Essential functions of Fibulin proteins in outflow tract morphogenesis.

Fibulin 蛋白在流出道形态发生中的基本功能。

基本信息

批准号：
10700864
负责人：
ANGELIKA G. Aleman
金额：
$ 4.77万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10700864
关键词：
Anterior Aortic Valve Stenosis Apoptosis Arteries Biological Assay Biological Models Biology Cardiac Cardiac Myocytes Cardiovascular system Cell Lineage Child Complement Congenital Abnormality Cues Data Defect Development Disease Distal Double Outlet Right Ventricle Elastic Fiber Elasticity Elements Embryo Endothelial Cells Etiology Evolution Exhibits Extracellular Matrix Extracellular Matrix Proteins Genes Genetic Growth Heart Heart Valve Prosthesis Human Impairment Investigation Lead Mediating Medical Modeling Molecular Morphogenesis Natural regeneration Operative Surgical Procedures Pathway interactions Patients Phosphorylation Play Polysaccharides Population Proliferating Proteins Protocols documentation Pulmonary valve structure Reporter Role Signal Transduction Smooth Muscle Smooth Muscle Myocytes Specific qualifier value Testing Tetralogy of Fallot Therapeutic Tissue Engineering Tissues Transforming Growth Factor beta Transgenes Transposition of Great Vessels cardiogenesis cell type cohort congenital heart disorder conotruncal heart defect experimental study fibulin gain of function gene function improved innovation insight loss of function malformation mortality neonate new therapeutic target novel pediatric patients progenitor risk stratification small molecular inhibitor surgical risk

项目摘要

PROJECT SUMMARY Congenital heart disease (CHD) is the most common birth defect in humans and patients with conotruncal defects comprise 20% of this population. Malformations of the distal aspect of the ventricle, the infundibulum, and the proximal aspect of the great arteries lead to conotruncal anomalies. Specific examples include transposition of the great vessels, double outlet right ventricle, and Tetralogy of Fallot. Moreover, in comparison to other types of CHDs, conotruncal defects are more frequently associated with genetic and syndromic abnormalities. Among this patient cohort, the mortality rate of 17% indicates a dire need for improvement in our understanding of the early developmental cues guiding these aberrations in outflow tract (OFT) morphogenesis. The purpose of this application is to uncover the molecular and cellular mechanisms that account for disruptions in OFT development and underlie human conotruncal CHDs. We identified Fibulin (Fbln) proteins as novel regulators of the extracellular matrix (ECM) essential for OFT morphogenesis. Our preliminary data demonstrate that Fblns are required for smooth muscle addition to the OFT and for TGF-β signaling in the late-differentiating progenitors that contribute to the arterial pole. In Aim 1, I will dissect the cellular and molecular mechanisms mediated by fbln genes during OFT growth. I will assess proliferation of the anterior SHF progenitor population in the Fbln loss-of-function model and evaluate differentiation and proliferation of the cardiomyocyte, endothelial cell, and smooth muscle cell contribution to anterior SHF-derived lineages at the arterial pole. I will employ EdU, apoptosis, and developmental timing assays to dissect these cell type-specific functions of Fbln genes. In Aim 2, I will probe cardiomyocyte, endothelial cell, and smooth muscle cell-type specific expression of pSmad3 in the OFT of Fbln loss-of-function embryos. Moreover, I will perform gain-of-function and loss-of-function experiments with a constitutively active Alk5 transgene and a small molecular inhibitor of Smad3 phosphorylation, respectively. These strategies will illuminate how Fbln proteins mediate OFT development via TGF-β signaling. Insights gained from these proposed studies will shed light on the mechanisms responsible for arterial compliance and elastic deformation at the arterial pole with implications for diseases involving stenosis of the aortic and pulmonary valves. Furthermore, probing the cell type-specific roles of Fbln proteins will augment our ability to identify novel therapeutic targets and protocols for tissue engineering of OFT conduits and artificial valves.

项目概要先天性心脏病 (CHD) 是人类和圆锥动脉干患者中最常见的出生缺陷占该人群 20% 的缺陷为心室远端、漏斗部畸形。大动脉近端导致圆锥干异常的具体例子包括。大血管转位、右心室双出口和法洛四联症。与其他类型的先心病相比，圆锥干缺陷更常与遗传和综合征相关在该患者队列中，17% 的死亡率表明我们迫切需要改进。了解引导流出道（OFT）形态发生中这些畸变的早期发育线索。该应用的目的是揭示造成破坏的分子和细胞机制我们发现 Fibulin (Fbln) 蛋白是一种新型蛋白。我们的初步数据表明，细胞外基质 (ECM) 的调节因子对 OFT 形态发生至关重要。 Fblns 是平滑肌添加到 OFT 以及晚期分化中的 TGF-β 信号传导所必需的在目标 1 中，我将剖析细胞和分子机制。 OFT 生长过程中由 fbln 基因介导，我将评估前 SHF 祖细胞群的增殖。在 Fbln 功能丧失模型中评估心肌细胞、内皮细胞的分化和增殖细胞和平滑肌细胞对动脉极前部 SHF 衍生谱系的贡献，我将采用 EdU，细胞凋亡和发育计时分析来剖析 Fbln 基因的这些细胞类型特异性功能。 2、我将探测pSmad3在心肌细胞、内皮细胞和平滑肌细胞类型中的特异性表达 Fbln 功能丧失胚胎的 OFT 此外，我将进行功能获得和功能丧失实验。具有组成型活性 Alk5 转基因和 Smad3 磷酸化小分子抑制剂，这些策略将阐明 Fbln 蛋白如何通过 TGF-β 信号传导介导 OFT 发育。从这些拟议的研究中获得的见解将揭示动脉粥样硬化的机制。动脉极的顺应性和弹性变形对涉及动脉狭窄的疾病的影响此外，探索 Fbln 蛋白的细胞类型特异性作用将增强我们的研究成果。能够确定 OFT 导管和人工组织工程的新治疗靶点和方案阀门。