Peptide-Mediated Disruption of Calmodulin–Cyclin E Interactions Inhibits Proliferation of Vascular Smooth Muscle Cells and Neointima Formation
| Autor: | Abdul Momen, Syed H.E. Zaidi, Al-Muktafi Sadi, Mansoor Husain, Jaehyun Choi, Sonya Hui, Kiwon Ban, Sarah K. Steinbach |
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| Rok vydání: | 2011 |
| Předmět: |
Neointima
Cyclin E Vascular smooth muscle Calmodulin Physiology Biology Muscle Smooth Vascular S Phase Coronary Restenosis Mice Animals Phosphorylation Aorta Cyclin Binding Sites Cell growth Cyclin-Dependent Kinase 2 Cyclin-dependent kinase 2 Blood Proteins Cell cycle Extracellular Matrix Cell biology Biochemistry biology.protein Peptides Cardiology and Cardiovascular Medicine Hydrophobic and Hydrophilic Interactions Protein Kinases |
| Zdroj: | Circulation Research. 108:1053-1062 |
| ISSN: | 1524-4571 0009-7330 |
| DOI: | 10.1161/circresaha.110.239483 |
| Popis: | Rationale: Cell cycle progression in vascular smooth muscle cells (VSMCs) is a therapeutic target for restenosis. Objective: Having discovered that calmodulin (CaM)-dependent cyclin E/CDK2 activity underlies Ca 2+ -sensitive G 1 -to-S phase transitions in VSMCs, we sought to explore the physiological importance of the CaM–cyclin E interaction. Methods and Results: A peptide based on the CaM binding sequence (CBS) of cyclin E was designed to interfere with CaM–cyclin E binding. Compared with control peptides, CBS blocked activating Thr160 phosphorylation of CDK2, decreased basal cyclin E/CDK2 activity, and eliminated Ca 2+ -sensitive cyclin E/CDK2 activity in nuclear extracts from mouse VSMCs. Nucleofection with CBS, or treatment with CBS conjugated to the HIV-1 TAT protein transduction domain to improve bioavailability, inhibited G 1 -to-S cell cycle progression in a dose-dependent manner. These effects were not observed with control peptides. TAT-CBS inhibited 3 H-thymidine incorporation in primary human aortic SMCs (HA-SMCs) in vitro, manifested greater transduction into HA-SMCs compared with endothelial cells in vitro, and limited decreased SM22α expression, neointima formation, and medial thickening without affecting collagen deposition or reendothelialization in a mouse model of carotid artery injury in vivo. The antiproliferative effects of CBS remained evident in mouse embryonic fibroblasts derived from wild-type mice but not cyclin E1/E2 double knockout mice. Conclusions: A synthetic peptide designed to disrupt CaM–cyclin E binding inhibits Ca 2+ /CaM-dependent CDK2 activity, cell cycle progression, and proliferation in VSMCs and limits arterial remodeling following injury. Importantly, this effect appears to be cyclin E–dependent and may form the basis of a potentially novel therapeutic approach for restenosis. |
| Databáze: | OpenAIRE |
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