Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1
Myotonic dystrophy type 1 (DM1) is the most complex and variable trinucleotide repeat disorder caused by an unstable CTG repeat expansion, reaching up to 4000 CTG in the most severe cases. The genetic and clinical variability of DM1 depend on the sex and age of the transmitting parent, but also on t...
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| Format: | Article |
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MDPI AG
2021-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/5/2616 |
| _version_ | 1797413926670958592 |
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| author | Antoine Mangin Laure de Pontual Yu-Chih Tsai Laetitia Monteil Mathilde Nizon Pierre Boisseau Sandra Mercier Janet Ziegle John Harting Cheryl Heiner Geneviève Gourdon Stéphanie Tomé |
| author_facet | Antoine Mangin Laure de Pontual Yu-Chih Tsai Laetitia Monteil Mathilde Nizon Pierre Boisseau Sandra Mercier Janet Ziegle John Harting Cheryl Heiner Geneviève Gourdon Stéphanie Tomé |
| author_sort | Antoine Mangin |
| collection | DOAJ |
| description | Myotonic dystrophy type 1 (DM1) is the most complex and variable trinucleotide repeat disorder caused by an unstable CTG repeat expansion, reaching up to 4000 CTG in the most severe cases. The genetic and clinical variability of DM1 depend on the sex and age of the transmitting parent, but also on the CTG repeat number, presence of repeat interruptions and/or on the degree of somatic instability. Currently, it is difficult to simultaneously and accurately determine these contributing factors in DM1 patients due to the limitations of gold standard methods used in molecular diagnostics and research laboratories. Our study showed the efficiency of the latest PacBio long-read sequencing technology to sequence large CTG trinucleotides, detect multiple and single repeat interruptions and estimate the levels of somatic mosaicism in DM1 patients carrying complex CTG repeat expansions inaccessible to most methods. Using this innovative approach, we revealed the existence of de novo CCG interruptions associated with CTG stabilization/contraction across generations in a new DM1 family. We also demonstrated that our method is suitable to sequence the DM1 locus and measure somatic mosaicism in DM1 families carrying more than 1000 pure CTG repeats. Better characterization of expanded alleles in DM1 patients can significantly improve prognosis and genetic counseling, not only in DM1 but also for other tandem DNA repeat disorders. |
| first_indexed | 2024-03-09T05:25:47Z |
| format | Article |
| id | doaj.art-e92175841b064964bae67769779c1a8c |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T05:25:47Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e92175841b064964bae67769779c1a8c2023-12-03T12:37:30ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-03-01225261610.3390/ijms22052616Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1Antoine Mangin0Laure de Pontual1Yu-Chih Tsai2Laetitia Monteil3Mathilde Nizon4Pierre Boisseau5Sandra Mercier6Janet Ziegle7John Harting8Cheryl Heiner9Geneviève Gourdon10Stéphanie Tomé11Centre de Recherche en Myologie, Inserm, Institut de Myologie, Sorbonne Université, F-75013 Paris, FranceCentre de Recherche en Myologie, Inserm, Institut de Myologie, Sorbonne Université, F-75013 Paris, FrancePacific Biosciences, Menlo Park, CA 94025, USAGenetics Department of the Hospital of Toulouse, F-31059 Toulouse, FranceCHU de Nantes, Service de Génétique Médicale, Laboratoire de Génétique Moléculaire, F-44000 Nantes, FranceCHU de Nantes, Service de Génétique Médicale, Laboratoire de Génétique Moléculaire, F-44000 Nantes, FranceCHU Nantes, Service de Génétique Médicale, Centre de Référence des Maladies Neuromusculaires AOC, F-44000 Nantes, FrancePacific Biosciences, Menlo Park, CA 94025, USAPacific Biosciences, Menlo Park, CA 94025, USAPacific Biosciences, Menlo Park, CA 94025, USACentre de Recherche en Myologie, Inserm, Institut de Myologie, Sorbonne Université, F-75013 Paris, FranceCentre de Recherche en Myologie, Inserm, Institut de Myologie, Sorbonne Université, F-75013 Paris, FranceMyotonic dystrophy type 1 (DM1) is the most complex and variable trinucleotide repeat disorder caused by an unstable CTG repeat expansion, reaching up to 4000 CTG in the most severe cases. The genetic and clinical variability of DM1 depend on the sex and age of the transmitting parent, but also on the CTG repeat number, presence of repeat interruptions and/or on the degree of somatic instability. Currently, it is difficult to simultaneously and accurately determine these contributing factors in DM1 patients due to the limitations of gold standard methods used in molecular diagnostics and research laboratories. Our study showed the efficiency of the latest PacBio long-read sequencing technology to sequence large CTG trinucleotides, detect multiple and single repeat interruptions and estimate the levels of somatic mosaicism in DM1 patients carrying complex CTG repeat expansions inaccessible to most methods. Using this innovative approach, we revealed the existence of de novo CCG interruptions associated with CTG stabilization/contraction across generations in a new DM1 family. We also demonstrated that our method is suitable to sequence the DM1 locus and measure somatic mosaicism in DM1 families carrying more than 1000 pure CTG repeats. Better characterization of expanded alleles in DM1 patients can significantly improve prognosis and genetic counseling, not only in DM1 but also for other tandem DNA repeat disorders.https://www.mdpi.com/1422-0067/22/5/2616myotonic dystrophy type 1long read sequencingsomatic mosaicisminterrupted CTG repeat expansion |
| spellingShingle | Antoine Mangin Laure de Pontual Yu-Chih Tsai Laetitia Monteil Mathilde Nizon Pierre Boisseau Sandra Mercier Janet Ziegle John Harting Cheryl Heiner Geneviève Gourdon Stéphanie Tomé Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1 International Journal of Molecular Sciences myotonic dystrophy type 1 long read sequencing somatic mosaicism interrupted CTG repeat expansion |
| title | Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1 |
| title_full | Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1 |
| title_fullStr | Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1 |
| title_full_unstemmed | Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1 |
| title_short | Robust Detection of Somatic Mosaicism and Repeat Interruptions by Long-Read Targeted Sequencing in Myotonic Dystrophy Type 1 |
| title_sort | robust detection of somatic mosaicism and repeat interruptions by long read targeted sequencing in myotonic dystrophy type 1 |
| topic | myotonic dystrophy type 1 long read sequencing somatic mosaicism interrupted CTG repeat expansion |
| url | https://www.mdpi.com/1422-0067/22/5/2616 |
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