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microRNAs (miRNAs) coordinate several physiological and pathological processes by regulating the fate of mRNAs. Studies conducted  in vitro  indicate a role of microRNAs in the control of host-microbe interactions. However, there is limited understanding of miRNA functions in  in vivo  models of bacterial infections. In this study, we systematically explored changes in miRNA expression levels of  Galleria mellonella  larvae (greater-wax moth), a model system that recapitulates the vertebrate innate immunity, following infection with  L. monocytogenes . Using an insect-specific miRNA microarray with more than 2000 probes, we found differential expression of 90 miRNAs (39 upregulated and 51 downregulated) in response to infection with  L. monocytogenes . We validated the expression of a subset of miRNAs which have mammalian homologs of known or predicted function. In contrast, non-pathogenic  L. innocua  failed to induce these miRNAs, indicating a virulence-dependent miRNA deregulation. To predict miRNA targets using established algorithms, we generated a publically available  G. mellonella  transcriptome database. We identified miRNA targets involved in innate immunity, signal transduction and autophagy, including spätzle, MAP kinase, and optineurin, respectively, which exhibited a virulence-specific differential expression. Finally,  in silico  estimation of minimum free energy of miRNA-mRNA duplexes of validated microRNAs and target transcripts revealed a regulatory network of the host immune response to  L. monocytogenes . In conclusion, this study provides evidence for a role of miRNAs in the regulation of the innate immune response following bacterial infection in a simple, rapid and scalable  in vivo  model that may predict host-microbe interactions in higher vertebrates.

Listeria monocytogenes Induces a Virulence-Dependent microRNA Signature That Regulates the Immune Response in Galleria mellonella