Department of Cell Biology, State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, 12501Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Department of Physiology, 26441Zhejiang University School of Medicine, Hangzhou, Zhejiang, China.
J Cardiovasc Pharmacol Ther. 2021 Nov;26(6):677-689. doi: 10.1177/10742484211015535. Epub 2021 May 14.
Pulmonary veno-occlusive disease (PVOD) is a fatal disease of pulmonary vascular lesions leading to right heart failure. Heritable PVOD (hPVOD) is related to biallelic mutation of (encoding GCN2), but its molecular mechanism remains unclear. In this study, we aimed to investigate the pathogenesis of PVOD and to find potential drug targets for PVOD. GCN2 dysfunction led to an enhanced transcription of collagen I gene ( and ) through decreasing ATF3-dependent p38 phosphorylation inhibition in PVOD, which promotes the collagen I synthesis in pulmonary arterial smooth muscle cells (PASMCs) and eventually leads to increased collagen deposition in pulmonary artery. Four GCN2 knockout (KO) cell lines (exon 15 or 33 mutation) were successfully constructed by epiCRISPR system. Two induced pluripotent stem cells (iPSCs) were generated by reprogramming peripheral blood mononuclear cells (PBMCs) of PVOD patient. It was also comfirmed that GCN2 dysfunction could lead to increased expression of collagen I in lateral plate mesoderm lineage-smooth muscle cells (LM-SMCs) differentiated from both GCN2 KO cell lines and iPSCs. SB203580 (a specific inhibitor of p38) improved hemodynamics and pulmonary vascular remodeling in mitomycin C (MMC)-induced PVOD rats by right ventricle echocardiography. On the whole, we proposed that GCN2 deficiency decreased ATF3-dependent p38 phosphorylation inhibition in PVOD development and suggested a potential therapeutic reagent of SB203580 for the treatment of the disease.
肺静脉闭塞病（PVOD）是一种致命的肺血管病变疾病，可导致右心衰竭。遗传性 PVOD（hPVOD）与双等位基因突变有关，编码 GCN2），但其分子机制尚不清楚。在这项研究中，我们旨在探讨 PVOD 的发病机制，并寻找潜在的 PVOD 药物靶点。GCN2 功能障碍通过降低 ATF3 依赖性 p38 磷酸化抑制，导致胶原 I 基因（和）的转录增强，从而促进肺动脉平滑肌细胞（PASMCs）中的胶原 I 合成，并最终导致肺动脉中胶原沉积增加。通过 epiCRISPR 系统成功构建了 4 个 GCN2 敲除（KO）细胞系（外显子 15 或 33 突变）。通过重编程 PVOD 患者的外周血单核细胞（PBMC）产生了 2 个诱导多能干细胞（iPSC）。还证实 GCN2 功能障碍可导致从 GCN2 KO 细胞系和 iPSC 分化而来的侧中胚层谱系-平滑肌细胞（LM-SMCs）中胶原 I 的表达增加。SB203580（p38 的特异性抑制剂）通过右心室超声心动图改善了丝裂霉素 C（MMC）诱导的 PVOD 大鼠的血液动力学和肺血管重塑。总的来说，我们提出 GCN2 缺乏会降低 PVOD 发展过程中 ATF3 依赖性 p38 磷酸化抑制，并提出了 SB203580 作为治疗该疾病的潜在治疗试剂。
