Mechanobiology of Cardiac Outflow Tract Morphogenesis

心脏流出道形态发生的力学生物学

基本信息

批准号：
10592432
负责人：
Jonathan Talbot Butcher
金额：
$ 74.32万
依托单位：
CORNELL UNIVERSITY
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：
2022-03-15 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10592432
关键词：
Ablation Acute Affect Automobile Driving Back Birds Birth Cardiac Cells Chick Chronic Circulation Clinical Collaborations Complement Complex Computer Analysis Congenital Abnormality Congenital Heart Defects Data Defect Distal Echocardiography Embryo Endocardium Environment Esthesia Etiology Event Failure Felis catus Fetal Death Fetal Growth Future Genes Genetic Growth Human In Situ Lasers Liquid substance Live Birth Lung Mechanical Stress Mechanics Mesenchymal Mesenchyme Modeling Molecular Morphogenesis Morphology Mutant Strains Mice Organ Culture Techniques Pattern Phase Phenotype Physical condensation Process Proliferating RNA Regulator Genes Resolution Resources Role Severities Shapes Signal Transduction Slide Stratification Stress Technology Testing Tissues Visualization Zebrafish antagonist aortic valve arm clinically relevant cohort fetal genetic approach hemodynamics in utero in vivo innovation innovative technologies insight malformation mechanical force mechanotransduction morphogens novel operation pressure programs protein expression response shear stress spatiotemporal stress state tissue stress transcriptomics ultrasound

项目摘要

Proper growth, septation, and maturation of the cardiac outflow tract (OFT) into valved aortic and pulmonary outlets are essential for oxygenated circulation after birth. 1-2% of live births and up to 30% of pre-term fetal deaths have congenital heart defects, many of which affect the remodeling of the valvuloseptal primordial tissues, called the proximal and distal outflow cushions. Despite much effort uncovering the genetic basis of early OFT cushion formation, this understanding has not explained the clinically relevant phases of growth, condensation and elongation into valves and septa. One reason for this appears to be the domination of conditional and collective signaling mechanisms that are well accessible by genetic approaches. Mechanical forces (shear stress, pressure, tension) are ever present during this complex period of OFT growth and remodeling, but to date no studies have investigated these key interactions, especially for their contributions to OFT defects. We believe that clinically relevant OFT remodeling arise from improper cushion endocardial and/or mesenchymal sensation of and/or response to their local mechanical environment, which in turn drives the incorrect signaling programs. The Butcher lab has pioneered innovative technology 1) to quantify local in vivo mechanical forces within this OFT region and register them with local in situ gene/protein expression, 2) to not-invasively visualize and precisely ablate intracardiac tissues without collateral damage in vivo, and 3) to directly test mechanobiological mechanisms of endocardial cushion growth and remodeling ex vivo. The preliminary data in this proposal present evidence of two mechanoregulated molecular switches that potentiate between OFT cushion proliferation and differentiation, which motivates the novel hypothesis that local mechanosensaton operates molecular switches to control sizing, shape, and stratification of the outflow valves and septa. Aim 1 will implement innovative non- invasive laser photoablations of the formed proximal or distal cushions of the avian OFT to create genetically unbiased clinically relevant outflow tract malformations. We will then quantitatively analyze and register their hemodynamic, morphological and phenotypic changes. We will further apply novel deconvolution integration of sc-Seq and slide-seq to reveal unprecedented spatio-temporal resolution of the cellular course of malformation, and elaborate how known and newly discovered molecular regulatory programs associate with local mechanical stress changes. Aim 2 will test the mechanistic causailty of the mechanotransduction operated molecular switches in the OFT cushion endocardium via shear stress patterns. Aim 3 will test the operation of different mechanobiogical switches in cushion mesenchyme via tension/compression. using high throughput ex vivo organ cultures. The findings from these studies will substantally advance our understanding of mechanoregulation and conditional signaling in outflow tract valuvloseptal maturation, paving the way for strategies to manipulate such signaling programs to reduce or even rescue CHD severity in utero.

心脏流出道 (OFT) 正常生长、分隔和成熟，进入带瓣膜的主动脉和肺动脉出口对于出生后的氧循环至关重要。 1-2% 的活产儿和高达 30% 的早产儿死亡有先天性心脏缺陷，其中许多影响瓣膜间隔原始组织的重塑，称为近端流出垫和远端流出垫。尽管付出了很多努力来揭示早期 OFT 的遗传基础缓冲垫的形成，这种理解并没有解释临床相关的生长阶段、凝结阶段以及阀门和隔膜的伸长。造成这种情况的原因之一似乎是有条件和可以通过遗传方法轻松访问的集体信号机制。机械力（剪切力在 OFT 成长和重塑的这个复杂时期，压力、压力、紧张）始终存在，但迄今为止没有研究调查这些关键的相互作用，特别是它们对 OFT 缺陷的影响。我们相信临床相关的 OFT 重塑是由不适当的心内膜和/或间质感觉垫引起的和/或对其本地机械环境的响应，这反过来又驱动了不正确的信号程序。 Butcher 实验室开创了创新技术 1) 量化体内局部机械力 OFT 区域并将其与局部原位基因/蛋白质表达进行登记，2) 以非侵入性方式可视化和精确消融心内组织而不会造成体内附带损伤，3）直接测试机械生物学离体心内膜垫生长和重塑的机制。本提案中的初步数据两个机械调节分子开关的证据可以增强 OFT 垫增殖和分化，激发了局部机械感觉操作分子开关的新假设控制流出阀和隔垫的尺寸、形状和分层。目标 1 将实施创新的非对禽类 OFT 形成的近端或远端垫进行侵入性激光光消融，以产生基因公正的临床相关流出道畸形。然后我们将定量分析并记录他们的血流动力学、形态和表型的变化。我们将进一步应用新颖的反卷积积分 sc-Seq和slide-seq揭示了细胞畸形过程前所未有的时空分辨率，并详细阐述已知和新发现的分子调控程序如何与局部机械相关联压力变化。目标 2 将测试机械传导操作分子的机械因果关系通过剪切应力模式在 OFT 缓冲心内膜中进行开关。目标3将测试不同的操作通过张力/压缩在缓冲间充质中进行机械生物学开关。使用高通量离体器官培养。这些研究的结果将极大地增进我们对流出道瓣膜成熟中的机械调节和条件信号传导，为操纵此类信号程序以减轻甚至挽救子宫内冠心病严重程度的策略。