Studies in vitro e in vivo of Phage Therapy Medical Products (PTMPs) targeting clinical strains of Klebsiella pneumoniae belonging to the clone ST512

Bleriot, I.; Blasco, L.; Fernandez-Grela, P.; Fernandez-Garcia, L.; Arman, L.; Ibaguren-Quiles, C.; Ortiz-Cartagena, C.; Barrio-Pujante, A.; Pano Pardo, J. R.; Oteo-Iglesias, J.; Tomas, M.

doi:10.1101/2024.11.20.624559

bioRxiv · 10.1101/2024.11.20.624559

Studies in vitro e in vivo of Phage Therapy Medical Products (PTMPs) targeting clinical strains of Klebsiella pneumoniae belonging to the clone ST512

Bleriot, I.·Blasco, L.·Fernandez-Grela, P.·Fernandez-Garcia, L.·Arman, L.·Ibaguren-Quiles, C.·Ortiz-Cartagena, C.·Barrio-Pujante, A.·Pano Pardo, J. R.·Oteo-Iglesias, J.·Tomas, M.

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Abstract

The widespread incidence of antimicrobial resistance has created renewed interest in the use of alternative antimicrobial treatments such as phage therapy. Phages are viruses that infect bacteria and generally have a narrow bacteria host-range. Combining phages with antibiotics can prevent the emergence of bacterial resistance. The aim of the present study was to develop phage therapy medical products (PTMPs) targeting clinical isolates of carbapenems-producing Klebsiella pneumoniae belonging to the high-risk clone ST512. From a collection of twenty-two seed of lytic phages sequenced belonging to MePRAM collection, four were used to generate PTMPs (CAC_Kpn1 and CAC_Kpn2). These PTMPs were partly active against three of the clinical strains of clone ST512 (A, B and C). The use of Appelmans method in the CAC_Kpn1_ad (adapted CAC_Kpn1) yielded a significant increase in the efficacy against strain A, while adapted CAC_Kpn2 (CAC_Kpn2_ad) only effectively reduced bacterial survival when combined with [1/2] x MIC {beta}-lactam antibiotic meropenem for 24 h in clinical strains B and C, showed after this time, resistance to PTMPs. In addition, the amounts of endotoxin released by the PTMPs were quantified and subsequently reduced in preparation for in vivo use of the PTMPs in Galleria mellonella infection model confirming the in vitro results from the CAC_Kpn1_ad and CAC_Kpn2_ad.

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Bleriot, I., Blasco, L., Fernandez-Grela, P., Fernandez-Garcia, L., Arman, L., Ibaguren-Quiles, C., Ortiz-Cartagena, C., Barrio-Pujante, A., Pano Pardo, J. R., Oteo-Iglesias, J., Tomas, M.. 2024-11-22. Studies in vitro e in vivo of Phage Therapy Medical Products (PTMPs) targeting clinical strains of Klebsiella pneumoniae belonging to the clone ST512. https://doi.org/10.1101/2024.11.20.624559

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