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.

项目概要 包括阿霉素在内的化疗用于治疗 50% 的儿科癌症患者，同时高度 对肿瘤有效，儿童时期接受阿霉素治疗的患者中有 11% 出现心功能障碍/衰竭 作为成年人。迟发性阿霉素心脏毒性机制尚不清楚，且无心脏保护作用 该剂目前已获得 FDA 批准用于儿童。我们独特的阿霉素心脏毒性大鼠模型结果 七个月潜伏期后的心脏功能障碍，允许研究儿科心脏保护治疗 免疫功能正常的大鼠中的癌症（骨肉瘤）。在这个模型中，我们确定了一组不同的 使用甲基测序目标富集，在阿霉素治疗的大鼠心脏中进行 DNA 甲基化区域 (DMR) 系统平台。我们假设阿霉素治疗改变了 DNA 甲基化模式 易感个体（包括动物和人类患者）的心肌细胞。这种异常的DNA甲基化 可以作为阿霉素后损伤的 DNA 生物标志物。我们进一步假设DNA甲基化 通过有效的心脏保护剂治疗可以最大限度地减少关键基因调控区域的变化， 减少心脏功能缺陷、心肌退化、间质纤维化和氧化应激 DNA 损害。在目标 1 中，我们将确定 DNA 中差异甲基化区域 (DMR) 的时间特征 阿霉素在心脏中的诱导作用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了一个由 6 个 DMR 组成的强大面板，我们将定义 阿霉素诱导雄性和雌性大鼠 DNA 甲基化。我们还将测试心脏保护剂的效果 DNA甲基化，验证DNA甲基化与心脏毒性之间的关系，并将确定 通过超声心动图、肌钙蛋白 I 监测这些药物预防心脏病的潜在有效性 测量和组织学评分。我们还将确定海岸区域的 DNA 甲基化是否 代表可以将甲基化扩散到 CpG 岛并影响 RNA 表达的早期变化。提供 作为转化人类研究的桥梁，我们将检查阿霉素诱导的心脏 DNA 甲基化变化 患有骨肉瘤的免疫功能正常的大鼠。在目标 2 中，我们将确定人类心脏 DNA 是否来自 与年龄匹配的对照患者相比，阿霉素治疗的患者具有差异甲基化区域。 来自阿霉素治疗患者的冷冻心肌库将与年龄匹配的对照进行比较 使用 Illumina Infinium 人类甲基化 450 珠芯片阵列来确定 DMR 模式。以确定是否有这些 模式是阿霉素特异性的，最近发表的来自缺血性心脏病患者的 DNA 甲基化数据 （公开）将与阿霉素暴露患者的 DNA 甲基化进行比较。我们的长期 目标（PA-19-111）是识别新型心脏特异性生物标志物，以协助肿瘤学家和心脏病学家 早期诊断心脏毒性的方法，以便可以采取心脏保护策略。这是第一步 “概念验证”研究旨在确定心脏组织中的 DMR 候选者，为未来的血清 DMR 研究做好准备 在有记录的阿霉素心脏毒性的儿童或成人队列中，以确定诊断敏感性。