Interruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts Disease
Abstract Mutations in the gene ABCA4 coding for photoreceptor-specific ATP-binding cassette subfamily A member 4, are responsible for Stargardts Disease type 1 (STGD1), the most common form of inherited macular degeneration. STGD1 typically declares early in life and leads to severe visual handicap....
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2024-12-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-024-81869-y |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850103166882807808 |
|---|---|
| author | Fabiana Sassone Catherine Estay-Ahumada Michel J. Roux Dominique Ciocca Paola Rossolillo Marie-Christine Birling Janet R. Sparrow Diego Montenegro David Hicks |
| author_facet | Fabiana Sassone Catherine Estay-Ahumada Michel J. Roux Dominique Ciocca Paola Rossolillo Marie-Christine Birling Janet R. Sparrow Diego Montenegro David Hicks |
| author_sort | Fabiana Sassone |
| collection | DOAJ |
| description | Abstract Mutations in the gene ABCA4 coding for photoreceptor-specific ATP-binding cassette subfamily A member 4, are responsible for Stargardts Disease type 1 (STGD1), the most common form of inherited macular degeneration. STGD1 typically declares early in life and leads to severe visual handicap. Abca4 gene-deletion mouse models of STGD1 accumulate lipofuscin, a hallmark of the disease, but unlike the human disease show no or only moderate structural changes and no functional decline. The human macula is highly enriched in cones, and reasoning that the low cone percentage in mice retinas (< 3%) might compromise faithful modelling of human maculopathies, we performed sub-retinal injections of CRISPR/Cas9-abca4 Adeno-Associated Virus constructs into young Sand Rats (Psammomys obesus), a diurnal rodent containing > 30% cones. Compared to control injections of AAV-abca4-GFP, treated eyes exhibited extensive retinal degeneration by two months. Sanger sequencing of the CRISPR targeted sequence show a clear edition of Abca4 gene. Non-invasive fundus imaging showed widespread photoreceptor loss, confirmed by ocular coherence tomography. Functional recording by single flash and flicker electroretinography showed significant decline in photopic (cone) light responses. Post-mortem real-time PCR, immunohistochemistry and western blotting showed significant decrease of cone-specific (MW cone opsin) but not rod-specific (rhodopsin) markers. Transmission electron microscopy showed large numbers of lipid inclusions in treated but not control retinal pigmented epithelium. Finally, ultra-high performance liquid chromatography analysis of whole P. obesus eyes showed the presence of all-trans retinal-dimer, not detected in rod-rich rat eyes. In conclusion, Abca4 knockout in P. obesus results in a predominantly cone degeneration phenotype, more accurately reflecting the etiology of human STGD1, and should be valuable for characterizing pathogenic pathways and exploring treatment options. |
| format | Article |
| id | doaj-art-349e2bbcd20448baa0c754039b60d54c |
| institution | DOAJ |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-349e2bbcd20448baa0c754039b60d54c2025-08-20T02:39:37ZengNature PortfolioScientific Reports2045-23222024-12-0114111610.1038/s41598-024-81869-yInterruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts DiseaseFabiana Sassone0Catherine Estay-Ahumada1Michel J. Roux2Dominique Ciocca3Paola Rossolillo4Marie-Christine Birling5Janet R. Sparrow6Diego Montenegro7David Hicks8INCI-UPR3212-CNRSINCI-UPR3212-CNRSIGBMC/Institut Clinique de La Souris - CNRS UMR 7104 Inserm U 1258Chronobiotron UAR3415-CNRSIGBMC/Institut Clinique de La Souris - CNRS UMR 7104 Inserm U 1258Université de Strasbourg, CNRS, INSERM, CELPHEDIA, PHENOMIN-Institut Clinique de La Souris (ICS)Departments of Ophthalmology, and Pathology and Cell Biology, Columbia University Medical CenterDepartments of Ophthalmology, and Pathology and Cell Biology, Columbia University Medical CenterINCI-UPR3212-CNRSAbstract Mutations in the gene ABCA4 coding for photoreceptor-specific ATP-binding cassette subfamily A member 4, are responsible for Stargardts Disease type 1 (STGD1), the most common form of inherited macular degeneration. STGD1 typically declares early in life and leads to severe visual handicap. Abca4 gene-deletion mouse models of STGD1 accumulate lipofuscin, a hallmark of the disease, but unlike the human disease show no or only moderate structural changes and no functional decline. The human macula is highly enriched in cones, and reasoning that the low cone percentage in mice retinas (< 3%) might compromise faithful modelling of human maculopathies, we performed sub-retinal injections of CRISPR/Cas9-abca4 Adeno-Associated Virus constructs into young Sand Rats (Psammomys obesus), a diurnal rodent containing > 30% cones. Compared to control injections of AAV-abca4-GFP, treated eyes exhibited extensive retinal degeneration by two months. Sanger sequencing of the CRISPR targeted sequence show a clear edition of Abca4 gene. Non-invasive fundus imaging showed widespread photoreceptor loss, confirmed by ocular coherence tomography. Functional recording by single flash and flicker electroretinography showed significant decline in photopic (cone) light responses. Post-mortem real-time PCR, immunohistochemistry and western blotting showed significant decrease of cone-specific (MW cone opsin) but not rod-specific (rhodopsin) markers. Transmission electron microscopy showed large numbers of lipid inclusions in treated but not control retinal pigmented epithelium. Finally, ultra-high performance liquid chromatography analysis of whole P. obesus eyes showed the presence of all-trans retinal-dimer, not detected in rod-rich rat eyes. In conclusion, Abca4 knockout in P. obesus results in a predominantly cone degeneration phenotype, more accurately reflecting the etiology of human STGD1, and should be valuable for characterizing pathogenic pathways and exploring treatment options.https://doi.org/10.1038/s41598-024-81869-yStargardt’s diseaseRetinal degenerationAnimal modelGene editingCone photoreceptorsElectrophysiology |
| spellingShingle | Fabiana Sassone Catherine Estay-Ahumada Michel J. Roux Dominique Ciocca Paola Rossolillo Marie-Christine Birling Janet R. Sparrow Diego Montenegro David Hicks Interruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts Disease Scientific Reports Stargardt’s disease Retinal degeneration Animal model Gene editing Cone photoreceptors Electrophysiology |
| title | Interruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts Disease |
| title_full | Interruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts Disease |
| title_fullStr | Interruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts Disease |
| title_full_unstemmed | Interruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts Disease |
| title_short | Interruption of the visual cycle in a novel animal model induces progressive vision loss resembling Stargardts Disease |
| title_sort | interruption of the visual cycle in a novel animal model induces progressive vision loss resembling stargardts disease |
| topic | Stargardt’s disease Retinal degeneration Animal model Gene editing Cone photoreceptors Electrophysiology |
| url | https://doi.org/10.1038/s41598-024-81869-y |
| work_keys_str_mv | AT fabianasassone interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT catherineestayahumada interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT micheljroux interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT dominiqueciocca interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT paolarossolillo interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT mariechristinebirling interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT janetrsparrow interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT diegomontenegro interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease AT davidhicks interruptionofthevisualcycleinanovelanimalmodelinducesprogressivevisionlossresemblingstargardtsdisease |