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Targeting Type IV pili as an antivirulence strategy against invasive meningococcal disease

Abstract

Bacterial virulence factors are attractive targets for the development of therapeutics. Type IV pili, which are associated with a remarkable array of properties including motility, the interaction between bacteria and attachment to biotic and abiotic surfaces, represent particularly appealing virulence factor targets. Type IV pili are present in numerous bacterial species and are critical for their pathogenesis. In this study, we report that trifluoperazine and related phenothiazines block functions associated with Type IV pili in different bacterial pathogens, by affecting piliation within minutes. Using Neisseria meningitidis as a paradigm of Gram-negative bacterial pathogens that require Type IV pili for pathogenesis, we show that piliation is sensitive to altered activity of the Na+ pumping NADH–ubiquinone oxidoreductase (Na+−NQR) complex and that these compounds probably altered the establishment of the sodium gradient. In vivo, these compounds exert a strong protective effect. They reduce meningococcal colonization of the human vessels and prevent subsequent vascular dysfunctions, intravascular coagulation and overwhelming inflammation, the hallmarks of invasive meningococcal infections. Finally, they reduce lethality. This work provides a proof of concept that compounds with activity against bacterial Type IV pili could beneficially participate in the treatment of infections caused by Type IV pilus-expressing bacteria.

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Fig. 1: Trifluoperazine inhibits bacterial aggregation.
Fig. 2: Trifluoperazine and thioridazine regulate the piliation status of meningococci.
Fig. 3: Trifluoperazine and thioridazine release bacteria from compact microcolonies formed on human endothelial cells.
Fig. 4: Trifluoperazine and thioridazine reduce bacteria-induced endothelial cell injury and matrix degradation.
Fig. 5: Trifluoperazine and thioridazine induce bacterial clearance from human blood vessels in vivo and reduce signs of thrombosis, vascular injury and inflammation.
Fig. 6: Thioridazine improves the outcome of meningococcal infection alone or in combination with antibiotics.

Data availability

Data supporting the findings of this study are available within the paper and the Supplementary Information. The sequences of the SNP variants resistant to the action of phenothiazines are available at NCBI’s BioProject database under accession number PRJNA481885.

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Acknowledgements

We thank M. Robert-Genthon and F. Chretien for providing the P. aeruginosa PAO1 strain and brain tissues; M. Favier of the histology facility, A. Schmitt and T. Guilbert of the imaging facility, and K. Bailly and M. Andrieu of the cytometry facility of the Institut Cochin for their expert technical help. K.D. and L.LG. were supported by a doctoral fellowship from la Region Ile de France and the Fondation pour la Recherche Médicale, respectively. This work was supported by collaborative research grants from the Agence Nationale de la Recherche of France (grant no. ANR-14-IFEC14-0006) to S.B. and X.N. in the framework of the Infect-ERA joint transnational call (European funding for infectious diseases research), by the Société d’Accélération du Transfert de Technologie (grant no. ANR-10-SATT-05-01) to S.B. and by grant no. ANR-10-EQPX-04-01 to F.L.

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Contributions

K.D. and M.L.B. conducted the in vitro experiments and the histological and immunofluorescence analysis of the skin grafts; L.L.G., J.P.B., C.F. and O.J.-L. conducted the in vivo experiments; A.G. performed the qPCR and multiplex assays; H.B.-S. and B.D. performed dSTORM analysis; A.J. and D.E. performed the whole-genome sequencing analysis of the bacterial resistant strains; P.P. provided the human skin tissues; M.C. provided bacterial mutants; N.B. and F.L. conducted the SEM assays; P.C.M. discussed the results and helped to write the manuscript; X.N. provided bacterial strains and discussed the results; and S.B. designed the experiments, supervised the project and wrote the manuscript.

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Correspondence to Sandrine Bourdoulous.

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Supplementary information

Supplementary Information

Supplementary Videos, Supplementary Tables 1–4 and Supplementary Figures 1–14b.

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Supplementary Video 1

Sample movie of meningococcal aggregates treated with trifluoperazine.

Supplementary Video 2

Sample movie of meningococcal aggregates treated with trifluoperazine.

Supplementary Video 3

Sample movie of gonococcal aggregates treated with trifluoperazine.

Supplementary Video 4

Sample movie of PilT meningococcal aggregates treated with trifluoperazine.

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Denis, K., Le Bris, M., Le Guennec, L. et al. Targeting Type IV pili as an antivirulence strategy against invasive meningococcal disease. Nat Microbiol 4, 972–984 (2019). https://doi.org/10.1038/s41564-019-0395-8

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