Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses
Abstract Background Diabetic foot ulcers (DFU) are among the fastest-growing diseases worldwide. Recent evidence has emphasized the critical role of microRNA (miRNA)-mRNA networks in various chronic wounds, including DFU. In this study, we aimed to clarify the miRNA-mRNA axes associated with the occ...
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| Format: | Article |
| Language: | English |
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BMC
2023-12-01
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| Series: | BMC Medical Genomics |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12920-023-01741-2 |
| _version_ | 1797413721493995520 |
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| author | Yuanyuan Xu Jianchang Xu Sirong Chen Anbang Zhou Guangjing Huang Shidao Huang Dianbo Yu Biaoliang Wu |
| author_facet | Yuanyuan Xu Jianchang Xu Sirong Chen Anbang Zhou Guangjing Huang Shidao Huang Dianbo Yu Biaoliang Wu |
| author_sort | Yuanyuan Xu |
| collection | DOAJ |
| description | Abstract Background Diabetic foot ulcers (DFU) are among the fastest-growing diseases worldwide. Recent evidence has emphasized the critical role of microRNA (miRNA)-mRNA networks in various chronic wounds, including DFU. In this study, we aimed to clarify the miRNA-mRNA axes associated with the occurrence of DFU. Methods Expression profiles of miRNAs and mRNAs were extracted from the Gene Expression Omnibus. Differentially expressed genes and differentially expressed miRNAs were identified, and miRNA-mRNA regulatory axes were constructed through integrated bioinformatics analyses. We validated the miRNA-mRNA axes using quantitative real-time PCR (qPCR) and dual-luciferase reporter assays. We conducted an immune infiltration analysis and confirmed the bioinformatics results using immunofluorescence staining. Single-sample gene set enrichment analysis (ssGSEA) was used to analyze the metabolic mechanisms. Results miR-182-5p-CHL1/MITF and miR-338-3p-NOVA1 interactions were identified using in silico analysis. The qPCR results showed apparent dysregulation of these miRNA-mRNA axes in DFU. The dual-luciferase reporter assay confirmed that miR-182-5p targeted CHL1 and MITF, and miR-338-3p targeted NOVA1. We conducted an immune infiltration analysis and observed that key genes correlated with decreased infiltration of M1 macrophages and resting mast cells in DFU. Immunofluorescence staining verified the co-localization of CHL1 and tryptase, while MITF and CD68 showed weak positive correlations. Metabolic pathways related to these three genes were identified using ssGSEA. Conclusions In summary, the miR-182-5p-CHL1/MITF and miR-338-3p-NOVA1 pathway interactions and decreased infiltration of M1 macrophages and resting mast cells may provide novel clues to the pathogenesis of DFU. Trial registration The clinical trial included in this study was registered in the Chinese Clinical Trial Registry ( ChiCTR2200066660 ) on December 13, 2022. |
| first_indexed | 2024-03-09T05:22:19Z |
| format | Article |
| id | doaj.art-8dfc5f8bd1f246b5afc2eb9117ca4656 |
| institution | Directory Open Access Journal |
| issn | 1755-8794 |
| language | English |
| last_indexed | 2024-03-09T05:22:19Z |
| publishDate | 2023-12-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Medical Genomics |
| spelling | doaj.art-8dfc5f8bd1f246b5afc2eb9117ca46562023-12-03T12:39:43ZengBMCBMC Medical Genomics1755-87942023-12-0116111410.1186/s12920-023-01741-2Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analysesYuanyuan Xu0Jianchang Xu1Sirong Chen2Anbang Zhou3Guangjing Huang4Shidao Huang5Dianbo Yu6Biaoliang Wu7Graduate School, Youjiang Medical University for NationalitiesThe First Clinical College of Wuhan UniversityGraduate School, Youjiang Medical University for NationalitiesGraduate School, Youjiang Medical University for NationalitiesGraduate School, Youjiang Medical University for NationalitiesGraduate School, Youjiang Medical University for NationalitiesGraduate School, Youjiang Medical University for NationalitiesDepartment of Endocrinology, The Affiliated Hospital of Youjiang Medical University for NationalitiesAbstract Background Diabetic foot ulcers (DFU) are among the fastest-growing diseases worldwide. Recent evidence has emphasized the critical role of microRNA (miRNA)-mRNA networks in various chronic wounds, including DFU. In this study, we aimed to clarify the miRNA-mRNA axes associated with the occurrence of DFU. Methods Expression profiles of miRNAs and mRNAs were extracted from the Gene Expression Omnibus. Differentially expressed genes and differentially expressed miRNAs were identified, and miRNA-mRNA regulatory axes were constructed through integrated bioinformatics analyses. We validated the miRNA-mRNA axes using quantitative real-time PCR (qPCR) and dual-luciferase reporter assays. We conducted an immune infiltration analysis and confirmed the bioinformatics results using immunofluorescence staining. Single-sample gene set enrichment analysis (ssGSEA) was used to analyze the metabolic mechanisms. Results miR-182-5p-CHL1/MITF and miR-338-3p-NOVA1 interactions were identified using in silico analysis. The qPCR results showed apparent dysregulation of these miRNA-mRNA axes in DFU. The dual-luciferase reporter assay confirmed that miR-182-5p targeted CHL1 and MITF, and miR-338-3p targeted NOVA1. We conducted an immune infiltration analysis and observed that key genes correlated with decreased infiltration of M1 macrophages and resting mast cells in DFU. Immunofluorescence staining verified the co-localization of CHL1 and tryptase, while MITF and CD68 showed weak positive correlations. Metabolic pathways related to these three genes were identified using ssGSEA. Conclusions In summary, the miR-182-5p-CHL1/MITF and miR-338-3p-NOVA1 pathway interactions and decreased infiltration of M1 macrophages and resting mast cells may provide novel clues to the pathogenesis of DFU. Trial registration The clinical trial included in this study was registered in the Chinese Clinical Trial Registry ( ChiCTR2200066660 ) on December 13, 2022.https://doi.org/10.1186/s12920-023-01741-2Diabetic foot ulcersmiRNAs-mRNAImmune infiltrationDual-luciferase reporter assayqPCRImmunofluorescence staining |
| spellingShingle | Yuanyuan Xu Jianchang Xu Sirong Chen Anbang Zhou Guangjing Huang Shidao Huang Dianbo Yu Biaoliang Wu Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses BMC Medical Genomics Diabetic foot ulcers miRNAs-mRNA Immune infiltration Dual-luciferase reporter assay qPCR Immunofluorescence staining |
| title | Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses |
| title_full | Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses |
| title_fullStr | Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses |
| title_full_unstemmed | Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses |
| title_short | Identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses |
| title_sort | identifying potential pathogenesis and immune infiltration in diabetic foot ulcers using bioinformatics and in vitro analyses |
| topic | Diabetic foot ulcers miRNAs-mRNA Immune infiltration Dual-luciferase reporter assay qPCR Immunofluorescence staining |
| url | https://doi.org/10.1186/s12920-023-01741-2 |
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