Magnetic resonance imaging of infarct-induced canonical wingless/integrated (Wnt)/β-catenin/T-cell factor pathway activation, in vivo

Cardiovasc Res. 2016 Dec;112(3):645-655. doi: 10.1093/cvr/cvw214. Epub 2016 Sep 26.

Abstract

Aims: Combined magnetic resonance imaging (MRI) of molecular and morpho-functional changes might prove highly valuable for the elucidation of pathological processes involved in the development of cardiac diseases. Our aim was to test a novel MRI reporter gene for in vivo assessment of the canonical Wnt/β-catenin/TCF pathway activation, an important regulator of post-ischaemic cardiac remodelling.

Methods and results: We designed and developed a chimeric construct encoding for both of iron-binding human ferritin heavy chain (hFTH) controlled by the β-catenin-responsive TCF/lymphoid-enhancer binding factor (Lef) promoter and constitutively expressed green fluorescent protein (GFP). It was carried by adeno-associated virus serotype 9 (rAAV9) vectors and delivered to the peri-infarct myocardium of rats subjected to coronary ligation (n = 11). By 1.5 T MRI and a multiecho T2* gradient echo sequence, we detected iron accumulation only in the border zone of the transduced infarcted hearts. In the same cardiac area, post-mortem histological analysis confirmed the co-existence of iron accumulation and GFP. The iron signal was absent when rats (n = 6) were chronically treated with SEN195 (10 mg/kg/day), a small-molecular inhibitor of β-catenin/TCF-dependent gene transcription. Canonical Wnt pathway inhibition attenuated the post-ischaemic remodelling process, as demonstrated by the significant preservation of cardiac function, the 42 ± 1% increase of peri-infarct arteriolar density and 43 ± 3% reduction in infarct scar size compared with untreated animals.

Conclusions: The TCF/Lef promoter-hFTH construct is a novel and reliable MRI reporter gene for in vivo detection of the canonical Wnt/β-catenin/TCF activation state in response to cardiac injury and therapeutic interventions.

Keywords: Ferritin; MRI; Myocardial infarction; Reporter gene; Wnt.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoferritins / biosynthesis
  • Apoferritins / genetics
  • Dependovirus / genetics
  • Disease Models, Animal
  • Genes, Reporter*
  • Genetic Vectors
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • HEK293 Cells
  • Humans
  • Iron / metabolism
  • Magnetic Resonance Imaging, Cine / methods*
  • Male
  • Molecular Imaging / methods*
  • Myocardial Infarction / diagnostic imaging*
  • Myocardial Infarction / metabolism
  • Myocardial Infarction / pathology
  • Myocardial Infarction / physiopathology
  • Myocardium / metabolism*
  • Myocardium / pathology
  • Predictive Value of Tests
  • Promoter Regions, Genetic
  • Rats, Wistar
  • Reproducibility of Results
  • TCF Transcription Factors / genetics
  • TCF Transcription Factors / metabolism*
  • Transfection
  • Ventricular Function, Left*
  • Ventricular Remodeling*
  • Wnt Signaling Pathway*

Substances

  • TCF Transcription Factors
  • Green Fluorescent Proteins
  • Apoferritins
  • Iron