Published online before print May 31, 2007, doi: 10.1161/CIRCRESAHA.107.153916
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
© 2007 American Heart Association, Inc.
Molecular Medicine |
Role of Dicer and Drosha for Endothelial MicroRNA Expression and Angiogenesis
From Molecular Cardiology, Department of Internal Medicine III, University of Frankfurt, Frankfurt, Germany.
Correspondence to Stefanie Dimmeler, PhD, Molecular Cardiology, Department of Internal Medicine III, University of Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany. E-mail dimmeler{at}em.uni-frankfurt.de
MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression by binding to the cellular transcript leading to translational repression or degradation of the target mRNA. Dicer and Drosha are the miRNA processing enzymes that are required for the maturation of miRNAs. Here, we investigated the role of Dicer and Drosha for angiogenesis. Endothelial cells were transfected with siRNA against Dicer and Drosha to inhibit miRNA biogenesis. Genetic silencing of Dicer and Drosha significantly reduced capillary sprouting of endothelial cells and tube forming activity. Migration of endothelial cells was significantly decreased in Dicer siRNA–transfected cells, whereas Drosha siRNA had no effect. Silencing of Dicer but not of Drosha reduced angiogenesis in vivo. Next, we attempted to identify miRNAs expressed in endothelial cells. A screening analysis of 168 human miRNAs using real-time PCR revealed that members of the let-7 family, mir-21, mir-126, mir-221, and mir-222 are highly expressed in endothelial cells. Dicer and Drosha siRNA reduced lef-7f and mir-27b expression. Inhibitors against let-7f and mir-27b also reduced sprout formation indicating that let-7f and mir-27b promote angiogenesis by targeting antiangiogenic genes. In silico analysis of predicted targets for let-7 cluster identified the endogenous angiogenesis inhibitor thrombospondin-1. Indeed, Dicer and Drosha siRNA significantly increased the expression of thrombospondin-1. Taken together, transient reduction of the miRNA-regulating enzyme Dicer impairs angiogenesis in vitro and in vivo, whereas Drosha siRNA induced a minor antiangiogenic effect in vitro and was not effective in vivo. The let-7 family and mir-27b appear to be attractive targets for modulating angiogenesis.
Key Words: angiogenesis • endothelial cells • gene expression
This article has been cited by other articles:
 |
|
 |
|
  R. Menghini, V. Casagrande, M. Cardellini, E. Martelli, A. Terrinoni, F. Amati, M. Vasa-Nicotera, A. Ippoliti, G. Novelli, G. Melino, et al. MicroRNA 217 Modulates Endothelial Cell Senescence via Silent Information Regulator 1 Circulation, October 13, 2009; 120(15): 1524 - 1532. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Catalucci, P. Gallo, and G. Condorelli MicroRNAs in Cardiovascular Biology and Heart Disease Circ Cardiovasc Genet, August 1, 2009; 2(4): 402 - 408. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Sotiropoulou, G. Pampalakis, E. Lianidou, and Z. Mourelatos Emerging roles of microRNAs as molecular switches in the integrated circuit of the cancer cell RNA, August 1, 2009; 15(8): 1443 - 1461. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Bonauer, G. Carmona, M. Iwasaki, M. Mione, M. Koyanagi, A. Fischer, J. Burchfield, H. Fox, C. Doebele, K. Ohtani, et al. MicroRNA-92a Controls Angiogenesis and Functional Recovery of Ischemic Tissues in Mice Science, June 26, 2009; 324(5935): 1710 - 1713. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Urbich, L. Rossig, D. Kaluza, M. Potente, J.-N. Boeckel, A. Knau, F. Diehl, J.-G. Geng, W.-K. Hofmann, A. M. Zeiher, et al. HDAC5 is a repressor of angiogenesis and determines the angiogenic gene expression pattern of endothelial cells Blood, May 28, 2009; 113(22): 5669 - 5679. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. J. Matkovich, D. J. Van Booven, K. A. Youker, G. Torre-Amione, A. Diwan, W. H. Eschenbacher, L. E. Dorn, M. A. Watson, K. B. Margulies, and G. W. Dorn II Reciprocal Regulation of Myocardial microRNAs and Messenger RNA in Human Cardiomyopathy and Reversal of the microRNA Signature by Biomechanical Support Circulation, March 10, 2009; 119(9): 1263 - 1271. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Izzotti, G. A. Calin, P. Arrigo, V. E. Steele, C. M. Croce, and S. De Flora Downregulation of microRNA expression in the lungs of rats exposed to cigarette smoke FASEB J, March 1, 2009; 23(3): 806 - 812. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Suarez and W. C. Sessa MicroRNAs As Novel Regulators of Angiogenesis Circ. Res., February 27, 2009; 104(4): 442 - 454. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-H. Tsai, M.-F. Wu, Y.-H. Wu, S.-J. Chang, S.-F. Lin, T. V. Sharp, and H.-W. Wang The M Type K15 Protein of Kaposi's Sarcoma-Associated Herpesvirus Regulates MicroRNA Expression via Its SH2-Binding Motif To Induce Cell Migration and Invasion J. Virol., January 15, 2009; 83(2): 622 - 632. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. E. Fish and D. Srivastava MicroRNAs: Opening a New Vein in Angiogenesis Research Sci. Signal., January 6, 2009; 2(52): pe1 - pe1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Kuhnert, M. R. Mancuso, J. Hampton, K. Stankunas, T. Asano, C.-Z. Chen, and C. J. Kuo Attribution of vascular phenotypes of the murine Egfl7 locus to the microRNA miR-126 Development, December 15, 2008; 135(24): 3989 - 3993. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Asada, T. Takahashi, K. Isodono, A. Adachi, H. Imoto, T. Ogata, T. Ueyama, H. Matsubara, and H. Oh Downregulation of Dicer expression by serum withdrawal sensitizes human endothelial cells to apoptosis Am J Physiol Heart Circ Physiol, December 1, 2008; 295(6): H2512 - H2521. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Thum Cardiac Dissonance Without Conductors: How Dicer Depletion Provokes Chaos in the Heart Circulation, October 7, 2008; 118(15): 1524 - 1527. [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Suarez, C. Fernandez-Hernando, J. Yu, S. A. Gerber, K. D. Harrison, J. S. Pober, M. L. Iruela-Arispe, M. Merkenschlager, and W. C. Sessa Dicer-dependent endothelial microRNAs are necessary for postnatal angiogenesis PNAS, September 16, 2008; 105(37): 14082 - 14087. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Thum, D. Catalucci, and J. Bauersachs MicroRNAs: novel regulators in cardiac development and disease Cardiovasc Res, September 1, 2008; 79(4): 562 - 570. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Urbich, A. Kuehbacher, and S. Dimmeler Role of microRNAs in vascular diseases, inflammation, and angiogenesis Cardiovasc Res, September 1, 2008; 79(4): 581 - 588. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-H. Wang, C.-I Kung, Y.-Y. Tseng, Y.-C. Lin, C.-H. Chen, C.-H. Tsai, and H.-I Yeh Activation of endothelial cells to pathological status by down-regulation of connexin43 Cardiovasc Res, August 1, 2008; 79(3): 509 - 518. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. C. Matouk and P. A. Marsden Epigenetic Regulation of Vascular Endothelial Gene Expression Circ. Res., April 25, 2008; 102(8): 873 - 887. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Zhang MicroRNomics: a newly emerging approach for disease biology Physiol Genomics, April 1, 2008; 33(2): 139 - 147. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Shilo, S. Roy, S. Khanna, and C. K. Sen Evidence for the Involvement of miRNA in Redox Regulated Angiogenic Response of Human Microvascular Endothelial Cells Arterioscler Thromb Vasc Biol, March 1, 2008; 28(3): 471 - 477. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Chen and D. H. Gorski Regulation of angiogenesis through a microRNA (miR-130a) that down-regulates antiangiogenic homeobox genes GAX and HOXA5 Blood, February 1, 2008; 111(3): 1217 - 1226. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. V.G. Latronico, D. Catalucci, and G. Condorelli Emerging Role of MicroRNAs in Cardiovascular Biology Circ. Res., December 7, 2007; 101(12): 1225 - 1236. [Abstract] [Full Text] [PDF]
|
|