鉴定与蒽环类抗生素引起的心肌病相关的新型高度甲基化或低度甲基化CpG位点和基因。
Identification of novel hypermethylated or hypomethylated CpG sites and genes associated with anthracycline-induced cardiomyopathy.
发表日期：2023 Aug 04
作者：
Purnima Singh, Liting Zhou, Disheet A Shah, Romina B Cejas, David K Crossman, Mariam Jouni, Tarek Magdy, Xuexia Wang, Noha Sharafeldin, Lindsey Hageman, Donald E McKenna, Steve Horvath, Saro H Armenian, Frank M Balis, Douglas S Hawkins, Frank G Keller, Melissa M Hudson, Joseph P Neglia, A Kim Ritchey, Jill P Ginsberg, Wendy Landier, Paul W Burridge, Smita Bhatia
来源：
Epigenetics & Chromatin
摘要：
蒽环类抗生素引起的心肌病是儿童癌症幸存者晚期发生的主要致残原因。错配的DNA甲基化在新发心血管疾病中起到一定作用。表观遗传过程可能在蒽环类抗生素引起的心肌病中发挥作用，但尚未进行研究。本研究旨在检验外周血DNA的基因组范围CpG位点差异性甲基化是否与蒽环类抗生素引起的心肌病相关联。本研究采用配对病例对照研究的参与者，52名非西班牙裔白人的蒽环类抗生素暴露的儿童癌症幸存者与心肌病患者1:1配对。使用集成参考大人外周血DNA甲基化分析流程（ChAMP）进行芯片分析和解卷积的Illumina HumanMethylation EPIC BeadChip芯片阵列数据分析。进行表观基因组关联研究（EWAS），模型经修正以考虑GrimAge、性别、年龄入组的交互项、胸腔放射照射、年龄的二次项，以及心血管风险因素（CVRFs：糖尿病、高血压、脂质代谢紊乱）。通过CRISPR/Cas9技术在人诱导多能干细胞心肌细胞（hiPSC-CMs）中进行基因敲除来对优先考虑的基因进行功能验证。DNA甲基化EPIC芯片分析确定了32个差异甲基化探针（DMP：15个高甲基化和17个低甲基化探针），重叠于23个基因和9个基因间区域。还发现了354个差异甲基化区域（DMRs）。其中几个基因与心功能异常相关。在hiPSC-CMs中敲除EXO6CB、FCHSD2、NIPAL2和SYNPO2基因会增加对多柔比星的敏感性。此外，EWAS分析发现基因RORA中探针'cg15939386'的低甲基化与蒽环类抗生素引起的心肌病显著相关。在这一基因组范围的DNA甲基化分析研究中，我们观察到蒽环类抗生素暴露的儿童癌症幸存者中患有心肌病与无心肌病者之间的CpG位点的DNA甲基化存在显著差异，提示某些基因的差异性DNA甲基化可能在蒽环类抗生素引起的心肌病的发病机制中发挥作用。© 2023. Springer Nature Limited.
Anthracycline-induced cardiomyopathy is a leading cause of late morbidity in childhood cancer survivors. Aberrant DNA methylation plays a role in de novo cardiovascular disease. Epigenetic processes could play a role in anthracycline-induced cardiomyopathy but remain unstudied. We sought to examine if genome-wide differential methylation at 'CpG' sites in peripheral blood DNA is associated with anthracycline-induced cardiomyopathy. This report used participants from a matched case-control study; 52 non-Hispanic White, anthracycline-exposed childhood cancer survivors with cardiomyopathy were matched 1:1 with 52 survivors with no cardiomyopathy. Paired ChAMP (Chip Analysis Methylation Pipeline) with integrated reference-based deconvolution of adult peripheral blood DNA methylation was used to analyze data from Illumina HumanMethylation EPIC BeadChip arrays. An epigenome-wide association study (EWAS) was performed, and the model was adjusted for GrimAge, sex, interaction terms of age at enrollment, chest radiation, age at diagnosis squared, and cardiovascular risk factors (CVRFs: diabetes, hypertension, dyslipidemia). Prioritized genes were functionally validated by gene knockout in human induced pluripotent stem cell cardiomyocytes (hiPSC-CMs) using CRISPR/Cas9 technology. DNA-methylation EPIC array analyses identified 32 differentially methylated probes (DMP: 15 hyper-methylated and 17 hypo-methylated probes) that overlap with 23 genes and 9 intergenic regions. Three hundred and fifty-four differential methylated regions (DMRs) were also identified. Several of these genes are associated with cardiac dysfunction. Knockout of genes EXO6CB, FCHSD2, NIPAL2, and SYNPO2 in hiPSC-CMs increased sensitivity to doxorubicin. In addition, EWAS analysis identified hypo-methylation of probe 'cg15939386' in gene RORA to be significantly associated with anthracycline-induced cardiomyopathy. In this genome-wide DNA methylation profile study, we observed significant differences in DNA methylation at the CpG level between anthracycline-exposed childhood cancer survivors with and without cardiomyopathy, implicating differential DNA methylation of certain genes could play a role in pathogenesis of anthracycline-induced cardiomyopathy.© 2023. Springer Nature Limited.