Cyclic adenosine monophosphate response-element binding protein activation by mitogen-activated protein kinase-activated protein kinase 3 and four-and-a-half LIM domains 5 plays a key role for vein graft intimal hyperplasia

J Vasc Surg. 2013 Jan;57(1):182-93, 193.e1-10. doi: 10.1016/j.jvs.2012.06.082. Epub 2012 Nov 3.


Objective: Intimal hyperplasia (IH) is the main cause of vein graft stenosis or failure after bypass surgery. Basic investigations are proceeding in an animal model of mechanically desquamated arteries, and numerous molecules for potential IH treatments have been identified; however, neither insights into the mechanism of IH nor substantially effective treatments for its suppression have been developed. The goals of the present study are to use human vein graft samples to identify therapeutic target genes that control IH and to investigate the therapeutic efficacy of these candidate molecules in animal models.

Methods: Using microarray analysis of human vein graft samples, we identified two previously unrecognized IH-related genes, mitogen-activated protein kinase-activated protein kinase 3 (MAPKAPK3) and four-and-a-half LIM domains 5 (FHL5).

Results: Transfer of either candidate gene resulted in significantly elevated vascular smooth muscle cell (VSMC) proliferation and migration. Interestingly, cotransfection of both genes increased VSMC proliferation in an additive manner. These genes activated cyclic adenosine monophosphate response-element (CRE) binding protein (CREB), but their mechanisms of activation were different. MAPKAPK3 phosphorylated CREB, but FHL5 bound directly to CREB. A CREB dominant-negative protein, KCREB, which blocks its ability to bind CRE, repressed VSMC proliferation and migration. In a wire-injury mouse model, gene transfer of KCREB plasmid significantly repressed IH. In this vessel tissue, CRE-activated gene expression was repressed. Furthermore, we confirmed the changes in MAPKAPK3 and FHL5 expression using vein graft samples from eight patients.

Conclusions: We successively identified two previously unrecognized IH activators, MAPKAPK3 and FHL5, using human vein graft samples. Gene transfer of KCREB repressed IH in an animal model. Inhibition of CREB function is a promising gene therapy strategy for IH.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Animals
  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Constriction, Pathologic
  • Cyclic AMP Response Element-Binding Protein / genetics
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Disease Models, Animal
  • Gene Expression Profiling / methods
  • Genetic Therapy
  • Graft Occlusion, Vascular / enzymology*
  • Graft Occlusion, Vascular / etiology
  • Graft Occlusion, Vascular / genetics
  • Graft Occlusion, Vascular / pathology
  • Graft Occlusion, Vascular / prevention & control
  • Humans
  • Hyperplasia
  • Intracellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • LIM Domain Proteins / genetics
  • LIM Domain Proteins / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Muscle, Smooth, Vascular / enzymology
  • Muscle, Smooth, Vascular / pathology
  • Mutation
  • Myocytes, Smooth Muscle / enzymology
  • Myocytes, Smooth Muscle / pathology
  • Neointima
  • Oligonucleotide Array Sequence Analysis
  • Phosphorylation
  • Protein Binding
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism*
  • Time Factors
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transfection
  • Vascular Grafting / adverse effects*
  • Vascular System Injuries / enzymology
  • Vascular System Injuries / genetics
  • Vascular System Injuries / pathology
  • Vascular System Injuries / prevention & control
  • Veins / enzymology
  • Veins / injuries
  • Veins / pathology
  • Veins / transplantation*


  • CREB1 protein, human
  • Cyclic AMP Response Element-Binding Protein
  • FHL5 protein, human
  • Fhl5 protein, mouse
  • Intracellular Signaling Peptides and Proteins
  • LIM Domain Proteins
  • Transcription Factors
  • MAP-kinase-activated kinase 3
  • Protein-Serine-Threonine Kinases