Division of Cardiology, Department of Medicine, Center for Pulmonary Vascular Biology and Medicine, Pittsburgh Heart, Lung, Blood, and Vascular Medicine Institute, University of Pittsburgh School of Medicine and University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania.
Physician Scientist Training Program, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.
Am J Physiol Lung Cell Mol Physiol. 2021 May 1;320(5):L726-L738. doi: 10.1152/ajplung.00407.2020. Epub 2021 Feb 10.
Pulmonary arterial hypertension (PAH) refers to a set of heterogeneous vascular diseases defined by elevation of pulmonary arterial pressure (PAP) and pulmonary vascular resistance (PVR), leading to right ventricular (RV) remodeling and often death. Early increases in pulmonary artery stiffness in PAH drive pathogenic alterations of pulmonary arterial endothelial cells (PAECs), leading to vascular remodeling. Dysregulation of microRNAs can drive PAEC dysfunction. However, the role of vascular stiffness in regulating pathogenic microRNAs in PAH is incompletely understood. Here, we demonstrated that extracellular matrix (ECM) stiffening downregulated miR-7 levels in PAECs. The RNA-binding protein quaking (QKI) has been implicated in the biogenesis of miR-7. Correspondingly, we found that ECM stiffness upregulated QKI, and QKI knockdown led to increased miR-7. Downstream of the QKI-miR-7 axis, the serine and arginine-rich splicing factor 1 (SRSF1) was identified as a direct target of miR-7. Correspondingly, SRSF1 was reciprocally upregulated in PAECs exposed to stiff ECM and was negatively correlated with miR-7. Decreased miR-7 and increased QKI and SRSF1 were observed in lungs from patients with PAH and PAH rats exposed to SU5416/hypoxia. Lastly, miR-7 upregulation inhibited human PAEC migration, whereas forced SRSF1 expression reversed this phenotype, proving that miR-7 depended upon SRSF1 to control migration. In aggregate, these results define the QKI-miR-7-SRSF1 axis as a mechanosensitive mechanism linking pulmonary arterial vascular stiffness to pathogenic endothelial function. These findings emphasize implications relevant to PAH and suggest the potential benefit of developing therapies that target this miRNA-dependent axis in PAH.
肺动脉高压(PAH)是一组异质性血管疾病,其特征为肺动脉压力(PAP)和肺血管阻力(PVR)升高,导致右心室(RV)重构,常导致死亡。PAH 中肺动脉僵硬度的早期增加会导致肺动脉内皮细胞(PAEC)的病理性改变,导致血管重构。microRNA 的失调可导致 PAEC 功能障碍。然而,血管僵硬在调节 PAH 中致病 microRNA 中的作用尚不完全清楚。在这里,我们证明细胞外基质(ECM)变硬会使 PAEC 中的 miR-7 水平降低。RNA 结合蛋白 quaking(QKI)已被牵涉到 miR-7 的生物发生中。相应地,我们发现 ECM 僵硬会上调 QKI,而 QKI 敲低会导致 miR-7 增加。在 QKI-miR-7 轴的下游,发现丝氨酸/精氨酸丰富剪接因子 1(SRSF1)是 miR-7 的直接靶标。相应地,在暴露于硬 ECM 的 PAEC 中,SRSF1 被反向上调,并且与 miR-7 呈负相关。在患有 PAH 的患者的肺部和暴露于 SU5416/低氧的 PAH 大鼠的肺部中观察到 miR-7 减少、QKI 和 SRSF1 增加。最后,miR-7 的上调抑制了人 PAEC 的迁移,而强制表达 SRSF1 则逆转了这种表型,证明 miR-7 依赖于 SRSF1 来控制迁移。总的来说,这些结果定义了 QKI-miR-7-SRSF1 轴作为将肺血管僵硬与致病内皮功能联系起来的机械敏感机制。这些发现强调了与 PAH 相关的意义,并表明靶向该 miRNA 依赖轴的治疗方法在 PAH 中具有潜在的益处。