title: Molecular Basis and Therapeutic Strategies to Rescue Factor IX Variants That Affect Splicing and Protein Function.
abstract: Mutations that result in amino acid changes can affect both pre-mRNA splicing and protein function. Understanding the combined effect is essential for correct diagnosis and for establishing the most appropriate therapeutic strategy at the molecular level. We have identified a series of disease-causing splicing mutations in coagulation factor IX (FIX) exon 5 that are completely recovered by a modified U1snRNP particle, through an SRSF2-dependent enhancement mechanism. We discovered that synonymous mutations and missense substitutions associated to a partial FIX secretion defect represent targets for this therapy as the resulting spliced-corrected proteins maintains normal FIX coagulant specific activity. Thus, splicing and protein alterations contribute to define at the molecular level the disease-causing effect of a number of exonic mutations in coagulation FIX exon 5. In addition, our results have a significant impact in the development of splicing-switching therapies in particular for mutations that affect both splicing and protein function where increasing the amount of a correctly spliced protein can circumvent the basic functional defects.
authors.affiliation: Human Molecular Genetics, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
authors.forename: Mojca
authors.initials: M
authors.lastname: Tajnik
authors.affiliation: Human Molecular Genetics, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
authors.forename: Malgorzata Ewa
authors.identifier: 0000-0002-6331-9815
authors.initials: ME
authors.lastname: Rogalska
authors.affiliation: Human Molecular Genetics, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
authors.forename: Erica
authors.initials: E
authors.lastname: Bussani
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Barbon
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
authors.forename: Dario
authors.identifier: 0000-0002-6675-9429
authors.initials: D
authors.lastname: Balestra
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy.
authors.forename: Mirko
authors.initials: M
authors.lastname: Pinotti
authors.affiliation: Human Molecular Genetics, International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
authors.forename: Franco
authors.initials: F
authors.lastname: Pagani
mesh_terms: D001778:Blood Coagulation Disorders / Q000235:genetics* / Q000473:pathology; D018929:Cell Culture Techniques; D005091:Exons / Q000235:genetics; D005164:Factor IX / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D009154:Mutation; D012322:RNA Precursors / Q000235:genetics; D022821:RNA Splice Sites / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics*; D017411:Ribonucleoproteins, Small Nuclear / Q000235:genetics; D000068103:Serine-Arginine Splicing Factors / Q000235:genetics; D014162:Transfection
chemical_list: D012322:RNA Precursors; D022821:RNA Splice Sites; D017411:Ribonucleoproteins, Small Nuclear; C552127:SNRNP35 protein, human; C073288:SRSF2 protein, human; D000068103:Serine-Arginine Splicing Factors; D005164:Factor IX