Dissecting mechanisms of anthracycline-induced cardiotoxicity

剖析蒽环类药物引起的心脏毒性机制

• 批准号: 10683784
• 负责人: Rene R.S. Packard
• 金额: $ 39万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10683784
• 关键词: ANXA5 gene; Address; Adolescence; Adult; Affect; Anthracycline; Apoptosis; Biochemical; Biological Assay; Breast Cancer Cell; CASP3 gene; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiotoxicity; Caspase; Cell Death; Cell Proliferation; Cessation of life; Child; Childhood; Chronic; Clinical; Co-Immunoprecipitations; Cre-LoxP; DNA Damage; Databases; Dependovirus; Development; Disease; Doxorubicin; EFRAC; Excision; Exhibits; Exposure to; Failure; Family; Genes; Goals; Heart; Heart Injuries; Histone H2A; In Situ Nick-End Labeling; Incidence; Injury; Lymphoma cell; Malignant Childhood Neoplasm; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Molecular; Molecular Biology; Mus; Muscle Cells; Neonatal; Pathway interactions; Patients; Persons; Play; Process; Protein Binding Domain; Proteins; Public Health; Pump; RNA Binding; Rattus; Recording of previous events; Regimen; Regulation; Research; Role; Serine; Stress; Systems Biology; Tamoxifen; Topoisomerase; Variant; Ventricular; Vertebral column; Whole Organism; cancer cell; cancer therapy; cardiac repair; cardioprotection; caveolin-3; chemotherapy; childhood cancer survivor; ds-DNA; enhanced green fluorescent protein; epidemiologic data; experience; gene network; heart function; imaging approach; improved; in vivo; insight; knock-down; notch protein; novel; overexpression; protective effect; side effect; transcriptome sequencing; transcriptomics

ABSTRACT While cancer affects more than one in three people over their lifetime, improved long-term survival has led to an increase in the incidence of adverse cardiac side-effects of cancer treatments. Anthracyclines such as doxorubicin (Dox) are a cornerstone of chemotherapy in various cancers, however, their use is complicated by anthracycline-induced cardiotoxicity, most commonly cardiomyopathy. The mechanisms at play in Dox cardiotoxicity are multifactorial and include Topoisomerase (Top)-2β-mediated DNA damage that may culminate in apoptosis. To identify novel actors in the disease process, we first conducted a comprehensive search in biomedical databases of Dox cardiotoxicity transcriptomic studies. Candidate genes were screened for responsiveness to Dox treatment, association with the Notch and Hippo pathways implicated in cardiac repair, and the control of cell death. This systematic approach led to the identification of serine incorporator-3 (serinc- 3). W e hypothesize that in the setting of Dox-induced cardiotoxicity, serinc-3 exhibits protective effects by opposing Top-2β-mediated cardiomyocyte apoptosis that we will explore in 2 Specific Aims . In Aim 1, we will determine the role of serinc-3 in Dox injury in cardiomyocytes and cancer cells. We will (i) dissect the protective role of serinc-3 on apoptosis pathways implicated in Dox cardiotoxicity, (ii) determine an association between serinc-3 and the cardioprotective Notch and Hippo pathways in cardiomyocytes, and (iii) perform transcriptome sequencing and quantification, gene network construction, and determine biochemical and functional pathway enrichment in Dox treatment following serinc-3 expression modulation, (iv) establish protein interactions with serinc-3 using a 3xFLAG tagging strategy, co-immunoprecipitation, and mass spectrometry, and (v) evaluate serinc-3 expression in breast cancer and lymphoma cells, and quantify cancer cell proliferation following Dox treatment and serinc-3 modulation. In Aim 2, we will scrutinize in vivo serinc-3 function in cardiomyocytes following adult mouse Dox treatment. We will (i) develop tamoxifen-inducible, cardiomyocyte-specific Cre-Lox mice with enhanced green fluorescent protein for successful serinc-3 Cre excision, (ii) quantify global (ejection fraction, fractional shortening) and segmental (strain, displacement) cardiac function following serinc-3 Cre-Lox knockdown or adeno-associated virus-9-mediated overexpression in the setting of chronic Dox injury, (iii) assess the effect of serinc-3 overexpression and knockdown on Dox-induced cell death pathways in vivo, and (iv) isolate cardiomyocytes from Dox treated mice to establish correlation matrices by transcriptome sequencing following modulation of serinc-3 expression. The successful implementation of the proposed research will provide novel mechanistic insights into the pathobiology of anthracycline-induced cardiotoxicity, with translational implications.

抽象的 虽然癌症在一生中影响了三分之一的人，但长期生存的改善导致了 癌症治疗的不良心脏副作用事件增加。蒽环类药物等 阿霉素（DOX）是各种癌症的化学疗法的基石，但是，它们的使用变得复杂 蒽环诱导的心脏毒性，最常见的是心肌病。 DOX的机制 心脏毒性是多因素的，包括拓扑异构酶（顶部）-2β介导的DNA损伤，可能最终 在凋亡中。为了确定疾病过程中的新型演员，我们首先进行了全面的搜索 DOX心脏毒性转录组学研究的生物医学数据库。筛选候选基因 对DOX治疗的反应，与心脏修复中实施的缺口和河马途径相关， 以及控制细胞死亡的控制。这种系统的方法导致了丝氨酸Incomporator-3的识别（Serinc-- 3）。 w 假设在DOX诱导的心脏毒性的情况下，Serinc-3表现出保护作用 我们将在2个特定目标中探索的相对2β介导的心肌细胞凋亡 。在AIM 1中，我们将 确定Serinc-3在DOX损伤中的作用在心肌细胞和癌细胞中。我们将（i）剖析受保护的 Serinc-3在DOX心脏毒性中实现的凋亡途径中的作用，（ii）确定 Serinc-3和心肌细胞中的心脏保护缺口和河马途径，（iii）执行转录组 测序和定量，基因网络构建并确定生化和功能途径 Serinc-3表达调制后DOX处理的富集，（iv）与 Serinc-3使用3XFLAG标记策略，共免疫沉淀和质谱法，以及（V）评估 乳腺癌和淋巴瘤细胞中的Serinc-3表达，并在DOX之后定量癌细胞增殖 处理和Serinc-3调节。在AIM 2中，我们将在心肌细胞中仔细检查体内Serinc-3功能 成年小鼠DOX治疗。我们将（i）开发可诱使的，可诱导的，心肌细胞特异性的Cre-lox 具有增强的绿色荧光蛋白的小鼠，用于成功的Serinc-3 CRE惊喜，（ii）量化全球 分数，分数缩短）和节段（应变，位移）心脏功能Serinc-3 Cre-lox之后 在慢性DOX损伤的情况下（III）评估，敲低或与腺相关的病毒-9介导的过表达 Serinc-3过表达和敲低对DOX诱导的细胞死亡途径的影响，（iv）分离株 来自DOX处理的小鼠的心肌细胞，通过转录组测序以下来建立相关物质 Serins-3表达的调节。拟议研究的成功实施将提供新颖 对蒽环类诱导的心脏毒性的病理生物学的机械洞察，具有翻译意义。