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MicroRNA‐181b inhibits thrombin‐mediated endothelial activation and arterial thrombosis by targeting caspase recruitment domain family member 10
Author(s) -
Lin Jibin,
He Shaolin,
Sun Xinghui,
Franck Gregory,
Deng Yihuan,
Yang Dafeng,
Haemmig Stefan,
Wara A. K. M.,
Icli Basak,
Li Dazhu,
Feinberg Mark W.
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.201500163r
Subject(s) - small interfering rna , microrna , signal transduction , microbiology and biotechnology , thrombin , gene silencing , gene knockdown , chemistry , endothelial activation , cancer research , biology , immunology , inflammation , transfection , apoptosis , platelet , biochemistry , gene
Thrombogenic and inflammatory mediators, such as thrombin, induce NF‐kB–mediated endothelial cell (EC) activation and dysfunction, which contribute to pathogenesis of arterial thrombosis. The role of anti‐inflammatory microRNA‐181b (miR‐181b) on thrombosis remains unknown. Our previous study demonstrated that miR‐181b inhibits downstream NF‐kB signaling in response to TNF‐a. Here, we demonstrate that miR‐181b uniquely inhibits upstream NF‐kB signaling in response to thrombin. Overexpression of miR‐181b inhibited thrombin‐induced activation of NF‐kB signaling, demonstratedbyreductionofphospho‐IKK‐b,‐IkB‐a, and p65 nuclear translocation in ECs. MiR‐181b also reduced expression of NF‐kB target genes VCAM‐1, intercellular adhesion molecule‐1, E‐selectin, and tissue factor. Mechanistically, miR‐181b targets caspase recruitment domain family member 10 (Card10), an adaptor protein that participates in activation of the IKK complex in response to signals transduced from protease‐activated receptor‐1. miR‐181b reduced expression of Card10 mRNA and protein, but not protease‐activated receptor‐1. 39‐Untranslated region reporter assays, argonaute‐2 microribonucleoprotein immunoprecipitation studies, and Card10 rescue studies revealed that Card10 is a bona fide direct miR‐181b target. Small interfering RNA–mediated knockdown of Card10 expression phenocopied effects of miR‐181b on NF‐kB signaling and targets. Card10 deficiency did not affect TNF‐a–induced activation of NF‐kB signaling, which suggested stimulus‐specific regulation of NF‐kB signaling and endothelial responses by miR‐181b in ECs. Finally, in response to photochemical injury‐induced arterial thrombosis, systemic delivery of miR‐181b reduced thrombus formation by 73% in carotid arteries and prolonged time to occlusion by 1.6‐fold, effects recapitulated by Card10 small interfering RNA. These data demonstrate that miR‐181b and Card10 are important regulators of thrombin‐induced EC activation and arterial thrombosis. These studies highlight the relevance of microRNA‐dependent targets in response to ligand‐specific signaling in ECs.—Lin, J., He, S., Sun, X., Franck, G., Deng, Y., Yang, D., Haemmig, S., Wara, A. K. M., Icli, B., Li, D., Feinberg, M. W. MicroRNA‐181b inhibits thrombin‐mediated endothelial activation and arterial thrombosis by targeting caspase recruitment domain family member 10. FASEB J. 30, 3216–3226 (2016). www.fasebj.org