Cardiovascular Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Department of Pathology, University Hospital of Giessen and Marburg (UKGM), Langhansstrasse 10, 35392 Giessen, Germany.
Int J Mol Sci. 2021 Aug 24;22(17):9105. doi: 10.3390/ijms22179105.
Pulmonary arterial hypertension (PAH) is a devastating lung disease characterized by the progressive obstruction of the distal pulmonary arteries (PA). Structural and functional alteration of pulmonary artery smooth muscle cells (PASMC) and endothelial cells (PAEC) contributes to PA wall remodeling and vascular resistance, which may lead to maladaptive right ventricular (RV) failure and, ultimately, death. Here, we found that decreased expression of sarcoplasmic/endoplasmic reticulum Ca ATPase 2a (SERCA2a) in the lung samples of PAH patients was associated with the down-regulation of bone morphogenetic protein receptor type 2 (BMPR2) and the activation of signal transducer and activator of transcription 3 (STAT3). Our results showed that the antiproliferative properties of SERCA2a are mediated through the STAT3/BMPR2 pathway. At the molecular level, transcriptome analysis of PASMCs co-overexpressing SERCA2a and BMPR2 identified STAT3 amongst the most highly regulated transcription factors. Using a specific siRNA and a potent pharmacological STAT3 inhibitor (STAT3i, HJC0152), we found that SERCA2a potentiated BMPR2 expression by repressing STAT3 activity in PASMCs and PAECs. In vivo, we used a validated and efficient model of severe PAH induced by unilateral left pneumonectomy combined with monocrotaline (PNT/MCT) to further evaluate the therapeutic potential of single and combination therapies using adeno-associated virus (AAV) technology and a STAT3i. We found that intratracheal delivery of AAV1 encoding SERCA2 or BMPR2 alone or STAT3i was sufficient to reduce the mean PA pressure and vascular remodeling while improving RV systolic pressures, RV ejection fraction, and cardiac remodeling. Interestingly, we found that combined therapy of AAV1.hSERCA2a with AAV1.hBMPR2 or STAT3i enhanced the beneficial effects of SERCA2a. Finally, we used cardiac magnetic resonance imaging to measure RV function and found that therapies using AAV1.hSERCA2a alone or combined with STAT3i significantly inhibited RV structural and functional changes in PNT/MCT-induced PAH. In conclusion, our study demonstrated that combination therapies using SERCA2a gene transfer with a STAT3 inhibitor could represent a new promising therapeutic alternative to inhibit PAH and to restore BMPR2 expression by limiting STAT3 activity.
肺动脉高压（PAH）是一种破坏性的肺部疾病，其特征是远端肺动脉渐进性阻塞。肺动脉平滑肌细胞（PASMC）和内皮细胞（PAEC）的结构和功能改变导致肺血管壁重塑和血管阻力增加，从而导致右心室（RV）失代偿和最终死亡。在这里，我们发现 PAH 患者肺组织样本中肌浆/内质网 Ca2+-ATP 酶 2a（SERCA2a）的表达降低与骨形态发生蛋白受体 2 型（BMPR2）下调和信号转导和转录激活因子 3（STAT3）激活有关。我们的结果表明，SERCA2a 的抗增殖特性是通过 STAT3/BMPR2 途径介导的。在分子水平上，共过表达 SERCA2a 和 BMPR2 的 PASMC 的转录组分析表明 STAT3 是最受调节的转录因子之一。使用特异性 siRNA 和一种有效的 STAT3 抑制剂（STAT3i，HJC0152），我们发现 SERCA2a 通过抑制 PASMC 和 PAEC 中的 STAT3 活性增强了 BMPR2 的表达。在体内，我们使用单侧左肺切除术联合单克隆抗体（PNT/MCT）诱导的已验证和有效的严重 PAH 模型来进一步评估使用腺相关病毒（AAV）技术和 STAT3i 的单一和联合治疗的治疗潜力。我们发现，经气管内递送编码 SERCA2a 或 BMPR2 的 AAV1 或单独或 STAT3i 足以降低平均肺动脉压和血管重塑，同时改善 RV 收缩压、RV 射血分数和心脏重塑。有趣的是，我们发现 AAV1.hSERCA2a 与 AAV1.hBMPR2 或 STAT3i 的联合治疗增强了 SERCA2a 的有益作用。最后，我们使用心脏磁共振成像测量 RV 功能，发现 AAV1.hSERCA2a 单独或与 STAT3i 联合治疗显著抑制了 PNT/MCT 诱导的 PAH 中的 RV 结构和功能变化。总之，我们的研究表明，SERCA2a 基因转移与 STAT3 抑制剂联合使用的联合治疗可能代表一种新的有前途的治疗选择，可抑制 PAH 并通过限制 STAT3 活性来恢复 BMPR2 表达。
