Upregulation of YAP attenuates doxorubicin-induced cardiotoxicity through inhibiting ferroptosis.

Background

Doxorubicin (DOX) is a widely utilized anthracycline chemotherapeutic agent in clinical settings; however, its application is significantly constrained by the cardiotoxic effects it induces, the underlying mechanisms of which remain inadequately understood. Ferroptosis, a recently identified form of iron-dependent programmed cell death, has been implicated in cardiovascular diseases, yet its involvement in doxorubicin-induced cardiotoxicity (DIC) requires further elucidation. Yes-associated protein (YAP), a pivotal component of the Hippo signaling pathway, plays a crucial role in myocardial cell survival and the preservation of cardiac function. Elucidating the alterations and mechanisms of YAP in the regulation of ferroptosis within the context of DIC may facilitate the identification of novel therapeutic targets for its prevention and treatment.

Methods

A DIC model was established both in vivo and in vitro using doxorubicin (DOX). Myocardial tissue damage and morphological changes in mice were assessed using echocardiography, lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) assays, hematoxylin and eosin (HE) staining, and transmission electron microscopy. Additionally, dihydroethidium (DHE) and C11-BODIPY fluorescence staining was employed to evaluate alterations in reactive oxygen species (ROS) and lipid peroxidation levels in mice and HL-1 cardiomyocytes. Utilizing the DIC model, YAP was overexpressed in HL-1 cardiomyocytes. Cell viability, along with levels of LDH, GSH, and Fe²⁺ in HL-1 cardiomyocytes, were quantified using specific commercial assay kits. The expression levels of GPX4, ferritin, FSP1, DHODH, xCT, YAP, phosphorylated YAP, and nuclear YAP were evaluated in mouse models and HL-1 cardiomyocytes using RT-qPCR analysis, Western blot analysis, and immunofluorescence techniques.

Results

Results indicated that within the DIC model, there was an increase in ferroptosis in both murine myocardial tissue and HL-1 cardiomyocytes, accompanied by a decrease in YAP protein expression and the translocation of nuclear YAP. Notably, YAP overexpression in HL-1 cardiomyocytes was found to inhibit ferroptosis and mitigate doxorubicin-induced cardiotoxicity. However, this protective effect was partially reversed by inhibition of YAP, GPX4, FSP1, and DHODH.

Conclusion

Our findings suggested that YAP overexpression attenuate DIC by inhibiting ferroptosis. These observations point to YAP as a potential therapeutic target for reducing DOX-induced myocardial injury.