合，本质上负责升主动脉 MFS 诱导的内侧变性最后，根据我们的生物信息学输出，我们将表型、转录组和蛋白质组结果作为输入。在基于人类 iPSC 的体外模型上测试我们的干预措施，并与氯沙坦治疗进行比较。常用的抗高血压药物，还表现出独特的抗重塑特性，并已显示该奖学金的培训计划将致力于管理动物和患者的 MFS 症状。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，以及这将通过定期的导师会议、期刊俱乐部、会议演讲、双向交流来实现。年度委员会会议和高级课程作业将在 Costa 实验室进行。 ISMMS 心血管研究中心是一个高度活跃和协作的环境，与我的研究主题和职业目标相一致的导师和学生将在其中进行额外的培训。拉米雷斯实验室拥有马凡综合征和生化研究技术的高级研究人员。