New insights into the antimicrobial mechanism of LEAP2 mutant zebrafish under Aeromonas hydrophila infection using transcriptome analysis.

Yueyue Fei,Qin Wang,Jigang Lu, Linyue Ouyang, Quiqin Hu,Liangbiao Chen

Fish & shellfish immunology(2023)

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摘要
Liver-expressed antimicrobial peptide 2 (LEAP2) is a blood-derived antimicrobial peptide expressed predominantly in the liver. Although LEAP2 has been reported to exert antimicrobial effects in various fish species, its antimicrobial mechanism is not entirely understood. Zebrafish is an intensively developing animal model for studying bacterial diseases. In this study, we used zebrafish to identify the role of LEAP2 in bacterial infection. We found that knockout of LEAP2 in zebrafish led to a higher bacterial burden and mortality. To further investigate the effect of LEAP2 mutation on the immune system, we conducted a comparative transcriptome analysis of zebrafish with a mutant of LEAP2. Based on gene ontologies (GO) enrichment, LEAP2 mutant zebrafish revealed that, compared to wild-type zebrafish, robust responses to bacteria, inflammatory factors, and disrupt immune homeostasis and induct hyperinflammation. Furthermore, based on Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, six immune pathways were identified: Phagosome, NOD-like receptor, ferroptosis, Cytokine-cytokine receptor, Toll-like receptor, and FOXO signalling pathways. Interestingly, besides the liver, muscle, intestine, and eggs are also significantly enriched to the ferroptosis pathway, as revealed using quantitative polymerase chain reaction (qPCR), further confirmed that the effect of LEAP2 mutations on inflammatory factors and ferroptosis-related genes. Most importantly, this is the first report of the zebrafish LEAP2 mutant transcriptome obtained using high-throughput sequencing. Our study employed comparative transcriptome analysis to reveal the inflammatory response and ferroptosis-signalling pathway as a novel potential mechanism of LEAP2 antibacterial activity, laying the foundation for future studies of LEAP2 immune functions.
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