Loss of p47phox subunit enhances susceptibility to biomechanical stress and heart failure because of dysregulation of cortactin and actin filaments

Circ Res. 2013 Jun 7;112(12):1542-56. doi: 10.1161/CIRCRESAHA.111.300299. Epub 2013 Apr 3.

Abstract

Rationale: The classic phagocyte nicotinamide adenine dinucleotide phosphate oxidase (gp91(phox) or Nox2) is expressed in the heart. Nox2 activation requires membrane translocation of the p47(phox) subunit and is linked to heart failure. We hypothesized that loss of p47(phox) subunit will result in decreased reactive oxygen species production and resistance to heart failure.

Objective: To define the role of p47(phox) in pressure overload-induced biomechanical stress.

Methods and results: Eight-week-old male p47(phox) null (p47(phox) knockout [KO]), Nox2 null (Nox2KO), and wild-type mice were subjected to transverse aortic constriction-induced pressure overload. Contrary to our hypothesis, p47(phox)KO mice showed markedly worsened systolic dysfunction in response to pressure overload at 5 and 9 weeks after transverse aortic constriction compared with wild-type-transverse aortic constriction mice. We found that biomechanical stress upregulated N-cadherin and β-catenin in p47(phox)KO hearts but disrupted the actin filament cytoskeleton and reduced phosphorylation of focal adhesion kinase. p47(phox) interacts with cytosolic cortactin by coimmunoprecipitation and double immunofluorescence staining in murine and human hearts and translocated to the membrane on biomechanical stress where cortactin interacted with N-cadherin, resulting in adaptive cytoskeletal remodeling. However, p47(phox)KO hearts showed impaired interaction of cortactin with N-cadherin, resulting in loss of biomechanical stress-induced actin polymerization and cytoskeletal remodeling. In contrast, Nox2 does not interact with cortactin, and Nox2-deficient hearts were protected from pressure overload-induced adverse myocardial and intracellular cytoskeletal remodeling.

Conclusions: We showed a novel role of p47(phox) subunit beyond and independent of nicotinamide adenine dinucleotide phosphate oxidase activity as a regulator of cortactin and adaptive cytoskeletal remodeling, leading to a paradoxically enhanced susceptibility to biomechanical stress and heart failure.

Keywords: actin cytoskeleton; cortactin; heart failure; mechanotransduction; p47phox; pressure overload; remodeling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism*
  • Actins / metabolism*
  • Animals
  • Biomechanical Phenomena
  • Cadherins / metabolism
  • Cells, Cultured
  • Cortactin / metabolism*
  • Disease Models, Animal
  • Echocardiography, Doppler
  • Fluorescent Antibody Technique
  • Focal Adhesion Kinase 1 / metabolism
  • Heart Failure / diagnostic imaging
  • Heart Failure / enzymology*
  • Heart Failure / genetics
  • Heart Failure / physiopathology
  • Heart Failure / prevention & control
  • Humans
  • Immunoprecipitation
  • Male
  • Mechanotransduction, Cellular*
  • Membrane Glycoproteins / deficiency
  • Membrane Glycoproteins / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Myocardium / enzymology*
  • Myocardium / pathology
  • Myocytes, Cardiac / enzymology
  • Myocytes, Cardiac / pathology
  • NADPH Oxidase 2
  • NADPH Oxidases / deficiency*
  • NADPH Oxidases / genetics
  • Oxidative Stress
  • Phosphorylation
  • Polymerization
  • Reactive Oxygen Species / metabolism
  • Stress, Mechanical
  • Time Factors
  • Ventricular Remodeling
  • beta Catenin / metabolism

Substances

  • Actins
  • CTNNB1 protein, mouse
  • Cadherins
  • Cdh2 protein, mouse
  • Cortactin
  • Membrane Glycoproteins
  • Reactive Oxygen Species
  • beta Catenin
  • Cybb protein, mouse
  • NADPH Oxidase 2
  • NADPH Oxidases
  • neutrophil cytosolic factor 1
  • Focal Adhesion Kinase 1
  • Ptk2 protein, mouse