Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injury
Abstract Background The opening of pannexin1 channels is considered as a key event in inflammation. Pannexin1 channel-mediated release of adenosine triphosphate triggers inflammasome signaling and activation of immune cells. By doing so, pannexin1 channels play an important role in several inflammat...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-10-01
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| Series: | Journal of Nanobiotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12951-023-02137-1 |
| _version_ | 1797451619495837696 |
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| author | Raf Van Campenhout Timo W. M. De Groof Prashant Kadam Brenda R. Kwak Serge Muyldermans Nick Devoogdt Mathieu Vinken |
| author_facet | Raf Van Campenhout Timo W. M. De Groof Prashant Kadam Brenda R. Kwak Serge Muyldermans Nick Devoogdt Mathieu Vinken |
| author_sort | Raf Van Campenhout |
| collection | DOAJ |
| description | Abstract Background The opening of pannexin1 channels is considered as a key event in inflammation. Pannexin1 channel-mediated release of adenosine triphosphate triggers inflammasome signaling and activation of immune cells. By doing so, pannexin1 channels play an important role in several inflammatory diseases. Although pannexin1 channel inhibition could represent a novel clinical strategy for treatment of inflammatory disorders, therapeutic pannexin1 channel targeting is impeded by the lack of specific, potent and/or in vivo-applicable inhibitors. The goal of this study is to generate nanobody-based inhibitors of pannexin1 channels. Results Pannexin1-targeting nanobodies were developed as potential new pannexin1 channel inhibitors. We identified 3 cross-reactive nanobodies that showed affinity for both murine and human pannexin1 proteins. Flow cytometry experiments revealed binding capacities in the nanomolar range. Moreover, the pannexin1-targeting nanobodies were found to block pannexin1 channel-mediated release of adenosine triphosphate. The pannexin1-targeting nanobodies were also demonstrated to display anti-inflammatory effects in vitro through reduction of interleukin 1 beta amounts. This anti-inflammatory outcome was reproduced in vivo using a human-relevant mouse model of acute liver disease relying on acetaminophen overdosing. More specifically, the pannexin1-targeting nanobodies lowered serum levels of inflammatory cytokines and diminished liver damage. These effects were linked with alteration of the expression of several NLRP3 inflammasome components. Conclusions This study introduced for the first time specific, potent and in vivo-applicable nanobody-based inhibitors of pannexin1 channels. As demonstrated for the case of liver disease, the pannexin1-targeting nanobodies hold great promise as anti-inflammatory agents, yet this should be further tested for extrahepatic inflammatory disorders. Moreover, the pannexin1-targeting nanobodies represent novel tools for fundamental research regarding the role of pannexin1 channels in pathological and physiological processes. Graphical Abstract |
| first_indexed | 2024-03-09T14:57:16Z |
| format | Article |
| id | doaj.art-8b9134db02464ebd9b9cd759de219169 |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-03-09T14:57:16Z |
| publishDate | 2023-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-8b9134db02464ebd9b9cd759de2191692023-11-26T14:08:36ZengBMCJournal of Nanobiotechnology1477-31552023-10-0121112010.1186/s12951-023-02137-1Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injuryRaf Van Campenhout0Timo W. M. De Groof1Prashant Kadam2Brenda R. Kwak3Serge Muyldermans4Nick Devoogdt5Mathieu Vinken6Entity of In Vitro Toxicology and Dermato-Cosmetology, Department of Pharmaceutical and Pharmacological Sciences, Vrije Universiteit BrusselIn Vivo Cellular and Molecular Imaging Laboratory, Department of Molecular Imaging, Vrije Universiteit BrusselEntity of In Vitro Toxicology and Dermato-Cosmetology, Department of Pharmaceutical and Pharmacological Sciences, Vrije Universiteit BrusselDepartment of Pathology and Immunology, Faculty of Medicine, University of GenevaLaboratory of Cellular and Molecular Immunology, Bioengineering Sciences Department, Vrije Universiteit BrusselIn Vivo Cellular and Molecular Imaging Laboratory, Department of Molecular Imaging, Vrije Universiteit BrusselEntity of In Vitro Toxicology and Dermato-Cosmetology, Department of Pharmaceutical and Pharmacological Sciences, Vrije Universiteit BrusselAbstract Background The opening of pannexin1 channels is considered as a key event in inflammation. Pannexin1 channel-mediated release of adenosine triphosphate triggers inflammasome signaling and activation of immune cells. By doing so, pannexin1 channels play an important role in several inflammatory diseases. Although pannexin1 channel inhibition could represent a novel clinical strategy for treatment of inflammatory disorders, therapeutic pannexin1 channel targeting is impeded by the lack of specific, potent and/or in vivo-applicable inhibitors. The goal of this study is to generate nanobody-based inhibitors of pannexin1 channels. Results Pannexin1-targeting nanobodies were developed as potential new pannexin1 channel inhibitors. We identified 3 cross-reactive nanobodies that showed affinity for both murine and human pannexin1 proteins. Flow cytometry experiments revealed binding capacities in the nanomolar range. Moreover, the pannexin1-targeting nanobodies were found to block pannexin1 channel-mediated release of adenosine triphosphate. The pannexin1-targeting nanobodies were also demonstrated to display anti-inflammatory effects in vitro through reduction of interleukin 1 beta amounts. This anti-inflammatory outcome was reproduced in vivo using a human-relevant mouse model of acute liver disease relying on acetaminophen overdosing. More specifically, the pannexin1-targeting nanobodies lowered serum levels of inflammatory cytokines and diminished liver damage. These effects were linked with alteration of the expression of several NLRP3 inflammasome components. Conclusions This study introduced for the first time specific, potent and in vivo-applicable nanobody-based inhibitors of pannexin1 channels. As demonstrated for the case of liver disease, the pannexin1-targeting nanobodies hold great promise as anti-inflammatory agents, yet this should be further tested for extrahepatic inflammatory disorders. Moreover, the pannexin1-targeting nanobodies represent novel tools for fundamental research regarding the role of pannexin1 channels in pathological and physiological processes. Graphical Abstracthttps://doi.org/10.1186/s12951-023-02137-1Acute liver diseaseInflammationNanobodyPannexin1Therapy |
| spellingShingle | Raf Van Campenhout Timo W. M. De Groof Prashant Kadam Brenda R. Kwak Serge Muyldermans Nick Devoogdt Mathieu Vinken Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injury Journal of Nanobiotechnology Acute liver disease Inflammation Nanobody Pannexin1 Therapy |
| title | Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injury |
| title_full | Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injury |
| title_fullStr | Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injury |
| title_full_unstemmed | Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injury |
| title_short | Nanobody-based pannexin1 channel inhibitors reduce inflammation in acute liver injury |
| title_sort | nanobody based pannexin1 channel inhibitors reduce inflammation in acute liver injury |
| topic | Acute liver disease Inflammation Nanobody Pannexin1 Therapy |
| url | https://doi.org/10.1186/s12951-023-02137-1 |
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