Impairing Gasdermin D-mediated pyroptosis is protective against retinal degeneration
Abstract Background Inflammasome activation and the subsequent release of pro-inflammatory cytokines including Interleukin 1β (IL-1β) have been widely reported to contribute to the progression of retinal degenerations, including age-related macular degeneration (AMD), the leading cause of blindness...
| 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 Neuroinflammation |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12974-023-02927-2 |
| _version_ | 1797557529741361152 |
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| author | Rakshanya Sekar Yvette Wooff Adrian V. Cioanca Melan Kurera Chinh Ngo Si Ming Man Riccardo Natoli |
| author_facet | Rakshanya Sekar Yvette Wooff Adrian V. Cioanca Melan Kurera Chinh Ngo Si Ming Man Riccardo Natoli |
| author_sort | Rakshanya Sekar |
| collection | DOAJ |
| description | Abstract Background Inflammasome activation and the subsequent release of pro-inflammatory cytokines including Interleukin 1β (IL-1β) have been widely reported to contribute to the progression of retinal degenerations, including age-related macular degeneration (AMD), the leading cause of blindness in the Western World. The role of Gasdermin D (GSDMD), a key executioner of pyroptosis following inflammasome activation, however, is less well-established. In this study we aimed to characterise the role of GSDMD in the healthy and degenerating retina, and uncover its role as a conduit for IL-1β release, including via extracellular vesicle (EV)-mediated release. Methods GSDMD mutant and knockout mice, in vitro models of inflammation and a well-established in vivo model of retinal degeneration (photo-oxidative damage; PD) were utilised to explore the role and pathological contribution of GSDMD in regulating IL-1β release and propagating retinal inflammation. RNA sequencing of whole retinas was used to investigate GSDMD-mediated inflammation during degeneration. The role of EVs in GSDMD-mediated IL-1β release was investigated using nanoparticle tracking analysis, ELISA and EV inhibition paradigms. Finally, the therapeutic efficacy of targeting GSDMD was examined using GSDMD-specific siRNA. Results We identified in this work that mice deficient in GSDMD had better-preserved retinal function, increased photoreceptor survivability and reduced inflammation. RNA-Seq analysis revealed that GSDMD may propagate inflammation in the retina via NF-κB signalling cascades and release of pro-inflammatory cytokines. We also showed that IL-1β was packaged and released via EV in a GSDMD-dependent manner. Finally, we demonstrated that impairing GSDMD function using RNAi or blocking EV release was able to reduce IL-1β content in cell-free supernatant and EV. Conclusions Taken together, these results suggest that pyroptotic pore-forming protein GSDMD plays a key role in the propagation of retinal inflammation, in particular via the release of EV-encapsulated IL-1β. Targeting GSDMD using genetic or pharmacological inhibitors may pose a therapeutic opportunity to dampen inflammatory cascades and delay the progression of retinal degeneration. |
| first_indexed | 2024-03-10T17:19:14Z |
| format | Article |
| id | doaj.art-61edecadb83e4d179b792c98e67bf4ba |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-03-10T17:19:14Z |
| publishDate | 2023-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-61edecadb83e4d179b792c98e67bf4ba2023-11-20T10:24:23ZengBMCJournal of Neuroinflammation1742-20942023-10-0120112410.1186/s12974-023-02927-2Impairing Gasdermin D-mediated pyroptosis is protective against retinal degenerationRakshanya Sekar0Yvette Wooff1Adrian V. Cioanca2Melan Kurera3Chinh Ngo4Si Ming Man5Riccardo Natoli6The John Curtin School of Medical Research, The Australian National UniversityThe John Curtin School of Medical Research, The Australian National UniversityThe John Curtin School of Medical Research, The Australian National UniversityThe John Curtin School of Medical Research, The Australian National UniversityThe John Curtin School of Medical Research, The Australian National UniversityThe John Curtin School of Medical Research, The Australian National UniversityThe John Curtin School of Medical Research, The Australian National UniversityAbstract Background Inflammasome activation and the subsequent release of pro-inflammatory cytokines including Interleukin 1β (IL-1β) have been widely reported to contribute to the progression of retinal degenerations, including age-related macular degeneration (AMD), the leading cause of blindness in the Western World. The role of Gasdermin D (GSDMD), a key executioner of pyroptosis following inflammasome activation, however, is less well-established. In this study we aimed to characterise the role of GSDMD in the healthy and degenerating retina, and uncover its role as a conduit for IL-1β release, including via extracellular vesicle (EV)-mediated release. Methods GSDMD mutant and knockout mice, in vitro models of inflammation and a well-established in vivo model of retinal degeneration (photo-oxidative damage; PD) were utilised to explore the role and pathological contribution of GSDMD in regulating IL-1β release and propagating retinal inflammation. RNA sequencing of whole retinas was used to investigate GSDMD-mediated inflammation during degeneration. The role of EVs in GSDMD-mediated IL-1β release was investigated using nanoparticle tracking analysis, ELISA and EV inhibition paradigms. Finally, the therapeutic efficacy of targeting GSDMD was examined using GSDMD-specific siRNA. Results We identified in this work that mice deficient in GSDMD had better-preserved retinal function, increased photoreceptor survivability and reduced inflammation. RNA-Seq analysis revealed that GSDMD may propagate inflammation in the retina via NF-κB signalling cascades and release of pro-inflammatory cytokines. We also showed that IL-1β was packaged and released via EV in a GSDMD-dependent manner. Finally, we demonstrated that impairing GSDMD function using RNAi or blocking EV release was able to reduce IL-1β content in cell-free supernatant and EV. Conclusions Taken together, these results suggest that pyroptotic pore-forming protein GSDMD plays a key role in the propagation of retinal inflammation, in particular via the release of EV-encapsulated IL-1β. Targeting GSDMD using genetic or pharmacological inhibitors may pose a therapeutic opportunity to dampen inflammatory cascades and delay the progression of retinal degeneration.https://doi.org/10.1186/s12974-023-02927-2Gasdermin DAge-related macular degeneration (AMD)Retinal degenerationsNeuroinflammationInflammasomeIL-1β |
| spellingShingle | Rakshanya Sekar Yvette Wooff Adrian V. Cioanca Melan Kurera Chinh Ngo Si Ming Man Riccardo Natoli Impairing Gasdermin D-mediated pyroptosis is protective against retinal degeneration Journal of Neuroinflammation Gasdermin D Age-related macular degeneration (AMD) Retinal degenerations Neuroinflammation Inflammasome IL-1β |
| title | Impairing Gasdermin D-mediated pyroptosis is protective against retinal degeneration |
| title_full | Impairing Gasdermin D-mediated pyroptosis is protective against retinal degeneration |
| title_fullStr | Impairing Gasdermin D-mediated pyroptosis is protective against retinal degeneration |
| title_full_unstemmed | Impairing Gasdermin D-mediated pyroptosis is protective against retinal degeneration |
| title_short | Impairing Gasdermin D-mediated pyroptosis is protective against retinal degeneration |
| title_sort | impairing gasdermin d mediated pyroptosis is protective against retinal degeneration |
| topic | Gasdermin D Age-related macular degeneration (AMD) Retinal degenerations Neuroinflammation Inflammasome IL-1β |
| url | https://doi.org/10.1186/s12974-023-02927-2 |
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