<i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse Model
The vancomycin-resistance associated sensor/regulator, <i>VraSR</i> two-component regulatory-system (<i>VraSR</i>), regulates virulence and the response of <i>Staphylococcus aureus</i> (SA) to environmental stress. To investigate the role of <i>VraSR</i&g...
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MDPI AG
2021-12-01
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| Online Access: | https://www.mdpi.com/2227-9059/10/1/35 |
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| author | Nilakshi Barua Ying Yang Lin Huang Margaret Ip |
| author_facet | Nilakshi Barua Ying Yang Lin Huang Margaret Ip |
| author_sort | Nilakshi Barua |
| collection | DOAJ |
| description | The vancomycin-resistance associated sensor/regulator, <i>VraSR</i> two-component regulatory-system (<i>VraSR</i>), regulates virulence and the response of <i>Staphylococcus aureus</i> (SA) to environmental stress. To investigate the role of <i>VraSR</i> in SA skin and soft tissue infections (SSTI), we inactivated the <i>VraSR</i> of a clinical CA-MRSA ST30 strain by insertional mutation in <i>vraR</i> gene using the TargeTron-Gene Knockout System. We constructed an organotypic keratinocyte fibroblast co-culture (3D-skin model) and a humanized mouse as SSTI infection models. In the 3D-skin model, inactivation of <i>VraSR</i> in the strains ST30 and USA300 showed 1-log reduction in adhesion and internalization (<i>p <</i> 0.001) compared to the respective wildtype. The mutant strains of ST30 (<i>p <</i> 0.05) and USA300-LAC (<i>p <</i> 0.001) also exhibited reduced apoptosis. The wildtype ST30 infection in the humanized mouse model demonstrated increased skin lesion size and bacterial burden compared to BALB/c mice (<i>p <</i> 0.01). The response of the humanized mouse towards the MRSA infection exhibited human similarity indicating that the humanized mouse SSTI model is more suitable for evaluating the role of virulence determinants. Inactivation of <i>VraSR</i> in ST30 strain resulted in decreased skin lesion size in the humanized mouse SSTI model (<i>p <</i> 0.05) and reduction in apoptotic index (<i>p <</i> 0.01) when compared with the wildtype. Our results reveal that inactivating the <i>VraSR</i> system may be a potent anti-virulence approach to control MRSA infection. |
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| spelling | doaj.art-d480e3b36d0543fe889e3d65e0bd2aea2023-11-23T13:02:41ZengMDPI AGBiomedicines2227-90592021-12-011013510.3390/biomedicines10010035<i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse ModelNilakshi Barua0Ying Yang1Lin Huang2Margaret Ip3Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Sha Tin, N.T., Hong Kong 999077, ChinaDepartment of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Sha Tin, N.T., Hong Kong 999077, ChinaDivision of Plastic, Reconstructive and Aesthetic Surgery, Department of Surgery, The Chinese University of Hong Kong, Prince of Wales Hospital, Sha Tin, N.T., Hong Kong 999077, ChinaDepartment of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Sha Tin, N.T., Hong Kong 999077, ChinaThe vancomycin-resistance associated sensor/regulator, <i>VraSR</i> two-component regulatory-system (<i>VraSR</i>), regulates virulence and the response of <i>Staphylococcus aureus</i> (SA) to environmental stress. To investigate the role of <i>VraSR</i> in SA skin and soft tissue infections (SSTI), we inactivated the <i>VraSR</i> of a clinical CA-MRSA ST30 strain by insertional mutation in <i>vraR</i> gene using the TargeTron-Gene Knockout System. We constructed an organotypic keratinocyte fibroblast co-culture (3D-skin model) and a humanized mouse as SSTI infection models. In the 3D-skin model, inactivation of <i>VraSR</i> in the strains ST30 and USA300 showed 1-log reduction in adhesion and internalization (<i>p <</i> 0.001) compared to the respective wildtype. The mutant strains of ST30 (<i>p <</i> 0.05) and USA300-LAC (<i>p <</i> 0.001) also exhibited reduced apoptosis. The wildtype ST30 infection in the humanized mouse model demonstrated increased skin lesion size and bacterial burden compared to BALB/c mice (<i>p <</i> 0.01). The response of the humanized mouse towards the MRSA infection exhibited human similarity indicating that the humanized mouse SSTI model is more suitable for evaluating the role of virulence determinants. Inactivation of <i>VraSR</i> in ST30 strain resulted in decreased skin lesion size in the humanized mouse SSTI model (<i>p <</i> 0.05) and reduction in apoptotic index (<i>p <</i> 0.01) when compared with the wildtype. Our results reveal that inactivating the <i>VraSR</i> system may be a potent anti-virulence approach to control MRSA infection.https://www.mdpi.com/2227-9059/10/1/35<i>VraSR</i> regulatory system3D Skin modelhumanized mouse model<i>Staphylococcus aureus</i>MRSAskin and soft-tissue infections |
| spellingShingle | Nilakshi Barua Ying Yang Lin Huang Margaret Ip <i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse Model Biomedicines <i>VraSR</i> regulatory system 3D Skin model humanized mouse model <i>Staphylococcus aureus</i> MRSA skin and soft-tissue infections |
| title | <i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse Model |
| title_full | <i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse Model |
| title_fullStr | <i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse Model |
| title_full_unstemmed | <i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse Model |
| title_short | <i>VraSR</i> Regulatory System Contributes to the Virulence of Community-Associated Methicillin-Resistant <i>Staphylococcus aureus</i> (CA-MRSA) in a 3D-Skin Model and Skin Infection of Humanized Mouse Model |
| title_sort | i vrasr i regulatory system contributes to the virulence of community associated methicillin resistant i staphylococcus aureus i ca mrsa in a 3d skin model and skin infection of humanized mouse model |
| topic | <i>VraSR</i> regulatory system 3D Skin model humanized mouse model <i>Staphylococcus aureus</i> MRSA skin and soft-tissue infections |
| url | https://www.mdpi.com/2227-9059/10/1/35 |
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