Modeling Susceptibility to Chemotherapy-Induced Cardiotoxicity Using Human iPSCs

使用人类 iPSC 模拟化疗引起的心脏毒性的易感性

• 批准号: 8904716
• 负责人: Joseph C. Wu
• 金额: $ 48.52万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-05 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8904716
• 关键词: Acute; Adult; Adverse effects; Affect; Aftercare; Animal Model; Animals; Anthracyclines; Antineoplastic Protocols; Biochemical; Biological; Biological Assay; Black Panther; Breast Cancer Patient; Breast Cancer Treatment; British Columbia; Calcium; Cancer Patient; Cardiac; Cardiac Myocytes; Cardiology; Cardiotoxicity; Cardiovascular Agents; Cardiovascular Diseases; Cell Line; Cell Survival; Clinic; Collaborations; Communities; Complication; Data; Dilated Cardiomyopathy; Dose; Doxorubicin; Familial Hypertrophic Cardiomyopathy; Future; Generations; Genetic; Genetic Markers; Genetic Transcription; Genomics; Gold; Health; Heart failure; Human; Image; In Vitro; Incidence; Individual; Knowledge; Lead; Left Ventricular Ejection Fraction; Long QT Syndrome; Long-Term Survivors; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Maps; Measures; Medicine; Methods; Mitochondria; Modeling; Molecular; National Heart, Lung, and Blood Institute; Patients; Pediatrics; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Physiological; Pilot Projects; Predisposition; Prevention; Production; Quantitative Trait Loci; Reactive Oxygen Species; Recovery; Recruitment Activity; Report (document); Resistance; Risk; Sampling; Science; Single Nucleotide Polymorphism; Somatic Cell; Source; Syndrome; System; Technology; Timothy syndrome; Toxic effect; Transcription Coactivator; Translational Research; Treatment Protocols; Work; chemotherapy; clinically relevant; cohort; experience; genome editing; high risk; human old age (65+); improved; in vitro Model; individualized medicine; induced pluripotent stem cell; interest; leukemia/lymphoma; malignant breast neoplasm; new technology; novel; nuclease; oncology; patch clamp; personalized medicine; pluripotency; prevent; professor; response; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): Doxorubicin is a well-established and highly effective chemotherapy drug commonly used to treat multiple cancers such as lymphoma, leukemia, ovary, lung, and breast cancer, but its use is limited by cardiotoxicity. Cardiotoxicity can range from asymptomatic reduction in left ventricular ejection fraction to highly symptomatic heart failure (Class III to IV). Acute doxorubicin-induced cardiotoxicity (DIC) occurs in ~11% of patients and long-term cardiotoxic side effects, which can manifest up to 10 years after treatment, are observed in up to 36% of patients. Currently, we cannot predict which patients will develop cardiotoxicity. A major hurdle in filling the significant gaps in our knowledge about the mechanisms of DIC and how best to prevent it is the lack of good human models, due to the inaccessibility of patient-specific human cardiomyocyte samples, and the difficulty in isolating and maintaining human cardiomyocytes in vitro. Animal models are limited by significant functional disparities between animal and human cardiomyocytes. This hurdle has now been overcome by the recent advances in the generation of human induced pluripotent stem cells (iPSCs), in which a patient's somatic cells can be reprogrammed to pluripotency and maintained indefinitely in vitro. These iPSCs can then be efficiently differentiated into iPSC-derived cardiomyocytes (iPSC-CMs) and further studied in detail. In our preliminary studies, we have developed and validated a set of tools for assessing DIC in human iPSC-CMs. We have established that iPSC-CMs, derived from patients who have developed DIC, accurately recapitulate the susceptibility phenotype in vitro. Single nucleotide polymorphism (SNP) studies have identified several SNPs that are predicted to be highly associated with DIC (P=10-9 to 10-5). Hence in Aim 1, we will generate iPSC lines from 100 cancer patients treated with doxorubicin, 50 of whom experienced cardiotoxicity and 50 did not. In Aim 2, we will use the assays established in our pilot studies to assess the susceptibility to DIC and perform RNA-seq and eQTL mapping to discover novel SNPs. In Aim 3, we will introduce a very well established DIC-related SNP into five control iPSC lines using transcription activator-like effector nucleases (TALENs) and assess the effect on DIC susceptibility. Hence the overall aim of this R01 proposal is to use patient-specific iPSC-CMs to help elucidate the molecular mechanisms of DIC.

描述（由申请人提供）：阿霉素是一种公认​​且高效的化学疗法药物，通常用于治疗多种癌症，例如淋巴瘤，白血病，卵巢，肺癌和乳腺癌，但其使用受心脏毒性的限制。心脏毒性的范围从左心室射血分数的无症状减少到高度症状的心力衰竭（III级至IV）。急性阿霉素诱导的心脏毒性（DIC）发生在约11％的患者中，长期心脏毒性副作用可在多达36％的患者中观察到可以表现出长达10年后的长期副作用。目前，我们无法预测哪些患者会发展出心脏毒性。由于患者特异性人类心肌细胞样本的无法访问，以及在隔离和维持体内体内的人类心肌细胞的困难，填补我们对DIC机制的了解以及如何最好地预防它的知识的重大差距是一个重大障碍。动物模型受动物和人类心肌细胞之间的显着功能差异的限制。现在，通过人类诱导的多能干细胞（IPSC）的最新进展克服了这一障碍，其中患者的体细胞可以重编程为多能性并无限期地维持体外。然后可以将这些IPSC有效地分化为IPSC衍生的心肌细胞（IPSC-CMS），并详细研究。在我们的初步研究中，我们开发了并验证了一组评估人IPSC-CM中DIC的工具。我们已经确定，源自开发DIC的患者的IPSC-CMS准确地概括了体外的敏感性表型。单核苷酸多态性（SNP）研究已经鉴定出了几种与DIC高度相关的SNP（p = 10-9至10-5）。因此，在AIM 1中，我们将从100例接受阿霉素治疗的癌症患者中生成IPSC系，其中50例患有心脏毒性，而50例则没有。在AIM 2中，我们将使用试点研究中建立的测定方法来评估DIC的敏感性并执行RNA-Seq和EQTL映射以发现新型SNP。在AIM 3中，我们将使用转录激活剂样效应子核酸酶（Talens）（TALENS）将非常合适的DIC相关SNP引入五个对照IPSC线，并评估对DIC敏感性的影响。因此，该R01提案的总体目的是使用患者特异性的IPSC-CMS来帮助阐明DIC的分子机制。