Practical approach to the genetic diagnosis of unsolved dystrophinopathies : a stepwise strategy in the genomic era
© Author(s) (or their employer(s)) 2021. No commercial re-use. See rights and permissions. Published by BMJ..
OBJECTIVE: To investigate the diagnostic value of implementing a stepwise genetic testing strategy (SGTS) in genetically unsolved cases with dystrophinopathies.
METHODS: After routine genetic testing in 872 male patients with highly suspected dystrophinopathies, we identified 715 patients with a pathogenic DMD variant. Of the 157 patients who had no pathogenic DMD variants and underwent a muscle biopsy, 142 patients were confirmed to have other myopathies, and 15 suspected dystrophinopathies remained genetically undiagnosed. These 15 patients underwent a more comprehensive evaluation as part of the SGTS pipeline, which included the stepwise analysis of dystrophin mRNA, short-read whole-gene DMD sequencing, long-read whole-gene DMD sequencing and in silico bioinformatic analyses.
RESULTS: SGTS successfully yielded a molecular diagnosis of dystrophinopathy in 11 of the 15 genetically unsolved cases. We identified 8 intronic and 2 complex structural variants (SVs) leading to aberrant splicing in 10 of 11 patients, of which 9 variants were novel. In one case, a molecular defect was detected on mRNA and protein level only. Aberrant splicing mechanisms included 6 pseudoexon inclusions and 4 alterations of splice sites and splicing regulatory elements. We showed for the first time the exonisation of a MER48 element as a novel pathogenic mechanism in dystrophinopathies.
CONCLUSION: Our study highlights the high diagnostic utility of implementing a SGTS pipeline in dystrophinopathies with intronic variants and complex SVs.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:58 |
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| Enthalten in: |
Journal of medical genetics - 58(2021), 11 vom: 05. Nov., Seite 743-751 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Xie, Zhiying [VerfasserIn] |
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| Links: |
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| Themen: |
DMD protein, human |
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| Anmerkungen: |
Date Completed 21.02.2022 Date Revised 21.02.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1136/jmedgenet-2020-107113 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM315490144 |
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| 245 | 1 | 0 | |a Practical approach to the genetic diagnosis of unsolved dystrophinopathies |b a stepwise strategy in the genomic era |
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| 500 | |a Date Revised 21.02.2022 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © Author(s) (or their employer(s)) 2021. No commercial re-use. See rights and permissions. Published by BMJ. | ||
| 520 | |a OBJECTIVE: To investigate the diagnostic value of implementing a stepwise genetic testing strategy (SGTS) in genetically unsolved cases with dystrophinopathies | ||
| 520 | |a METHODS: After routine genetic testing in 872 male patients with highly suspected dystrophinopathies, we identified 715 patients with a pathogenic DMD variant. Of the 157 patients who had no pathogenic DMD variants and underwent a muscle biopsy, 142 patients were confirmed to have other myopathies, and 15 suspected dystrophinopathies remained genetically undiagnosed. These 15 patients underwent a more comprehensive evaluation as part of the SGTS pipeline, which included the stepwise analysis of dystrophin mRNA, short-read whole-gene DMD sequencing, long-read whole-gene DMD sequencing and in silico bioinformatic analyses | ||
| 520 | |a RESULTS: SGTS successfully yielded a molecular diagnosis of dystrophinopathy in 11 of the 15 genetically unsolved cases. We identified 8 intronic and 2 complex structural variants (SVs) leading to aberrant splicing in 10 of 11 patients, of which 9 variants were novel. In one case, a molecular defect was detected on mRNA and protein level only. Aberrant splicing mechanisms included 6 pseudoexon inclusions and 4 alterations of splice sites and splicing regulatory elements. We showed for the first time the exonisation of a MER48 element as a novel pathogenic mechanism in dystrophinopathies | ||
| 520 | |a CONCLUSION: Our study highlights the high diagnostic utility of implementing a SGTS pipeline in dystrophinopathies with intronic variants and complex SVs | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
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| 650 | 4 | |a neuromuscular diseases | |
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| 650 | 7 | |a Dystrophin |2 NLM | |
| 700 | 1 | |a Sun, Chengyue |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Yilin |e verfasserin |4 aut | |
| 700 | 1 | |a Yu, Meng |e verfasserin |4 aut | |
| 700 | 1 | |a Zheng, Yiming |e verfasserin |4 aut | |
| 700 | 1 | |a Meng, Lingchao |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Gao |e verfasserin |4 aut | |
| 700 | 1 | |a Cornejo-Sanchez, Diana M |e verfasserin |4 aut | |
| 700 | 1 | |a Bharadwaj, Thashi |e verfasserin |4 aut | |
| 700 | 1 | |a Yan, Jin |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Lingxiang |e verfasserin |4 aut | |
| 700 | 1 | |a Pineda-Trujillo, Nicolas |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Wei |e verfasserin |4 aut | |
| 700 | 1 | |a Leal, Suzanne M |e verfasserin |4 aut | |
| 700 | 1 | |a Schrauwen, Isabelle |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Zhaoxia |e verfasserin |4 aut | |
| 700 | 1 | |a Yuan, Yun |e verfasserin |4 aut | |
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