Endothelial Bmp4 is induced during arterial remodeling: effects on smooth muscle cell migration and proliferation.
BACKGROUND
Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-beta superfamily of proteins that have multiple functional roles in mammalian development. A role for BMP4 in adult vascular remodeling has recently been suggested. We evaluated the expression of Bmp4 during neointimal lesion development in vivo.


METHODS
Heterozygous Bmp4(lacZ/+) mice were used to evaluate in vivo Bmp4 expression after carotid ligation. beta-galactosidase (beta-gal) activity was evaluated in histological sections 1 to 14 d after carotid ligation and this was compared with control carotid arteries. The effects of recombinant human (rh) BMP4 on smooth muscle cell (SMC) migration and proliferation were evaluated using a rat aortic SMC line. We next assessed the effects of BMP4 signaling by over-expressing a constitutively active BMP receptor (BMPR-IA/Alk-3) using adenovirus-mediated gene transfer. SMC proliferation, migration, and apoptosis were evaluated in adenovirus transfected cells.


RESULTS
Ligated carotid arteries expressed endothelium-specific beta-gal staining after 1 d. Staining intensity increased at both 3 d and 1 wk after ligation and remained stable at 2 weeks while no beta-gal staining was observed in control vessels. Endothelial-specific expression of beta-galactosidase was confirmed through positive staining for PECAM-1. When human recombinant BMP4 was added to cultured SMCs, it inhibited migration but did not affect cultured SMC proliferation. SMCs infected with adenovirus encoding for the active BMP receptor Alk-3 demonstrated dose-dependent receptor expression. Alk-3 over-expressing cells showed a dose-dependent decrease in proliferation and migration but no effect on apoptosis.


CONCLUSIONS
These results demonstrate that endothelial Bmp4 expression is upregulated after carotid ligation in vivo, and furthermore, that activating the BMP signaling cascade results in decreased SMC proliferation and migration. This suggests that BMPs may counterbalance the effect of mitogen up-regulation observed during the development of neointimal hyperplasia.
Corriere
Matthew A
MA
Division of Vascular Surgery, Vanderbilt University Medical Center, Nashville, Tennessee 37232, USA.
Rogers
Chris M
CM
Eliason
Jonathan L
JL
Faulk
Jimbob
J
Kume
Tsutomu
T
Hogan
Brigid L M
BL
Guzman
Raul J
RJ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D055415:Bone Morphogenetic Protein 4; D052004:Bone Morphogenetic Protein Receptors / Q000235:genetics / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002339:Carotid Arteries / Q000378:metabolism / Q000473:pathology*; D002465:Cell Movement / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D008026:Ligation; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000378:metabolism; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
C525809:BMP4 protein, human; C525810:Bmp4 protein, mouse; C525811:Bmp4 protein, rat; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D052004:Bone Morphogenetic Protein Receptors; D001616:beta-Galactosidase
Mol Biol Cell. 1999 Nov;10(11):3801-13
10564272
Curr Top Dev Biol. 2006;76:1-48
17118262
Ann N Y Acad Sci. 2000 May;902:230-9; discussion 239-40
10865843
Am J Physiol Endocrinol Metab. 2003 May;284(5):E972-9
12527559
Am J Physiol Lung Cell Mol Physiol. 2003 Sep;285(3):L740-54
12740218
J Biol Chem. 2003 Aug 15;278(33):31128-35
12766166
Genes Dev. 2003 Oct 1;17(19):2362-7
12975322
Arterioscler Thromb Vasc Biol. 2003 Dec;23(12):2185-91
14576075
Am J Pathol. 2004 Jan;164(1):43-8
14695317
J Am Coll Cardiol. 2004 Aug 18;44(4):733-9
15312851
Science. 1986 Jan 24;231(4736):405-7
3941904
Science. 1988 Dec 16;242(4885):1528-34
3201241
J Clin Invest. 1993 Apr;91(4):1800-9
8473518
Circ Res. 1993 Nov;73(5):792-6
8403250
J Clin Invest. 1993 Oct;92(4):2013-21
8408655
Circulation. 1993 Dec;88(6):2838-48
8252697
Atherosclerosis. 1995 Mar;113(2):153-6
7605353
Genes Dev. 1996 Jul 1;10(13):1580-94
8682290
Can J Physiol Pharmacol. 1996 Jul;74(7):834-41
8946070
Genes Dev. 1997 Feb 15;11(4):451-62
9042859
Arterioscler Thromb Vasc Biol. 1997 Oct;17(10):2238-44
9351395
J Clin Invest. 1997 Dec 1;100(11):2824-32
9389748
Cell. 1998 Jun 12;93(6):985-96
9635428
Calcif Tissue Int. 1998 Sep;63(3):221-9
9701626
Genes Dev. 1999 Feb 15;13(4):424-36
10049358
J Vasc Res. 1999 Mar-Apr;36(2):120-5
10213907
J Vasc Surg. 2005 Feb;41(2):321-31
15768016
Circ Res. 2005 May 27;96(10):1053-63
15845886
Gene. 2005 Aug 29;357(1):1-8
16125875
J Vasc Surg. 2006 Jan;43(1):150-8
16414402
Curr Med Res Opin. 2006;22 Suppl 1:S7-11
16882364
J Cell Physiol. 2000 Jul;184(1):37-45
10825232