Doxorubicin induces trans-differentiation and MMP1 expression in cardiac fibroblasts via cell death-independent pathways.

Akane Nagasako,Kouichi Tamura,Takayuki Fujita,Masanari Umemura,Masatoshi Narikawa,Kazuo Kimura,Partha Mukhopadhyay,Tomoaki Ishigami,Mayu Hikichi,Yoshihiro Ishikawa,Ryo Tanaka,Utako Yokoyama

doi:10.1371/journal.pone.0221940

Abstract

Although doxorubicin (DOX)-induced cardiomyopathy causes lethal heart failure (HF), no early detection or effective treatment methods are available. The principal mechanisms of cardiotoxicity are considered to involve oxidative stress and apoptosis of cardiomyocytes. However, the effect of DOX on cardiac fibroblasts at non-lethal concentrations remains unknown. The aim of this study was to investigate the direct effect of doxorubicin on the activation of cardiac fibroblasts independent of cell death pathways. We first found that DOX induced α-SMA expression (marker of trans-differentiation) at a low concentration range, which did not inhibit cell viability. DOX also increased MMP1, IL-6, TGF-β and collagen expression in human cardiac fibroblasts (HCFs). In addition, DOX promoted Akt and Smad phosphorylation. A Smad inhibitor prevented DOX-induced α-SMA and IL-6 protein expression. An PI3K inhibitor also prevented MMP1 mRNA expression in HCFs. These findings suggest that DOX directly induces fibrotic changes in HCFs via cell death-independent pathways. Furthermore, we confirmed that these responses are organ- and species-specific for HCFs based on experiments using different types of human and murine fibroblast cell lines. These results suggest potentially new mechanisms of DOX-induced cardiotoxicity from the viewpoint of fibrotic changes in cardiac fibroblasts.

Highlights

Associated with the remarkable developments in cancer treatment and the aging of patients with cancer, anticancer drugs have increased mortality due to adverse effects [1]
DOX markedly induced cytokine expression in human cardiac fibroblasts and normal human dermal fibroblasts compared with murine cardiac fibroblasts
We focused on IL-6 expression at the early phase in human cardiac fibroblasts (HCFs) and Normal human dermal fibroblasts (NHDFs) under DOX stimulation and MMP1 and MMP2 expression at the late phase in HCFs and NHDFs exposed to DOX

Summary

Introduction

Associated with the remarkable developments in cancer treatment and the aging of patients with cancer, anticancer drugs have increased mortality due to adverse effects [1]. Cardiac complications are the most lethal side effects, and the cardiotoxicity of anticancer drugs has recently received much attention [2]. Because doxorubicin (DOX) causes a high incidence of congestive heart failure (HF), many mechanisms of DOX-induced cardiotoxicity have been reported [3]. Doxorubicin induces trans-differentiation and MMP1 expression in cardiac fibroblasts (AMED) (66890005, 66890011, 66890001, 66890023 to Y.I.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

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