Differentially methylated gene regions (DMRs) induced by doxorubicin in heart: significance and clinical application

阿霉素诱导心脏差异甲基化基因区（DMR）：意义及临床应用

• 批准号: 10463850
• 负责人: KATHLEEN Louise GABRIELSON
• 金额: $ 20.47万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10463850
• 关键词: Aberrant DNA Methylation; Acute; Adult; Affect; Aftercare; Age; Aging; Animal Model; Animals; Area; Biological Markers; Blood; Cancer Patient; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiotonic Agents; Cardiotoxicity; Child; Childhood; Cohort Studies; Cytosine; DNA; DNA Damage; DNA Methylation; DNA Repair; DNA methylation profiling; Data; Data Set; Development; Diagnostic Sensitivity; Dinucleoside Phosphates; Disease; Dose; Doxorubicin; Doxorubicin-DNA Complex; Early Diagnosis; Echocardiography; Effectiveness; Event; FDA approved; Failure; Female; Fibrosis; Freezing; Functional disorder; Future; Gene Expression; Genes; Genetic Transcription; Goals; Guanine; Health; Heart; Heart Diseases; Heart Injuries; Heart failure; Histology; Human; Immunocompetent; Individual; Island; Label; Malignant Childhood Neoplasm; Malignant Neoplasms; Measurement; Methods; Methylation; Modeling; Monitor; Mutation; Myocardial Degeneration; Myocardial Ischemia; Myocardial dysfunction; Myocardium; Nucleic Acid Regulatory Sequences; Oncogenes; Oncologist; Oxidative Stress; Patients; Pattern; Pharmacotherapy; Process; Promoter Regions; Publishing; Rattus; Regulator Genes; Science; Serum; Study of serum; System; Testing; Time; Tissues; Toxic effect; Troponin; Troponin I; Tumor Suppressor Genes; base; bead chip; cancer cell; cardioprotection; chemotherapy; clinical application; cohort; genome wide methylation; heart function; heart imaging; imaging modality; inorganic phosphate; interstitial; male; methylation pattern; myocardial injury; novel; novel strategies; novel therapeutic intervention; osteosarcoma; potential biomarker; pre-clinical; prevent; protein biomarkers; repaired; response; specific biomarkers; tissue preparation; tumor; tumor progression

Project Summary Chemotherapy that includes doxorubicin is used to treat 50% of all pediatric cancer patients, and while highly effective against tumors, 11% of patients treated as children with doxorubicin develop heart dysfunction/ failure as adults. Mechanisms of delayed doxorubicin cardiotoxicity are not clearly understood and no cardioprotective agent is currently FDA-approved for use in children. Our unique rat model of doxorubicin cardiotoxicity results in cardiac dysfunction after a seven-month latency, allowing the study of cardioprotective treatments in pediatric cancer (osteosarcoma) in immunocompetent rats. In this model, we have identified a set of differentially methylated regions of DNA (DMRs) in hearts of doxorubicin-treated rats, using a Methyl-Seq Target Enrichment System platform. We hypothesize that doxorubicin treatments alter patterns of DNA methylation in cardiomyocytes of susceptible individuals, including animals and human patients. This aberrant DNA methylation could serve as a DNA biomarker for post- doxorubicin damage. We further hypothesize that DNA methylation changes in critical gene-regulatory regions can be minimized by effective cardioprotective agent therapy, diminishing heart function deficits, myocardial degeneration, interstitial fibrosis and oxidative stress DNA damage. In Aim 1, we will determine the temporal features of differentially methylated regions (DMRs) in DNA induced by doxorubicin in heart and how cardioprotective agents modulate the DNA methylation pattern. Using a robust panel of 6 DMRs that we identified in preliminary studies, we will define the temporal development of doxorubicin-induced DNA methylation in male and female rats. We will also test effects of cardioprotective agents on DNA methylation, to verify relationships between DNA methylation and cardiotoxicity, and will determine potential effectiveness of these agents for preventing heart disease, monitored by echocardiography, troponin I measurements, and histology scores. We will also determine whether DNA methylation in shore regions represent early changes that can spread methylation into the CpG islands and affect RNA expression. To provide a bridge to translational human studies, we will examine doxorubicin-induced DNA methylation changes in hearts of immunocompetent rats bearing osteosarcoma. In Aim 2, we will determine whether human heart DNA from doxorubicin treated patients have differentially methylated regions compared to age matched control patients. Banked frozen myocardium from doxorubicin treated patients will be compared to age matched controls using the Illumina Infinium human methylation 450 beadchip array to determine DMR patterns. To determine if these patterns are doxorubicin specific, recently published DNA methylation data from ischemic heart disease patients (publically available) will be compared to DNA methylation from the doxorubicin-exposed patients. Our long-term goal (PA-19-111) is to identify novel heart specific biomarkers to assist oncologists and cardiologists with a method for early diagnosis of heart toxicity so that cardioprotection strategies can be used. This is the first step “proof of concept” study to identify DMRs candidates in heart tissues to prepare for future serum DMRs studies in pediatric or adult cohorts with documented heart doxorubicin toxicity to establish diagnostic sensitivity.

项目摘要 包括阿霉素在内的化学疗法用于治疗所有儿科癌症患者的50％，虽然高度 有效防止肿瘤，有11％的患者患有阿霉素患有心脏功能障碍/失败的儿童 作为成年人。延迟的阿霉素心脏毒性的机制尚不清楚，也没有心脏保护 目前，代理商已批准用于儿童。我们独特的大鼠阿霉素心脏毒性结果 七个月的潜伏期后心脏功能障碍，允许研究儿科治疗 免疫能力大鼠的癌症（骨肉瘤）。在此模型中，我们已经确定了一组差异性 用甲基 - 季节靶富集 系统平台。我们假设阿霉素治疗改变了DNA甲基化模式 易感人群的心肌细胞，包括动物和人类患者。这种异常的DNA甲基化 可以用作无毒素后损伤的DNA生物标志物。我们进一步假设DNA甲基化 有效的心脏保护剂治疗可以将关键基因调节区域的变化最小化， 心脏功能不足，心肌变性，间质性纤维化和氧化应激DNA 损害。在AIM 1中，我们将确定DNA中不同甲基化区域（DMR）的临时特征 阿霉素在心脏中诱导，以及心脏保护剂如何调节DNA甲基化模式。使用 我们在初步研究中确定的6个DMR的强大面板，我们将定义 阿霉素诱导的雄性和雌性大鼠的DNA甲基化。我们还将测试心脏保护剂的效果 关于DNA甲基化，以验证DNA甲基化和心脏毒性之间的关系，并将确定 这些药物对预防心脏病的潜在有效性，通过超声心动图，肌钙蛋白I监测 测量和组织学评分。我们还将确定是否在海岸区域中的DNA甲基化 表示可以将甲基化扩散到CpG岛并影响RNA表达的早期变化。提供 翻译人类研究的桥梁，我们将检查阿霉素诱导的心脏中的DNA甲基化变化 具有骨肉瘤的免疫能力大鼠。在AIM 2中，我们将确定人心DNA是否来自 与年龄相匹配的对照患者相比，接受阿霉素治疗的患者的甲基化区域不同。 将接受阿霉素治疗的患者的库存冷冻心肌与使用年龄匹配的对照组相比 Illumina Infinium人甲基化450 Beadchip阵列以确定DMR模式。确定这些是否 模式是阿霉素特异性的，最近发表的缺血性心脏病患者的DNA甲基化数据 （公开可用）将与暴露阿霉素暴露的患者的DNA甲基化进行比较。我们的长期 目标（PA-19-111）是确定新颖的心脏特异性生物标志物，以帮助肿瘤学家和心脏病学家 早期诊断心脏毒性的方法，因此可以使用心脏保护策略。这是第一步 “概念验证”研究以识别心脏组织中的DMR候选者，以准备未来的血清DMRS研究 在具有记录的心脏阿霉素毒性的小儿或成人队列中，以建立诊断敏感性。