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Lung microRNA Profiling Across the Estrous Cycle in Ozone-exposed Mice
Published on: January 7, 2019
Investigation of the miRNA-mRNA Regulatory Circuits and Immune Signatures Associated with Bronchopulmonary Dysplasia
Sen Li1, Shuling Liang2, Shunyu Xie1
1Guangzhou Women and Children's Medical Center, State Key Laboratory of Respiratory Disease and Guangzhou Medical University, Guangzhou, Guangdong Province, People's Republic of China.
Background:
Bronchopulmonary dysplasia (BPD) has become a major cause of morbidity and mortality in preterm infants worldwide, yet its pathogenesis and underlying mechanisms remain poorly understood. The present study sought to explore microRNA-mRNA regulatory networks and immune cells involvement in BPD through a combination of bioinformatic analysis and experimental validation.
Methods:
MicroRNA and mRNA microarray datasets were obtained from the Gene Expression Omnibus (GEO) database. Differentially expressed microRNAs (DEMs) were identified in BPD patients compared to control subjects, and their target genes were predicted using miRWalk, miRNet, miRDB, and TargetScan databases. Subsequently, protein-protein interaction (PPI) and functional enrichment analyses were conducted on the target genes. 30 hub genes were screened using the Cytohubba plugin of the Cytoscape software. Additionally, mRNA microarray data was utilized to validate the expression of hub genes and to perform immune infiltration analysis. Finally, real-time PCR (RT-PCR), immunohistochemistry (IHC), and flow cytometry were conducted using a mouse model of BPD to confirm the bioinformatics findings.
Results:
Two DEMs (miR-15b-5p and miR-20a-5p) targeting genes primarily involved in the regulation of cell cycle phase transition, ubiquitin ligase complex, protein serine/threonine kinase activity, and MAPK signaling pathway were identified. APP and four autophagy-related genes (DLC1, PARP1, NLRC4, and NRG1) were differentially expressed in the mRNA microarray dataset. Analysis of immune infiltration revealed significant differences in levels of neutrophils and naive B cells between BPD patients and control subjects. RT-PCR and IHC confirmed reduced expression of APP in a mouse model of BPD. Although the proportion of total neutrophils did not change appreciably, the activation of neutrophils, marked by loss of CD62L, was significantly increased in BPD mice.
Conclusion:
Downregulation of APP mediated by miR-15b-5p and miR-20a-5p may be associated with the development of BPD. Additionally, increased CD62L- neutrophil subset might be important for the immune-mediated injury in BPD.
Insights
MicroRNA-mRNA networks and immune cell changes are implicated in bronchopulmonary dysplasia (BPD). Reduced APP expression and activated neutrophils may drive BPD pathogenesis in preterm infants.
Area of Science:
- Pulmonary Medicine
- Genomics
- Immunology
Background:
- Bronchopulmonary dysplasia (BPD) is a significant cause of morbidity and mortality in preterm infants.
- The underlying pathogenesis and mechanisms of BPD remain poorly understood.
- This study investigates microRNA-mRNA regulatory networks and immune cell involvement in BPD.
Purpose of the Study:
- To explore microRNA-mRNA regulatory networks in BPD.
- To identify the role of immune cells in BPD pathogenesis.
- To validate bioinformatic findings through experimental methods.
Main Methods:
- Utilized Gene Expression Omnibus (GEO) datasets for microRNA and mRNA expression.
- Predicted microRNA targets and performed protein-protein interaction analysis.
- Screened hub genes and analyzed immune cell infiltration.
- Validated findings using a mouse model with RT-PCR, IHC, and flow cytometry.
Main Results:
- Identified two microRNAs (miR-15b-5p, miR-20a-5p) targeting genes in cell cycle and signaling pathways.
- Found differential expression of APP and autophagy-related genes.
- Observed significant differences in neutrophils and naive B cells in BPD patients.
- Confirmed reduced APP expression and increased neutrophil activation in a BPD mouse model.
Conclusions:
- Downregulation of APP, potentially mediated by miR-15b-5p and miR-20a-5p, may contribute to BPD development.
- An increased CD62L-negative neutrophil subset could be crucial for immune-mediated injury in BPD.
