Single-Cell RNA Sequencing of Cardiac Organoids to Determine the Genetic Basis for Cell-Specific Responses to Anticancer Drugs

心脏类器官的单细胞 RNA 测序以确定抗癌药物细胞特异性反应的遗传基础

基本信息

批准号：
10679493
负责人：
Erik McIntire
金额：
$ 4.77万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-30 至 2026-09-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10679493
关键词：
Acute Adverse effects Affect Alleles Anthracycline Antineoplastic Agents Arrhythmia Biological Markers Biological Models Blood Vessels Cancer Patient Cancer Survivor Candidate Disease Gene Cardiac Cardiac Myocytes Cardiotoxicity Cardiovascular Physiology Cardiovascular system Catalogs Cell Culture Techniques Cell Differentiation process Cell Line Cells Cessation of life Classification Data Disease Doxorubicin Drug usage Endoderm Endothelial Cells Environment Fibroblasts Fluorouracil Functional disorder Future Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Genotype Heart Heart failure Heritability Hypertension Individual Intuition Long-Term Effects Malignant Neoplasms Maps Measures Mediating Mesoderm Cell Modeling Morbidity - disease rate Neural Crest Cell Oncology Organoids Pathway interactions Patients Pharmaceutical Preparations Pharmacogenomics Pharmacotherapy Predisposition Primitive foregut structure Procedures Reporting Reproducibility Risk Risk Factors Sample Size Sampling Supporting Cell Symptoms System Testing Time Tissues Toxic effect Transcript Treatment Protocols Variant Vascular Endothelial Cell Vascular System Work anticancer treatment bevacizumab cancer therapy cardiovascular risk factor cell type clinical phenotype differentiation protocol disorder risk effective therapy experimental study genetic architecture genetic variant genotyped patients improved individual patient induced pluripotent stem cell inter-individual variation mortality novel personalized medicine premature response side effect single-cell RNA sequencing stem cells trait transcriptomics

项目摘要

PROJECT SUMMARY Anticancer drug-induced cardiovascular toxicity (CT) is a major side effect for many patients undergoing treatment for oncological disorders. CT symptoms vary widely across individuals, both in presentation and in time to onset. Risk of CT complicates treatment protocols and places cancer patients under additional duress. Genetic background is broadly understood to be a component of CT susceptibility, but specific variants and mechanisms remain largely unknown. To understand the genetic basis for drug-induced CT, we need to understand how anticancer drugs stimulate transcriptomic responses in multiple cardiovascular cell types from individuals of different genetic backgrounds. Inter-individual variability in response to anticancer drugs is mediated by genetic variants that affect gene regulation in a drug-dependent manner (response eQTLs). In other words, genetic variants respond to anticancer drugs by regulating the activity of specific genes. Notably, different cell types can vary in their response eQTLs to anticancer drugs. I propose to determine the genetic basis for transcriptomic responses to the anticancer drugs doxorubicin (DOX), 5-fluorouracil (5-FU), and bevacizumab (BVC) in multiple cardiac cell types from a genetically diverse panel of 70 individuals. Identification of CT-associated response eQTLs necessitates a high-throughput model system comprised of multiple cardiac cell types. For Aim 1 of my proposal, I have developed a culture environment and guided differentiation protocol conducive to cardiac lineage and supporting cell types. This procedure reproducibly transforms induced pluripotent stem cell (iPSC)-derived embryoid bodies (EBs) into cardiac organoids. Preliminary data demonstrate that the cardiac organoids harbor cardiomyocytes, fibroblasts, vascular endothelial cells, and other mesodermal cell types. In Aim 2, I will perform single-cell RNA sequencing (scRNA-seq) on a panel of 70 cardiac organoids cultured in control and drug-treated conditions. Repeating this experiment across multiple individuals will allow me to identify the response eQTLs that regulate how different cardiovascular cell types respond to each drug. In Aim 3, I will quantify gene expression levels and identify response eQTLs that regulate transcriptional changes to each anticancer drug in cardiovascular cell types. Response eQTLs (which are anchored by genotype) provide a catalog of loci that interact either directly or indirectly with the treatment. These response eQTLs may reveal specific genes and pathways important for normal cardiovascular function. Elucidating the genetic architecture underlying CT risk will provide intuition on cardiotoxic mechanisms and associated genes and inform future studies that aim to classify individual patient susceptibility.

项目概要抗癌药物引起的心血管毒性（CT）是许多接受化疗的患者的主要副作用治疗肿瘤疾病。 CT 症状因人而异，无论是表现形式还是表现形式发病时间。 CT 的风险使治疗方案变得复杂，并使癌症患者面临额外的压力。人们广泛认为遗传背景是 CT 易感性的一个组成部分，但特定变异和机制仍然很大程度上未知。为了了解药物诱导 CT 的遗传基础，我们需要了解抗癌药物如何刺激多种心血管细胞类型的转录组反应不同遗传背景的个体。抗癌药物反应存在个体差异由以药物依赖性方式影响基因调控的遗传变异介导（反应 eQTL）。在换句话说，基因变异通过调节特定基因的活性来响应抗癌药物。尤其，不同的细胞类型对抗癌药物的 eQTL 反应可能有所不同。我建议确定遗传抗癌药物阿霉素（DOX）、5-氟尿嘧啶（5-FU）和贝伐珠单抗 (BVC) 对来自 70 名遗传多样性个体的多种心脏细胞类型进行了研究。 CT 相关响应 eQTL 的识别需要一个高通量模型系统，其中包括多种心肌细胞类型。对于我提案的目标 1，我开发了一种文化环境并指导有利于心脏谱系和支持细胞类型的分化方案。此过程可重复将诱导多能干细胞 (iPSC) 衍生的胚状体 (EB) 转化为心脏类器官。初步数据表明，心脏类器官含有心肌细胞、成纤维细胞、血管细胞内皮细胞和其他中胚层细胞类型。在目标 2 中，我将进行单细胞 RNA 测序 (scRNA-seq) 对在对照和药物处理条件下培养的 70 个心脏类器官进行分析。重复这个针对多个个体的实验将使我能够识别响应 eQTL，从而调节如何不同的心血管细胞类型对每种药物有反应。在目标 3 中，我将量化基因表达水平并识别调节心血管细胞中每种抗癌药物转录变化的反应 eQTL 类型。响应 eQTL（由基因型锚定）提供了直接相互作用的基因座目录或间接与治疗有关。这些反应 eQTL 可能揭示对某些重要的特定基因和途径心血管功能正常。阐明 CT 风险背后的遗传结构将为以下方面提供直觉：心脏毒性机制和相关基因，并为未来旨在对个体患者进行分类的研究提供信息易感性。