Biomechanically induced gene iex-1 inhibits vascular smooth muscle cell proliferation and neointima formation

Circ Res. 2003 Dec 12;93(12):1210-7. doi: 10.1161/01.RES.0000103635.38096.2F. Epub 2003 Oct 30.

Abstract

Mechanotransduction plays a prominent role in vascular pathophysiology but is incompletely understood. In this study, we report the biomechanical induction of the immediate early response gene iex-1 in vascular smooth muscle cells (SMCs). Mechanical induction of iex-1 was confirmed by Northern (30-fold induction after 2 hours) and Western (6-fold induction after 24 hours) analyses. Expression of iex-1 was regulated by mechanical activation of nuclear factor (NF)-kappaB and abolished by overexpression of IkappaB in SMCs. The function of iex-1 in SMCs was explored by gene transfer using adenoviral vectors overexpressing iex-1. After 48 hours of 4% cyclic mechanical strain, adenoviral vectors overexpressing iex-1-infected cells had lower 3[H]-thymidine incorporation compared with AdGFP-infected controls (71.3+/-8.5% versus 180.2+/-19.4% in controls; P<0.001). Overexpression of iex-1 suppressed mitogenesis induced by platelet-derived growth factor (208.1+/-108.3% versus 290.0+/-120.5% in controls; P<0.05). This was accompanied by reduced degradation of p27kip1, inhibition of Rb hyperphosphorylation, and reduced cell cycle progression. To investigate functional effects of iex-1 in vivo, we performed carotid artery mechanical injury and endothelial denudation in low-density lipoprotein receptor-deficient mice followed by intraluminal injection of adenoviral vectors (3x10(9) pfu in 50 microL) for overexpression of iex-1 or gfp (control). Vascular overexpression of iex-1 reduced neointima formation 2 weeks after injury (intima/media ratio, 0.23+/-0.04 versus 0.5+/-0.24 in controls; P<0.05). Our findings demonstrate that biomechanical strain induces iex-1 with subsequent antiproliferative effects in SMCs and that selective gene transfer of iex-1 inhibits the local vascular response after injury. These findings suggest that the induction of iex-1 represents a novel negative biomechanical feedback mechanism limiting the vascular response to injury.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Apolipoproteins E / genetics
  • Apoptosis Regulatory Proteins
  • Arteriosclerosis / genetics
  • Arteriosclerosis / physiopathology
  • Carotid Artery Injuries / genetics
  • Carotid Artery Injuries / physiopathology
  • Cell Cycle / physiology
  • Cell Division / physiology
  • Cells, Cultured
  • Gene Expression Regulation
  • Green Fluorescent Proteins
  • Humans
  • Immediate-Early Proteins / genetics*
  • Immediate-Early Proteins / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mechanotransduction, Cellular / physiology
  • Membrane Proteins
  • Mice
  • Mice, Knockout
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / metabolism*
  • NF-kappa B / metabolism
  • Neoplasm Proteins / genetics*
  • Neoplasm Proteins / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Receptors, LDL / genetics
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Stress, Mechanical
  • Tunica Intima / physiopathology*

Substances

  • Apolipoproteins E
  • Apoptosis Regulatory Proteins
  • IER3 protein, human
  • Immediate-Early Proteins
  • Luminescent Proteins
  • Membrane Proteins
  • NF-kappa B
  • Neoplasm Proteins
  • RNA, Messenger
  • Receptors, LDL
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins