The Wnt7a/ROR2 axis in the pathogenesis of pulmonary arterial hypertension

Wnt7a/ROR2轴在肺动脉高压发病机制中的作用

基本信息

批准号：
10444951
负责人：
VINICIO A DE JESUS PEREZ
金额：
$ 57.71万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-02-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10444951
关键词：
Adherens Junction Affect Animal Model Blood Vessels Blood capillaries Cardiopulmonary Cardiovascular system Cell Differentiation process Cell Extracts Cell model Cells Clinical Complex Cytoplasmic Tail DNA DNA Methylation Disease Echocardiography Endothelial Cells Endothelium Epigenetic Process Equilibrium Female of child bearing age Fluorescence Resonance Energy Transfer Focal Adhesions Genes Genetic Genetic Transcription Goals Histone Acetylation Histone Deacetylation Human Hypermethylation Hypoxia Hypoxia Inducible Factor Integrins KDR gene Knockout Mice Life Ligands Lung Measures Messenger RNA Microfluidics Modeling Molecular Morphogenesis Mus Outcome Output Pathogenesis Pathologic Pathway interactions Patients Permeability Process Proteins Proteomics Pulmonary Hypertension Rattus Receptor Protein-Tyrosine Kinases Recombinants Recovery Regulation Repression Research Project Grants Right Ventricular Function Role Signal Transduction Supplementation Testing Therapeutic Tissues VEGFA gene Vascular Endothelial Growth Factors Vascular Endothelium Vascular remodeling Ventricular Work angiogenesis base cell motility chromatin immunoprecipitation density endothelial dysfunction epigenetic regulation exosome improved lung microvascular endothelial cells lung pressure novel planar cell polarity prevent pulmonary arterial hypertension pyrosequencing response restoration right ventricular failure sensor therapeutic evaluation

项目摘要

Pulmonary arterial hypertension (PAH) is a life-threatening disease characterized by abnormally elevated pulmonary pressures and right ventricular (RV) failure. Inappropriate angiogenesis is a key pathological feature of PAH associated with endothelial dysfunction and progressive loss of pulmonary and RV microvessels. Angiogenesis is the process by which new vessels arise from existing vessels and is mainly driven by VEGF signaling. In response to VEGF-A, endothelial cells differentiate into tip cells, highly motile cells that direct vessel sprouting and elongation. Our previous R01 was built on the hypothesis that tip cell formation by PMVECs requires crosstalk between the VEGF and the Wnt/planar cell polarity (Wnt/PCP), a pathway responsible for coordinating cell movements during tissue morphogenesis. We demonstrated that Wnt/PCP activation in PMVECs is driven by the interaction between the ligand Wnt7a and ROR2, a tyrosine kinase receptor that phosphorylates residues in the endothelial VEGFR2 cytoplasmic domain to augment VEGF signaling output. We found that, compared to healthy donors, tip cell formation and angiogenesis in response to VEGF-A was significantly reduced in pulmonary microvascular endothelial cells (PMVEC) from PAH patients. Most importantly, supplementation with recombinant Wnt7a or restoration of ROR2 expression in PAH PMVECs results in recovery of the VEGF-A response, leading us to conclude that Wnt7a/ROR2 signaling is required for appropriate VEGF signaling activation and angiogenic response in PMVECs. This renewal will focus on elucidating the transcriptional and epigenetic mechanisms that regulate Wnt7a/ROR2 expression in healthy and PAH PMVECs (Aim 1), how interaction between ROR2 and integrins is required to establish a functional lung endothelial barrier (Aim 2), and the critical role of Wnt7a/ROR2 as a key pro- angiogenic mechanism that supports compensatory angiogenesis during RV adaptation to PAH (Aim 3). The studies in this renewal will confirm the role of Wnt7a/ROR2 signaling as a master regulator of cardiopulmonary angiogenesis and demonstrate the therapeutic potential of restoring Wnt7a/ROR2 signaling to prevent small vessel loss and improve RV function in PAH.

肺动脉高压（PAH）是一种威胁生命的疾病，其特征是异常升高肺部压力和右心（RV）衰竭。不适当的血管生成是关键病理特征与内皮功能障碍以及肺部和RV微血管的逐渐丧失相关的PAH。血管生成是新容器由现有血管产生的过程，主要由VEGF驱动信号。为了响应VEGF-A，内皮细胞分化为尖端细胞，高度运动细胞，这些细胞引导血管发芽和伸长。我们以前的R01建立在PMVEC尖端形成的假设上需要在VEGF和Wnt/Planar Cell Pallation（Wnt/PCP）之间进行串扰，该途径负责组织形态发生过程中的细胞运动。我们证明了Wnt/PCP激活 PMVEC是由配体Wnt7a和ROR2之间的相互作用驱动的，ROR2是一种酪氨酸激酶受体在内皮VEGFR2细胞质结构域中磷酸化的残基以增加VEGF信号输出。我们发现，与健康的供体相比，对VEGF-A的尖端细胞形成和血管生成是 PAH患者的肺微血管内皮细胞（PMVEC）的肺部微血管内皮细胞（PMVEC）显着降低。最多重要的是，补充重组Wnt7a或在PAH PMVEC中恢复ROR2表达导致VEGF-A响应的恢复，使我们得出结论，Wnt7a/ror2信号是需要的 PMVEC中适当的VEGF信号传导激活和血管生成反应。这种更新将着重于阐明调节WNT7A/ROR2的转录和表观遗传机制在健康和PAH PMVEC中的表达（AIM 1），ROR2与整合素之间的相互作用如何建立功能性的肺内皮屏障（AIM 2），以及WNT7A/ROR2作为关键方面的关键作用在RV适应PAH期间支持补偿性血管生成的血管生成机制（AIM 3）。这该续约的研究将证实Wnt7a/ROR2信号传导作为主调节器的作用心肺血管生成并证明了恢复WNT7A/ROR2的治疗潜力发出信号以防止小血管损失并改善PAH中的RV功能。