Ex vivo Improvement of a von Willebrand Disease Type 2A Phenotype Using an Allele-Specific Small-Interfering RNA
| Autor: | Ferdows Atiq, Richard J. Dirven, Seyed Yahya Anvar, Jeroen Eikenboom, Johan Boender, Annika de Jong, Frank W.G. Leebeek, Bart J.M. van Vlijmen |
|---|---|
| Přispěvatelé: | Hematology |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
congenital hereditary and neonatal diseases and abnormalities Small interfering RNA small-interfering RNAs Mutant Mutation Missense von Willebrand Disease Type 2 030204 cardiovascular system & hematology Transfection Polymorphism Single Nucleotide Endothelial activation 03 medical and health sciences 0302 clinical medicine Von Willebrand factor hemic and lymphatic diseases New Technologies Diagnostic Tools and Drugs von Willebrand Factor Von Willebrand disease medicine Humans Platelet RNA Small Interfering Alleles biology Chemistry Endothelial Cells Hematology medicine.disease Molecular biology von Willebrand Diseases Phenotype HEK293 Cells 030104 developmental biology Amino Acid Substitution Hemostasis hemostasis cardiovascular system biology.protein RNA Interference von Willebrand disease circulatory and respiratory physiology |
| Zdroj: | Thrombosis and Haemostasis, 120(11), 1569-1579. GEORG THIEME VERLAG KG Thrombosis and Haemostasis, 120(11), 1569-1579. Georg Thieme Verlag Thrombosis and Haemostasis |
| ISSN: | 2567-689X 0340-6245 |
| DOI: | 10.1055/s-0040-1715442 |
| Popis: | Von Willebrand disease (VWD) is the most common inherited bleeding disorder and is mainly caused by dominant-negative mutations in the multimeric protein von Willebrand factor (VWF). These mutations may either result in quantitative or qualitative defects in VWF. VWF is an endothelial protein that is secreted to the circulation upon endothelial activation. Once secreted, VWF multimers bind platelets and chaperone coagulation factor VIII in the circulation. Treatment of VWD focuses on increasing VWF plasma levels, but production and secretion of mutant VWF remain uninterrupted. Presence of circulating mutant VWF might, however, still affect normal hemostasis or functionalities of VWF beyond hemostasis. We hypothesized that inhibition of the production of mutant VWF improves the function of VWF overall and ameliorates VWD phenotypes. We previously proposed the use of allele-specific small-interfering RNAs (siRNAs) that target frequent VWF single nucleotide polymorphisms to inhibit mutant VWF. The aim of this study is to prove the functionality of these allele-specific siRNAs in endothelial colony-forming cells (ECFCs). We isolated ECFCs from a VWD type 2A patient with an intracellular multimerization defect, reduced VWF collagen binding, and a defective processing of proVWF to VWF. After transfection of an allele-specific siRNA that specifically inhibited expression of mutant VWF, we showed amelioration of the laboratory phenotype, with normalization of the VWF collagen binding, improvement in VWF multimers, and enhanced VWF processing. Altogether, we prove that allele-specific inhibition of the production of mutant VWF by siRNAs is a promising therapeutic strategy to improve VWD phenotypes. |
| Databáze: | OpenAIRE |
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