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

• 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
• 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

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相关的药物毒性已得到充分证实。然而，在这方面， 基于单克隆抗体的免疫检查点引起的心脏不良反应的新报告 抑制剂（ICI）是一个日益关注的问题。这些ICI作用于程序性细胞死亡蛋白1（PD-1）受体或 其配体程序性细胞死亡配体1（PD-L1）增强抗肿瘤反应，这些通常导致 免疫相关不良事件（iRAE）。几项研究指出，肿瘤之间的同源性 抗原和心脏蛋白作为结果的原因之一，如心肌炎，心脏的变化， 心律失常和血管炎。 由于罕见，早期检测iRAE和患者样本稀缺。ICI诱导的细胞凋亡机制 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 具有免疫细胞的iPSC衍生的3-D心脏类器官（CO）可用于对ICI诱导的心脏类器官进行建模。 损伤用还原论方法揭示ICI治疗引起心脏损伤的疾病机制 可能导致发现新的靶点，可以为免疫疗法提供心脏保护。为了 为了实现这一目标，我确定了三个具体目标：（1）我将通过自组装生成CO。 iPSC-心肌细胞、iPSC-内皮细胞和iPSC-心脏成纤维细胞，并测试两种常用的ICI 派姆单抗（PD-1抑制剂）和阿特珠单抗（PD-L1抑制剂）。我将评估CO的变化 在存在和不存在用于iPSC的外周血单核细胞（PBMC）时的功能 衍生（2）我将在单细胞水平上对CO进行深入的分子分析，并确定潜在的 目标负责超免疫反应可以心脏功能障碍（3）最后，我将使用尖端 基因组编辑技术，以敲除有助于在ICI后拯救CO功能的基因靶标 治疗目标2中确定的靶点将通过向小鼠注射ICI在小鼠模型中再次验证 不同的免疫背景，然后评估心脏功能和分子变化， 组织. 在这个多学科的项目中，在我尊敬的导师，顾问和 与合作者，我相信我将接受卓越的培训，以完成科学和职业生涯 发展目标鉴于我迄今为止的科学记录，以及通过这一点寻求的补充培训， K99/R 00奖，一起将帮助我发展和领导几个研究领域的建设走向成功 独立的职业生涯。