A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes Identification
The management of unsolved inherited retinal dystrophies (IRD) cases is challenging since no standard pipelines have been established. This study aimed to define a diagnostic algorithm useful for the diagnostic routine and to address unsolved cases. Here, we applied a Next-Generation Sequencing-base...
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doaj-2b053a0b33c5476ea0a2f0338acdd6262020-12-09T00:03:13ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-12-01219355935510.3390/ijms21249355A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes IdentificationMarta Martín-Sánchez0Nereida Bravo-Gil1María González-del Pozo2Cristina Méndez-Vidal3Elena Fernández-Suárez4Enrique Rodríguez-de la Rúa5Salud Borrego6Guillermo Antiñolo7Department of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, SpainDepartment of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, SpainDepartment of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, SpainDepartment of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, SpainDepartment of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, SpainDepartment of Ophthalmology, University Hospital Virgen Macarena, 41013 Seville, SpainDepartment of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, SpainDepartment of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, University Hospital Virgen del Rocío/CSIC/University of Seville, 41013 Seville, SpainThe management of unsolved inherited retinal dystrophies (IRD) cases is challenging since no standard pipelines have been established. This study aimed to define a diagnostic algorithm useful for the diagnostic routine and to address unsolved cases. Here, we applied a Next-Generation Sequencing-based workflow, including a first step of panel sequencing (PS) followed by clinical-exome sequencing (CES) and whole-exome sequencing (WES), in 46 IRD patients belonging to 42 families. Twenty-six likely causal variants in retinal genes were found by PS and CES. CES and WES allowed proposing two novel candidate <i>loci</i> (<i>WDFY3 </i>and a X-linked region including <i>CITED1</i>), both abundantly expressed in human retina according to RT-PCR and immunohistochemistry. After comparison studies, PS showed the best quality and cost values, CES and WES involved similar analytical efforts and WES presented the highest diagnostic yield. These results reinforce the relevance of panels as a first step in the diagnostic routine and suggest WES as the next strategy for unsolved cases, reserving CES for the simultaneous study of multiple conditions. Standardizing this algorithm would enhance the efficiency and equity of clinical genetics practice. Furthermore, the identified candidate genes could contribute to increase the diagnostic yield and expand the mutational spectrum in these disorders.https://www.mdpi.com/1422-0067/21/24/9355next generation sequencinginherited retinal dystrophiesgenetic diagnosis<i>WDFY3</i><i>CITED1</i> |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Marta Martín-Sánchez Nereida Bravo-Gil María González-del Pozo Cristina Méndez-Vidal Elena Fernández-Suárez Enrique Rodríguez-de la Rúa Salud Borrego Guillermo Antiñolo |
spellingShingle |
Marta Martín-Sánchez Nereida Bravo-Gil María González-del Pozo Cristina Méndez-Vidal Elena Fernández-Suárez Enrique Rodríguez-de la Rúa Salud Borrego Guillermo Antiñolo A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes Identification International Journal of Molecular Sciences next generation sequencing inherited retinal dystrophies genetic diagnosis <i>WDFY3</i> <i>CITED1</i> |
author_facet |
Marta Martín-Sánchez Nereida Bravo-Gil María González-del Pozo Cristina Méndez-Vidal Elena Fernández-Suárez Enrique Rodríguez-de la Rúa Salud Borrego Guillermo Antiñolo |
author_sort |
Marta Martín-Sánchez |
title |
A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes Identification |
title_short |
A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes Identification |
title_full |
A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes Identification |
title_fullStr |
A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes Identification |
title_full_unstemmed |
A Multi-Strategy Sequencing Workflow in Inherited Retinal Dystrophies: Routine Diagnosis, Addressing Unsolved Cases and Candidate Genes Identification |
title_sort |
multi-strategy sequencing workflow in inherited retinal dystrophies: routine diagnosis, addressing unsolved cases and candidate genes identification |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2020-12-01 |
description |
The management of unsolved inherited retinal dystrophies (IRD) cases is challenging since no standard pipelines have been established. This study aimed to define a diagnostic algorithm useful for the diagnostic routine and to address unsolved cases. Here, we applied a Next-Generation Sequencing-based workflow, including a first step of panel sequencing (PS) followed by clinical-exome sequencing (CES) and whole-exome sequencing (WES), in 46 IRD patients belonging to 42 families. Twenty-six likely causal variants in retinal genes were found by PS and CES. CES and WES allowed proposing two novel candidate <i>loci</i> (<i>WDFY3 </i>and a X-linked region including <i>CITED1</i>), both abundantly expressed in human retina according to RT-PCR and immunohistochemistry. After comparison studies, PS showed the best quality and cost values, CES and WES involved similar analytical efforts and WES presented the highest diagnostic yield. These results reinforce the relevance of panels as a first step in the diagnostic routine and suggest WES as the next strategy for unsolved cases, reserving CES for the simultaneous study of multiple conditions. Standardizing this algorithm would enhance the efficiency and equity of clinical genetics practice. Furthermore, the identified candidate genes could contribute to increase the diagnostic yield and expand the mutational spectrum in these disorders. |
topic |
next generation sequencing inherited retinal dystrophies genetic diagnosis <i>WDFY3</i> <i>CITED1</i> |
url |
https://www.mdpi.com/1422-0067/21/24/9355 |
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