M. A. Zimmermann

Characterization of duplication breakpoints in the factor VIII gene.

See also Goodeve AC. Another step towards understanding hemophilia A molecular pathogenesis. This issue, pp 2693–5.

Identification and characterization of mutations in the promoter region of the factor VIII gene

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Expression studies of mutant factor VIII alleles with premature termination codons with regard to inhibitor formation.

About 10% of mutations in haemophilia A cases generate a premature termination codon in the factor VIII gene (F8). Upon therapeutic FVIII substitution, it was noted that the risk of developing inhibitors is higher when the nonsense mutation is located in the light chain (LC) of the factor VIII (FVIII) protein than in the heavy chain (HC). We analysed the impact of six different nonsense mutations distributed over the six FVIII domains on recombinant FVIII expression to elucidate the process of inhibitor formation in haemophilic patients. Full‐length F8 mRNA was transcribed from all constructs despite the presence of nonsense mutations. Polyclonal antigen assays revealed high antigen levels in transfection experiments with constructs truncated in LC whereas low antigen was detected from constructs truncated in HC. Those results were supported by FVIII localization experiments. These findings suggest that F8 transcription occurs in a usual way despite nonsense mutations, whereas translation appears to be interrupted by the premature stop codon. We hypothesize that the inclusion of the B domain enables proteins truncated in LC to accumulate in the ER. Proteins truncated in HC are mainly degraded or may pass through the ER and be secreted into the blood circulation, thus presumably preventing inhibitor formation after therapeutic FVIII substitution. The LC is known to have higher immunogenicity than the HC. Moreover, translation of the F8B gene comprising F8 exons 23–26 may be dependent on the position of the premature stop codon and thus contributes to the immune response of truncated FVIII proteins.

Analysis of F8 mRNA in haemophilia A patients with silent mutations or presumptive splice site mutations

Analysis of F8 mRNA in haemophilia A patients with silent mutations or presumptive splice site mutations -