A Bioluminescence-Based Ex Vivo Burn Wound Model for Real-Time Assessment of Novel Phage-Inspired Enzybiotics

Overview

Journal Pharmaceutics

Publisher MDPI

Date 2022 Dec 23

PMID 36559047

Authors

Vincent De Maesschalck

Diana Gutierrez

Jan Paeshuyse

Yves Briers

Greetje Vande Velde

Rob Lavigne

Affiliations

Soon will be listed here.

Abstract

The silent pandemic of antibiotic resistance is thriving, prompting the urgent need for the development of new antibacterial drugs. However, within the preclinical pipeline, in vitro screening conditions can differ significantly from the final in vivo settings. To bridge the gap between in vitro and in vivo assays, we developed a pig-skin-based bioluminescent ex vivo burn wound infection model, enabling real-time assessment of antibacterials in a longitudinal, non-destructive manner. We provide a proof-of-concept for NCTC13423, a multidrug-resistant clinical isolate, which was equipped with the operon as a reporter using a Tn7-based tagging system. This bioluminescence model provided a linear correlation between the number of bacteria and a broad dynamic range (10 to 10 CFU). This longitudinal model was subsequently validated using a fast-acting enzybiotic, 1D10. Since this model combines a realistic, clinically relevant yet strictly controlled environment with real-time measurement of bacterial burden, we put forward this ex vivo model as a valuable tool to assess the preclinical potential of novel phage-inspired enzybiotics.

Citing Articles

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References

Abdullahi A, Amini-Nik S, Jeschke M . Animal models in burn research. Cell Mol Life Sci. 2014; 71(17):3241-55. PMC: 4134422. DOI: 10.1007/s00018-014-1612-5. View

Turton J, Kaufmann M, Gill M, Pike R, Scott P, Fishbain J . Comparison of Acinetobacter baumannii isolates from the United Kingdom and the United States that were associated with repatriated casualties of the Iraq conflict. J Clin Microbiol. 2006; 44(7):2630-4. PMC: 1489513. DOI: 10.1128/JCM.00547-06. View

Theuretzbacher U, Gottwalt S, Beyer P, Butler M, Czaplewski L, Lienhardt C . Analysis of the clinical antibacterial and antituberculosis pipeline. Lancet Infect Dis. 2018; 19(2):e40-e50. DOI: 10.1016/S1473-3099(18)30513-9. View

Matsumoto A, Schluter T, Melkonian K, Takeda A, Nakagami H, Mine A . A versatile Tn transposon-based bioluminescence tagging tool for quantitative and spatial detection of bacteria in plants. Plant Commun. 2022; 3(1):100227. PMC: 8760037. DOI: 10.1016/j.xplc.2021.100227. View

Andersson M, Madsen L, Schmidtchen A, Puthia M . Development of an Experimental Ex Vivo Wound Model to Evaluate Antimicrobial Efficacy of Topical Formulations. Int J Mol Sci. 2021; 22(9). PMC: 8126222. DOI: 10.3390/ijms22095045. View

Totte J, van Doorn M, Pasmans S . Successful Treatment of Chronic Related Dermatoses with the Topical Endolysin Staphefekt SA.100: A Report of 3 Cases. Case Rep Dermatol. 2017; 9(2):19-25. PMC: 5465516. DOI: 10.1159/000473872. View

Bertesteanu S, Triaridis S, Stankovic M, Lazar V, Chifiriuc M, Vlad M . Polymicrobial wound infections: pathophysiology and current therapeutic approaches. Int J Pharm. 2013; 463(2):119-26. DOI: 10.1016/j.ijpharm.2013.12.012. View

Wang H, Agrawal A, Wang Y, Crawford D, Siler Z, Peterson M . An ex vivo model of medical device-mediated bacterial skin translocation. Sci Rep. 2021; 11(1):5746. PMC: 7952406. DOI: 10.1038/s41598-021-84826-1. View

Czaplewski L, Bax R, Clokie M, Dawson M, Fairhead H, Fischetti V . Alternatives to antibiotics-a pipeline portfolio review. Lancet Infect Dis. 2016; 16(2):239-51. DOI: 10.1016/S1473-3099(15)00466-1. View

10.

Choi K, Gaynor J, White K, Lopez C, Bosio C, Karkhoff-Schweizer R . A Tn7-based broad-range bacterial cloning and expression system. Nat Methods. 2005; 2(6):443-8. DOI: 10.1038/nmeth765. View

11.

Alves D, Booth S, Scavone P, Schellenberger P, Salvage J, Dedi C . Development of a High-Throughput Burn Wound Model Using Porcine Skin, and Its Application to Evaluate New Approaches to Control Wound Infection. Front Cell Infect Microbiol. 2018; 8:196. PMC: 6013584. DOI: 10.3389/fcimb.2018.00196. View

12.

Brackman G, Coenye T . In Vitro and In Vivo Biofilm Wound Models and Their Application. Adv Exp Med Biol. 2015; 897:15-32. DOI: 10.1007/5584_2015_5002. View

13.

Bachman N, Biery M, Boeke J, Craig N . Tn7-mediated mutagenesis of Saccharomyces cerevisiae genomic DNA in vitro. Methods Enzymol. 2002; 350:230-47. DOI: 10.1016/s0076-6879(02)50966-6. View

14.

Silva-Rocha R, Martinez-Garcia E, Calles B, Chavarria M, Arce-Rodriguez A, de Las Heras A . The Standard European Vector Architecture (SEVA): a coherent platform for the analysis and deployment of complex prokaryotic phenotypes. Nucleic Acids Res. 2012; 41(Database issue):D666-75. PMC: 3531073. DOI: 10.1093/nar/gks1119. View

15.

Theuretzbacher U, Outterson K, Engel A, Karlen A . The global preclinical antibacterial pipeline. Nat Rev Microbiol. 2019; 18(5):275-285. PMC: 7223541. DOI: 10.1038/s41579-019-0288-0. View

16.

Shivak D, MacKenzie K, Watson N, Pasternak J, Jones B, Wang Y . A Modular, Tn7-Based System for Making Bioluminescent or Fluorescent Salmonella and Escherichia coli Strains. Appl Environ Microbiol. 2016; 82(16):4931-43. PMC: 4968541. DOI: 10.1128/AEM.01346-16. View

17.

Avci P, Karimi M, Sadasivam M, Antunes-Melo W, Carrasco E, Hamblin M . In-vivo monitoring of infectious diseases in living animals using bioluminescence imaging. Virulence. 2017; 9(1):28-63. PMC: 6067836. DOI: 10.1080/21505594.2017.1371897. View

18.

Gerstmans H, Grimon D, Gutierrez D, Lood C, Rodriguez A, van Noort V . A VersaTile-driven platform for rapid hit-to-lead development of engineered lysins. Sci Adv. 2020; 6(23):eaaz1136. PMC: 7269649. DOI: 10.1126/sciadv.aaz1136. View

19.

Lewis K . Platforms for antibiotic discovery. Nat Rev Drug Discov. 2013; 12(5):371-87. DOI: 10.1038/nrd3975. View

20.

Figurski D, Helinski D . Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979; 76(4):1648-52. PMC: 383447. DOI: 10.1073/pnas.76.4.1648. View