Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt disease
Abstract Background Stargardt disease type 1 (STGD1) is a progressive retinal disorder caused by bi-allelic variants in the ABCA4 gene. A recurrent variant at the exon-intron junction of exon 6, c.768G>T, causes a 35-nt elongation of exon 6 that leads to premature termination of protein synthesis...
Saved in:
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Communications Medicine |
| Online Access: | https://doi.org/10.1038/s43856-024-00712-7 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832585453184745472 |
|---|---|
| author | Dyah W. Karjosukarso Femke Bukkems Lonneke Duijkers Tomasz Z. Tomkiewicz Julia Kiefmann Andrei Sarlea Sander Bervoets Irene Vázquez-Domínguez Laurie L. Molday Robert S. Molday Mihai G. Netea Carel B. Hoyng Alejandro Garanto Rob W. J. Collin |
| author_facet | Dyah W. Karjosukarso Femke Bukkems Lonneke Duijkers Tomasz Z. Tomkiewicz Julia Kiefmann Andrei Sarlea Sander Bervoets Irene Vázquez-Domínguez Laurie L. Molday Robert S. Molday Mihai G. Netea Carel B. Hoyng Alejandro Garanto Rob W. J. Collin |
| author_sort | Dyah W. Karjosukarso |
| collection | DOAJ |
| description | Abstract Background Stargardt disease type 1 (STGD1) is a progressive retinal disorder caused by bi-allelic variants in the ABCA4 gene. A recurrent variant at the exon-intron junction of exon 6, c.768G>T, causes a 35-nt elongation of exon 6 that leads to premature termination of protein synthesis. Methods To correct this aberrant splicing, twenty-five 2′-O-methoxyethyl antisense oligonucleotides (AONs) were designed, spanning the entire exon elongation. Results Testing of these AONs in patient-derived photoreceptor precursor cells and retinal organoids allow the selection of a lead candidate AON (A7 21-mer) that rescues on average 52% and 50% expression of wild-type ABCA4 transcript and protein, respectively. In situ hybridization and probe-based ELISA demonstrate its distribution and stability in vitro and in vivo. No major safety concerns regarding off-targets, immunostimulation and toxicity are observed in transcriptomics analysis, cytokine stimulation assays in human primary immune cells, and cytotoxicity assays. Conclusions Additional optimization and in vivo studies will be performed to further investigate the lead candidate. Considering the high prevalence of this variant, a substantial number of patients are likely to benefit from a successful further development and implementation of this therapy. |
| format | Article |
| id | doaj-art-c8d71f82247c413f9d01a8bd284f4ed1 |
| institution | Kabale University |
| issn | 2730-664X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Medicine |
| spelling | doaj-art-c8d71f82247c413f9d01a8bd284f4ed12025-01-26T12:49:57ZengNature PortfolioCommunications Medicine2730-664X2025-01-015111310.1038/s43856-024-00712-7Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt diseaseDyah W. Karjosukarso0Femke Bukkems1Lonneke Duijkers2Tomasz Z. Tomkiewicz3Julia Kiefmann4Andrei Sarlea5Sander Bervoets6Irene Vázquez-Domínguez7Laurie L. Molday8Robert S. Molday9Mihai G. Netea10Carel B. Hoyng11Alejandro Garanto12Rob W. J. Collin13Department of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Internal Medicine and Radboud Center for Infectious Diseases, Research Institute for Medical Innovation, Radboud University Medical CentreRadboudumc Technology Center Bioinformatics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Biochemistry and Molecular Biology, The University of British ColumbiaDepartment of Biochemistry and Molecular Biology, The University of British ColumbiaDepartment of Internal Medicine and Radboud Center for Infectious Diseases, Research Institute for Medical Innovation, Radboud University Medical CentreAstherna B.V.Department of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterDepartment of Human Genetics, Research Institute for Medical Innovation, Radboud University Medical CenterAbstract Background Stargardt disease type 1 (STGD1) is a progressive retinal disorder caused by bi-allelic variants in the ABCA4 gene. A recurrent variant at the exon-intron junction of exon 6, c.768G>T, causes a 35-nt elongation of exon 6 that leads to premature termination of protein synthesis. Methods To correct this aberrant splicing, twenty-five 2′-O-methoxyethyl antisense oligonucleotides (AONs) were designed, spanning the entire exon elongation. Results Testing of these AONs in patient-derived photoreceptor precursor cells and retinal organoids allow the selection of a lead candidate AON (A7 21-mer) that rescues on average 52% and 50% expression of wild-type ABCA4 transcript and protein, respectively. In situ hybridization and probe-based ELISA demonstrate its distribution and stability in vitro and in vivo. No major safety concerns regarding off-targets, immunostimulation and toxicity are observed in transcriptomics analysis, cytokine stimulation assays in human primary immune cells, and cytotoxicity assays. Conclusions Additional optimization and in vivo studies will be performed to further investigate the lead candidate. Considering the high prevalence of this variant, a substantial number of patients are likely to benefit from a successful further development and implementation of this therapy.https://doi.org/10.1038/s43856-024-00712-7 |
| spellingShingle | Dyah W. Karjosukarso Femke Bukkems Lonneke Duijkers Tomasz Z. Tomkiewicz Julia Kiefmann Andrei Sarlea Sander Bervoets Irene Vázquez-Domínguez Laurie L. Molday Robert S. Molday Mihai G. Netea Carel B. Hoyng Alejandro Garanto Rob W. J. Collin Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt disease Communications Medicine |
| title | Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt disease |
| title_full | Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt disease |
| title_fullStr | Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt disease |
| title_full_unstemmed | Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt disease |
| title_short | Preclinical assessment of splicing modulation therapy for ABCA4 variant c.768G>T in Stargardt disease |
| title_sort | preclinical assessment of splicing modulation therapy for abca4 variant c 768g t in stargardt disease |
| url | https://doi.org/10.1038/s43856-024-00712-7 |
| work_keys_str_mv | AT dyahwkarjosukarso preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT femkebukkems preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT lonnekeduijkers preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT tomaszztomkiewicz preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT juliakiefmann preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT andreisarlea preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT sanderbervoets preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT irenevazquezdominguez preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT laurielmolday preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT robertsmolday preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT mihaignetea preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT carelbhoyng preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT alejandrogaranto preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease AT robwjcollin preclinicalassessmentofsplicingmodulationtherapyforabca4variantc768gtinstargardtdisease |