MR Imaging and Electrophysiological Features of Doxorubicin-Induced Fibrosis: Protocol Development in a Small Preclinical Pig Study with Histological Validation

Abstract

A critical chemotherapeutic complication is cardiotoxicity, often leading, in time, to heart failure. In this work, we developed a novel animal protocol using magnetic resonance (MR) imaging and electrophysiology (EP) tests, designed to detect subtle structural and functional changes associated with myocardial damage in sub-chronic phases post-chemotherapy. A weekly dose of doxorubicin (DOX) was injected in four juvenile swine throughout a four-week plan, using an intravenous approach that mimics the treatment in cancer patients. We performed cardiac MR imaging as follows: in all four pigs pre-DOX; at 1 and 5 weeks post-DOX in a group of two pigs; and, at 1 and 9 weeks post-DOX in the other two pigs, using Cine imaging to assess ejection fraction (EF) and late gadolinium enhancement to quantify collagen density in the left ventricle. Additionally, X-ray-guided voltage mapping and arrhythmia tests were conducted in the group at 9 weeks post-DOX and in a healthy pig. Tissue samples were collected for histology. The results showed that EF decreased from ~46% pre-DOX to ~34% within the first 9 weeks post-DOX. This decline in LV function was explained by a gradual increase in collagen density, especially noticeable at week 9 post-DOX as derived from MRI analysis. Furthermore, ventricular fibrillation was induced via rapid pacing at 9 weeks post-DOX, most likely caused by fibrotic patches identified in voltage maps, as confirmed by MRI and collagen-sensitive histological stains. Overall, our novel preclinical protocol was able to reveal key signs of potentially-irreversible tissue changes, along with electrical remodeling and arrhythmia risk in the early months following DOX therapy. Future work will include more datasets to statistically power the study, and will use the protocol to test cardioprotective strategies.