Multi-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted region

Abstract Background Beckwith–Wiedemann syndrome (BWS) is a congenital imprinting disorder (ID) caused by molecular defects in the 11p15.5 imprinted region, such as hypomethylation of the KCNQ1OT1:TSS-differentially methylated region (KCNQ1OT1-DMR) and hypermethylation of the H19/IGF2:IG-DMR, and mat...

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Main Authors: Tatsuki Urakawa, Yuri Kanamaru, Naoko Amano, Akira Uchida, Maki Fukami, Masayo Kagami
Format: Article
Language:English
Published: BMC 2025-06-01
Series:Clinical Epigenetics
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Online Access:https://doi.org/10.1186/s13148-025-01907-y
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author Tatsuki Urakawa
Yuri Kanamaru
Naoko Amano
Akira Uchida
Maki Fukami
Masayo Kagami
author_facet Tatsuki Urakawa
Yuri Kanamaru
Naoko Amano
Akira Uchida
Maki Fukami
Masayo Kagami
author_sort Tatsuki Urakawa
collection DOAJ
description Abstract Background Beckwith–Wiedemann syndrome (BWS) is a congenital imprinting disorder (ID) caused by molecular defects in the 11p15.5 imprinted region, such as hypomethylation of the KCNQ1OT1:TSS-differentially methylated region (KCNQ1OT1-DMR) and hypermethylation of the H19/IGF2:IG-DMR, and maternal CDKN1C pathogenic variants, with various clinical characteristics, including overgrowth and macroglossia. Recently, the concept of Beckwith–Wiedemann spectrum (BWSp) and a clinical scoring system for BWS have been proposed, and cases with four or more points are diagnosed with classic BWS, and 20% of cases with BWS have no molecular defects in the 11p15.5 imprinted region. Pseudohypoparathyroidism type 1B (PHP1B, alias inactivating parathyroid hormone (PTH)/PTH-related protein signaling disorder 3) has characteristics of hormone resistance, particularly PTH, caused by methylation defects in DMRs at the GNAS locus (GNAS-DMRs). Some cases with PHP1B show postnatal overgrowth, which overlaps the BWS-phenotype. However, no studies have conducted a multi-locus methylation analysis for the ID-responsible DMRs other than the DMRs in 11p15.5 in cases with the BWS-phenotype and without molecular defects in the 11p15.5 imprinted region. Results We conducted methylation analysis using pyrosequencing in 77 patients showing the BWS-phenotype without molecular defects in the 11p15.5 imprinted region. Consequently, we identified three patients with methylation defects in the GNAS-DMRs. Patients 1, 2, and 3 had 9, 5, and 4 points in a BWSp score, respectively. All three patients had macroglossia and postnatal overgrowth. Further analyses, methylation-specific multiple ligation-dependent probe amplification for multiple DMRs, array-based methylation analysis, exome sequencing, array comparative genome hybridization analysis, and microsatellite marker analysis showed 9p deletion in Patient 1 and paternal uniparental isodisomy of chromosome 20 in Patient 2 together with multiple methylation defects in DMRs other than the GNAS-DMRs. Patient 3 had methylation defects in only the GNAS-DMRs. Conclusion Methylation defects in the GNAS-DMRs can cause the BWS-phenotype. For cases with the BWS-phenotype but no molecular defects in the 11p15.5 imprinted region, methylation analysis for the DMRs at the GNAS locus should be considered.
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spelling doaj-art-c1330e862c7a4983ae907df6561c0bbb2025-08-20T03:21:03ZengBMCClinical Epigenetics1868-70832025-06-0117111310.1186/s13148-025-01907-yMulti-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted regionTatsuki Urakawa0Yuri Kanamaru1Naoko Amano2Akira Uchida3Maki Fukami4Masayo Kagami5Department of Molecular Endocrinology, National Research Institute for Child Health and DevelopmentDepartment of Molecular Endocrinology, National Research Institute for Child Health and DevelopmentDepartment of Pediatrics, Saitama City HospitalDepartment of Neonatology, Hiroshima City Hiroshima Citizens HospitalDepartment of Molecular Endocrinology, National Research Institute for Child Health and DevelopmentDepartment of Molecular Endocrinology, National Research Institute for Child Health and DevelopmentAbstract Background Beckwith–Wiedemann syndrome (BWS) is a congenital imprinting disorder (ID) caused by molecular defects in the 11p15.5 imprinted region, such as hypomethylation of the KCNQ1OT1:TSS-differentially methylated region (KCNQ1OT1-DMR) and hypermethylation of the H19/IGF2:IG-DMR, and maternal CDKN1C pathogenic variants, with various clinical characteristics, including overgrowth and macroglossia. Recently, the concept of Beckwith–Wiedemann spectrum (BWSp) and a clinical scoring system for BWS have been proposed, and cases with four or more points are diagnosed with classic BWS, and 20% of cases with BWS have no molecular defects in the 11p15.5 imprinted region. Pseudohypoparathyroidism type 1B (PHP1B, alias inactivating parathyroid hormone (PTH)/PTH-related protein signaling disorder 3) has characteristics of hormone resistance, particularly PTH, caused by methylation defects in DMRs at the GNAS locus (GNAS-DMRs). Some cases with PHP1B show postnatal overgrowth, which overlaps the BWS-phenotype. However, no studies have conducted a multi-locus methylation analysis for the ID-responsible DMRs other than the DMRs in 11p15.5 in cases with the BWS-phenotype and without molecular defects in the 11p15.5 imprinted region. Results We conducted methylation analysis using pyrosequencing in 77 patients showing the BWS-phenotype without molecular defects in the 11p15.5 imprinted region. Consequently, we identified three patients with methylation defects in the GNAS-DMRs. Patients 1, 2, and 3 had 9, 5, and 4 points in a BWSp score, respectively. All three patients had macroglossia and postnatal overgrowth. Further analyses, methylation-specific multiple ligation-dependent probe amplification for multiple DMRs, array-based methylation analysis, exome sequencing, array comparative genome hybridization analysis, and microsatellite marker analysis showed 9p deletion in Patient 1 and paternal uniparental isodisomy of chromosome 20 in Patient 2 together with multiple methylation defects in DMRs other than the GNAS-DMRs. Patient 3 had methylation defects in only the GNAS-DMRs. Conclusion Methylation defects in the GNAS-DMRs can cause the BWS-phenotype. For cases with the BWS-phenotype but no molecular defects in the 11p15.5 imprinted region, methylation analysis for the DMRs at the GNAS locus should be considered.https://doi.org/10.1186/s13148-025-01907-yBeckwith–Wiedemann syndromeImprinting disordersGNASPseudohypoparathyroidismMethylation analysis9p deletion
spellingShingle Tatsuki Urakawa
Yuri Kanamaru
Naoko Amano
Akira Uchida
Maki Fukami
Masayo Kagami
Multi-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted region
Clinical Epigenetics
Beckwith–Wiedemann syndrome
Imprinting disorders
GNAS
Pseudohypoparathyroidism
Methylation analysis
9p deletion
title Multi-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted region
title_full Multi-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted region
title_fullStr Multi-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted region
title_full_unstemmed Multi-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted region
title_short Multi-locus methylation analyses reveal GNAS methylation defects in three patients with the Beckwith–Wiedemann syndrome phenotype and no molecular defects in the 11p15.5 imprinted region
title_sort multi locus methylation analyses reveal gnas methylation defects in three patients with the beckwith wiedemann syndrome phenotype and no molecular defects in the 11p15 5 imprinted region
topic Beckwith–Wiedemann syndrome
Imprinting disorders
GNAS
Pseudohypoparathyroidism
Methylation analysis
9p deletion
url https://doi.org/10.1186/s13148-025-01907-y
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