Abstract
Factor VIII gene, F8, mutations cause haemophilia A (HA), an X-linked recessive disorder. Expression in heterozygous females has been ascribed to skewed X-chromosome inactivation (XCI). To investigate the cause of HA in three heterozygous females within an Atlantic Canadian kindred, the proband (severely affected girl, FVIII activity: 2%) and 17 relatives across three generations were studied. F8 genotype, FVIII activity, XCI ratio (XCIR) (paternal active X: maternal active X), karyotype, submegabase resolution tiling set array competitive genome hybridization (competitive genomic hybridization (SMRT)), and microsatellite analyses were utilized. A positive linear relationship between FVIII activity and percentage-activated normal X-chromosome was found in HA heterozygous females (R2=0.87). All affected, but no unaffected females, had an XCIR skewed toward activation of the mutant X-chromosome (proband 92:8, SD 2). Unexpectedly, high numbers of females have dramatically skewed XCIRs (>80:20 or <20:80) (P<0.05). The distribution of XCIR frequencies within this family was significantly different than predicted by normal population data or models of random XCI (P<0.025), with more females having higher degrees of skewing. Known causes of skewing, such as chromosomal abnormalities, selection against deleterious alleles, and X-inactive-specific transcript mutations, are not consistent with our results. This study shows that FVIII activity in HA heterozygous females can be directly related to XCI skewing, and that low FVIII activity in females in this family is due to unfavourable XCI skewing. Further, the findings suggest that these XCI ratios are genetically influenced, consistent with a novel heritable human X controlling element (XCE) functioning similarly to the mouse Xce.
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Acknowledgements
This work was supported by grants from the CDHA, Halifax, Nova Scotia (WG, SD), and the Canadian Hemophilia Society (WG, SD). NR was supported by scholarships from the Canadian Institutes of Health Research (CIHR), and the Killam Foundation. Part of this work was completed by the Australian Genome Research Facility (AGRF), Victoria, Australia. We acknowledge their contribution and the support that the AGRF receives from the Commonwealth. We thank Dr D Lillicrap and J Leggo at the National Program for Hemophilia Mutation Testing and the AHCDC for their support. We thank Dr R Howell for critical reading of the paper, Dr B Morash for technical guidance, and P Steele for patient recruitment and correspondence.
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Renault, N., Dyack, S., Dobson, M. et al. Heritable skewed X-chromosome inactivation leads to haemophilia A expression in heterozygous females. Eur J Hum Genet 15, 628–637 (2007). https://doi.org/10.1038/sj.ejhg.5201799
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DOI: https://doi.org/10.1038/sj.ejhg.5201799
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