Titrating haemophilia B phenotypes using siRNA strategy: evidence that antithrombotic activity is separated from bleeding liability
| Autor: | Lizbeth M. Hoos, Laura Sepp-Lorenzino, Walter Strapps, John Levorse, Dietmar Seiffert, Karen R. Leander, Marti DiPietro, Yuchen Zhou, Anil Thankappan, Weizhen Wu, Tian-Quan Cai, Zuo Zhang, Zhu Chen, Yiming Xu, Marija Tadin-Strapps, Myung K. Shin, Patrick Andre, Joseph M. Metzger, Kunal Desai, KehDih Lai, Nina Jochnowitz, Ross Bentley |
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| Rok vydání: | 2014 |
| Předmět: |
Male
medicine.medical_specialty Small interfering RNA Time Factors Genotype Haemophilia A Hemorrhage 030204 cardiovascular system & hematology Pharmacology Transfection Ferric Compounds Hemophilia B Cell Line Factor IX Rats Sprague-Dawley 03 medical and health sciences 0302 clinical medicine Chlorides Bleeding time Antithrombotic medicine Gene silencing Animals Haemophilia B RNA Messenger RNA Small Interfering Gene knockdown Hemostasis medicine.diagnostic_test business.industry Thrombosis Hematology medicine.disease Surgery Rats Disease Models Animal Phenotype RNAi Therapeutics Gene Expression Regulation Liver 030220 oncology & carcinogenesis RNA Interference Rats Transgenic business medicine.drug |
| Zdroj: | Thrombosis and haemostasis. 113(6) |
| ISSN: | 2567-689X |
| Popis: | SummaryHaemophilia A and B are characterised by a life-long bleeding predisposition, and several lines of evidence suggest that risks of atherothrombotic events may also be reduced. Establishing a direct correlation between coagulation factor levels, thrombotic risks and bleeding propensity has long been hampered by an inability to selectively and specifically inhibit coagulation factor levels. Here, the exquisite selectivity of gene silencing combined with a gene knockout (KO) approach was used to define the relative contribution of factor IX (fIX) to thrombosis and primary haemostasis in the rat. Using a lipid nanoparticle (LNP) formulation, we successfully delivered fIX siRNAs to the liver by intravenous administration. The knockdown (KD) of target gene mRNA was achieved rapidly (within 24 hour post-siRNA dosing), sustained (maintained for at least 7 days post dosing) and not associated with changes in mRNA expression levels of other coagulation factors. We found that intermediate levels of liver fIX mRNA silencing (60–95 %) translating into a 50–99 % reduction of plasma fIX activity provided protection from thrombosis without prolonging the cuticle bleeding time. Over 99 % inhibition of fIX activity was required to observe increase in bleeding, a phenotype confirmed in fIX KO rats. These data provide substantial evidence of a participation of fIX in the mechanisms regulating thrombosis prior to those regulating primary haemostasis, therefore highlighting the potential of fIX as a therapeutic target. In addition, hepatic mRNA silencing using LNP-encapsulated siRNAs may represent a promising novel approach for the chronic treatment and prevention of coagulation-dependent thrombotic disorders in humans. |
| Databáze: | OpenAIRE |
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