on August 3, 2006
Published online before print August 3, 2006, doi: 10.1161/01.ATV.0000239441.29687.97
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Submitted on January 5, 2006
Accepted on July 5, 2006
l-Caldesmon Regulates Proliferation and Migration of Vascular Smooth Muscle Cells and Inhibits Neointimal Formation After Angioplasty
Kazuhiko Yokouchi ;
From Nagoya University Graduate School of Medicine (K.Y., Y.N., R.K., R.M., Y.O., T.K., K.O., T.M.), Department of Cardiology; Nagoya University School of Medicine (Y.N., M.I.), Department of Medical Science of Proteases, Showa, Nagoya, Japan; Komaki Municipal Hospital, Department of Cardiology (H.I.), Komaki, Aichi, Japan; Vascular Biology Program (D.E.I.), Departments of Pathology and Surgery, Children’s Hospital & Harvard Medical School, Boston, Mass.
* To whom correspondence should be addressed. E-mail: numa2{at}med.nagoya-u.ac.jp.
Objective--Light-type caldesmon (l-CaD) is a potent cytostatic and antiangiogenic protein that regulates cell growth and survival via modulation of the cell shape and cytoskeleton. The aim of this study is to explore the potential value of l-CaD for use as a cytostatic agent to inhibit neointimal formation after angioplasty by suppressing vascular smooth muscle cell (VSMC) growth and migration.
Methods and Results--We tested the cytostatic function of l-CaD in cultured VSMCs using assays for apoptosis, cell proliferation, and migration, and evaluated the expression pattern of relevant signaling proteins (focal adhesion kinase (FAK) and mitogen-activated protein kinases) in VSMCs. Transfection of adenoviral vector encoding l-CaD (Ad-l-CaD) resulted in progressive loss of actin stress fibers and cell retraction. Enzyme-linked immunosorbent assay demonstrated that Ad-l-CaD transfection increased the apoptosis rate by 75% and reduced BrdU uptake by 49%. Furthermore, transfection of Ad-l-CaD inhibited migration of VSMCs induced by platelet-derived growth factor-BB (PDGF) by 36% (P<0.05). Immunoblotting analysis revealed that l-CaD overexpression reduced PDGF-induced phosphorylation of both FAK and extracellular signal regulated-kinase (extracellular signal-regulated kinase [ERK]). In balloon-injured rat carotid arteries, Ad-l-CaD transfection inhibited neointimal formation by 37% (P<0.05) without delaying re-endothelialization at 14 days.
Conclusions--Overexpression of l-CaD suppressed cell growth and survival in VSMCs and inhibited neointimal formation after experimental angioplasty, partly by regulating the cytoskeletal tension-FAK-ERK axis.
Key words: animal models of human disease • cell biology/structural biology • gene therapy • restenosis • smooth muscle proliferation and differentiation
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  P.-P. Zheng, L.-A. Severijnen, M. van der Weiden, R. Willemsen, and J. M. Kros A crucial role of caldesmon in vascular development in vivo Cardiovasc Res, February 1, 2009; 81(2): 362 - 369. [Abstract] [Full Text] [PDF]
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