Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation Sequencing
Mucopolysaccharidosis type II is an X-linked lysosomal storage disorder caused by mutations in the IDS gene that encodes the iduronate-2-sulfatase enzyme. The IDS gene is located on the long arm of the X-chromosome, comprising 9 exons, spanning approximately 24 kb. The analysis of carriers, in addit...
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| Format: | Article |
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Frontiers Media S.A.
2021-11-01
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| Series: | Frontiers in Molecular Biosciences |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2021.789350/full |
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| author | Alice Brinckmann Oliveira Netto Alice Brinckmann Oliveira Netto Ana Carolina Brusius-Facchin Ana Carolina Brusius-Facchin Ana Carolina Brusius-Facchin Sandra Leistner-Segal Francyne Kubaski Francyne Kubaski Francyne Kubaski Francyne Kubaski Juliana Josahkian Juliana Josahkian Roberto Giugliani Roberto Giugliani Roberto Giugliani Roberto Giugliani |
| author_facet | Alice Brinckmann Oliveira Netto Alice Brinckmann Oliveira Netto Ana Carolina Brusius-Facchin Ana Carolina Brusius-Facchin Ana Carolina Brusius-Facchin Sandra Leistner-Segal Francyne Kubaski Francyne Kubaski Francyne Kubaski Francyne Kubaski Juliana Josahkian Juliana Josahkian Roberto Giugliani Roberto Giugliani Roberto Giugliani Roberto Giugliani |
| author_sort | Alice Brinckmann Oliveira Netto |
| collection | DOAJ |
| description | Mucopolysaccharidosis type II is an X-linked lysosomal storage disorder caused by mutations in the IDS gene that encodes the iduronate-2-sulfatase enzyme. The IDS gene is located on the long arm of the X-chromosome, comprising 9 exons, spanning approximately 24 kb. The analysis of carriers, in addition to detecting mutations in patients, is essential for genetic counseling, since the risk of recurrence for male children is 50%. Mosaicism is a well-known phenomenon described in many genetic disorders caused by a variety of mechanisms that occur when a mutation arises in the early development of an embryo. Sanger sequencing is limited in detecting somatic mosaicism and sequence change levels of less than 20% may be missed. The Next Generation Sequencing (NGS) has been increasingly used in diagnosis. It is a sensitive and fast method for the detection of somatic mosaicism. Compared to Sanger sequencing, which represents a cumulative signal, NGS technology analyzes the sequence of each DNA read in a sample. NGS might therefore facilitate the detection of mosaicism in mothers of MPS II patients. The aim of this study was to reanalyze, by NGS, all MPS II mothers that showed to be non-carriers by Sanger analysis. Twelve non-carriers were selected for the reanalysis on the Ion PGM and Ion Torrent S5 platform, using a custom panel that includes the IDS gene. Results were visualized in the Integrative Genomics Viewer (IGV). We were able to detected the presence of the variant previously found in the index case in three of the mothers, with frequencies ranging between 13 and 49% of the reads. These results suggest the possibility of mosaicism in the mothers. The use of a more sensitive technology for detecting low-level mosaic mutations is essential for accurate recurrence-risk estimates. In our study, the NGS analysis showed to be an effective methodology to detect the mosaic event. |
| first_indexed | 2024-12-19T04:20:44Z |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-12-19T04:20:44Z |
| publishDate | 2021-11-01 |
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| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-3b22f180790947778d87aeca9c971d992022-12-21T20:36:09ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2021-11-01810.3389/fmolb.2021.789350789350Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation SequencingAlice Brinckmann Oliveira Netto0Alice Brinckmann Oliveira Netto1Ana Carolina Brusius-Facchin2Ana Carolina Brusius-Facchin3Ana Carolina Brusius-Facchin4Sandra Leistner-Segal5Francyne Kubaski6Francyne Kubaski7Francyne Kubaski8Francyne Kubaski9Juliana Josahkian10Juliana Josahkian11Roberto Giugliani12Roberto Giugliani13Roberto Giugliani14Roberto Giugliani15Laboratory of Molecular Genetics, Medical Genetics Service, HCPA, Porto Alegre, BrazilPostgraduate Program in Genetics and Molecular Biology, UFRGS, Porto Alegre, BrazilLaboratory of Molecular Genetics, Medical Genetics Service, HCPA, Porto Alegre, BrazilNational Institute on Population Medical Genetics, INAGEMP, Porto Alegre, BrazilBioDiscovery Laboratory, Experimental Research Center, HCPA, Porto Alegre, BrazilLaboratory of Molecular Genetics, Medical Genetics Service, HCPA, Porto Alegre, BrazilLaboratory of Molecular Genetics, Medical Genetics Service, HCPA, Porto Alegre, BrazilPostgraduate Program in Genetics and Molecular Biology, UFRGS, Porto Alegre, BrazilNational Institute on Population Medical Genetics, INAGEMP, Porto Alegre, BrazilBioDiscovery Laboratory, Experimental Research Center, HCPA, Porto Alegre, BrazilPostgraduate Program in Genetics and Molecular Biology, UFRGS, Porto Alegre, BrazilDepartment of Clinical Medicine, Hospital Universitario de Santa Maria (HUSM), Santa Maria, BrazilLaboratory of Molecular Genetics, Medical Genetics Service, HCPA, Porto Alegre, BrazilNational Institute on Population Medical Genetics, INAGEMP, Porto Alegre, BrazilBioDiscovery Laboratory, Experimental Research Center, HCPA, Porto Alegre, BrazilDepartment of Genetics, UFRGS, Porto Alegre, BrazilMucopolysaccharidosis type II is an X-linked lysosomal storage disorder caused by mutations in the IDS gene that encodes the iduronate-2-sulfatase enzyme. The IDS gene is located on the long arm of the X-chromosome, comprising 9 exons, spanning approximately 24 kb. The analysis of carriers, in addition to detecting mutations in patients, is essential for genetic counseling, since the risk of recurrence for male children is 50%. Mosaicism is a well-known phenomenon described in many genetic disorders caused by a variety of mechanisms that occur when a mutation arises in the early development of an embryo. Sanger sequencing is limited in detecting somatic mosaicism and sequence change levels of less than 20% may be missed. The Next Generation Sequencing (NGS) has been increasingly used in diagnosis. It is a sensitive and fast method for the detection of somatic mosaicism. Compared to Sanger sequencing, which represents a cumulative signal, NGS technology analyzes the sequence of each DNA read in a sample. NGS might therefore facilitate the detection of mosaicism in mothers of MPS II patients. The aim of this study was to reanalyze, by NGS, all MPS II mothers that showed to be non-carriers by Sanger analysis. Twelve non-carriers were selected for the reanalysis on the Ion PGM and Ion Torrent S5 platform, using a custom panel that includes the IDS gene. Results were visualized in the Integrative Genomics Viewer (IGV). We were able to detected the presence of the variant previously found in the index case in three of the mothers, with frequencies ranging between 13 and 49% of the reads. These results suggest the possibility of mosaicism in the mothers. The use of a more sensitive technology for detecting low-level mosaic mutations is essential for accurate recurrence-risk estimates. In our study, the NGS analysis showed to be an effective methodology to detect the mosaic event.https://www.frontiersin.org/articles/10.3389/fmolb.2021.789350/fullmosaicismmucopolysaccharidosis type IIhunter syndromenext-generation sequencingIDS genex-linked disease |
| spellingShingle | Alice Brinckmann Oliveira Netto Alice Brinckmann Oliveira Netto Ana Carolina Brusius-Facchin Ana Carolina Brusius-Facchin Ana Carolina Brusius-Facchin Sandra Leistner-Segal Francyne Kubaski Francyne Kubaski Francyne Kubaski Francyne Kubaski Juliana Josahkian Juliana Josahkian Roberto Giugliani Roberto Giugliani Roberto Giugliani Roberto Giugliani Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation Sequencing Frontiers in Molecular Biosciences mosaicism mucopolysaccharidosis type II hunter syndrome next-generation sequencing IDS gene x-linked disease |
| title | Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation Sequencing |
| title_full | Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation Sequencing |
| title_fullStr | Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation Sequencing |
| title_full_unstemmed | Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation Sequencing |
| title_short | Detection of Mosaic Variants in Mothers of MPS II Patients by Next Generation Sequencing |
| title_sort | detection of mosaic variants in mothers of mps ii patients by next generation sequencing |
| topic | mosaicism mucopolysaccharidosis type II hunter syndrome next-generation sequencing IDS gene x-linked disease |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2021.789350/full |
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