癌症治疗的最新进展大大提高了患者的生存率，但当前蒽环类药物 (AC) 治疗方案的长期疗效往往包括严重的长期并发症，特别是蒽环类药物引起的心脏毒性 (AIC)。尽管 AC 治疗和 AIC 之间存在已知的关联，但对潜在分子途径的全面了解仍然难以实现。可靠的治疗干预措施的稀缺凸显了这一差距，右雷佐生是 FDA 批准的唯一用于减轻 AIC 风险的药物。本研究旨在通过分析先前生成的心脏球体 RNA 测序数据集（经受临床相关剂量的 AC 后），阐明人类心肌细胞 (hCM) 对 AC 暴露的转录反应。分析揭示了在不同时间点发现的强大转录反应。我们的目标是通过采用预测算法突出关键 TF 以进行进一步实验验证，从而识别介导 AIC 的重要转录因子 (TF)。使用 shRNA 构建体，我们进一步评估了这些 TF 对 hCM 对阿霉素 (DOX) 反应的影响，并发现这些 TF 在 DOX 暴露后对 hCM 存活率具有显着影响。进一步研究了 TF FOXO3、GATA2、AHR 和 NFE2L2 在 AIC 中的作用，包括细胞活力、DOX 摄取、DNA 损伤修复和通过 Cleaved-Caspase 3 诱导细胞凋亡。我们的研究表明，消除 FOXO3 和 GATA2 会使 hCM 更容易受到 DOX 的影响GATA2、NFE2L2和AHR的缺乏导致细胞内DOX水平显着升高。此外，FOXO3 在修复 hCM DNA 损伤中发挥了作用，因为我们观察到 CDKN1A 水平显着增强。我们还注意到，当 FOXO3、GATA2、NFE2L2 和 AHR 不存在时，通过 COMET 检测，DNA 损伤显着增加。此外，我们通过将我们的结果与基于阿霉素诱导心脏毒性患者的 hiPSC-CM 的研究结果进行比较，研究了临床相关性，确定了重叠的 TF 及其在细胞周期和 DNA 修复等关键细胞过程中的调节作用。这种方法不仅增进了对 AIC 背后分子机制的理解，还为新的治疗方法打开了可能的窗口，以减轻患者 AC 治疗的负面副作用。版权所有 © 2024。由 Elsevier B.V. 出版。

Recent advances in cancer therapy have substantially increased survival rates among patients, yet the prolonged effect of current treatment regimens with anthracyclines (ACs) often include severe long-term complications, notably in the form of anthracycline-induced cardiotoxicity (AIC). Despite known associations between AC treatment and AIC, a comprehensive understanding of the underlying molecular pathways remains elusive. This gap is highlighted by the scarcity of reliable therapeutic interventions, with dexrazoxane being the sole FDA-approved drug to mitigate AIC risks. This study aims at elucidating the transcriptional response of human cardiomyocytes (hCMs) to AC exposure by analyzing a previously generated RNA-sequencing dataset of cardiac spheroids subjected to clinically relevant doses of ACs. The analysis revealed a robust transcriptional response identified across various time points. We aimed at identifying important transcription factors (TFs) mediating AIC by employing predictive algorithms to highlight key TFs for further experimental validation. Using shRNA constructs, we further assessed the impact of these TFs on hCM response to doxorubicin (DOX) and revealed that these TFs had a notable impact on hCM survival upon DOX exposure. TFs FOXO3, GATA2, AHR and NFE2L2 were further investigated for their role in AIC including cell viability, DOX uptake, DNA damage repair and induction of apoptosis through Cleaved-Caspase 3. Our study demonstrated that eliminating FOXO3 and GATA2 made hCMs more vulnerable to DOX and the lack of GATA2, NFE2L2 and AHR led to significantly higher intracellular levels of DOX. Additionally, FOXO3 played a role in the repair of hCM DNA damage as we observed markedly enhanced levels of CDKN1A. We also noted significant increases in DNA damage through COMET-assays when FOXO3, GATA2, NFE2L2 and AHR were absent. Furthermore, we investigated the clinical relevance by comparing our results with those from a study based on hiPSC-CMs derived from patients with doxorubicin-induced cardiotoxicity, identifying overlapping TFs and their regulatory roles in critical cellular processes like the cell cycle and DNA repair. This approach not only advances the understanding of the molecular mechanisms behind AIC but also opens possible windows for new therapeutic approaches to mitigate the negative side-effects from patient AC treatment.Copyright © 2024. Published by Elsevier B.V.