Role of MicroRNAs 424 and 503 in Pulmonary Arterial Hypertension

MicroRNA 424 和 503 在肺动脉高压中的作用

• 批准号: 8534272
• 负责人: Hyung Joon Chun
• 金额: $ 39.63万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8534272
• 关键词: Blood Vessels; Cells; Cessation of life; Clinical; Data; Diagnosis; Disease; Endothelial Cells; Endothelium; Enhancers; Experimental Models; FGF2 gene; FGFR1 gene; Failure; Fibroblast Growth Factor; G Protein-Coupled Receptor Signaling; Homeostasis; Human; Hypoxia; In Vitro; Lead; Lesion; Ligands; Lung; MicroRNAs; Modeling; Molecular; Monocrotaline; Mus; Muscle Cells; Outcome; Pathway interactions; Patients; Play; Process; Pulmonary Hypertension; Pulmonary Vascular Resistance; Pulmonary artery structure; Rattus; Regulation; Rodent Model; Role; Severity of illness; Signal Pathway; Signal Transduction; Structure of parenchyma of lung; Syndrome; Transcriptional Regulation; Vascular remodeling; Ventricular; arteriole; base; cohort; human disease; improved; mortality; novel; pulmonary arterial hypertension; pulmonary artery endothelial cell; receptor; reconstitution; restoration; therapeutic target

DESCRIPTION (provided by applicant): Pulmonary arterial hypertension (PAH) is a disease characterized by the vascular remodeling of the pulmonary arterioles, including formation of plexiform and concentric lesions comprised of proliferative endothelial cells. We recently found that disruption of the G protein-coupled receptor signaling axis comprised of the ligand apelin and the receptor APJ leads to significant worsening of hypoxia induced pulmonary hypertension (PH) in mice, and found that the pathway is also perturbed in clinical PAH. To further characterize the downstream endothelial targets which may be utilized by apelin-APJ signaling to protect against the remodeling processes in PAH, we carried out a microRNA (miRNA) array analysis of pulmonary artery endothelial cells (PAECs) subjected to knockdown of apelin, APJ, or both. We identified two miRNAs (miR-424 and miR-503) that are highly expressed in PAECs, but are significantly downregulated in the context of apelin/APJ knockdown. Remarkably, these miRNAs are also markedly decreased in the experimental monocrotaline rat model of PH, as well as in PAECs isolated from patients with clinically diagnosed PAH. Our in vitro functional studies demonstrated that both miR-424 and miR-503 exert anti-proliferative, anti-angiogenic effects, via a mechanism that at least in part involves targeting of the FGF signaling pathway. We found that restoration of miR-424/503 expression in the monocrotaline induced model of PH by intranasal lentiviral delivery can significantly ameliorate the severity of the disease. Based o these preliminary data, we hypothesize that miR-424/503 play a critical role in maintaining pulmonary vascular homeostasis, and that disruption of their expression serves a critical contribution to the aberrant endothelial proliferation which is a hallmark of PAH. We propose the following aims to further evaluate the role of miR-424/503 in PAH: 1) Determine the transcriptional regulatory mechanisms of miR-424/503 in PAECs that involves targeting of the myocyte-specific enhancer factor 2 (MEF2) by apelin-APJ signaling, 2) Evaluate the efficacy of endothelial specific expression of miR-424/503 in experimental models of PH, and 3) Further validate the role for miR-424/503 in clinical PAH.

描述(申请人提供)：肺动脉高压(PAH)是一种以肺小动脉血管重构为特征的疾病，包括由增殖性内皮细胞组成的丛状和同心性病变。我们最近发现，由配体apelin和受体APJ组成的G蛋白偶联受体信号轴的破坏导致小鼠缺氧性肺动脉高压(PH)的显著恶化，并发现该通路在临床PAH中也受到干扰。为了进一步确定apelin-APJ信号可能用来保护PAH重塑过程的下游内皮靶点，我们对受apelin和/或APJ抑制的肺动脉内皮细胞(PAECs)进行了microRNA(MiRNA)阵列分析。我们发现了两个在PAEC中高表达的miRNAs(miR-424和miR-503)，但在apelin/APJ击倒的背景下显著下调。值得注意的是，在实验性野百合碱大鼠的PH模型中，以及从临床诊断为PAH的患者分离的PAEC中，这些miRNAs也显著减少。我们的体外功能研究表明，miR-424和miR-503都具有抗增殖、抗血管生成的作用，其机制至少部分涉及靶向成纤维细胞生长因子信号通路。我们发现，在野百合碱诱导的PH模型中，通过鼻腔慢病毒注射恢复miR-424/503的表达可以显著改善疾病的严重程度。根据这些初步数据，我们推测miR-424/503在维持肺血管内环境稳定中起关键作用，并且它们的表达中断在作为PAH标志的内皮细胞异常增殖中起关键作用。我们建议进一步评估miR-424/503在PAH中的作用：1)确定miR-424/503在PAEC中的转录调控机制，包括通过apelin-APJ信号靶向肌细胞特异性增强因子2(MEF2)；2)评估血管内皮细胞特异性表达miR-424/503在实验性PH模型中的有效性；以及3)进一步验证miR-424/503在临床PAH中的作用。