In Vivo Bactericidal Efficacy of GWH1 Antimicrobial Peptide Displayed on Protein Nanoparticles, a Potential Alternative to Antibiotics
Jose V. Carratalá,
Eric Brouillette,
Naroa Serna,
Alejandro Sánchez-Chardi,
Julieta M. Sánchez,
Antonio Villaverde,
Anna Arís,
Elena Garcia-Fruitós,
Neus Ferrer-Miralles,
François Malouin
Affiliations
Jose V. Carratalá
Institute for Biotechnology and Biomedicine, Autonomous University of Barcelona, Bellaterra, 08193 Barcelona, Spain
Eric Brouillette
Centre d’Étude et de Valorisation de la Diversité Microbienne (CEVDM), Département de Biologie, Université de Sherbrooke, 2500 Boul. Université, Sherbrooke, QC J1K 2R1, Canada
Naroa Serna
Institute for Biotechnology and Biomedicine, Autonomous University of Barcelona, Bellaterra, 08193 Barcelona, Spain
Alejandro Sánchez-Chardi
Microscopy Service, Autonomous University of Barcelona, Bellaterra, 08193 Barcelona, Spain
Julieta M. Sánchez
Institute for Biotechnology and Biomedicine, Autonomous University of Barcelona, Bellaterra, 08193 Barcelona, Spain
Antonio Villaverde
Institute for Biotechnology and Biomedicine, Autonomous University of Barcelona, Bellaterra, 08193 Barcelona, Spain
Anna Arís
Department of Ruminant Production, Institute of Agriculture and Agrifood Research and Technology (IRTA), Caldes de Montbui, 08140 Barcelona, Spain
Elena Garcia-Fruitós
Department of Ruminant Production, Institute of Agriculture and Agrifood Research and Technology (IRTA), Caldes de Montbui, 08140 Barcelona, Spain
Neus Ferrer-Miralles
Institute for Biotechnology and Biomedicine, Autonomous University of Barcelona, Bellaterra, 08193 Barcelona, Spain
François Malouin
Centre d’Étude et de Valorisation de la Diversité Microbienne (CEVDM), Département de Biologie, Université de Sherbrooke, 2500 Boul. Université, Sherbrooke, QC J1K 2R1, Canada
Oligomerization of antimicrobial peptides into nanosized supramolecular complexes produced in biological systems (inclusion bodies and self-assembling nanoparticles) seems an appealing alternative to conventional antibiotics. In this work, the antimicrobial peptide, GWH1, was N-terminally fused to two different scaffold proteins, namely, GFP and IFN-γ for its bacterial production in the form of such recombinant protein complexes. Protein self-assembling as regular soluble protein nanoparticles was achieved in the case of GWH1-GFP, while oligomerization into bacterial inclusion bodies was reached in both constructions. Among all these types of therapeutic proteins, protein nanoparticles of GWH1-GFP showed the highest bactericidal effect in an in vitro assay against Escherichia coli, whereas non-oligomerized GWH1-GFP and GWH1-IFN-γ only displayed a moderate bactericidal activity. These results indicate that the biological activity of GWH1 is specifically enhanced in the form of regular multi-display configurations. Those in vitro observations were fully validated against a bacterial infection using a mouse mastitis model, in which the GWH1-GFP soluble nanoparticles were able to effectively reduce bacterial loads.