Activation of the Amino Acid Response Pathway Blunts the Effects of Cardiac Stress

J Am Heart Assoc. 2017 May 9;6(5):e004453. doi: 10.1161/JAHA.116.004453.

Abstract

Background: The amino acid response (AAR) is an evolutionarily conserved protective mechanism activated by amino acid deficiency through a key kinase, general control nonderepressible 2. In addition to mobilizing amino acids, the AAR broadly affects gene and protein expression in a variety of pathways and elicits antifibrotic, autophagic, and anti-inflammatory activities. However, little is known regarding its role in cardiac stress. Our aim was to investigate the effects of halofuginone, a prolyl-tRNA synthetase inhibitor, on the AAR pathway in cardiac fibroblasts, cardiomyocytes, and in mouse models of cardiac stress and failure.

Methods and results: Consistent with its ability to inhibit prolyl-tRNA synthetase, halofuginone elicited a general control nonderepressible 2-dependent activation of the AAR pathway in cardiac fibroblasts as evidenced by activation of known AAR target genes, broad regulation of the transcriptome and proteome, and reversal by l-proline supplementation. Halofuginone was examined in 3 mouse models of cardiac stress: angiotensin II/phenylephrine, transverse aortic constriction, and acute ischemia reperfusion injury. It activated the AAR pathway in the heart, improved survival, pulmonary congestion, left ventricle remodeling/fibrosis, and left ventricular function, and rescued ischemic myocardium. In human cardiac fibroblasts, halofuginone profoundly reduced collagen deposition in a general control nonderepressible 2-dependent manner and suppressed the extracellular matrix proteome. In human induced pluripotent stem cell-derived cardiomyocytes, halofuginone blocked gene expression associated with endothelin-1-mediated activation of pathologic hypertrophy and restored autophagy in a general control nonderepressible 2/eIF2α-dependent manner.

Conclusions: Halofuginone activated the AAR pathway in the heart and attenuated the structural and functional effects of cardiac stress.

Keywords: amino acid response; fibrosis; halofuginone; heart failure; hypertrophy.

MeSH terms

  • Amino Acids / deficiency
  • Amino Acids / metabolism*
  • Amino Acyl-tRNA Synthetases / antagonists & inhibitors
  • Amino Acyl-tRNA Synthetases / metabolism
  • Animals
  • Autophagy / drug effects
  • Cells, Cultured
  • Disease Models, Animal
  • Dose-Response Relationship, Drug
  • Enzyme Inhibitors / pharmacology*
  • Fibroblasts / drug effects*
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Fibrosis
  • Heart Failure / metabolism
  • Heart Failure / pathology
  • Heart Failure / physiopathology
  • Heart Failure / prevention & control*
  • Humans
  • Hypertrophy, Left Ventricular / metabolism
  • Hypertrophy, Left Ventricular / pathology
  • Hypertrophy, Left Ventricular / physiopathology
  • Hypertrophy, Left Ventricular / prevention & control
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Male
  • Mice, Inbred C57BL
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / pathology
  • Piperidines / pharmacology*
  • Protein Serine-Threonine Kinases / metabolism
  • Protein Synthesis Inhibitors / pharmacology*
  • Quinazolinones / pharmacology*
  • Stress, Physiological*
  • Time Factors
  • Ventricular Function, Left / drug effects
  • Ventricular Remodeling / drug effects

Substances

  • Amino Acids
  • Enzyme Inhibitors
  • Piperidines
  • Protein Synthesis Inhibitors
  • Quinazolinones
  • EIF2AK4 protein, human
  • Eif2ak4 protein, mouse
  • Protein Serine-Threonine Kinases
  • Amino Acyl-tRNA Synthetases
  • halofuginone