Galleria mellonella Larvae as an Infection Model to Investigate sRNA-Mediated Pathogenesis in Staphylococcus aureus. Ménard, G., Rouillon, A., Ghukasyan, G., Emily, M., Felden, B., & Donnio, P. Front. Cell. Infect. Microbiol., April, 2021. Publisher: Frontiers
Galleria mellonella Larvae as an Infection Model to Investigate sRNA-Mediated Pathogenesis in Staphylococcus aureus [link]Paper  doi  abstract   bibtex   
\textlessp\textgreaterSmall regulatory RNAs (sRNAs) are key players in bacterial regulatory networks. Monitoring their expression inside living colonized or infected organisms is essential for identifying sRNA functions, but few studies have looked at sRNA expression during host infection with bacterial pathogens. Insufficient \textlessitalic\textgreaterin vivo\textless/italic\textgreater studies monitoring sRNA expression attest to the difficulties in collecting such data, we therefore developed a non-mammalian infection model using larval \textlessitalic\textgreaterGalleria mellonella\textless/italic\textgreater to analyze the roles of \textlessitalic\textgreaterStaphylococcus aureus\textless/italic\textgreater sRNAs during larval infection and to quickly determine possible sRNA involvement in staphylococcal virulence before proceeding to more complicated animal testing. We began by using the model to test infected larvae for immunohistochemical evidence of infection as well as host inflammatory responses over time. To monitor sRNA expression during infection, total RNAs were extracted from the larvae and invading bacteria at different time points. The expression profiles of the tested sRNAs were distinct and they fluctuated over time, with expression of both \textlessitalic\textgreatersprD\textless/italic\textgreater and \textlessitalic\textgreatersprC\textless/italic\textgreater increased during infection and associated with mortality, while \textlessitalic\textgreaterrnaIII\textless/italic\textgreater expression remained barely detectable over time. A strong correlation was observed between \textlessitalic\textgreatersprD\textless/italic\textgreater expression and the mortality. To confirm these results, we used sRNA-knockout mutants to investigate sRNA involvement in \textlessitalic\textgreaterStaphylococcus\textless/italic\textgreater \textlessitalic\textgreateraureus\textless/italic\textgreater pathogenesis, finding that the decrease in death rates is delayed when either \textlessitalic\textgreatersprD\textless/italic\textgreater or \textlessitalic\textgreatersprC\textless/italic\textgreater was lacking. These results demonstrate the relevance of this \textlessitalic\textgreaterG. mellonella\textless/italic\textgreater model for investigating the role of sRNAs as transcriptional regulators involved in staphylococcal virulence. This insect model provides a fast and easy method for monitoring sRNA (and mRNA) participation in \textlessitalic\textgreaterS. aureus\textless/italic\textgreater pathogenesis, and can also be used for other human bacterial pathogens.\textless/p\textgreater
@article{menard_galleria_2021-1,
	title = {Galleria mellonella {Larvae} as an {Infection} {Model} to {Investigate} {sRNA}-{Mediated} {Pathogenesis} in {Staphylococcus} aureus},
	volume = {11},
	issn = {2235-2988},
	url = {https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2021.631710/full},
	doi = {10.3389/fcimb.2021.631710},
	abstract = {{\textless}p{\textgreater}Small regulatory RNAs (sRNAs) are key players in bacterial regulatory networks. Monitoring their expression inside living colonized or infected organisms is essential for identifying sRNA functions, but few studies have looked at sRNA expression during host infection with bacterial pathogens. Insufficient {\textless}italic{\textgreater}in vivo{\textless}/italic{\textgreater} studies monitoring sRNA expression attest to the difficulties in collecting such data, we therefore developed a non-mammalian infection model using larval {\textless}italic{\textgreater}Galleria mellonella{\textless}/italic{\textgreater} to analyze the roles of {\textless}italic{\textgreater}Staphylococcus aureus{\textless}/italic{\textgreater} sRNAs during larval infection and to quickly determine possible sRNA involvement in staphylococcal virulence before proceeding to more complicated animal testing. We began by using the model to test infected larvae for immunohistochemical evidence of infection as well as host inflammatory responses over time. To monitor sRNA expression during infection, total RNAs were extracted from the larvae and invading bacteria at different time points. The expression profiles of the tested sRNAs were distinct and they fluctuated over time, with expression of both {\textless}italic{\textgreater}sprD{\textless}/italic{\textgreater} and {\textless}italic{\textgreater}sprC{\textless}/italic{\textgreater} increased during infection and associated with mortality, while {\textless}italic{\textgreater}rnaIII{\textless}/italic{\textgreater} expression remained barely detectable over time. A strong correlation was observed between {\textless}italic{\textgreater}sprD{\textless}/italic{\textgreater} expression and the mortality. To confirm these results, we used sRNA-knockout mutants to investigate sRNA involvement in {\textless}italic{\textgreater}Staphylococcus{\textless}/italic{\textgreater} {\textless}italic{\textgreater}aureus{\textless}/italic{\textgreater} pathogenesis, finding that the decrease in death rates is delayed when either {\textless}italic{\textgreater}sprD{\textless}/italic{\textgreater} or {\textless}italic{\textgreater}sprC{\textless}/italic{\textgreater} was lacking. These results demonstrate the relevance of this {\textless}italic{\textgreater}G. mellonella{\textless}/italic{\textgreater} model for investigating the role of sRNAs as transcriptional regulators involved in staphylococcal virulence. This insect model provides a fast and easy method for monitoring sRNA (and mRNA) participation in {\textless}italic{\textgreater}S. aureus{\textless}/italic{\textgreater} pathogenesis, and can also be used for other human bacterial pathogens.{\textless}/p{\textgreater}},
	language = {English},
	urldate = {2024-09-11},
	journal = {Front. Cell. Infect. Microbiol.},
	author = {Ménard, Guillaume and Rouillon, Astrid and Ghukasyan, Gevorg and Emily, Mathieu and Felden, Brice and Donnio, Pierre-Yves},
	month = apr,
	year = {2021},
	note = {Publisher: Frontiers},
	keywords = {Galleria mellonella, regulation, sRNA, Staphylococcus aureus, Virulence},
	file = {Texte intégral:D\:\\Home\\maguillout\\Zotero\\Donnees\\storage\\J8QQNEVU\\Ménard et al. - 2021 - Galleria mellonella Larvae as an Infection Model t.pdf:application/pdf},
}
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