Doxorubicin induces cardiotoxicity in a pluripotent stem cell model of aggressive B cell lymphoma cancer patients

Abstract

Cancer therapies with anthracyclines have been shown to induce cardiovascular complications. The aims of this study were to establish an in vitro induced pluripotent stem cell model (iPSC) of anthracycline-induced cardiotoxicity (ACT) from patients with an aggressive form of B-cell lymphoma and to examine whether doxorubicin (DOX)-treated ACT-iPSC cardiomyocytes (CM) can recapitulate the clinical features exhibited by patients, and thus help uncover a DOX-dependent pathomechanism. ACT-iPSC CM generated from individuals with CD20+ B-cell lymphoma who had received high doses of DOX and suffered cardiac dysfunction were studied and compared to control-iPSC CM from cancer survivors without cardiac symptoms. In cellular studies, ACT-iPSC CM were persistently more susceptible to DOX toxicity including augmented disorganized myofilament structure, changed mitochondrial shape, and increased apoptotic events. Consistently, ACT-iPSC CM and cardiac fibroblasts isolated from fibrotic human ACT myocardium exhibited higher DOX-dependent reactive oxygen species. In functional studies, Ca2+ transient amplitude of ACT-iPSC CM was reduced compared to control cells, and diastolic sarcoplasmic reticulum Ca2+ leak was DOX-dependently increased. This could be explained by overactive CaMKIIδ in ACT CM. Together with DOX-dependent augmented proarrhythmic cellular triggers and prolonged action potentials in ACT CM, this suggests a cellular link to arrhythmogenic events and contractile dysfunction especially found in ACT engineered human myocardium. CamKIIδ inhibition prevented proarrhythmic triggers in ACT. In contrast, control CM upregulated SERCA2a expression in a DOX-dependent manner, possibly to avoid heart failure conditions. In conclusion, we developed the first human patient-specific stem cell model of DOX-induced cardiac dysfunction from patients with B-cell lymphoma. Our results suggest that DOX-induced stress resulted in arrhythmogenic events associated with contractile dysfunction and finally in heart failure after persistent stress activation in ACT patients.

Raman and fluorescence micro-spectroscopy applied for the monitoring of sunitinib-loaded porous silicon nanocontainers in cardiac cells

“…The cells were stained with MitoTracker ® Orange (λ ex = 552 nm) and incubated with SU or SU-loaded nanoparticles in concentrations between 5 and 50 μM for 24 h ( Supplementary Figure S1 ). The images were analyzed as described in Haupt et al (2022 ). These analyses suggest that the mitochondrial network might be affected by SU.…”

“…The images format was 1024 × 1024 pixels and the speed of image acquisition was 100 Hz. For the quantitative analysis of the mitochondrial structures, Fiji program was used to preprocess the measurements, to create a segmentation map of the mitochondria, and to perform a binary skeleton analysis, following the protocol presented by Haupt et al (2022) ( Schindelin et al, 2012 ). Based on this, the percentage of branched mitochondrial networks was calculated and compared through the different concentrations.…”

Tachycardiomyopathy entails a dysfunctional pattern of interrelated mitochondrial functions

“…Human iPSC–CM were differentiated from four healthy individuals by sequential targeting of the WNT pathway as described previously [ 12 , 32 , 65 ]. Following differentiation, purity and quality of iPSC–CM were determined by flow cytometry (~ 90% cardiac TNT +), cardiac immunofluorescence, morphology, and qPCR for cardiac sub-type marker (data not shown).…”

Membrane remodelling triggers maturation of excitation–contraction coupling in 3D-shaped human-induced pluripotent stem cell-derived cardiomyocytes

“…The technology to generate cardiomyocytes from human-induced pluripotent stem cells (hiPSC-CM) represents a great breakthrough for pharmacological tests and disease modelling in cardiovascular research [ 12 , 22 ] and fuels new hope for cell-based therapy of patients with heart failure [ 28 , 42 ]. Human iPSC-CM reveal all essential cardiac features at the electro-mechanical level but retain a premature phenotype compared to ventricular cardiomyocytes of the adult heart.…”