Targeted Disruption of JCAD (Junctional Protein Associated With Coronary Artery Disease)/KIAA1462, a Coronary Artery Disease-Associated Gene Product, Inhibits Angiogenic Processes In Vitro and In Vivo

Arterioscler Thromb Vasc Biol. 2017 Sep;37(9):1667-1673. doi: 10.1161/ATVBAHA.117.309721. Epub 2017 Jul 13.

Abstract

Objective: Recent genome-wide association studies newly identified the human KIAA1462 gene as a new locus for coronary artery disease. However, the function of the gene product, named JCAD (junctional protein associated with coronary artery disease), is unknown. Because JCAD is expressed at cell-cell junctions in endothelial cells, we hypothesized and tested whether JCAD regulates angiogenic processes in vitro and in vivo.

Approach and results: Cell culture experiments revealed impaired angiogenic ability (proliferation, migration, and cord formation) by the knockdown of JCAD with siRNA (P<0.05 versus control siRNA). We have generated mice lacking JCAD (mKIAA1462-/-) by gene-targeted deletion of JCAD to address in vivo angiogenic function. mKIAA1462-/- mice did not show morphological differences in development of retinal vasculature. Ex vivo aortic ring model demonstrated impaired neovascularization in aorta from mKIAA1462-/- mice than control wild-type mice (P<0.05). Tumor growth was assessed by monitoring tumor volume after the subcutaneous injection of melanoma, LLC (Lewis lung carcinoma), and E0771 cells into the mice. mKIAA1462-/- mice exhibited significantly smaller tumor volume compared with wild-type mice (P<0.001). Histological assessment of the tumor exhibited less smooth muscle actin-positive neovascularization determined by CD31-positive vascular structure in tumor of mKIAA1462-/- mice than wild-type mice, indicating that knockdown of JCAD inhibited the vascular maturation in pathological angiogenic process.

Conclusions: These in vitro and in vivo studies suggest that JCAD has a redundant functional role in physiological angiogenesis but serves a pivotal role in pathological angiogenic process after birth.

Keywords: adherens junctions; angiogenesis; coronary artery disease.

MeSH terms

  • Animals
  • Carcinoma, Lewis Lung / blood supply
  • Carcinoma, Lewis Lung / genetics
  • Carcinoma, Lewis Lung / metabolism
  • Cell Adhesion Molecules / deficiency
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / metabolism*
  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Endothelial Cells / metabolism*
  • Genotype
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Intercellular Junctions / metabolism*
  • Melanoma, Experimental / blood supply
  • Melanoma, Experimental / genetics
  • Melanoma, Experimental / metabolism
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Neovascularization, Pathologic*
  • Neovascularization, Physiologic*
  • Phenotype
  • RNA Interference
  • Retinal Neovascularization*
  • Signal Transduction
  • Time Factors
  • Tissue Culture Techniques
  • Transfection
  • Tumor Burden

Substances

  • Cell Adhesion Molecules
  • JCAD protein, human
  • JCAD protein, mouse