Variable readthrough responsiveness of nonsense mutations in hemophilia A.

Variable readthrough responsiveness of nonsense mutations in hemophilia A. Haematologica. 2019 Jun 13;: Authors: Martorell L, Cortina V, Parra R, Barquinero J, Vidal F Abstract Readthrough therapy relies on the use of small molecules that enable premature termination codons in mRNA open reading frames to be misinterpreted by the translation machinery, thus allowing the generation of full-length, potentially functional proteins from mRNAs carrying nonsense mutations. In patients with hemophilia A, nonsense mutations potentially sensitive to readthrough agents represent about 16% of the point mutations. The aim of this study was to measure the readthrough effect of different compounds and to analyze the influence of premature termination codon context in selected nonsense mutations causing hemophilia A. To this end, primary fibroblasts from three patients with hemophilia A caused by nonsense mutations (p.W1586X, p.Q1636X and p.R1960X) and CHO cells transfected with 12 different plasmids encoding mutated F8 (p.Q462X, p.Q1705X, p.Q1764X, p.W274X, p.W1726X, p.W2015X, p.W2131X, p.R1715X, p.R1822X, p.R1960X, p.R2071X and p.R2228X) were treated with gentamicin, geneticin, PTC124, RTC13 or RTC14. Responses were assessed by analyzing not only F8 mRNA expression and FVIII biosynthesis (FVIII antigen by ELISA, Western blot and immunofluorescence) but also at the FVIII activity (by chromogenic assay). In the patients' fibroblasts, readthrough age...
Source: Haematologica - Category: Hematology Authors: Tags: Haematologica Source Type: research