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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

基本信息

批准号：
10059530
负责人：
THOMAS QUERTERMOUS
金额：
$ 8.36万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-05 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10059530
关键词：
ATAC-seq Acute Adult Anthracycline Antineoplastic Protocols Arrhythmia Breast Cancer Patient Breast Cancer Treatment CRISPR screen CRISPR/Cas technology Calcium Cancer Patient Cardiac Cardiac Myocytes Cardiotoxicity Cardiovascular Diseases Cell Survival Cells ChIP-seq Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Congestive Development Down-Regulation Doxorubicin Drug Screening Drug Targeting Elements Epigenetic Process Exhibits Gene Expression Profile Genes Genetic Genetic Transcription Genome Genomics Grant Heart Heart failure Histones Homeostasis Human Impairment Journals Knowledge Left Left Ventricular Ejection Fraction Libraries Malignant Neoplasms Mammalian Cell Maps Mass Spectrum Analysis Measures Mediating Medicine Metabolic Metabolism Methods Mitochondria Modeling Molecular Nature Paper Patients Pharmaceutical Preparations Pharmacogenomics Phenotype Predisposition Publishing Qi Quantitative Trait Loci RNA Interference Reactive Oxygen Species Recovery Regulator Genes Research Personnel Role Single Nucleotide Polymorphism Stress Techniques Technology Testing Therapeutic Time Topoisomerase II Topoisomerase II inhibition Treatment Efficacy Tyrosine Kinase Inhibitor Up-Regulation Ventricular base cancer type chemotherapy clinical phenotype cost efficient design and construction disease phenotype effective therapy experience experimental study gene function genome editing genome-wide improved in vitro Model induced pluripotent stem cell interest new therapeutic target novel screening side effect stem cells therapeutic target transcriptome transcriptome sequencing

项目摘要

PROJECT SUMMARY Doxorubicin is a well-established and highly effective chemotherapy drug commonly used to treat multiple cancer types, 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 are observed in up to 36% of patients. However, the underlying mechanisms of DIC remain largely unknown, hampering the development of effective therapeutics for DIC. To that end, in this proposal we aim to use state-of-the-art approaches in genomics and epigenetics to identify the genetic and molecular mechanisms of DIC. In Aim 1, we will generate iPSC lines from 100 cancer patients treated with doxorubicin, 50 of whom experienced cardiotoxicity and 50 did not. From these lines, we will perform RNA-seq and eQTL mapping to discover novel single nucleotide polymorphisms (SNPs) responsible for DIC, and the relevant SNPs will be introduced to or deleted from non- DIC or DIC patient iPSC lines, respectively, with the CRISPR gene editing technique. We will investigate the functional and transcriptional changes and determine whether the identified SNP is responsible for the disease phenotype. In Aim 2, we will perform a combination of epigenetic techniques in ATAC-seq, ChIP-seq, and IP- mass spectrometry to identify genes regulated by topoisomerase II-beta (TOP2B), a transcriptional regulator known to be inhibited by doxorubicin. We hypothesize that TOP2B controls genes critical for cardiomyocyte contraction, metabolism, and homeostasis, the expression of which is disrupted upon TOP2B inhibition by doxorubicin. Results from these experiments will reveal the specific genes regulated by TOP2B, which can be used as potential therapeutic targets of DIC. In Aim 3, we will identify genes suitable as drug targets that are related to DIC using the CRISPR genome screening approach. This novel technique offers a unique and cost- efficient opportunity to systemically screen for drug target genes dysregulated by doxorubicin treatment in iPSC- CMs. Using patient-specific iPSC-CMs, the proposed aims will allow us to elucidate for the first time the genetic and molecular basis for DIC.

项目总结阿霉素是一种久负盛名的高效化疗药物，常用于治疗多发性癌症类型，但其使用受到心脏毒性的限制。心脏毒性可从左侧无症状的减少心室射血分数到高度症状性心力衰竭(III至IV级)。急性阿霉素所致约11%的患者发生心脏毒性(DIC)，高达36%的患者观察到长期心脏毒性副作用病人的数量。然而，DIC的潜在机制在很大程度上仍然未知，阻碍了DIC的发展治疗DIC的有效疗法。为此，在这项提案中，我们旨在使用最先进的方法来基因组学和表观遗传学，以确定DIC的遗传和分子机制。在目标1中，我们将生成来自100名接受阿霉素治疗的癌症患者的IPSC株，其中50人经历了心脏毒性，50人发生了不。从这些品系中，我们将进行rna-seq和eqtl定位，以发现新的单核苷酸。导致DIC的多态(SNP)，相关SNP将引入或删除非DIC DIC或DIC患者的IPSC系，分别采用CRISPR基因编辑技术。我们将调查功能和转录变化，并确定已识别的SNP是否导致疾病表型。在目标2中，我们将在atac-seq、Chip-seq和ip-seq中执行表观遗传学技术的组合。用质谱法鉴定转录调节因子拓扑异构酶II-β调控基因已知可被阿霉素抑制。我们假设TOP2B控制着对心肌细胞至关重要的基因收缩、新陈代谢和动态平衡，其表达在TOP2B被抑制时被破坏阿霉素。这些实验的结果将揭示TOP2B调控的特定基因，这可能是可作为DIC的潜在治疗靶点。在目标3中，我们将识别适合作为药物靶点的基因使用CRISPR基因组筛选方法与DIC相关。这项新技术提供了一种独特的成本- 在IPSC中系统筛选阿霉素治疗失调的药物靶基因的有效机会- 不育系CMS。使用患者特定的IPSC-CMS，拟议的目标将使我们能够首次阐明基因和DIC的分子基础。