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Transcriptome sequencing and ceRNA (lncRNA-miRNA-mRNA) network construction identified the key molecular markers and signaling pathways of chronic thromboembolic pulmonary hypertension

ABSTRACT: For patients with chronic thromboembolic pulmonary hypertension (CTEPH), it can lead to death if left untreated. In this study, we attempted to identify key genes and pathways that could help in the diagnosis and treatment of CTEPH. It also provides research direction for further understanding the molecular mechanism of CTEPH.Firstly, we extract RNA from blood samples to construct the library. Then, qualified libraries were sequenced using PE100 strategy on BGIseq platform. Subsequently, the DESeq2 package in R was used to screen differentially expressed mRNAs (DEmRNAs) and differentially expressed lncRNAs (DElncRNAs) of the CTEPH patient group and the normal control group. Afterwards, we performed functional enrichment and protein-protein interaction analysis of DEmRNAs. In addition, we also performed lncRNA-mRNA co-expression analysis and lncRNA-miRNA-mRNA network construction. A total of 437 DEmRNAs and 192 DElncRNAs were obtained. Subsequently, 205 pairs of interacting DEmRNAs and 232 pairs of lncRNA-mRNA relationship were obtained. Moreover, DEmRNAs were significantly enriched in chemokine signaling pathway, metabolic pathways, arachidonic acid metabolism and MAPK signaling pathway. In addition, only one regulation pathway of SOBP-has-miR-320b-LINC00472 was found through ceRNA network construction. In diagnostic analysis, the area under curve (AUC) of LINC00472, PIK3R6, SCN3A and TCL6 were greater than 0.7, which were 0.964, 0.893, 0.750 and 0.732, respectively. It is indicated that LINC00472, PIK3R6, SCN3A and TCL6 may as the potential diagnostic gene markers in CTEPH.

ORGANISM(S): Homo sapiens

PROVIDER: GSE188938 | GEO | 2022/08/17

REPOSITORIES: GEO

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Project description:Adiponectin (APN) is an endogenous adipokine secreted from adipocytes that exerts an anti-inflammation property. AdipoAI is an orally active adiponectin receptor agonist identified by our group, which can emulate APN's anti-inflammatory properties through mechanisms not fully understood. To explore AdipoAI function, we used lncRNA microarray and got differential lncRNA/mRNA expression medicated by AdipoAI. Identified as one kind of non-coding RNA with more than 200bp length, lncRNA has been demonstrated to have abundance biological functions, including anti-inflammatory response. In the current study, we performed an lncRNA microarray in LPS-induced Raw264.7 cells which pre-stimulated with AdipoAI, and screened 110 DElncRNAs and 190 DEmRNAs. Enrichment analyses were conducted to total mRNAs and DEmRNAs, including GSVA, ssGSEA, GO/KEGG, GSEA and PPI analysis. Among all these processes, endocytosis was significantly enriched. A co-expression analysis was built based on DElncRNAs and DEmRNAs. Then, using Targetscan and miRwalk to predict related microRNAs of DElncRNAs and DEmRNAs respectively, we established competing endogenous RNA (ceRNA) networks including 54 mRNAs from 8 GO items. Furthermore, 33 m6A methylation related marker genes were obtained from previous study and used for the construction of m6A related-lncRNA network using the co-expression analysis. We identified FTO as the hub gene of the network, and 14 lncRNAs that interacted with it. The expression levels of 10 lncRNAs selected from ceRNA and FTO- related lncRNAs networks were validated with qRT-PCR. Finally, Macrophage phenotype scores showed that AdipoAI could attenuate the M2b and M2c polarization of macrophage and correlate with the above lncRNAs. Our work reveals that lncRNA might involve in the anti-inflammation process of AdipoAI in LPS-induced macrophages through ceRNA network and epigenetic regulation of m6A. Mechanistically, these lncRNAs associated with AdipoAI might be related to endocytosis and polarization in macrophage, and provide new candidates for the anti-inflammatory application of APN and its receptor agonist.

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Transcriptome sequencing and ceRNA (lncRNA-miRNA-mRNA) network construction identified the key molecular markers and signaling pathways of chronic thromboembolic pulmonary hypertension

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