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



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.



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.
Laboratory of Medical Science, Institute of Life Sciences, Scuola Superiore Sant'Anna, Via G. Moruzzi, 1, 56124 Pisa, Italy.
Marco
M
Matteucci
Laboratory of Medical Science, Institute of Life Sciences, Scuola Superiore Sant'Anna, Via G. Moruzzi, 1, 56124 Pisa, Italy.
Valentina
V
Casieri
Laboratory of Medical Science, Institute of Life Sciences, Scuola Superiore Sant'Anna, Via G. Moruzzi, 1, 56124 Pisa, Italy.
Khatia
K
Gabisonia
Fondazione Toscana 'G. Monasterio', 56124 Pisa, Italy.
Giovanni Donato
GD
Aquaro
Laboratory of Medical Science, Institute of Life Sciences, Scuola Superiore Sant'Anna, Via G. Moruzzi, 1, 56124 Pisa, Italy.
Silvia
S
Agostini
Siena Biotech Medicine Research Centre, 53100 Siena, Italy.
Giuseppe
G
Pollio
Siena Biotech Medicine Research Centre, 53100 Siena, Italy.
Daniela
D
Diamanti
Siena Biotech Medicine Research Centre, 53100 Siena, Italy.
Marco
M
Rossi
Siena Biotech Medicine Research Centre, 53100 Siena, Italy.
Massimiliano
M
Travagli
Siena Biotech Medicine Research Centre, 53100 Siena, Italy.
Valentina
V
Porcari
Laboratory of Medical Science, Institute of Life Sciences, Scuola Superiore Sant'Anna, Via G. Moruzzi, 1, 56124 Pisa, Italy.|Cardiovascular Research Center, Lewis Katz School of Medicine, Temple University, 19140 Philadelphia, PA, USA.
Fabio A
FA
Recchia
Laboratory of Medical Science, Institute of Life Sciences, Scuola Superiore Sant'Anna, Via G. Moruzzi, 1, 56124 Pisa, Italy v.lionetti@sssup.it.|Fondazione Toscana 'G. Monasterio', 56124 Pisa, Italy.
Vincenzo
V
Lionetti
D000818:Animals; D001052:Apoferritins / Q000096:biosynthesis / Q000235:genetics; D000229:Dependovirus / Q000235:genetics; D004195:Disease Models, Animal; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D007501:Iron / Q000378:metabolism; D019028:Magnetic Resonance Imaging, Cine / Q000379:methods*; D008297:Male; D057054:Molecular Imaging / Q000379:methods*; D009203:Myocardial Infarction / Q000000981:diagnostic imaging* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D011237:Predictive Value of Tests; D011401:Promoter Regions, Genetic; D017208:Rats, Wistar; D015203:Reproducibility of Results; D051820:TCF Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D016277:Ventricular Function, Left; D020257:Ventricular Remodeling; D060449:Wnt Signaling Pathway
D051820:TCF Transcription Factors; D049452:Green Fluorescent Proteins; D001052:Apoferritins; D007501:Iron
Ferritin; MRI; Myocardial infarction; Reporter gene; Wnt
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