FVIII Immunogenicity—Bioinformatic Approaches to Evaluate Inhibitor Risk in Non-severe Hemophilia A

The life-long inhibitor risk in non-severe hemophilia A has been an important clinical and research focus in recent years. Non-severe hemophilia A is most commonly caused by point mutation, missense F8 genotypes, of which over 500 variants are described. The immunogenic potential of just a single amino acid change within a complex 2,332 amino acid protein is an important reminder of the challenges of protein replacement therapies in diverse, global populations. Although some F8 genotypes have been identified as “high risk” mutations in non-severe hemophilia A (e.g., R593C), this is likely, in part at least, a reporting bias and oversimplification of the underlying immunological mechanism. Bioinformatic approaches offer a strategy to dissect the contribution of F8 genotype in the context of the wider HLA diversity through which antigenic peptides will necessarily be presented. Extensive modeling of all permutations of FVIII-derived fifteen-mer peptides straddling all reported F8 genotype positions demonstrate the likely heterogeneity of peptide binding affinity to different HLA II grooves. For the majority of F8 genotypes it is evident that inhibitor risk prediction is dependent on the combination of F8 genotype and available HLA II. Only a minority of FVIII-derived peptides are predicted to bind to all candidate HLA molecules. In silico predictions still over call the risk of inhibitor occurrence, suggestive of mechanisms of “protection” against clinically meaningful inhibitor events. The structural homology between FVIII and FV provides an attractive mechanism by which some F8 genotypes may be afforded co-incidental tolerance through homology of FV and FVIII primary amino sequence. In silico strategies enable the extension of this hypothesis to analyse the extent to which co-incidental cross-matching exists between FVIII-derived primary peptide sequences and any other protein in the entire human proteome and thus potential central tolerance. This review of complimentary in vitro, in silico, and clinical epidemiology data documents incremental insights into immunological mechanism of inhibitor occurrence in non-severe hemophilia A over the last decade. However, complex questions remain about antigenic processing and presentation to truly understand and predict an individual person with hemophilia risk of inhibitor occurrence.