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International journal of molecular sciences
Sep 04, 2022;23 (17):

Liposomal Delivery of Newly Identified Prophage Lysins in a Model.

Diana Morais, Luís Tanoeiro, Andreia T Marques, Tiago Gonçalves, Aida Duarte, António Pedro Alves Matos, Joana S Vital, Maria Eugénia Meirinhos Cruz, Manuela Colla Carvalheiro, Elsa Anes, Jorge M B Vítor, Maria Manuela Gaspar, Filipa F Vale
Author Information
  1. Diana Morais: Pathogen Genome Bioinformatics and Computational Biology, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal.
  2. Luís Tanoeiro: Pathogen Genome Bioinformatics and Computational Biology, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal. ORCID
  3. Andreia T Marques: Pathogen Genome Bioinformatics and Computational Biology, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal. ORCID
  4. Tiago Gonçalves: Pathogen Genome Bioinformatics and Computational Biology, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal.
  5. Aida Duarte: Faculty of Pharmacy, Universidade de Lisboa, Av. Gama Pinto, 1649-003 Lisboa, Portugal. ORCID
  6. António Pedro Alves Matos: Centro de Investigação Interdisciplinar Egas Moniz (CiiEM), Instituto Superior Egas Moniz, Quinta da Granja, 2829-511 Monte da Caparica, Portugal.
  7. Joana S Vital: Pathogen Genome Bioinformatics and Computational Biology, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal.
  8. Maria Eugénia Meirinhos Cruz: Advanced Technologies for Drug Delivery, Research Institute for Medicines (iMed-ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal.
  9. Manuela Colla Carvalheiro: Advanced Technologies for Drug Delivery, Research Institute for Medicines (iMed-ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal. ORCID
  10. Elsa Anes: Host-Pathogen Interactions Unit, Research Institute for Medicines (iMed-ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal. ORCID
  11. Jorge M B Vítor: Pathogen Genome Bioinformatics and Computational Biology, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal. ORCID
  12. Maria Manuela Gaspar: Advanced Technologies for Drug Delivery, Research Institute for Medicines (iMed-ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal. ORCID
  13. Filipa F Vale: Pathogen Genome Bioinformatics and Computational Biology, Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, 1649-003 Lisboa, Portugal.
PMID: 36077542 DOI: 10.3390/ijms231710143

Abstract

is a Gram-negative opportunistic bacterium that presents resistance to several antibiotics, thus, representing a major threat to human and animal health. Phage-derived products, namely lysins, or peptidoglycan-hydrolyzing enzymes, can be an effective weapon against antibiotic-resistant bacteria. Whereas in Gram-positive bacteria, lysis from without is facilitated by the exposed peptidoglycan layer, this is not possible in the outer membrane-protected peptidoglycan of Gram-negative bacteria. Here, we suggest the encapsulation of lysins in liposomes as a delivery system against Gram-negative bacteria, using the model of . Bioinformatic analysis allowed for the identification of 38 distinct complete prophages within 66 genomes (16 of which newly sequenced) and led to the identification of 19 lysins of diverse sequence and function, 5 of which proceeded to wet lab analysis. The four purifiable lysins showed hydrolytic activity against Gram-positive bacterial lawns and, on zymogram assays, constituted of autoclaved cells. Additionally, lysins Pa7 and Pa119 combined with an outer membrane permeabilizer showed activity against cells. These two lysins were successfully encapsulated in DPPC:DOPE:CHEMS (molar ratio 4:4:2) liposomes with an average encapsulation efficiency of 33.33% and 32.30%, respectively. The application of the encapsulated lysins to the model led to a reduction in cell viability and resulted in cell lysis as observed in MTT cell viability assays and electron microscopy. In sum, we report here that prophages may be important sources of new enzybiotics, with prophage lysins showing high diversity and activity. In addition, these enzybiotics following their incorporation in liposomes were able to potentiate their antibacterial effect against the Gram-negative bacteria , used as the model.

Keywords

Pseudomonas aeruginosa antibiotic resistance bacteriophage gram-negative bacteria liposomes lysins phage therapy prophage

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Grants

  1. PTDC/BTM-SAL/28978/2017/Fundação para a Ciência e Tecnologia
  2. PTDC/BTM-TEC/3238/2020/Fundação para a Ciência e Tecnologia
  3. CEECIND/03023/2017/Fundação para a Ciência e Tecnologia
  4. UIDB/04138/2020/Fundação para a Ciência e Tecnologia
  5. UIDP/04138/2020/Fundação para a Ciência e Tecnologia

MeSH Term

Animals
Anti-Bacterial Agents
Gram-Negative Bacteria
Gram-Positive Bacteria
Humans
Liposomes
Peptidoglycan
Prophages
Pseudomonas aeruginosa

Chemicals

Anti-Bacterial Agents
Liposomes
Peptidoglycan
Journal Article
Article has an altmetric score of 3

Word Cloud

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