Pseudomonas aeruginosa outer membrane vesicle-packed sRNAs can enter host cells and regulate innate immune responses

• Published in: Microbial pathogenesis 2024-03, Vol.188, p.106562-106562, Article 106562
• Main Authors: Xie, Zhen, Wang, Xiao, Huang, Yangyang, Chen, Shukun, Liu, Mohua, Zhang, Fuhua, Li, Mengyuan, Gu, Yanchao, Yang, Yadong, Shen, Xihui, Wang, Yao, Xu, Yang, Xu, Lei
• Format: Article
• Language: English
• Subjects: Host-Pathogen Interactions; Humans; Immunity, Innate; Innate immune; Outer membrane vesicles (OMVs); Pathogen-host interactions; Pseudomonas aeruginosa; Pseudomonas aeruginosa - physiology; Pseudomonas Infections - microbiology; RNA, Bacterial - genetics; RNA, Bacterial - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Small RNA
• ISSN: 0882-4010; 1096-1208
• DOI: 10.1016/j.micpath.2024.106562

Bacterial outer membrane vesicles (OMVs) can package and deliver virulence factors into host cells, which is an important mechanism mediating host-pathogen interactions. It has been reported that small RNAs (sRNAs) can be packed into OMVs with varying relative abundance, which might affect the function and/or stability of host mRNAs. In this study, we used OptiPrep density gradient ultra-high-speed centrifugation to purify OMVs from Pseudomonas aeruginosa. Next, the sequences and abundance of sRNAs were detected by using Small RNA-Seq. In particular, sRNA4518698, sRNA2316613 and sRNA809738 were the three most abundant sRNAs in OMVs, which are all fragments of P. aeruginosa non-coding RNAs. sRNAs were shielded within the interior of OMVs and remained resistant to external RNase cleavage. The miRanda and RNAhybrid analysis demonstrated that those sRNAs could target a large number of host mRNAs, which were enriched in host immune responses by the functions of GO and KEGG enrichment. Experimentally, we demonstrated that the transfection of synthetic sRNA4518698, sRNA2316613, or sRNA809738 could reduce the expression of innate immune response genes in RAW264.7 cells. Together, we demonstrated that P. aeruginosa OMVs sRNAs can regulate innate immune responses. This study uncovered a mechanism in which the OMVs regulate host responses by transferring bacterial sRNAs. •Enrichment of small RNAs within outer membrane vesicles of Pseudomonas aeruginosa.•Outer membrane vesicles transfer sRNAs into host cells to target innate immune responses.•SRNA4518698, sRNA809738, and sRNA2316613 suppress expression of host innate immune genes.