Assessing Doxorubicin-Induced Cardiomyopathy by ^99mTc-3PRGD2 Scintigraphy Targeting Integrin αvβ3 in a Rat Model

Abstract

The present study evaluated interstitial alterations in doxorubicin-induced cardiomyopathy using a radiolabeled RGD peptide ^99mTc-3PRGD2 specific for integrin αvβ3 that targets myofibroblasts.

Cardiomyopathy was induced in 20 Sprague-Dawley rats by intraperitoneal doxorubicin injections (2.5 mg/kg/week) for up to six weeks. ^99mTc-3PRGD2 scintigraphy was performed in control rats (n = 6) at baseline and three, six, and nine weeks after first doxorubicin administration (n = 6, 6, and 5 for each time point). For another three rats of 6-week modeling, cold c(RGDyK) was co-injected with ^99mTc-3PRGD2 to evaluate specific radiotracer binding. Semi-quantitative parameters were acquired to compare radiotracer uptake among all groups. The biodistribution of ^99mTc-3PRGD2 was evaluated by a γ-counter after scintigraphy. Haematoxylin and eosin, and Masson’s staining were used to evaluate myocardial injury and fibrosis, while western blotting and immunofluorescence co-localization were used to analyze integrin αvβ3 expression in the myocardium.

The ^99mTc-3PRGD2 half-life in the cardiac region (Heartt _1/2) of the 9-week model and heart radioactivity percentage (%Heart_20 min, %Heart_40 min and %Heart_60 min) of the 6 and 9-week models were significantly increased compared to the control. Heart-to-background ratio (HBR_20 min, HBR_40 min and HBR_60 min) increase began in the third week, continued until the sixth week, and was reversed in the ninth week, which paralleled the changing trend of cardiac integrin αvβ3 expression. The myocardial biodistribution of ^99mTc-3PRGD2 was significantly correlated with integrin β3 expression.

The ^99mTc-3PRGD2 scintigraphy allows for non-invasive visualization of interstitial alterations during doxorubicin-induced cardiomyopathy.

Zusammenfassung

Die vorliegende Studie untersuchte interstitielle Veränderungen bei Doxorubicin-induzierter Kardiomyopathie unter Verwendung eines radioaktiv markierten RGD-Peptids ^99mTc-3PRGD2, das für Integrin αvβ3 spezifisch ist und auf Myofibroblasten abzielt.

Eine Kardiomyopathie wurde in 20 Sprague-Dawley-Ratten durch intraperitoneale Doxorubicin-Injektionen (2,5 mg/kg/Woche) für bis zu 6 Wochen induziert. Die ^99mTc-3PRGD2-Szintigrafie wurde bei Kontrollratten (n = 6) zu Studienbeginn und 3, 6 und 9 Wochen nach der ersten Doxorubicin-Injektion durchgeführt (n = 6, 6 und 5 für jeden Zeitpunkt). Bei weiteren 3 Ratten mit 6-Wochen-Modellierung wurde kaltes c(RGDyK) mit ^99mTc-3PRGD2 koinjiziert, um die spezifische Radiotracer-Bindung zu bewerten. Semiquantitative Parameter wurden erfasst, um die Radiotracer-Aufnahme zwischen allen Gruppen zu vergleichen. Die Biodistribution von ^99mTc-3PRGD2 wurde nach der Szintigrafie mit einem γ-Zähler ausgewertet. Hämatoxylin und Eosin sowie die Masson-Färbung wurden zur Beurteilung der myokardialen Schädigung und Fibrose verwendet, während Western-Blot und Immunfluoreszenz-Co-Lokalisation zur Analyse der Integrin-αvβ3-Expression im Myokard eingesetzt wurden.

Die ^99mTc-3PRGD2-Halbwertszeit in der Herzregion (Heart/_1/2) des 9-Wochen-Modells und die prozentuale Herzradioaktivität (%Heart_20 min, %Heart_40 min und %Heart_60 min) der 6- und 9-Wochen-Modelle waren im Vergleich zur Kontrolle signifikant erhöht. Der Anstieg der Herz-Background-Ratio (HBR_20 min, HBR_40 min und HBR_60 min) begann in der dritten Woche, setzte sich bis zur sechsten Woche fort und kehrte sich in der neunten Woche um, was mit dem sich ändernden Trend der kardialen Integrin-αvβ3-Expression einherging. Die myokardiale Biodistribution von ^99mTc-3PRGD2 korrelierte signifikant mit der Expression von Integrin β3.

Die ^99mTc-3PRGD2-Szintigrafie ermöglicht die nichtinvasive Visualisierung von interstitiellen Veränderungen während der Doxorubicin-induzierten Kardiomyopathie.

Key words

^99mTc-3PRGD2 - integrin αvβ3 - scintigraphy - doxorubicin - cardiomyopathy

Schlüsselwörter

^99mTc-3PRGD2 - Integrin αvβ3 - Szintigrafie - Doxorubicin - Kardiomyopathie

Publication History

Received: 20 June 2020

Accepted: 06 December 2020

Article published online:
26 February 2021

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