Clinical application of long-read nanopore sequencing in a preimplantation genetic testing pre-clinical workup to identify the junction for complex Xq chromosome rearrangement-related disease.

Autor: Mariya T; Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan.; Departments of Medical Genetics and Genomics, School of Medicine, Sapporo Medical University, Sapporo, Japan.; Department of Obstetrics and Gynecology, School of Medicine, Sapporo Medical University, Sapporo, Japan., Shichiri Y; Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan., Sugimoto T; Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan., Kawamura R; Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan., Miyai S; Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan., Inagaki H; Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan., Sugihara E; Open Facility Center, Research Promotion and Support Headquarters, Fujita Health University, Toyoake, Aichi, Japan., Ikeda K; Department of Obstetrics and Gynecology, School of Medicine, Sapporo Medical University, Sapporo, Japan., Baba T; Department of Obstetrics and Gynecology, School of Medicine, Sapporo Medical University, Sapporo, Japan., Ishikawa A; Departments of Medical Genetics and Genomics, School of Medicine, Sapporo Medical University, Sapporo, Japan., Ammae M; IVF Namba Clinic, Osaka, Japan., Nakaoka Y; IVF Namba Clinic, Osaka, Japan., Saito T; Department of Obstetrics and Gynecology, School of Medicine, Sapporo Medical University, Sapporo, Japan., Sakurai A; Departments of Medical Genetics and Genomics, School of Medicine, Sapporo Medical University, Sapporo, Japan., Kurahashi H; Division of Molecular Genetics, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan.
Jazyk: angličtina
Zdroj: Prenatal diagnosis [Prenat Diagn] 2023 Mar; Vol. 43 (3), pp. 304-313. Date of Electronic Publication: 2023 Feb 24.
DOI: 10.1002/pd.6334
Abstrakt: Objective: Xq chromosome duplication with complex rearrangements is generally acknowledged to be associated with neurodevelopmental disorders, such as Pelizaeus-Merzbacher disease (PMD) and MECP2 duplication syndrome. For couples who required a PGT-M (pre-implantation genetic testing for monogenic disease) for these disorders, junction-specific PCR is useful to directly detect pathogenic variants. Therefore, pre-clinical workup for PGT-M requires the identification of the junction of duplicated segments in PMD and MECP2 duplication syndrome, which is generally difficult.
Methods: In this report, we used nanopore long-read sequencing targeting the X chromosome using an adaptive sampling method to identify breakpoint junctions in disease-causing triplications.
Results: By long-read sequencing, we successfully identified breakpoint junctions in one PMD case with PLP1 triplication and in another MECP2 triplication case in a single sequencing run. Surprisingly, the duplicated region involving MECP2 was inserted 45 Mb proximal to the original position. This inserted region was confirmed by FISH analysis. With the help of precise mapping of the pathogenic variant, we successfully re-established STR haplotyping for PGT-M and avoided any potential misinterpretation of the pathogenic allele due to recombination.
Conclusion: Long-read sequencing with adaptive sampling in a PGT-M pre-clinical workup is a beneficial method for identifying junctions of chromosomal complex structural rearrangements.
(© 2023 John Wiley & Sons Ltd.)
Databáze: MEDLINE
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