Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial Cells
Human induced pluripotent stem cells (hiPSCs) generated from patients and the derivative retinal cells enable the investigation of pathological and novel variants in relevant cell populations. Biallelic pathogenic variants in RPE65 cause early-onset severe retinal dystrophy (EOSRD) or Leber congenit...
Saved in:
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2021-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2021/4536382 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832546030853292032 |
|---|---|
| author | Benjamin M. Nash To Ha Loi Milan Fernando Amin Sabri James Robinson Anson Cheng Steven S. Eamegdool Elizabeth Farnsworth Bruce Bennetts John R. Grigg Seo-Kyung Chung Anai Gonzalez-Cordero Robyn V. Jamieson |
| author_facet | Benjamin M. Nash To Ha Loi Milan Fernando Amin Sabri James Robinson Anson Cheng Steven S. Eamegdool Elizabeth Farnsworth Bruce Bennetts John R. Grigg Seo-Kyung Chung Anai Gonzalez-Cordero Robyn V. Jamieson |
| author_sort | Benjamin M. Nash |
| collection | DOAJ |
| description | Human induced pluripotent stem cells (hiPSCs) generated from patients and the derivative retinal cells enable the investigation of pathological and novel variants in relevant cell populations. Biallelic pathogenic variants in RPE65 cause early-onset severe retinal dystrophy (EOSRD) or Leber congenital amaurosis (LCA). Increasingly, regulatory-approved in vivo RPE65 retinal gene replacement therapy is available for patients with these clinical features, but only if they have biallelic pathological variants and sufficient viable retinal cells. In our cohort of patients, we identified siblings with early-onset severe retinal degeneration where genomic studies revealed compound heterozygous variants in RPE65, one a known pathogenic missense variant and the other a novel synonymous variant of uncertain significance. The synonymous variant was suspected to affect RNA splicing. Since RPE65 is very poorly expressed in all tissues except the retinal pigment epithelium (RPE), we generated hiPSC-derived RPE cells from the parental carrier of the synonymous variant. Sequencing of RNA obtained from hiPSC-RPE cells demonstrated heterozygous skipping of RPE65 exon 2 and the introduction of a premature stop codon in the mRNA. Minigene studies confirmed the splicing aberration. Results from this study led to reclassification of the synonymous variant to a pathogenic variant, providing the affected patients with access to RPE65 gene replacement therapy. |
| format | Article |
| id | doaj-art-efc0b3e3faab4cc08d30e555d2e914e1 |
| institution | Kabale University |
| issn | 1687-9678 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-efc0b3e3faab4cc08d30e555d2e914e12025-02-03T07:24:09ZengWileyStem Cells International1687-96782021-01-01202110.1155/2021/4536382Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial CellsBenjamin M. Nash0To Ha Loi1Milan Fernando2Amin Sabri3James Robinson4Anson Cheng5Steven S. Eamegdool6Elizabeth Farnsworth7Bruce Bennetts8John R. Grigg9Seo-Kyung Chung10Anai Gonzalez-Cordero11Robyn V. Jamieson12Eye Genetics Research UnitEye Genetics Research UnitStem Cell Medicine Group and Stem Cell and Organoid FacilityEye Genetics Research UnitDepartment of OphthalmologyEye Genetics Research UnitEye Genetics Research UnitSydney Genome DiagnosticsSpecialty of Genomic MedicineEye Genetics Research UnitTranslational Neurogenomics GroupStem Cell Medicine Group and Stem Cell and Organoid FacilityEye Genetics Research UnitHuman induced pluripotent stem cells (hiPSCs) generated from patients and the derivative retinal cells enable the investigation of pathological and novel variants in relevant cell populations. Biallelic pathogenic variants in RPE65 cause early-onset severe retinal dystrophy (EOSRD) or Leber congenital amaurosis (LCA). Increasingly, regulatory-approved in vivo RPE65 retinal gene replacement therapy is available for patients with these clinical features, but only if they have biallelic pathological variants and sufficient viable retinal cells. In our cohort of patients, we identified siblings with early-onset severe retinal degeneration where genomic studies revealed compound heterozygous variants in RPE65, one a known pathogenic missense variant and the other a novel synonymous variant of uncertain significance. The synonymous variant was suspected to affect RNA splicing. Since RPE65 is very poorly expressed in all tissues except the retinal pigment epithelium (RPE), we generated hiPSC-derived RPE cells from the parental carrier of the synonymous variant. Sequencing of RNA obtained from hiPSC-RPE cells demonstrated heterozygous skipping of RPE65 exon 2 and the introduction of a premature stop codon in the mRNA. Minigene studies confirmed the splicing aberration. Results from this study led to reclassification of the synonymous variant to a pathogenic variant, providing the affected patients with access to RPE65 gene replacement therapy.http://dx.doi.org/10.1155/2021/4536382 |
| spellingShingle | Benjamin M. Nash To Ha Loi Milan Fernando Amin Sabri James Robinson Anson Cheng Steven S. Eamegdool Elizabeth Farnsworth Bruce Bennetts John R. Grigg Seo-Kyung Chung Anai Gonzalez-Cordero Robyn V. Jamieson Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial Cells Stem Cells International |
| title | Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial Cells |
| title_full | Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial Cells |
| title_fullStr | Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial Cells |
| title_full_unstemmed | Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial Cells |
| title_short | Evaluation for Retinal Therapy for RPE65 Variation Assessed in hiPSC Retinal Pigment Epithelial Cells |
| title_sort | evaluation for retinal therapy for rpe65 variation assessed in hipsc retinal pigment epithelial cells |
| url | http://dx.doi.org/10.1155/2021/4536382 |
| work_keys_str_mv | AT benjaminmnash evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT tohaloi evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT milanfernando evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT aminsabri evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT jamesrobinson evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT ansoncheng evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT stevenseamegdool evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT elizabethfarnsworth evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT brucebennetts evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT johnrgrigg evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT seokyungchung evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT anaigonzalezcordero evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells AT robynvjamieson evaluationforretinaltherapyforrpe65variationassessedinhipscretinalpigmentepithelialcells |