Yao Maozhong, Zhong Keyan, Zheng Xinbin, Yang Zhaoxin, Li Chunying, Gu Yong, Chen Zhanjuan
Clinical Research Center, Affiliated Chinese Medicine Hospital of Hainan Medical University (Hainan Academy of Medical Sciences), Haikou, Hainan, China.
Clinical Research Center, Hainan Hospital, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou University of Chinese Medicine, Haikou, Hainan, China.
J Cell Mol Med. 2025 Jul;29(13):e70692. doi: 10.1111/jcmm.70692.
Endothelial-to-mesenchymal transition (EndMT) induced by dysfunctional pulmonary artery endothelial cells (PAECs) is regarded as an initiating and pivotal factor in pulmonary hypertension (PH). This study focuses on identifying a novel therapeutic target for regulating EndMT in PH. A comprehensive analysis of 2 hypoxic PAECs datasets yielded 310 overlapping upregulated and 229 downregulated differentially expressed genes (DEGs). These upregulated DEGs were primarily enriched in HIF-1 signalling pathway and glycolysis/gluconeogenesis, while downregulated only in spliceosome, as indicated by KEGG. Through PPI network analysis and the application of MCC algorithms, 5 hub genes were identified among these upregulated DEGs: GAPDH, LDHA, ALDOA, PFKL, and PFKP. Their enrichment in the 2 aforementioned pathways was confirmed by cross-pathway DEGs analysis and ClueGo. Among the hub genes, LDHA was chosen as the key gene based upon expression and correlation analysis of the validation set from PH patients. Subsequent GSEA also revealed the enrichment of LDHA in these 2 pathways. Additionally, the increased expression of LDHA protein in tissues and cells was confirmed, and the elevated enzymatic activity of LDHA in clinical serum samples was also verified. From 2 online databases, 4 LDHA inhibitors were filtered out, and the stable binding between the inhibitors and the LDHA protein was confirmed through molecular docking and molecular dynamics simulation. Finally, the experimental results indicated that one of the inhibitors FX11 reversed EndMT by inhibiting the lactate-SNAI1 axis, thereby alleviating hypoxia-induced PH. The potential of LDHA as a therapeutic target for PH by modulating EndMT was proposed in this study.
功能失调的肺动脉内皮细胞(PAECs)诱导的内皮-间充质转化(EndMT)被认为是肺动脉高压(PH)的起始和关键因素。本研究聚焦于确定一种调节PH中EndMT的新型治疗靶点。对2个缺氧PAECs数据集的综合分析产生了310个重叠上调和229个下调的差异表达基因(DEGs)。KEGG分析表明,这些上调的DEGs主要富集于HIF-1信号通路和糖酵解/糖异生,而下调的DEGs仅富集于剪接体。通过蛋白质-蛋白质相互作用(PPI)网络分析和应用MCC算法,在这些上调的DEGs中鉴定出5个枢纽基因:GAPDH、LDHA、ALDOA、PFKL和PFKP。通过交叉通路DEGs分析和ClueGo证实了它们在上述2条通路中的富集。在枢纽基因中,基于对PH患者验证集的表达和相关性分析,选择LDHA作为关键基因。随后的基因集富集分析(GSEA)也揭示了LDHA在这2条通路中的富集。此外,还证实了LDHA蛋白在组织和细胞中的表达增加,以及临床血清样本中LDHA酶活性的升高。从2个在线数据库中筛选出4种LDHA抑制剂,并通过分子对接和分子动力学模拟证实了抑制剂与LDHA蛋白之间的稳定结合。最后,实验结果表明,其中一种抑制剂FX11通过抑制乳酸-SNAI1轴逆转EndMT,从而减轻缺氧诱导的PH。本研究提出了LDHA作为通过调节EndMT治疗PH的潜在靶点。