ABSTRACT
Inhibitors are an impediment to the effective management of haemophilia B (HB), but there is limited understanding of the underlying genetic risk factors. In this study we aim to understand the role of F9 gene mutations on inhibitor development in patients with HB. Mutations in the F9 gene were identified and HLA typing performed for five boys with severe HB. Data from the CDC Haemophilia B Mutation Project (CHBMP) database were used to assess association between F9 gene mutation type and inhibitor development. Analysis of the CHBMP database showed that larger disruptions in the F9 gene are associated with a higher life-time prevalence of inhibitors. We detected the following mutations in the five subjects, including four novel mutations: Nonsense in three patients (c.223 C>T; p.Arg75* in two siblings, c.553 C>T; p.Glu185*); Splice site in two patients (c.723 + 1 G>A, c.278-27 A>G); Missense in one patient (c.580 A>G, p.Thr194Ala; c.723 G>T; p.Gln241His). Of the two siblings only one responded to immune tolerance induction (ITI). These siblings have identical F9 gene mutations but differ with respect to the HLA alleles. Interestingly, an analysis of peptide-MHC binding affinities shows a significantly higher (one-sided unpaired t-test, P = 0.0018) median affinity for FIX-derived peptides in the sibling that responded to ITI. We conclude that the nature of the F9 gene mutation may be an important risk factor for the development of inhibitors. In addition, the HLA alleles of the individual patients, in conjunction with the mutation type, could be a predictor for the development of inhibitors as well as the response to ITI.
Subject(s)
Factor IX/genetics , Factor IX/immunology , Hemophilia B/genetics , Hemophilia B/immunology , Isoantibodies/immunology , Adolescent , Child , Computational Biology , Databases, Factual , Exons , Factor IX/therapeutic use , Genetic Association Studies , Genetic Markers , HLA-DRB1 Chains/genetics , HLA-DRB1 Chains/immunology , Hemophilia B/diagnosis , Hemophilia B/drug therapy , Humans , Male , Mutation , Odds Ratio , RNA Splicing , Severity of Illness Index , Young AdultABSTRACT
The possibility of alloimmunization in patients receiving protein replacement therapy depends on (at least) three risk factors, which are necessary concomitantly but insufficient alone. The first is the degree of structural difference between the therapeutic protein and the patient's own endogenous protein, if expressed. Such differences depend on the nature of the disease mutation and the pre-mutation endogenous protein structure as well as on post-translational changes and sequence-engineered alterations in the therapeutic protein. Genetic variations in the recipients' immune systems comprise the second set of risk determinants for deleterious immune responses. For example, the limited repertoire of MHC class II isomers encoded by a given person's collection of HLA genes may or may not be able to present a 'foreign' peptide(s) produced from the therapeutic protein - following its internalization and proteolytic processing - on the surface of their antigen-presenting cells (APCs). The third (and least characterized) variable is the presence or absence of immunologic 'danger signals' during the display of foreign-peptide/MHC-complexes on APCs. A choice between existing therapeutic products or the manufacture of new proteins, which may be less immunogenic in some patients or patient populations, may require prior definition of the first two of these variables. This leads then to the possibility of developing personalized therapies for disorders due to genetic deficiencies in endogenous proteins, such as haemophilia A and B. [Correction made after online publication 11 July 2011: several critical corrections have been made to the abstract].
Subject(s)
Factor VIII , Hemophilia A , Economics, Pharmaceutical , Factor VIII/genetics , Factor VIII/immunology , Factor VIII/therapeutic use , Genetic Predisposition to Disease , Hemophilia A/drug therapy , Hemophilia A/genetics , Hemophilia A/immunology , Histocompatibility Antigens Class II/immunology , Humans , Immune Tolerance/genetics , Isoantibodies/immunology , Risk FactorsABSTRACT
The completion of the yeast genome project enables an analysis of various phenomena for a whole eukaryotic genome. We aimed at characterizing a full spectrum of target genes for a transcription activator, and specifically characterized putative targets for GCN4 in the budding yeast. The results suggest that about 1% of the genes are regulated by GCN4 and that these genes code for proteins involved in amino-acid and nucleotide metabolism. Our analysis proposes that, when enough data about the binding nature of a transcription factor exists, it is possible to identify its putative targets and also to try and assign a physiological role for this transcription factor.
