Lamberto Federica, Shashikadze Bachuki, Elkhateib Radwa, Lombardo Salvo Danilo, Horánszky Alex, Balogh Andrea, Kistamás Kornél, Zana Melinda, Menche Jörg, Fröhlich Thomas, Dinnyés András
BioTalentum Ltd., Aulich Lajos Str. 26, Gödöllő, H-2100, Hungary; Department of Physiology and Animal Health, Institute of Physiology and Animal Nutrition, Hungarian University of Agriculture and Life Sciences, Páter Károly Str. 1, H-2100, Gödöllő, Hungary.
Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, LMU Munich, 81377, Munich, Germany.
Environ Pollut. 2023 Oct 15;335:122359. doi: 10.1016/j.envpol.2023.122359. Epub 2023 Aug 9.
Early embryonic development represents a sensitive time-window during which the foetus might be vulnerable to the exposure of environmental contaminants, potentially leading to heart diseases also later in life. Bisphenol A (BPA), a synthetic chemical widely used in plastics manufacturing, has been associated with heart developmental defects, even in low concentrations. This study aims to investigate the effects of environmentally relevant doses of BPA on developing cardiomyocytes using a human induced pluripotent stem cell (hiPSC)-derived model. Firstly, a 2D in vitro differentiation system to obtain cardiomyocytes from hiPSCs (hiPSC-CMs) have been established and characterised to provide a suitable model for the early stages of cardiac development. Then, the effects of a repeated BPA exposure, starting from the undifferentiated stage throughout the differentiation process, were evaluated. The chemical significantly decreased the beat rate of hiPSC-CMs, extending the contraction and relaxation time in a dose-dependent manner. Quantitative proteomics analysis revealed a high abundance of basement membrane (BM) components (e.g., COL4A1, COL4A2, LAMC1, NID2) and a significant increase in TNNC1 and SERBP1 proteins in hiPSC-CMs treated with BPA. Network analysis of proteomics data supported altered extracellular matrix remodelling and provided a disease-gene association with well-known pathological conditions of the heart. Furthermore, upon hypoxia-reoxygenation challenge, hiPSC-CMs treated with BPA showed higher rate of apoptotic events. Taken together, our results revealed that a long-term treatment, even with low doses of BPA, interferes with hiPSC-CMs functionality and alters the surrounding cellular environment, providing new insights about diseases that might arise upon the toxin exposure. Our study contributes to the current understanding of BPA effects on developing human foetal cardiomyocytes, in correlation with human clinical observations and animal studies, and it provides a suitable model for New Approach Methodologies (NAMs) for environmental chemical hazard and risk assessment.
早期胚胎发育代表着一个敏感的时间窗口，在此期间胎儿可能易受环境污染物暴露的影响，这可能在日后生活中导致心脏病。双酚A（BPA）是一种广泛用于塑料制造的合成化学物质，即使在低浓度下也与心脏发育缺陷有关。本研究旨在使用人诱导多能干细胞（hiPSC）衍生模型，研究环境相关剂量的BPA对发育中心肌细胞的影响。首先，建立并表征了一种从hiPSC获得心肌细胞（hiPSC-CMs）的二维体外分化系统，为心脏发育的早期阶段提供了合适的模型。然后，评估了从未分化阶段开始贯穿整个分化过程的重复BPA暴露的影响。该化学物质显著降低了hiPSC-CMs的搏动率，以剂量依赖的方式延长了收缩和舒张时间。定量蛋白质组学分析显示，在用BPA处理的hiPSC-CMs中，基底膜（BM）成分（如COL4A1、COL4A2、LAMC1、NID2）含量很高，TNNC1和SERBP1蛋白显著增加。蛋白质组学数据的网络分析支持细胞外基质重塑的改变，并提供了与心脏著名病理状况的疾病-基因关联。此外，在缺氧-复氧挑战下，用BPA处理的hiPSC-CMs显示出更高的凋亡事件发生率。综上所述，我们的结果表明，即使是低剂量的BPA长期处理也会干扰hiPSC-CMs的功能并改变周围的细胞环境，为毒素暴露后可能出现的疾病提供了新的见解。我们的研究有助于当前对BPA对发育中的人类胎儿心肌细胞影响的理解，与人类临床观察和动物研究相关，并为环境化学危害和风险评估的新方法学（NAMs）提供了合适的模型。
