Unraveling Adverse Effects of Checkpoint Inhibitors Using iPSC-derived Cardiac Organoids

使用 iPSC 衍生的心脏类器官揭示检查点抑制剂的副作用

基本信息

批准号：
10591918
负责人：
Dilip Thomas
金额：
$ 12.46万
依托单位：
STANFORD UNIVERSITY
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：
2023-01-13 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10591918
关键词：
3-Dimensional Adverse effects Affinity Animal Model Antibodies Antitumor Response Area Award Binding Biological Biomedical Engineering Biopsy CRISPR interference Cancer Patient Cardiac Cardiac Myocytes Cardiotoxicity Cardiovascular Models Cardiovascular system Cell Communication Cell Line Cell Proliferation Cells Chromatin Clinical Clinical Treatment Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Collaborations Cytometry D Cells Derivation procedure Disease Dose Early Diagnosis Early identification Echocardiography Electrophysiology (science)Endothelial Cells Evaluation Event Exposure to Female Fibroblasts Frequencies Functional disorder Gene Targeting Genomics Goals Half-Life Heart Heart Diseases Heart Injuries Histology Immune Immune checkpoint inhibitor Immune response Immune system Immunological Models Immunology Immunotherapy In Vitro Incidence Inflammation Lead Ligands MALAT1 gene Malignant Neoplasms Measures Mediating Mentors Molecular Molecular Analysis Molecular Profiling Molecular Target Monoclonal Antibodies Mus Myocardial dysfunction Myocarditis Organoids Outcome PDL1 inhibitors Pathway interactions Patients Peripheral Blood Mononuclear Cell Pharmaceutical Preparations Phase Phenotype Physiological Proteins Proteomics Qi Reaction Reporting Research Research Design Sampling Structure Symptoms Techniques Technology Testing Therapeutic Tissue Engineering Tissues Toxic effect Training Tumor Antigens Validation Vasculitis cardioprotection career career development cytokine drug development drug efficacy endothelial stem cell experimental study functional outcomes genome editing heart function heart preservation heart rhythm immune activation immune cell infiltrate immune-related adverse events in vivo induced pluripotent stem cell induced pluripotent stem cell technology inhibitor interdisciplinary approach knock-down male monolayer mouse model multidisciplinary pembrolizumab programmed cell death ligand 1 programmed cell death protein 1 receptor response self assembly single-cell RNA sequencing small molecule stem cell biology transcriptome sequencing transcriptomics

项目摘要

Project Summary Cardiotoxicities associated with small molecule cancer therapeutics are well documented. However, emerging reports of adverse cardiac reactions due to monoclonal antibody-based immune checkpoint inhibitors (ICIs) is a growing concern. These ICIs act on programmed cell death protein 1 (PD-1) receptor or its ligand programmed cell death-ligand 1 (PD-L1) enhancing antitumor response, and these often lead to such immune-related adverse events (iRAEs). Several studies point toward the homology between tumor antigens and cardiac proteins as one of the reasons for outcomes such as myocarditis, changes in cardiac rhythm, and vasculitis. Due to the rare, early detection iRAEs and scarcity of patient samples. The mechanism of ICI-induced cardiac disease remains elusive. Hence, utilizing a unique multi-disciplinary approach and expertise in induced pluripotent stem cell (iPSC) technology, immunology and bioengineering strategies, I hypothesize that, iPSC-derived 3-D cardiac organoids (COs) with immune cells can be used to model ICI-induced cardiac injury. Unraveling the disease mechanism of cardiac injury due to ICI treatment using a reductionist approach may lead to discovery of new targets that can confer cardioprotection for immunotherapies. In order to accomplish this goal, I have defined three specific aims: (1) I will generate COs through self-assembly of iPSC-cardiomyocytes, iPSC-endothelial cells and iPSC-cardiac fibroblasts and test two commonly used ICIs Pembrolizumab (PD-1 inhibitor) and Atezolizumab (PD-L1 inhibitor). I will for assess for changes in CO function in the presence and absence of peripheral blood mononuclear cells (PBMCs) used for iPSCs derivation (2) I will conduct in-depth molecular analyses of the COs at a single cell level and identify potential targets responsible to hyper immune response can cardiac dysfunction (3) Finally, I will use cutting-edge genome editing techniques to knockdown the gene targets that help rescue the CO function after ICI treatment. The targets identified in Aim 2 will be validated again in mouse models by injecting ICIs into mice with diverse immune backgrounds followed by evaluation of heart function and molecular changes within the tissue. In this multidisciplinary project, with the support from my esteemed panel of mentors, advisors and collaborators, I am confident that I will receive par excellence training to accomplish both scientific and career development goals. Given my scientific track record to date, and complementary training sought through this K99/R00 award, together will help me develop and lead several research areas building toward a successful independent career.

项目摘要与小分子癌症治疗剂有关的心脏毒性已充分记录。然而，基于单克隆抗体的免疫检查点引起的不良心脏反应的新兴报告抑制剂（ICIS）越来越关注。这些ICI对程序性细胞死亡蛋白1（PD-1）受体或它的配体编程的细胞死亡配体1（PD-L1）增强了抗肿瘤反应，这些反应通常会导致这种与免疫相关的不良事件（IRAE）。几项研究指出肿瘤之间的同源性抗原和心脏蛋白作为诸如心肌炎等结局的原因之一，心脏变化节奏和血管炎。由于罕见的早期检测和患者样本的稀缺性。 ICI诱导的机制心脏病仍然难以捉摸。因此，利用独特的多学科方法和专业知识诱导多能干细胞（IPSC）技术，免疫学和生物工程策略，我假设这是，具有免疫细胞的IPSC衍生的3-D心脏器官（COS）可用于建模ICI诱导的心脏受伤。使用还原主义方法来阐明因ICI治疗而导致心脏损伤的疾病机制可能导致发现可以为免疫疗法提供心脏保护的新靶标。为了完成这个目标，我定义了三个具体目标：（1）我将通过自我组装来生成cos IPSC-毛肌细胞，IPSC-内皮细胞和IPSC-cardiac成纤维细胞，并测试两个常用的ICI pembrolizumab（PD-1抑制剂）和atezolizumab（PD-L1抑制剂）。我将评估CO的更改在存在和不存在用于IPSCS的外周血单核细胞（PBMC）的情况下的功能推导（2）我将在单个细胞水平上对COS进行深入的分子分析，并确定电位负责超级免疫反应的靶标可能会心脏功能障碍（3）最后，我将使用尖端基因组编辑技术以敲除帮助挽救ICI后CO功能的基因靶标治疗。 AIM 2中确定的目标将在鼠标模型中再次验证，通过将ICI注射到小鼠中具有不同的免疫背景，然后评估心脏功能和分子变化组织。在这个多学科项目中，在我尊敬的导师，顾问和合作者，我有信心我将获得卓越的培训，以完成科学和职业发展目标。鉴于我迄今为止的科学记录，以及通过此培训寻求的补充培训 K99/R00奖，将共同帮助我开发并领导几个研究领域，建立成功独立职业。