The challenge of genetically unresolved ...
Document type :
Article dans une revue scientifique: Article original
DOI :
Title :
The challenge of genetically unresolved haemophilia A patients: Interest of the combination of whole F8 gene sequencing and functional assays
Author(s) :
Lassalle, Fanny [Auteur]
Récepteurs Nucléaires, Maladies Métaboliques et Cardiovasculaires - U 1011 [RNMCD]
Jourdy, Yohann [Auteur]
Hémostase, Inflammation et sepsis
Jouan, Loubna [Auteur]
Swystun, Laura [Auteur]
Gauthier, Julie [Auteur]
CHU Sainte Justine [Montréal]
Zawadzki, Christophe [Auteur]
Récepteurs nucléaires, maladies cardiovasculaires et diabète - U 1011 [RNMCD]
Goudemand, Jenny [Auteur]
Centre Hospitalier Régional Universitaire [CHU Lille] [CHRU Lille]
Susen, Sophie [Auteur]
Récepteurs Nucléaires, Maladies Métaboliques et Cardiovasculaires - U 1011 [RNMCD]
Rivard, Georges‐etienne [Auteur]
Lillicrap, David [Auteur]
Queen's University [Kingston, Canada]
Récepteurs Nucléaires, Maladies Métaboliques et Cardiovasculaires - U 1011 [RNMCD]
Jourdy, Yohann [Auteur]
Hémostase, Inflammation et sepsis
Jouan, Loubna [Auteur]
Swystun, Laura [Auteur]
Gauthier, Julie [Auteur]
CHU Sainte Justine [Montréal]
Zawadzki, Christophe [Auteur]

Récepteurs nucléaires, maladies cardiovasculaires et diabète - U 1011 [RNMCD]
Goudemand, Jenny [Auteur]
Centre Hospitalier Régional Universitaire [CHU Lille] [CHRU Lille]
Susen, Sophie [Auteur]
Récepteurs Nucléaires, Maladies Métaboliques et Cardiovasculaires - U 1011 [RNMCD]
Rivard, Georges‐etienne [Auteur]
Lillicrap, David [Auteur]
Queen's University [Kingston, Canada]
Journal title :
Haemophilia
Pages :
1056-1063
Publisher :
Wiley
Publication date :
2020-10-23
ISSN :
1351-8216
HAL domain(s) :
Sciences du Vivant [q-bio]
English abstract : [en]
Abstract Background The causative variant remains unidentified in 2%–5% of haemophilia A (HA) patients despite an exhaustive sequencing of the full F8 coding sequence, splice consensus sequences, 5’/3’ untranslated regions ...
Show more >Abstract Background The causative variant remains unidentified in 2%–5% of haemophilia A (HA) patients despite an exhaustive sequencing of the full F8 coding sequence, splice consensus sequences, 5’/3’ untranslated regions and copy number variant (CNV) analysis. Next‐generation sequencing (NGS) has provided significant improvements for a complete F8 analysis. Aim The aim of this study was to identify and characterize pathogenic non‐coding variants in F8 of 15 French and Canadian HA patients genetically unresolved, through the use of NGS, mRNA sequencing and functional confirmation of aberrant splicing. Methods We sequenced the entire F8 gene using an NGS capture method. We analysed F8 mRNA in order to detect aberrant transcripts. The pathogenic effect of candidate intronic variants was further confirmed using a minigene assay. Results After bioinformatic analysis, 11 deep intronic variants were identified in 13 patients (8 new variants and 3 previously reported). Three variants were confirmed to be likely pathogenic with the presence of an aberrant transcript during mRNA analysis and minigene assay. We also found a small intronic deletion in 6 patients, recently described as causing mild HA. Conclusion With this comprehensive work combining NGS and functional assays, we report new deep intronic variants that cause HA through splicing alteration mechanism. Functional analyses are critical to confirm the pathogenic effect of these variants and will be invaluable in the future to study the large number of variants of uncertain significance that may affect splicing that will be found in the human genome.Show less >
Show more >Abstract Background The causative variant remains unidentified in 2%–5% of haemophilia A (HA) patients despite an exhaustive sequencing of the full F8 coding sequence, splice consensus sequences, 5’/3’ untranslated regions and copy number variant (CNV) analysis. Next‐generation sequencing (NGS) has provided significant improvements for a complete F8 analysis. Aim The aim of this study was to identify and characterize pathogenic non‐coding variants in F8 of 15 French and Canadian HA patients genetically unresolved, through the use of NGS, mRNA sequencing and functional confirmation of aberrant splicing. Methods We sequenced the entire F8 gene using an NGS capture method. We analysed F8 mRNA in order to detect aberrant transcripts. The pathogenic effect of candidate intronic variants was further confirmed using a minigene assay. Results After bioinformatic analysis, 11 deep intronic variants were identified in 13 patients (8 new variants and 3 previously reported). Three variants were confirmed to be likely pathogenic with the presence of an aberrant transcript during mRNA analysis and minigene assay. We also found a small intronic deletion in 6 patients, recently described as causing mild HA. Conclusion With this comprehensive work combining NGS and functional assays, we report new deep intronic variants that cause HA through splicing alteration mechanism. Functional analyses are critical to confirm the pathogenic effect of these variants and will be invaluable in the future to study the large number of variants of uncertain significance that may affect splicing that will be found in the human genome.Show less >
Language :
Anglais
Peer reviewed article :
Oui
Audience :
Internationale
Popular science :
Non
Source :
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