l-Caldesmon regulates proliferation and migration of vascular smooth muscle cells and inhibits neointimal formation after angioplasty.
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.


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 (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.
Yokouchi
Kazuhiko
K
Institute for Laboratory Animal Research, Nagoya University, Japan.
Numaguchi
Yasushi
Y
Kubota
Ryuji
R
Ishii
Masakazu
M
Imai
Hajime
H
Murakami
Ryuichiro
R
Ogawa
Yasuhiro
Y
Kondo
Takahisa
T
Okumura
Kenji
K
Ingber
Donald E
DE
Murohara
Toyoaki
T
D017130:Angioplasty; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002148:Calmodulin-Binding Proteins / Q000494:pharmacology*; D020212:Carotid Artery Injuries / Q000209:etiology / Q000503:physiopathology; D002404:Catheterization; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000187:drug effects; D042783:Endothelial Cells / Q000166:cytology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000201:enzymology; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000201:enzymology / Q000502:physiology*; D017239:Paclitaxel / Q000494:pharmacology; D020033:Protein Isoforms / Q000494:pharmacology; D051381:Rats; D017539:Tunica Intima / Q000254:growth & development*
D002148:Calmodulin-Binding Proteins; D020033:Protein Isoforms; D051416:Focal Adhesion Protein-Tyrosine Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D017239:Paclitaxel