A Patient-Specific hiPSC Model of Nilotinib-Induced Peripheral Artery Disease Pharmacogenomics

尼罗替尼诱导的外周动脉疾病药物基因组学的患者特异性 hiPSC 模型

• 批准号: 10386766
• 负责人: Emily Pinheiro
• 金额: $ 4.25万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10386766
• 关键词: Address; Adverse effects; Affect; Amputation; Atherosclerosis; Biochemical; Biological Assay; Blinded; Blood Pressure; CRISPR/Cas technology; Cardiac Myocytes; Cell Line; Cells; Chronic Myeloid Leukemia; Clinical; Coronary heart disease; Cytogenetics; Data; Development; Disease remission; Drug Exposure; Endothelial Cells; Endothelium; Etiology; Experimental Designs; Future; Gene Expression; Genetic; Genetic Diseases; Genetic Predisposition to Disease; Genetic Screening; Genomics; Goals; Human; Hypertension; In Vitro; Incidence; Individual; Intervention; Investigation; Ischemic Stroke; Literature; Medical Genetics; Methodology; Modeling; Morbidity - disease rate; Myocardial Infarction; Pathologic; Patients; Peripheral arterial disease; Persons; Pharmaceutical Preparations; Pharmacogenomics; Phenotype; Physicians; Predisposition; Quantitative Trait Loci; Research Proposals; Risk; Risk Assessment; Risk Factors; Safety; Scientist; Single Nucleotide Polymorphism; Smooth Muscle Myocytes; System; Technical Expertise; Training; Tyrosine Kinase Inhibitor; United States; Variant; Vascular Smooth Muscle; adverse drug reaction; cardiovascular risk factor; case-by-case basis; causal variant; design; differential expression; disorder risk; drug discovery; effective therapy; experimental analysis; genetic association; genetic variant; genome editing; hypercholesterolemia; in vitro Model; in vitro testing; induced pluripotent stem cell; insight; inter-individual variation; mortality; mortality risk; novel; patient response; patient subsets; pre-doctoral; prevent; response; screening; side effect; skills; stem cell biology; stem cell model; tool; transcriptome; transcriptome sequencing; whole genome

PROJECT SUMMARY Peripheral artery disease (PAD), a subtype of atherosclerosis, affects approximately 8.5 million people in the United States. PAD confers a three-fold risk of all-cause mortality and is comparable to coronary heart disease in increased risk for mortality, myocardial infarction, and ischemic stroke. Nilotinib is a highly effective treatment for chronic myeloid leukemia (CML) but causes pathologic changes in blood pressure consistent with PAD in 26% and 35% of patients on first- and second-line nilotinib respectively. PAD occurs even in nilotinib-treated patients without pre-existing cardiovascular risk factors and in some cases is sufficiently severe to necessitate amputation. Currently no tools exist to understand the mechanism of nilotinib-induced PAD (N-PAD) or preemptively identify which patients may susceptible to this adverse effect, meaning that susceptible patients are identified only after they have developed irreversible complications. Human induced pluripotent stem cells (hiPSCs) constitute a unique and efficient system with which to study interindividual variability in adverse drug reactions. hiPSC derivatives, including cardiomyocytes, endothelial cells, and vascular smooth muscle cells, have previously been shown to recapitulate patient-specific susceptibility to both drug-induced and genetic phenotypes. Because hiPSCs and their derivatives are genetically identical to the patients from whom they are derived, they are well-suited to the study of the pharmacogenomics of N-PAD and can be used to both identify and validate causal variants. These variants will then inform clinical genetic screening as well as mechanistic understanding of N-PAD. In this study we will develop an in vitro model of N-PAD. In Aim 1 we will functionally and biochemically characterize response to nilotinib exposure in hiPSC-derived endothelial cells and vascular smooth muscle cells from patients with and without N-PAD. We predict that these cells will recapitulate patient- specific susceptibility to nilotinib treatment and provide a model with which to probe the mechanism of this effect. In Aim 2 we will assess the gene expression response of hiPSC-derived cells from patients with N-PAD to nilotinib in order to identify novel variants, which will then be validated through CRISPR/Cas9 editing. Accomplishment of these aims will establish an in vitro model for PAD and atherosclerosis in addition to elucidating the mechanism of N-PAD and identifying relevant variants for clinical screening. Additionally, the proposed project will provide a platform for the applicant's predoctoral training and allow for the development of expertise in experimental design and analysis, a broad repertoire of technical skills, and expertise in computational pharmacogenomics approaches while also enhancing clinical and professional skills.

项目摘要 动脉粥样硬化的亚型外周动脉疾病（PAD）影响约850万人 美国。垫子赋予了全因死亡率的三重风险，与冠心病相当 死亡率，心肌梗塞和缺血性中风的风险增加。尼洛替尼是一种高效的治疗方法 对于慢性髓样白血病（CML），但引起与PAD一致的血压的病理变化 一线和二线尼洛替尼的患者分别为26％和35％。垫即使在尼洛蒂尼治疗的 没有预先存在的心血管危险因素的患者，在某些情况下，患者足够严重，需要 截肢。目前尚无了解Nilotinib诱导的PAD（N-PAD）或 先发制地确定哪些患者可能易受这种不良反应的影响，这意味着易感患者 仅在出现不可逆的并发症后才确定。人诱导的多能干细胞 （HIPSC）构成了一个独特而有效的系统，可以研究不良药物的个体变异性 反应。 HIPSC衍生物，包括心肌细胞，内皮细胞和血管平滑肌细胞， 以前已显示出对药物诱导和遗传的患者特异性敏感性概括 表型。因为HIPSC及其衍生物在遗传上与他们的患者相同 得出的，它们非常适合研究N-PAD的药物基因组学，可用于识别 并验证因果变体。然后，这些变体将为临床基因筛查和机械提供信息 对n-pad的理解。在这项研究中，我们将开发N-PAD的体外模型。在AIM 1中，我们将在功能上 并在生物化学上表征了对HIPSC衍生的内皮细胞和血管中尼洛替尼暴露的反应 来自有或没有N-PAD患者的平滑肌细胞。我们预测这些细胞将概括患者 - 对尼洛替尼治疗的特定敏感性，并提供了一个模型，以探测这种作用的机制。 在AIM 2中，我们将评估hipsc衍生细胞从N-PAD患者到的基因表达反应 Nilotinib为了识别新型变体，然后通过CRISPR/CAS9编辑来验证。 这些目标的实现还将建立一个体外模型，除了 阐明N-PAD的机制，并确定相关的临床筛查变体。另外， 拟议的项目将为申请人的专业培训提供一个平台，并允许开发 实验设计和分析方面的专业知识，技术技能的广泛曲目以及专业知识 计算药物基因组学方法同时还提高了临床和专业技能。