Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo
Bacterial vaginosis is characterized by an imbalance of the vaginal microbiome and a characteristic biofilm formed on the vaginal epithelium, which is initiated and dominated by <i>Gardnerella</i> bacteria, and is frequently refractory to antibiotic treatment. We investigated endolysins...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/1/54 |
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| author | Christine Landlinger Lenka Tisakova Vera Oberbauer Timo Schwebs Abbas Muhammad Agnieszka Latka Leen Van Simaey Mario Vaneechoutte Alexander Guschin Gregory Resch Sonja Swidsinski Alexander Swidsinski Lorenzo Corsini |
| author_facet | Christine Landlinger Lenka Tisakova Vera Oberbauer Timo Schwebs Abbas Muhammad Agnieszka Latka Leen Van Simaey Mario Vaneechoutte Alexander Guschin Gregory Resch Sonja Swidsinski Alexander Swidsinski Lorenzo Corsini |
| author_sort | Christine Landlinger |
| collection | DOAJ |
| description | Bacterial vaginosis is characterized by an imbalance of the vaginal microbiome and a characteristic biofilm formed on the vaginal epithelium, which is initiated and dominated by <i>Gardnerella</i> bacteria, and is frequently refractory to antibiotic treatment. We investigated endolysins of the type 1,4-beta-N-acetylmuramidase encoded on <i>Gardnerella</i> prophages as an alternative treatment. When recombinantly expressed, these proteins demonstrated strong bactericidal activity against four different <i>Gardnerella</i> species. By domain shuffling, we generated several engineered endolysins with 10-fold higher bactericidal activity than any wild-type enzyme. When tested against a panel of 20 <i>Gardnerella</i> strains, the most active endolysin, called PM-477, showed minimum inhibitory concentrations of 0.13–8 µg/mL. PM-477 had no effect on beneficial lactobacilli or other species of vaginal bacteria. Furthermore, the efficacy of PM-477 was tested by fluorescence in situ hybridization on vaginal samples of fifteen patients with either first time or recurring bacterial vaginosis. In thirteen cases, PM-477 killed the <i>Gardnerella</i> bacteria and physically dissolved the biofilms without affecting the remaining vaginal microbiome. The high selectivity and effectiveness in eliminating <i>Gardnerella</i>, both in cultures of isolated strains as well as in clinically derived samples of natural polymicrobial biofilms, makes PM-477 a promising alternative to antibiotics for the treatment of bacterial vaginosis, especially in patients with frequent recurrence. |
| first_indexed | 2024-03-09T05:29:57Z |
| format | Article |
| id | doaj.art-181e14edbf6248f4804c0381b513d3f8 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-09T05:29:57Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-181e14edbf6248f4804c0381b513d3f82023-12-03T12:33:08ZengMDPI AGPathogens2076-08172021-01-011015410.3390/pathogens10010054Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex VivoChristine Landlinger0Lenka Tisakova1Vera Oberbauer2Timo Schwebs3Abbas Muhammad4Agnieszka Latka5Leen Van Simaey6Mario Vaneechoutte7Alexander Guschin8Gregory Resch9Sonja Swidsinski10Alexander Swidsinski11Lorenzo Corsini12PhagoMed Biopharma GmbH, Vienna Biocenter, 1110 Wien, AustriaPhagoMed Biopharma GmbH, Vienna Biocenter, 1110 Wien, AustriaPhagoMed Biopharma GmbH, Vienna Biocenter, 1110 Wien, AustriaPhagoMed Biopharma GmbH, Vienna Biocenter, 1110 Wien, AustriaPhagoMed Biopharma GmbH, Vienna Biocenter, 1110 Wien, AustriaLaboratory Bacteriology Research, Department of Diagnostic Sciences, Faculty of Medicine & Health Sciences, Ghent University, Flanders, 9000 Gent, BelgiumLaboratory Bacteriology Research, Department of Diagnostic Sciences, Faculty of Medicine & Health Sciences, Ghent University, Flanders, 9000 Gent, BelgiumLaboratory Bacteriology Research, Department of Diagnostic Sciences, Faculty of Medicine & Health Sciences, Ghent University, Flanders, 9000 Gent, BelgiumMoscow Scientific and Practical Center of Dermatovenerology and Cosmetology Moscow, 119071 Moscow, RussiaDepartment of Fundamental Microbiology, University of Lausanne, 1015 Lausanne, SwitzerlandMDI Limbach Berlin GmbH, 13407 Berlin, GermanyMedizinische Klinik, Charité CCM, Humboldt Universität, 10117 Berlin, GermanyPhagoMed Biopharma GmbH, Vienna Biocenter, 1110 Wien, AustriaBacterial vaginosis is characterized by an imbalance of the vaginal microbiome and a characteristic biofilm formed on the vaginal epithelium, which is initiated and dominated by <i>Gardnerella</i> bacteria, and is frequently refractory to antibiotic treatment. We investigated endolysins of the type 1,4-beta-N-acetylmuramidase encoded on <i>Gardnerella</i> prophages as an alternative treatment. When recombinantly expressed, these proteins demonstrated strong bactericidal activity against four different <i>Gardnerella</i> species. By domain shuffling, we generated several engineered endolysins with 10-fold higher bactericidal activity than any wild-type enzyme. When tested against a panel of 20 <i>Gardnerella</i> strains, the most active endolysin, called PM-477, showed minimum inhibitory concentrations of 0.13–8 µg/mL. PM-477 had no effect on beneficial lactobacilli or other species of vaginal bacteria. Furthermore, the efficacy of PM-477 was tested by fluorescence in situ hybridization on vaginal samples of fifteen patients with either first time or recurring bacterial vaginosis. In thirteen cases, PM-477 killed the <i>Gardnerella</i> bacteria and physically dissolved the biofilms without affecting the remaining vaginal microbiome. The high selectivity and effectiveness in eliminating <i>Gardnerella</i>, both in cultures of isolated strains as well as in clinically derived samples of natural polymicrobial biofilms, makes PM-477 a promising alternative to antibiotics for the treatment of bacterial vaginosis, especially in patients with frequent recurrence.https://www.mdpi.com/2076-0817/10/1/54bacterial vaginosis<i>Gardnerella</i> biofilmprophageendolysingenus-specificityantimicrobial resistance |
| spellingShingle | Christine Landlinger Lenka Tisakova Vera Oberbauer Timo Schwebs Abbas Muhammad Agnieszka Latka Leen Van Simaey Mario Vaneechoutte Alexander Guschin Gregory Resch Sonja Swidsinski Alexander Swidsinski Lorenzo Corsini Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo Pathogens bacterial vaginosis <i>Gardnerella</i> biofilm prophage endolysin genus-specificity antimicrobial resistance |
| title | Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo |
| title_full | Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo |
| title_fullStr | Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo |
| title_full_unstemmed | Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo |
| title_short | Engineered Phage Endolysin Eliminates <i>Gardnerella</i> Biofilm without Damaging Beneficial Bacteria in Bacterial Vaginosis Ex Vivo |
| title_sort | engineered phage endolysin eliminates i gardnerella i biofilm without damaging beneficial bacteria in bacterial vaginosis ex vivo |
| topic | bacterial vaginosis <i>Gardnerella</i> biofilm prophage endolysin genus-specificity antimicrobial resistance |
| url | https://www.mdpi.com/2076-0817/10/1/54 |
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