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Novel MicroRNA Involved in Host Response to Avian Pathogenic Escherichia coli Identified by Deep Sequencing and
Xinzheng Jia1,2, Qinghua Nie3,2, Xiquan Zhang1,2
1Department of Animal Genetics, Breeding and Reproduction, College of Animal Science, South China Agricultural University, Guangzhou, Guangdong, China.
Abstract:
Avian pathogenic Escherichia coli (APEC) causes one of the most common bacterial diseases of poultry worldwide. Effective control methods are therefore desirable and will be facilitated by a better understanding of the host response to the pathogen. Currently, microRNAs (miRNAs) involved in host resistance to APEC are unknown. Here, we applied RNA sequencing to explore the changed miRNAs and deregulated genes in the spleen of three groups of broilers: nonchallenged (NC), APEC-challenged with mild pathology (CM), and APEC-challenged with severe pathology (CS). Twenty-seven differentially expressed miRNAs (fold change >1.5; P value <0.01) were identified, including 13 miRNAs between the NC and CM, 17 between the NC and CS, and 14 between the CM and CS groups. Through functional analysis of these miRNA targets, 12 immune-related biological processes were found to be significantly enriched. Based on combined analyses of differentially expressed miRNAs and mRNAs within each of the three groups, 43 miRNA-mRNA pairs displayed significantly negative correlations (r < -0.8). Notably, gga-miR-429 was greatly increased in the CS group compared to levels in both the CM and NC groups. In vitro, gga-miR-429 directly repressed luciferase reporter gene activity via binding to 3' untranslated regions of TMEFF2, NTRK2, and SHISA2. Overexpression of gga-miR-429 in the HD11 macrophage cell line significantly inhibited TMEFF2 and SHISA2 expression, which are involved in the lipopolysaccharide-induced platelet-derived growth factor (PDGF) and Wnt signaling pathways. In summary, we provide the first report characterizing the miRNA changes during APEC infection, which may help to shed light on the roles of these recently identified genetic elements in the mechanisms of host resistance and susceptibility to APEC.
Insights
This study identifies microRNAs (miRNAs) changing in chicken spleens during avian pathogenic Escherichia coli (APEC) infection. These findings reveal potential new genetic targets for improving poultry disease resistance to APEC.
Area of Science:
- Veterinary Immunology
- Molecular Biology
- Poultry Science
Background:
- Avian pathogenic Escherichia coli (APEC) is a major bacterial pathogen causing significant disease in poultry globally.
- Understanding host responses, particularly genetic factors like microRNAs (miRNAs), is crucial for developing effective APEC control strategies.
- The role of miRNAs in host resistance and susceptibility to APEC remains largely unexplored.
Purpose of the Study:
- To characterize differential miRNA expression in broiler chicken spleens following APEC challenge.
- To identify immune-related biological processes regulated by these differentially expressed miRNAs.
- To investigate the relationship between specific miRNAs and their mRNA targets in the context of APEC infection.
Main Methods:
- RNA sequencing was employed to analyze miRNA and mRNA expression profiles in the spleens of nonchallenged (NC), mildly APEC-challenged (CM), and severely APEC-challenged (CS) broilers.
- Differential expression analysis identified significant changes in miRNAs and mRNAs.
- Functional enrichment analysis and correlation analysis were performed to understand miRNA-mRNA interactions and their biological relevance.
Main Results:
- Twenty-seven differentially expressed miRNAs were identified across the experimental groups.
- Functional analysis revealed enrichment of 12 immune-related biological processes.
- Forty-three negatively correlated miRNA-mRNA pairs were identified, with gga-miR-429 showing increased expression in severe APEC infection and repressing TMEFF2 and SHISA2 in vitro.
Conclusions:
- This study provides the first comprehensive characterization of miRNA expression changes during APEC infection in poultry.
- The identified miRNAs, particularly gga-miR-429, represent potential regulators of host immune responses to APEC.
- These findings offer insights into the molecular mechanisms underlying host resistance and susceptibility to APEC, paving the way for novel therapeutic targets.
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