title: miR-30e targets IGF2-regulated osteogenesis in bone marrow-derived mesenchymal stem cells, aortic smooth muscle cells, and ApoE-/- mice.
abstract: OBJECTIVE
  Activation of an osteogenic transcriptional program contributes to the initiation of aortic calcification in atherosclerosis. The role of microRNAs in regulating aortic calcification is understudied. We tested the hypothesis that miR-30e regulates an osteogenic program in bone marrow-derived mesenchymal stem cells (MSCs), aortic smooth muscle cells (SMCs), and ApoE(-/-) mice.
  
  
  RESULTS
  In aortas of wild-type mice, we found that miR-30e is highly expressed in medial SMCs. In aortas of old ApoE(-/-) mice, we found that miR-30e transcripts are down-regulated in an inverse relation to the osteogenic markers Runx2, Opn, and Igf2. In vitro, miR-30e over-expression reduced the proliferation of MSCs and SMCs while increasing adipogenic differentiation of MSCs and smooth muscle differentiation of SMCs. In MSCs and SMCs over-expressing miR-30e, microarrays and qPCR showed repression of an osteogenic gene panel including Igf2. Inhibiting miR-30e in MSCs increased Igf2 transcripts. In SMCs, IGF2 recombinant protein rescued miR-30e-repressed osteogenic differentiation. Luciferase and mutagenesis assays showed binding of miR-30e to a novel and essential site at the 3'UTR of Igf2. In ApoE(-/-) mice, injections of antimiR-30e oligos increased Igf2 expression in the aortas and livers and significantly enhanced OPN protein expression and calcium deposition in aortic valves.
  
  
  CONCLUSIONS
  miR-30e represses the osteogenic program in MSCs and SMCs by targeting IGF2 and drives their differentiation into adipogenic or smooth muscle lineage, respectively. Our data suggest that down-regulation of miR-30e in aortas with age and atherosclerosis triggers vascular calcification. The miR-30e pathway plays an important regulatory role in vascular diseases.
authors: Department of Molecular and Cellular Pharmacology, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA.
authors: Wen
authors: W
authors: Ding
authors: Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA.
authors: Jihe
authors: J
authors: Li
authors: Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA.
authors: Jayanti
authors: J
authors: Singh
authors: Department of Biochemistry, University of Miami, Coral Gables, FL 33136, USA.
authors: Razan
authors: R
authors: Alif
authors: Department of Surgery, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA Vascular Biology Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA.
authors: Roberto I
authors: RI
authors: Vazquez-Padron
authors: Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA.
authors: Samirah A
authors: SA
authors: Gomes
authors: Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA.
authors: Joshua M
authors: JM
authors: Hare
authors: Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA Vascular Biology Institute, University of Miami Leonard M. Miller School of Medicine, Miami, FL 33136, USA lshehadeh@med.miami.edu.
authors: Lina A
authors: LA
authors: Shehadeh
mesh_terms: D000375:Aging / Q000502:physiology; D000818:Animals; D001011:Aorta / Q000166:cytology; D001057:Apolipoproteins E / Q000172:deficiency* / Q000235:genetics / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology; D002114:Calcinosis / Q000503:physiopathology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D015536:Down-Regulation / Q000502:physiology; D007335:Insulin-Like Growth Factor II / Q000502:physiology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D035683:MicroRNAs / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology*; D010012:Osteogenesis / Q000502:physiology*
chemical_list: D001057:Apolipoproteins E; C513121:IGF2 protein, mouse; D035683:MicroRNAs; C534715:Mirn30d microRNA, mouse; D007335:Insulin-Like Growth Factor II
keywords: Calcification; Mesenchymal stem cells; MicroRNA-30e; Osteogenesis; Smooth muscle cells