Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment
BackgroundMacrophages play an important role in regulating the course of spinal tuberculosis within the bone microenvironment. This study aimed to investigate the differential expression of miRNA in macrophage-derived exosomes within the tuberculosis-infected bone microenvironment, to identify miRNA...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-08-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1236012/full |
| _version_ | 1797755377763221504 |
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| author | Zhicheng Sun Xiaoyang Pang Xiyang Wang Hao Zeng Hao Zeng |
| author_facet | Zhicheng Sun Xiaoyang Pang Xiyang Wang Hao Zeng Hao Zeng |
| author_sort | Zhicheng Sun |
| collection | DOAJ |
| description | BackgroundMacrophages play an important role in regulating the course of spinal tuberculosis within the bone microenvironment. This study aimed to investigate the differential expression of miRNA in macrophage-derived exosomes within the tuberculosis-infected bone microenvironment, to identify miRNAs that hold potential as diagnostic markers and therapeutic targets.MethodsWe established study cohorts for spinal tuberculosis, collected bone marrow blood samples, isolated macrophage exosomes, and performed exosome miRNA sequencing. A miRNA-mRNA co-expression network was constructed using WGCNA analysis. Gene GO analysis and KEGG pathway enrichment analysis were performed using KOBAS software. Target miRNAs were selected based on fold change, P-value, and false discovery rate, and their validation was carried out using qRT-PCR and ROC curve studies. Subsequently, we constructed a target gene network for these miRNAs and performed KEGG pathway enrichment analysis to explore the potential signaling mechanisms involved in regulating the disease course of spinal tuberculosis.ResultsOur findings revealed that macrophages from the tuberculosis-infected bone microenvironment exhibited an M1 phenotype. The successful extraction of exosomes from macrophage supernatants was confirmed through electron microscopy, particle size analysis, and protein blot analysis. Exosome miRNA-seq demonstrated that 28 miRNAs were up-regulated, while 34 miRNAs were down-regulated in individuals with spinal tuberculosis. GO analysis and KEGG pathway enrichment analysis indicated that the differentially expressed miRNAs were involved in various biological processes, cell components, molecular functions, and signaling pathways, which collectively contribute to the regulation of the disease course of spinal tuberculosis. Notably, miRNA-125b-5p was successfully selected based on fold change, p-value, and false discovery rate. qRT-PCR validation further confirmed the significant up-regulation of miRNA-125b-5p in spinal tuberculosis. The ROC curve revealed that miR-125b-5p is a potential diagnostic biomarker for spinal tuberculosis. Moreover, construction of the miRNA-125b-5p target gene network and subsequent KEGG enrichment analysis highlighted the importance of MAPK, TNF, Ras, Rap1, and the PI3K-Akt signaling pathways in the regulation of the disease course of spinal tuberculosis.ConclusionOur study demonstrates differential expression of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment. Specifically, MiRNA-125b-5p is significantly up-regulated in spinal tuberculosis and shows potential as a diagnostic biomarker for spinal tuberculosis. |
| first_indexed | 2024-03-12T17:47:01Z |
| format | Article |
| id | doaj.art-dd00e901656948739fc0080e8bc84bcb |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-03-12T17:47:01Z |
| publishDate | 2023-08-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-dd00e901656948739fc0080e8bc84bcb2023-08-03T16:49:31ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-08-011410.3389/fmicb.2023.12360121236012Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironmentZhicheng Sun0Xiaoyang Pang1Xiyang Wang2Hao Zeng3Hao Zeng4Department of Spinal Surgery, Xiangya Hospital of Central South University, Changsha, Hunan, ChinaDepartment of Spinal Surgery, Xiangya Hospital of Central South University, Changsha, Hunan, ChinaDepartment of Spinal Surgery, Xiangya Hospital of Central South University, Changsha, Hunan, ChinaDepartment of Spinal Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, ChinaGuangxi Key Laboratory of Regenerative Medicine, Nanning, Guangxi, ChinaBackgroundMacrophages play an important role in regulating the course of spinal tuberculosis within the bone microenvironment. This study aimed to investigate the differential expression of miRNA in macrophage-derived exosomes within the tuberculosis-infected bone microenvironment, to identify miRNAs that hold potential as diagnostic markers and therapeutic targets.MethodsWe established study cohorts for spinal tuberculosis, collected bone marrow blood samples, isolated macrophage exosomes, and performed exosome miRNA sequencing. A miRNA-mRNA co-expression network was constructed using WGCNA analysis. Gene GO analysis and KEGG pathway enrichment analysis were performed using KOBAS software. Target miRNAs were selected based on fold change, P-value, and false discovery rate, and their validation was carried out using qRT-PCR and ROC curve studies. Subsequently, we constructed a target gene network for these miRNAs and performed KEGG pathway enrichment analysis to explore the potential signaling mechanisms involved in regulating the disease course of spinal tuberculosis.ResultsOur findings revealed that macrophages from the tuberculosis-infected bone microenvironment exhibited an M1 phenotype. The successful extraction of exosomes from macrophage supernatants was confirmed through electron microscopy, particle size analysis, and protein blot analysis. Exosome miRNA-seq demonstrated that 28 miRNAs were up-regulated, while 34 miRNAs were down-regulated in individuals with spinal tuberculosis. GO analysis and KEGG pathway enrichment analysis indicated that the differentially expressed miRNAs were involved in various biological processes, cell components, molecular functions, and signaling pathways, which collectively contribute to the regulation of the disease course of spinal tuberculosis. Notably, miRNA-125b-5p was successfully selected based on fold change, p-value, and false discovery rate. qRT-PCR validation further confirmed the significant up-regulation of miRNA-125b-5p in spinal tuberculosis. The ROC curve revealed that miR-125b-5p is a potential diagnostic biomarker for spinal tuberculosis. Moreover, construction of the miRNA-125b-5p target gene network and subsequent KEGG enrichment analysis highlighted the importance of MAPK, TNF, Ras, Rap1, and the PI3K-Akt signaling pathways in the regulation of the disease course of spinal tuberculosis.ConclusionOur study demonstrates differential expression of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment. Specifically, MiRNA-125b-5p is significantly up-regulated in spinal tuberculosis and shows potential as a diagnostic biomarker for spinal tuberculosis.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1236012/fullspinal tuberculosisbone microenvironmentmacrophagesexosomesmiRNA-seqbiomarker |
| spellingShingle | Zhicheng Sun Xiaoyang Pang Xiyang Wang Hao Zeng Hao Zeng Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment Frontiers in Microbiology spinal tuberculosis bone microenvironment macrophages exosomes miRNA-seq biomarker |
| title | Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment |
| title_full | Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment |
| title_fullStr | Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment |
| title_full_unstemmed | Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment |
| title_short | Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment |
| title_sort | differential expression analysis of mirnas in macrophage derived exosomes in the tuberculosis infected bone microenvironment |
| topic | spinal tuberculosis bone microenvironment macrophages exosomes miRNA-seq biomarker |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1236012/full |
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