title: Endothelial miR-17∼92 cluster negatively regulates arteriogenesis via miRNA-19 repression of WNT signaling.
abstract: The contribution of endothelial-derived miR-17∼92 to ischemia-induced arteriogenesis has not been investigated in an in vivo model. In the present study, we demonstrate a critical role for the endothelial-derived miR-17∼92 cluster in shaping physiological and ischemia-triggered arteriogenesis. Endothelial-specific deletion of miR-17∼92 results in an increase in collateral density limbs and hearts and in ischemic limbs compared with control mice, and consequently improves blood flow recovery. Individual cluster components positively or negatively regulate endothelial cell (EC) functions in vitro, and, remarkably, ECs lacking the cluster spontaneously form cords in a manner rescued by miR-17a, -18a, and -19a. Using both in vitro and in vivo analyses, we identified FZD4 and LRP6 as targets of miR-19a/b. Both of these targets were up-regulated in 17∼92 KO ECs compared with control ECs, and both were shown to be targeted by miR-19 using luciferase assays. We demonstrate that miR-19a negatively regulates FZD4, its coreceptor LRP6, and WNT signaling, and that antagonism of miR-19a/b in aged mice improves blood flow recovery after ischemia and reduces repression of these targets. Collectively, these data provide insights into miRNA regulation of arterialization and highlight the importance of vascular WNT signaling in maintaining arterial blood flow.
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Shira
authors.initials: S
authors.lastname: Landskroner-Eiger
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Cong
authors.initials: C
authors.lastname: Qiu
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Paola
authors.initials: P
authors.lastname: Perrotta
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Mauro
authors.initials: M
authors.lastname: Siragusa
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Monica Y
authors.initials: MY
authors.lastname: Lee
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Victoria
authors.initials: V
authors.lastname: Ulrich
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Amelia K
authors.initials: AK
authors.lastname: Luciano
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Yale Cardiovascular Research Center, Department of Internal Medicine (Cardiology), Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Zhen W
authors.initials: ZW
authors.lastname: Zhuang
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Yale Cardiovascular Research Center, Department of Internal Medicine (Cardiology), Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Federico
authors.initials: F
authors.lastname: Corti
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Yale Cardiovascular Research Center, Department of Internal Medicine (Cardiology), Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Michael
authors.initials: M
authors.lastname: Simons
authors.affiliation: miRagen Therapeutics, Boulder, CO 80301.
authors.forename: Rusty L
authors.initials: RL
authors.lastname: Montgomery
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Dianqing
authors.initials: D
authors.lastname: Wu
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Yale Cardiovascular Research Center, Department of Internal Medicine (Cardiology), Yale University School of Medicine, New Haven, CT 06520;
authors.forename: Jun
authors.initials: J
authors.lastname: Yu
authors.affiliation: Vascular Biology and Therapeutics Program, Yale University School of Medicine, New Haven, CT 06520; Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520; william.sessa@yale.edu.
authors.forename: William C
authors.initials: WC
authors.lastname: Sessa
mesh_terms: D000818:Animals; D051157:Frizzled Receptors / Q000235:genetics / Q000378:metabolism*; D007511:Ischemia / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D060488:Low Density Lipoprotein Receptor-Related Protein-6 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family / Q000502:physiology*; D018919:Neovascularization, Physiologic / Q000502:physiology*; D060449:Wnt Signaling Pathway / Q000502:physiology*
chemical_list: D051157:Frizzled Receptors; C498581:Fzd4 protein, mouse; D060488:Low Density Lipoprotein Receptor-Related Protein-6; C555476:Lrp6 protein, mouse; D035683:MicroRNAs
keywords: arteriogenesis; endothelium; microRNA; vascular