Identification of potential diagnostic biomarkers and therapeutic targets in patients with hypoxia pulmonary hypertension

• Published in: International immunopharmacology 2024-12, Vol.142 (Pt A), p.113028, Article 113028
• Main Authors: Li, Hongyan, Liu, Yi, Zhang, Hongli, Shi, Xianbao, Luo, Yue, Fu, Gaoge, Zhao, Churong, Guo, Lixuan, Li, Xin, Shan, Lina
• Format: Article
• Language: English
• Subjects: Akt; Animals; Arginase - genetics; Arginase - metabolism; Biomarkers; Disease Models, Animal; EPHA3; Gene Expression Profiling; Humans; Hypertension, Pulmonary - genetics; Hypertension, Pulmonary - metabolism; Hypoxia - genetics; Hypoxia - metabolism; M2 macrophage; Macrophages - immunology; Macrophages - metabolism; Male; Mice; Proto-Oncogene Proteins c-akt - metabolism; Pulmonary hypertension; Rats; Rats, Sprague-Dawley; RAW 264.7 Cells; Signal Transduction; WGCNA
• ISSN: 1567-5769; 1878-1705; 1878-1705
• DOI: 10.1016/j.intimp.2024.113028

•We analyze the underlying molecular mechanisms of pulmonary hypertension (PH) and the immune cell infiltration environment within the PH.•WGCNA combines PCA, cibersort to make the results more accurate.•EPHA3, LRRC7, BICC1, and PAMR1 could be potential biomarkers and therapeutic targets for PH.•The activation of the P-Akt/Akt signaling pathway promoted by EPHA3 played an essential role in the progression of PH. Pulmonary hypertension is a serious disease. Emerging studies have shown that M2 macrophages play an essential role in pulmonary hypertension; however, their mechanism of action is uncertain. Four GEO datasets were downloaded. The differentially expressed genes (DEGs) were obtained using the limma package. Simultaneously, the Cell-type Identification By Estimating Relative Subsets Of RNA Transcripts (CIBERSORT) algorithm and weighted gene co-expression network analysis (WGCNA) were used to get the information about M2 macrophage–related modules. Potential key genes were obtained by intersecting DEGs with M2 macrophage–related module genes (M2MRGs), and finally the area under the curve (AUC) was calculated. Rats were exposed to hypoxia condition (10 % O2) for 4 weeks to induce PH. Subsequently, potential key genes with AUC>0.7 were analyzed by quantitative real-time polymerase chain reaction and Western blot using normoxia and hypoxia rat lungs. We knocked down EPHA3 in Raw264.7 cells and detected the protein expression of M2 macrophage markers including arginase 1 (ARG1) and interleukin 10 (IL-10), phospho-protein kinase B (P-Akt), and protein kinase B (Akt) to explore the downstream pathways of EPHA3. Seven potential hub genes were detected by intersecting M2MRGs and DEGs. Six genes with AUC values above 0.7 were used for further exploration. The expression of EPHA3 mRNA and protein was significantly more upregulated in rats with hypoxia than in rats with normoxia. The expression levels of IL10, ARG1, and P-Akt/Akt decreased after knocking down EPHA3. This study suggested that the activation of the P-Akt/Akt signaling pathway promoted by EPHA3 played an essential role in the progression of pulmonary hypertension.