Developmental basis for vascular smooth muscle cell dysfunction in Marfan syndrome aortic aneurysm

马凡综合征主动脉瘤血管平滑肌细胞功能障碍的发育基础

• 批准号: 10463538
• 负责人: Albert J. Pedroza
• 金额: $ 6.76万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-20 至 2022-08-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10463538
• 关键词: Affect; Aneurysm; Aorta; Aortic Aneurysm; Aortic Diseases; Aortic Segment; Automobile Driving; Behavior; Biochemical; Blood Vessels; Cardiovascular Surgical Procedures; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cells; Chest; Coculture Techniques; Computing Methodologies; Connective Tissue Diseases; Data; Data Set; Development; Disease; Dissection; Embryo; Event; Extracellular Matrix; FBN1; Fibroblasts; Functional disorder; Gene Expression; Gene Expression Profile; Genes; Growth Factor; Heart; Heterogeneity; Homeostasis; In Vitro; Individual; Inherited; Investigation; Lead; Life Expectancy; Marfan Syndrome; Medical; Medicine; Mentorship; Methods; Modeling; Modernization; Molecular; Mus; Mutation; Neural Crest; Operative Surgical Procedures; Pathologic; Pathology; Pathway interactions; Patients; Phenotype; Plant Roots; Predisposition; Process; Production; Proteins; Relaxation; Research Training; Resolution; Risk; Role; Rupture; Ruptured Aneurysm; Scientist; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Source; Specificity; Specimen; Stimulus; Syndrome; Synthetic Genes; Techniques; Thoracic Aortic Aneurysm; Thoracic aorta; Tissues; Training; Transforming Growth Factor beta; Transgenic Mice; Universities; Vascular Diseases; Vascular Smooth Muscle; Work; aortic valve; ascending aorta; career; cell behavior; experimental study; heritable connective tissue disorder; high throughput screening; in vivo; mouse model; novel; novel therapeutics; paracrine; prevent; repaired; response; response to injury; single-cell RNA sequencing; therapy development; tool; transcriptomics; transdifferentiation

Project Summary/Abstract Marfan syndrome (MFS), caused by mutations in the fibrillin-1 (FBN1) gene, is the most common inherited connective tissue disorder, affecting 1 in 5,000 individuals. Aortic root aneurysm leads to reduced life expectancy due to dissection or rupture of the aneurysm unless preventative aortic surgery is performed. Normally, vascular smooth muscle cells (SMCs) maintain homeostasis within the aorta via dynamic contraction/relaxation and extracellular matrix production, however these cells retain significant plasticity to alter their phenotype in response to injury, growth factors, or other stimuli. Dysfunctional SMC phenotype modulation is known to contribute to aneurysm development in MFS. Dysregulated transforming growth factor-beta (TGF-b) signaling also contributes to aortic aneurysm, though the precise role of this pathway remains controversial. Furthermore, mechanisms driving the tendency of the aortic root (the segment most proximal to the aortic valve) to develop focal aneurysm despite systemic effects of FBN1 mutations are poorly understood. Distinct embryonic origins of SMCs populating the aortic root (from the second heart field, SHF) and ascending aortic segments (from neural crest, NC) is hypothesized to contribute to aortic root-specific pathology, however it remains unclear how these embryonic origins affect propensity for aneurysm development. Recently, single-cell RNA sequencing (scRNA- seq) has permitted high-resolution analysis of individual SMC gene expression. My preliminary work applying scRNA-seq to a mouse model of MFS has identified a subset of SMCs with a severely modulated, pathologic phenotype. The proposed study will advance our current understanding of SMC development and dysfunction in MFS aortic aneurysm using two complementary aims. In Aim 1 I will define the distinct phenotypes of thoracic aortic SMCs derived from the second heart field and neural crest lineages by applying single-cell RNA sequencing to an embryonic lineage-tracing mouse model and in vitro studies of TGF-b dysregulation on SHF and NC-derived SMC phenotype. In Aim 2 I will characterize the source and pathologic effects of modulated SMCs in MFS aortic root aneurysm by lineage-tracing early SMCs in a murine MFS model and applying computational transcriptomic analysis tools to scRNAseq data to determine molecular mechanisms driving their phenotype changes. Co-culture experiments of phenotypically modulated SMCs with healthy aortic SMCs will model aortic aneurysm pathology in vitro. These studies will generate important data that will help pinpoint molecular mechanisms driving aortic pathology in MFS and other hereditary aneurysm disorders toward new therapy development. The proposed research training plan features direct mentorship from a committee of accomplished clinician-scientists and access to state-of-the art facilities and techniques. This plan also incorporates professional development and career planning strategies, employing the unique collaborative spirit between Cardiovascular Surgery and Medicine at Stanford University intended to maximize training potential.

项目摘要/摘要 由Fibrillin-1（FBN1）基因突变引起的Marfan综合征（MFS）是最常见的遗传 结缔组织障碍，影响5,000个人中有1个。主动脉根动脉瘤可降低预期寿命 除非进行预防性主动脉手术，否则由于动脉瘤的解剖或破裂。通常，血管 平滑肌细胞（SMC）通过动态收缩/放松和 细胞外基质产生，但是这些细胞保留了明显的可塑性以改变其表型 对伤害，生长因子或其他刺激的反应。已知功能失调的SMC表型调制已知 有助于MFS的动脉瘤开发。转化生长因子-BETA（TGF-B）信号的失调失调 尽管该途径的确切作用仍然引起争议，但也有助于主动脉瘤。此外， 驱动主动脉根（主动脉瓣近端的段）趋势的机制 尽管FBN1突变的全身作用，局灶性动脉瘤知之甚少。独特的胚胎起源 SMC填充主动脉根（来自第二个心脏场，SHF）和上升主动脉片段（来自神经 假设Crest，NC）有助于主动脉根特异性病理，但是尚不清楚这些如何 胚胎起源会影响动脉瘤发育的倾向。最近，单细胞RNA测序（SCRNA- SEQ允许对单个SMC基因表达的高分辨率分析。我的初步工作适用 SCRNA-SEQ到MFS的小鼠模型已确定了具有严重调节的SMC的子集 表型。拟议的研究将提高我们目前对SMC开发和功能障碍的理解 在MFS主动脉动脉瘤中，使用两个互补目标。在目标1中，我将定义胸腔的独特表型 通过施加单细胞RNA，源自第二个心脏场和神经rest谱系的主动脉smcs 对胚胎谱系追踪小鼠模型的测序和SHF上TGF-B失调的体外研究 和NC衍生的SMC表型。在AIM 2中，我将表征调制的来源和病理效应 MFS主动脉根动脉瘤中的SMC通过在鼠MFS模型中追踪谱系早期SMC并应用 计算转录组分析工具用于SCRNASEQ数据以确定驱动其分子机制 表型改变。具有健康主动脉SMC的表型调节SMC的共培养实验将 在体外模型主动脉瘤病理学。这些研究将产生重要的数据，这将有助于查明 驱动MFS和其他遗传性动脉瘤的分子机制向新的 治疗开发。拟议的研究培训计划以一个委员会的直接指导 出色的临床医生科学家，并获得了最先进的设施和技术。这个计划也是如此 纳入专业发展和职业规划策略，采用独特的协作精神 斯坦福大学的心血管外科手术和医学之间旨在最大程度地发挥培训潜力。

项目成果

Acute Induced Pressure Overload Rapidly Incites Thoracic Aortic Aneurysmal Smooth Muscle Cell Phenotype.

• DOI: 10.1161/hypertensionaha.121.18640
• 发表时间: 2022-04
• 期刊: Hypertension (Dallas, Tex. : 1979)
• 作者: Pedroza AJ;Shad R;Dalal AR;Yokoyama N;Nakamura K;Hiesinger W;Fischbein MP
• 通讯作者: Fischbein MP

Androgens Accentuate TGF-β Dependent Erk/Smad Activation During Thoracic Aortic Aneurysm Formation in Marfan Syndrome Male Mice.

• DOI: 10.1161/jaha.119.015773
• 发表时间: 2020-10-20
• 期刊: Journal of the American Heart Association
• 影响因子: 5.4
• 作者: Tashima Y;He H;Cui JZ;Pedroza AJ;Nakamura K;Yokoyama N;Iosef C;Burdon G;Koyano T;Yamaguchi A;Fischbein MP
• 通讯作者: Fischbein MP

Mosaicism for the smooth muscle cell (SMC)-specific knock-in of the Acta2 R179C pathogenic variant: Implications for gene editing therapies.

Acta2 R179C 致病性变异的平滑肌细胞 (SMC) 特异性敲入的嵌合现象：对基因编辑疗法的影响。

• DOI: 10.1016/j.yjmcc.2022.07.004
• 发表时间: 2022
• 期刊: Journal of molecular and cellular cardiology
• 影响因子: 5
• 作者: Kaw,Anita;Pedroza,AlbertJ;Chattopadhyay,Abhijnan;Pinard,Amelie;Guo,Dongchuan;Kaw,Kaveeta;Zhou,Zhen;Shad,Rohan;Fischbein,MichaelP;Kwartler,CallieS;Milewicz,DiannaM
• 通讯作者: Milewicz,DiannaM