Robinson Emma L, Ameri Pietro, Delrue Leen, Vanderheyden Marc, Bartunek Jozef, Altieri Paola, Heymans Stephane, Heggermont Ward A
Cardiovascular Research Institute Maastricht, Maastricht University, Maastricht, Netherlands.
Division of Cardiology, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, United States.
Front Cardiovasc Med. 2023 Mar 9;10:884174. doi: 10.3389/fcvm.2023.884174. eCollection 2023.
Anthracycline-induced cardiotoxicity is a well-known serious clinical entity. However, detailed mechanistic insights on how short-term administration leads to late and long-lasting cardiotoxicity, are still largely undiscovered. We hypothesize that chemotherapy provokes a memory effect at the level of epigenomic DNA modifications which subsequently lead to cardiotoxicity even years after cessation of chemotherapy.
We explored the temporal evolution of epigenetic modifiers in early and late cardiotoxicity due to anthracyclines by means of RNA-sequencing of human endomyocardial left ventricular biopsies and mass spectrometry of genomic DNA. Based on these findings, validation of differentially regulated genes was obtained by performing RT-qPCR. Finally, a proof-of-concept mechanistic study was performed to dissect some of the mechanistic aspects of epigenetic memory in anthracycline-induced cardiotoxicity.
Correlation of gene expression between late and early onset cardiotoxicity revealed an value of 0.98, demonstrating a total of 369 differentially expressed genes (DEGs, FDR < 0.05). of which 72% ( = 266) were upregulated, and 28% of genes, ( = 103) downregulated in later as compared to earlier onset cardiotoxicity. Gene ontology analysis showed significant enrichment of genes involved in methyl-CpG DNA binding, chromatin remodeling and regulation of transcription and positive regulation of apoptosis. Differential mRNA expression of genes involved in DNA methylation metabolism were confirmed by RT-qPCR in endomyocardial biopsies. In a larger biopsy cohort, it was shown that Tet2 was more abundantly expressed in cardiotoxicity biopsies vs. control biopsies and vs. non-ischemic cardiomyopathy patients. Moreover, an study was performed: following short-term doxorubicin treatment, H9c2 cells were cultured and passaged once they reached a confluency of 70%-80%. When compared to vehicle-only treated cells, in doxorubicin-treated cells, three weeks after short term treatment, and other genes involved in active DNA demethylation were markedly upregulated. These alterations coincided with a loss of DNA methylation and a gain in hydroxymethylation, reflecting the epigenetic changes seen in the endomyocardial biopsies.
Short-term administration of anthracyclines provokes long-lasting epigenetic modifications in cardiomyocytes both and , which explain in part the time lapse between the use of chemotherapy and the development of cardiotoxicity and, eventually, heart failure.
蒽环类药物引起的心脏毒性是一种众所周知的严重临床病症。然而，关于短期给药如何导致晚期和长期心脏毒性的详细机制仍 largely undiscovered（很大程度上未被揭示）。我们推测化疗会在表观基因组 DNA 修饰水平引发记忆效应，这随后会导致即使在化疗停止数年之后仍出现心脏毒性。
我们通过对人类左心室心内膜活检组织进行 RNA 测序以及对基因组 DNA 进行质谱分析，探究了蒽环类药物导致的早期和晚期心脏毒性中表观遗传修饰因子的时间演变。基于这些发现，通过进行 RT-qPCR 对差异调节基因进行了验证。最后，进行了一项概念验证机制研究，以剖析蒽环类药物诱导的心脏毒性中表观遗传记忆的一些机制方面。
晚期和早期心脏毒性之间的基因表达相关性显示相关值为 0.98，共揭示了 369 个差异表达基因（DEGs，FDR < 0.05）。其中，与早期心脏毒性相比，晚期有 72%（ = 266）的基因上调，28%（ = 103）的基因下调。基因本体分析表明，参与甲基化 CpG DNA 结合、染色质重塑以及转录调控和凋亡正调控的基因有显著富集。通过对心内膜活检组织进行 RT-qPCR，证实了参与 DNA 甲基化代谢的基因的差异 mRNA 表达。在一个更大的活检队列中，结果显示与对照活检组织以及非缺血性心肌病患者相比，Tet2 在心脏毒性活检组织中表达更为丰富。此外，进行了一项 研究：在短期给予阿霉素治疗后，培养 H9c2 细胞，当它们达到 70%-80%汇合度时传代。与仅用载体处理的细胞相比，在阿霉素处理的细胞中，短期治疗三周后， 以及其他参与活性 DNA 去甲基化的基因显著上调。这些改变与 DNA 甲基化的丧失和羟甲基化的增加相一致，反映了在心内膜活检组织中观察到的表观遗传变化。
短期给予蒽环类药物会在心肌细胞中引发长期的表观遗传修饰，这在一定程度上解释了化疗使用与心脏毒性以及最终心力衰竭发生之间的时间间隔。
