Detection of acute TLR-7 agonist-induced hemorrhagic myocarditis in mice by refined multi-parametric quantitative cardiac MRI

Abstract

BACKGROUND: Hemorrhagic myocarditis is a potentially fatal complication of excessive levels of systemic inflammation. It has been reported in viral infection, but is also possible in systemic autoimmunity. Cardiac magnetic resonance (CMR) imaging is the current gold standard for non-invasive detection of suspected inflammatory damage to the heart and changes in T 1 and T 2 relaxation times are commonly used to detect edema associated with immune cell infiltration and fibrosis. These measurements also form the basis of the Lake Louise Criteria, which define a framework for the CMR-based diagnosis of myocarditis.However, they do not take into account the possibility of hemorrhage leading to tissue iron deposition which strongly influences T 1 and T 2 measurements and may complicate interpretation based on these two parameters only. METHODS:Systemic inflammation was induced in CFN mice by application of the TLR-7 agonist Resiquimod. Histopathology was performed on heart sections to assess immune cell infiltration (Hematoxylin & Eosin), fibrosis (PicoSirius Red) and tissue iron deposition (Perl's Prussian Blue). A multi-parametric cardiac MRI tissue mapping approach measuring T 1 , T 2, and T 2 * relaxation times was established to non-invasively identify these parameters in this small rodent model. RESULTS:Resiquimod-treated mice developed severe thrombocytopenia and hemorrhagic myocarditis. We identified patches of cardiac hemorrhage based on the presence of two major MRI phenotypes. Increased T 2 with normal T 1 and T 2 * values correlated with CMR imaging for hemorrhagic myocarditis in mice Baxan et al. 2018 2 infiltration/edema only, while decreased T 1 , T 2 , and T 2 * values identify areas with infiltration/edema in the presence of iron indicating hemorrhagic myocarditis. CONCLUSION:We show that over-activation of the TLR-7 pathway by Resiquimod treatment of CFN mice induces an early immune response reminiscent of excessive systemic inflammation due to viral infection. This causes internal bleeding which manifests most prominently as severe hemorrhagic myocarditis. We optimized a comprehensive noninvasive in vivo MRI approach based on quantitative measurement of T 1 , T 2 and T 2 * to demonstrate the presence of diffuse myocardial edema, infiltration and iron deposition without the need of contrast agent administration. We propose that adding quantitative T 2 * mapping to CMR protocols for detection of myocarditis will improve diagnostic sensitivity and interpretation of disease mechanisms.