Melphalan induces cardiotoxicity through oxidative stress in cardiomyocytes derived from human induced pluripotent stem cells

Rui Liu,Yuhong Du,Fangxu Sun,Peter Fischbach,Ronghu Wu,Haian Fu,Joshua T Maxwell,Antonio Rampoldi,Dong Li,Sharon M Castellino,Chunhui Xu,Anant Mandawat

doi:10.1186/s13287-020-01984-1

Abstract

BackgroundTreatment-induced cardiotoxicity is a leading noncancer-related cause of acute and late onset morbidity and mortality in cancer patients on antineoplastic drugs such as melphalan—increasing clinical case reports have documented that it could induce cardiotoxicity including severe arrhythmias and heart failure. As the mechanism by which melphalan impairs cardiac cells remains poorly understood, here, we aimed to use cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) to investigate the cellular and molecular mechanisms of melphalan-induced cardiotoxicity.MethodshiPSC-CMs were generated and treated with clinically relevant doses of melphalan. To characterize melphalan-induced cardiotoxicity, cell viability and apoptosis were quantified at various treatment durations. Ca2+ transient and contractility analyses were used to examine the alterations of hiPSC-CM function. Proteomic analysis, reactive oxygen species detection, and RNA-Sequencing were conducted to investigate underlying mechanisms.ResultsMelphalan treatment of hiPSC-CMs induced oxidative stress, caused Ca2+ handling defects and dysfunctional contractility, altered global transcriptomic and proteomic profiles, and resulted in apoptosis and cell death. The antioxidant N-acetyl-l-cysteine attenuated these genomic, cellular, and functional alterations. In addition, several other signaling pathways including the p53 and transforming growth factor-β signaling pathways were also implicated in melphalan-induced cardiotoxicity according to the proteomic and transcriptomic analyses.ConclusionsMelphalan induces cardiotoxicity through the oxidative stress pathway. This study provides a unique resource of the global transcriptomic and proteomic datasets for melphalan-induced cardiotoxicity and can potentially open up new clinical mechanism-based targets to prevent and treat melphalan-induced cardiotoxicity.

Highlights

Treatment-induced cardiotoxicity is a leading noncancer-related cause of acute and late onset morbidity and mortality in cancer patients on antineoplastic drugs such as melphalan—increasing clinical case reports have documented that it could induce cardiotoxicity including severe arrhythmias and heart failure
Melphalan treatment induces cell death and apoptosis in hiPSC-CMs To investigate the cardiotoxicity of melphalan, we generated enriched hiPSC-CMs (Fig. S1) and treated them with melphalan at 4 doses ranging from 0 to 20 μM; the highest dose was slightly above the Cmax of melphalan (15.4 μM) [23]. hiPSC-CMs exposed to 20 μM melphalan contracted weakly after 24 h compared with other groups
With the use of hiPSC-CMs as a novel human cell-based model for the characterization of cardiac defects induced by melphalan treatment, we provide a unique resource of human global transcriptomic and proteomic datasets for melphalan-induced cardiotoxicity, which could be valuable for further investigation of the molecular mechanisms underlying melphalan-induced cardiotoxicity

Summary

Introduction

Treatment-induced cardiotoxicity is a leading noncancer-related cause of acute and late onset morbidity and mortality in cancer patients on antineoplastic drugs such as melphalan—increasing clinical case reports have documented that it could induce cardiotoxicity including severe arrhythmias and heart failure. Melphalan, a cytotoxic alkylating agent used in treatment for malignancies such as multiple myeloma, leukemia, and ovarian cancer [3,4,5], could induce cardiac complications including supraventricular tachycardia, atrial fibrillation, ventricular tachycardia, and left ventricular heart failure [6, 7]. Another study indicated that a rapid ventricular rate was associated with 91.6% of the patients who developed atrial fibrillation related to melphalan treatment [9] It remains unknown how melphalan causes the adverse cardiac effects. Since melphalan is a mainstay treatment for several malignancies and for bone marrow transplantation conditioning regimens, it is necessary to study the mechanism of melphalaninduced cardiotoxicity so that targeted treatment can be developed to ameliorate its cardiotoxicity

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