Antisense Locked Nucleic Acids Targeting agrA Inhibit Quorum Sensing and Pathogenesis of Community-Associated Methicillin-Resistant Staphylococcus aureus.
J Appl Microbiol. 2016 Oct 8;:
Authors: Da F, Yao L, Su Z, Hou Z, Li Z, Xue X, Meng J, Luo X
AIM: Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is commonly associated with non-nosocomial skin and soft tissue infections (SSTIs) due to its virulence, which is mainly controlled by the accessory gene regulator (agr) quorum sensing (QS) system. In this study (KFF)3 K peptide-conjugated Locked Nucleic Acids (PLNAs) targeting agrA mRNA were developed to inhibit agr activity and arrest the pathogenicity of CA-MRSA.
METHODS AND RESULTS: Two PLNAs were designed, and synthesized, after predicting the secondary structure of agrA mRNA. The influence on bacterial growth was tested using a growth curve assay. RT-qPCR, hemolysis assay, LDH release assay and chemotaxis assay were used to evaluate the effects of the PLNAs on inhibiting agr QS. A mouse skin infection model was employed to test the protective effect of the PLNAs in vivo. None of the PLNAs were found to be bacteriostatic or bactericidal in vitro. However, one PLNA, PLNA34, showed strong ability to suppress expression of agrA and the effector molecule RNAIII in USA300 LAC strain. Furthermore, PLNA34 inhibited the expression of virulence genes that are upregulated by agr, including hla, psmα, psmβ and pvl. The hemolytic activity of the supernatants from PLNA34-treated bacteria was also dramatically reduced, as well as the capacity to lyse and recruit neutrophils. Moreover, PLNA34 showed high levels of protection in the CA-MRSA mouse skin infection model.
CONCLUSIONS: The anti-agrA PLNA34 can effectively inhibit the agr QS and suppress CA-MRSA pathogenicity. This article is protected by copyright. All rights reserved.
PMID: 27718524 [PubMed - as supplied by publisher]