Electronic cigarette extract induced toxic effect in iPS-derived cardiomyocytes

Abstract

Background: Cigarette smoking is an important risk factor for cardiac diseases. In the current study, we sought to assess the effect of electronic cigarette extract (ECE) and conventional cigarette smoke extract (CSE) on cardiomyocytes. Methods: iPSCs-derived cardiomyocytes were used in the study to evaluate cellular toxicities. Cells were exposed to either ECE or CSE for two consecutive days as an acute exposure or every other day for 14 days. Concentration of nicotine in both ECE and CSE were measured by Mass-Spectrometry and Q-Exactive-HF was used to identify other ingredients in both extracts. Fluorescent microscopy was used to measure the oxidative stress after ECE and CSE exposure. Motility and beat frequency of cardiomyocytes were determined using the Sisson-Ammons Video Analysis system. Heart failure target panel genes of exposed cardiomyocytes were compared to control unexposed cells. Results: Despite nicotine concentration in CSE being six-fold higher than ECE (50 μg in CSE and 8 μg in ECE), ECE had similar toxic effect on cardiomyocytes. Both CSE and ECE generate significant cellular reactive oxygen species. The Sisson-Ammons Video Analysis (SAVA) analysis showed significant changes in myocyte function with both CSE and ECE slowing beating and increasing cell death. Chronic exposure of both ECE and CSE significantly decreased cardiomyocytes viability long term at all doses. Target panel gene expression profiles of both ECE and CSE exposed cardiomyocytes were different from controls with distinct pattern of genes that involved cell proliferation, inflammation, and apoptosis.

Chronic Cardiotoxicity Assays Using Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes (hiPSC-CMs)

“…Long-term exposure of nicotine reduced hESC-CMs cell viability, increased ROS and Ca 2+ signaling was affected, increasing the risk of arrhythmias. These results are further extended by Basma et al [ 37 ], where they exposed hiPSC-CMs (iCell CMs) to electronic cigarette extract (ECE) and conventional cigarette smoke extract (CSE) for acute and chronic (14 days) exposures. Both ECE and CSE generate ROS, cytotoxicity and slowed beating.…”

Cardiovascular consequences of vaping

“…The cellular response to E-cig exposure is vital to understand mechanisms of pathogenesis. RNA-sequencing data of iPSC-derived cardiomyocytes exposed to E-cig extract for 2 days in vitro altered gene expression signatures for proliferation and apoptosis [50]. Notably, natriuretic peptide B (NPPB) upregu-lation was observed alongside downregulated myosin light chain kinase (MYLK) and troponin I3 (TNNI3).…”

“…Disruption in the balance of prooxidant and antioxidant systems in vasculature is associated with arrhythmias and myocardial remodeling because of pathogenic hypertrophy and apoptotic signaling [49]. E-cig exposure increases ROS in cardiomyo-cytes and aortic tissue vas detected by dihydroethi-dium (DHE) fluorescence [30 ▪ ,42,50]. Increased ROS coincided with decreased endothelial nitric oxide synthase (eNOS) following 3 days of exposure in mice, suggesting that decreased antioxidant nitric oxide is involved in the increase in oxidative stress [42].…”

Environmental Alterations during Embryonic Development: Studying the Impact of Stressors on Pluripotent Stem Cell-Derived Cardiomyocytes

“…In addition, Ca2+ signalling was found to be affected in hESC-CMs upon nicotine exposure, increasing the propensity to Ca2+-associated arrhythmia [212]. Electronic and conventional cigarette smoking extract impaired hiPSC-CMs function, slowed beating and increased ROS-induced cell death [221]. Notably, RNA-seq revealed numerous altered genes essential for normal heart function and response to stress, including MYLK, NPPA, TNNT2 and TNNI3.…”

“…Notably, RNA-seq revealed numerous altered genes essential for normal heart function and response to stress, including MYLK, NPPA, TNNT2 and TNNI3. Most of them were downregulated, probably due to a significant increase in upstream methylation signals (for both DNMT3A and B pathways) [221]. Interestingly, pathological heart remodelling, transcriptional and epigenetic changes have been also observed in mice [213].…”