TY - JOUR
T1 - MicroRNA-181b inhibits thrombin-mediated endothelial activation and arterial thrombosis by targeting caspase recruitment domain family member 10
AU - Lin, Jibin
AU - He, Shaolin
AU - Sun, Xinghui
AU - Franck, Gregory
AU - Deng, Yihuan
AU - Yang, Dafeng
AU - Haemmig, Stefan
AU - Wara, A. K.M.
AU - Icli, Basak
AU - Li, Dazhu
AU - Feinberg, Mark W.
N1 - Funding Information:
This work was supported by the National Institutes of Health (NIH) Grants HL115141 and HL117994 (National Heart, Lung, and Blood Institute), and GM115605 (National Institute of General Medical Sciences) (all to M.W.F.); the Arthur K. Watson Charitable Trust (to M.W.F.); the Dr. Ralph and Marian Falk Medical Research Trust, Bank of America, N.A. Trustee (to M.W.F.); a State Scholarship Fund of the China Scholarship Council (to J.L.); a Jonathan Levy Research Fund (to M.W.F.), the American Heart Association (SDG#15SDG25400012) (to X.S.); and a Boston Nutrition Obesity Research Center Pilot and Feasibility award under NIH National Institute of Diabetes and Digestive and Kidney Diseases Grant P30KD046200 (to X.S.). Author contributions: X. Sun and M. W. Feinberg designed research; J. Lin, S. He, X. Sun, Y. Deng, D. Yang, S. Haemmig, and M. W. Feinberg analyzed data; J. Lin, S. He, X. Sun, G. Franck, Y. Deng, S. D. Yang, and Haemmig performed research; J. Lin, X. Sun, and M. W. Feinberg wrote the paper; and X. Sun, A. K. M. Wara, B. Icli, D. Li, and M. W. Feinberg contributed to experimental design.
Publisher Copyright:
© FASEB.
PY - 2016/9
Y1 - 2016/9
N2 - Thrombogenic and inflammatory mediators, such as thrombin, induce NF-κB'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-κB signaling in response to TNF-α. Here, we demonstrate that miR-181b uniquely inhibits upstreamNF-κB signaling in response to thrombin. Overexpression of miR-181b inhibited thrombin-induced activation ofNF-κB signaling, demonstratedby reductionof phospho- IKK-β, -IκB-α, and p65 nuclear translocation in ECs.MiR-181b also reduced expression of NF-κB 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-κB signaling and targets. Card10 deficiency did not affect TNF-α-induced activation of NF-κB signaling, which suggested stimulus-specific regulation of NF-κB signaling and endothelial responses by miR-181b in ECs. Finally, in response to photochemical injuryinduced 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.
AB - Thrombogenic and inflammatory mediators, such as thrombin, induce NF-κB'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-κB signaling in response to TNF-α. Here, we demonstrate that miR-181b uniquely inhibits upstreamNF-κB signaling in response to thrombin. Overexpression of miR-181b inhibited thrombin-induced activation ofNF-κB signaling, demonstratedby reductionof phospho- IKK-β, -IκB-α, and p65 nuclear translocation in ECs.MiR-181b also reduced expression of NF-κB 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-κB signaling and targets. Card10 deficiency did not affect TNF-α-induced activation of NF-κB signaling, which suggested stimulus-specific regulation of NF-κB signaling and endothelial responses by miR-181b in ECs. Finally, in response to photochemical injuryinduced 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.
KW - Card10
KW - Endothelial cells
KW - NF-κB
UR - http://www.scopus.com/inward/record.url?scp=84990857130&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84990857130&partnerID=8YFLogxK
U2 - 10.1096/fj.201500163R
DO - 10.1096/fj.201500163R
M3 - Article
C2 - 27297585
AN - SCOPUS:84990857130
VL - 30
SP - 3216
EP - 3226
JO - FASEB Journal
JF - FASEB Journal
SN - 0892-6638
IS - 9
ER -