Research Paper

Viral Vector-Mediated 12/15-Lipoxygenase Overexpression in Vascular Smooth Muscle Cells Enhances Inflammatory Gene Expression and Migration
Roopashree S. Dwarakanath^a, Saurabh Sahar^a, Linda Lanting^a, Nanping Wang^c, Michael B. Stemerman^b, Rama Natarajan^a, Marpadga A. Reddy<sup>a

a</sup>Department of Diabetes, Beckman Research Institute of City of Hope, Duarte, Calif., and
^bAlbert Einstein College of Medicine, New York, N.Y., USA;
^cInstitute of Cardiovascular Science, Peking University Health Science Center, Beijing, China

Address of Corresponding Author

J Vasc Res 2008;45:132-142 (DOI: 10.1159/000109966)

  Key Words

• 12/15-Lipoxygenase
• Adenoviruses
• Atherosclerosis
• Baculoviruses
• Diabetes
• Inflammatory genes
• Nuclear factor-B
• Vascular smooth muscle cells
• Viral vectors

  Abstract

Increased expression and activity of 12/15-lipoxygenase (12/15-LO) in vascular smooth muscle cells (VSMCs) play a key role in the pathogenesis of diabetes and vascular complications. However, the consequences of 12/15-LO overexpression for VSMC migration and inflammatory gene expression are not known. In this study, 12/15-LO was overexpressed using adeno- and baculoviral vectors in human VSMC (HVSMCs) and proatherogenic responses compared with control enhanced green fluorescent protein (EGFP)-expressing cells. HVSMCs transduced with 12/15-LO viruses expressed high levels of enzymatically active protein and produced increased levels of the LO product, 12(S)-hydroxyeicosatetraenoic acid. 12/15-LO-overexpressing HVSMCs exhibited increased oxidant stress, activation of p38 mitogen-activated protein kinase, migration and inflammatory gene expression relative to HVSMCs expressing EGFP. Furthermore, inflammatory gene expression induced by 12/15-LO overexpression was abolished by anti-oxidants, siRNAs targeting p65 (nuclear factor-B), or new-generation baculoviruses expressing inhibitory IB or IB superrepressor mutant. Thus, we have used novel viral vector delivery systems, including baculoviruses, for the first time to deliver foreign genes into VSMCs and thereby demonstrated that 12/15-LO overexpression increases oxidant stress, mitogen-activated protein kinase activation, migration and inflammatory genes in VSMCs and that NF-B is a key downstream effector. Enhanced proatherogenic responses in VSMCs triggered by increased 12/15-LO levels under pathological conditions may contribute to vascular dysfunction.

Copyright © 2007 S. Karger AG, Basel

  Author Contacts

Dr. Marpadga A. Reddy
Department of Diabetes, Beckman Research Institute of City of Hope
Gonda Bldg 2019, 1500 E. Duarte Rd
Duarte, CA 91010 (USA)
Tel. +1 626 256 4673, ext. 63671, Fax +1 626 301 8136, E-Mail mreddy@coh.org

  Article Information

Received: April 2, 2007
Accepted after revision: July 5, 2007
Published online: October 17, 2007
Number of Print Pages : 11
Number of Figures : 6, Number of Tables : 0, Number of References : 51