Developing a patient-specific organoid model of pulmonary fibrosis using iPSCs

使用 iPSC 开发患者特异性肺纤维化类器官模型

• 批准号: 10026360
• 负责人: Darrell N. Kotton
• 金额: $ 50.86万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10026360
• 关键词: ABCA3 gene; Aging; Basal Cell; Biological Assay; Biological Models; Cells; Collaborations; Data; Development; Disease; Engineering; Epithelial; Epithelial Cells; Event; Fibroblasts; Fibrosis; Functional disorder; Genes; Genetic Polymorphism; Genetic Recombination; Goals; Human; Hybrids; In Vitro; Individual; Interstitial Lung Diseases; Interstitial Pneumonia; Lead; Literature; Lung; MUC5B gene; Mesenchymal; Mesenchyme; Modeling; Morbidity - disease rate; Mutation; Myofibroblast; Organoids; Pathogenesis; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Play; Publishing; Pulmonary Fibrosis; Recombinants; Reporter; Role; Sampling; Source; System; TP53 gene; Telomerase; Telomere Shortening; Testing; Tissues; alveolar epithelium; base; biobank; cell repository; disorder risk; fibrogenesis; genetic variant; genome wide association study; high-throughput drug screening; idiopathic pulmonary fibrosis; in vitro Model; in vivo; induced pluripotent stem cell; knock-down; lung basal segment; mitochondrial dysfunction; mortality; mutant; novel; novel therapeutics; open source; overexpression; repository; response; scale up; single-cell RNA sequencing; telomere; three dimensional cell culture

Project Summary/Abstract Idiopathic pulmonary fibrosis (IPF) remains a deadly interstitial lung disease (ILD) with treatment options limited by an incomplete understanding of the mechanisms that initiate and perpetuate disease. A growing literature now implicates lung epithelial dysfunction as playing a role in the events that lead to downstream fibroblast activation, culminating in relentless fibrosis. These studies, together with the observation that lung epithelial cells in all forms of IPF display shortened telomeres, suggests that lung epithelial dysfunction may initiate IPF, and accelerated aging phenotypes or telomerase pathway abnormalities likely contribute to this pathogenesis. However, without access to patient-specific human epithelial-mesenchymal model systems, there are limited options for testing hypotheses of how epithelial changes induced by gene polymorphisms or telomerase perturbations might mechanistically contribute to IPF. Here we propose to develop a human organoid-based in vitro model system for the study of IPF. We have established a biorepository of induced pluripotent stem cells (iPSCs) generated from individuals with sporadic or familial pulmonary fibrosis. In aim 1 we apply this repository by directing the in vitro differentiation of banked IPF iPSCs carrying telomerase mutations (vs normal iPSCs) into various airway and alveolar lung epithelial cells for the purpose of generating a reductionist, epithelial-only 3D culture model of the intrinsic epithelial dysfunction that we posit may initiate pulmonary fibrosis. In aim 2 we augment the complexity of this model by introducing human organoids composed of iPSC-derived lung epithelia juxtaposed with human mesenchymal lineages in order to model the epithelial-mesenchymal interactions hypothesized to perpetuate IPF. Finally, in aim 3 we test the hypothesis that different telomerase pathway mutations result in shared lung epithelial perturbations, including short telomeres and p53 activation, that then leads to downstream mesenchymal activation.

项目总结/摘要 特发性肺纤维化（IPF）仍然是一种致命的间质性肺病（ILD）， 由于对启动和延续的机制不完全了解，选择受到限制 疾病现在越来越多的文献暗示肺上皮功能障碍在这些事件中发挥作用 导致下游成纤维细胞活化最终导致无情的纤维化这些研究， 连同观察到所有形式的IPF中的肺上皮细胞显示缩短的端粒， 提示肺上皮功能障碍可能引发IPF，加速老化表型或 端粒酶途径异常可能有助于这种发病机制。然而，由于无法获得 患者特异性人上皮-间充质模型系统，测试的选择有限 基因多态性或端粒酶扰动如何诱导上皮变化的假说 可能在机制上导致IPF。在这里，我们建议开发一种基于人类类器官的体外 用于IPF研究的模型系统。我们建立了诱导多能干细胞的生物贮藏库 细胞（iPSC），其产生自具有散发性或家族性肺纤维化的个体。在目标1中， 通过指导携带端粒酶的库存IPF iPSC的体外分化来应用该储存库 突变（相对于正常iPSC）到各种气道和肺泡肺上皮细胞中， 产生固有上皮功能障碍的简化的仅上皮3D培养模型， 我们可能会引发肺纤维化。在aim 2中，我们通过以下方式增加了该模型的复杂性： 引入由iPSC衍生的肺上皮与人类器官并置组成的人类器官 间充质谱系，以模拟上皮-间充质相互作用， 使IPF永久化。最后，在aim 3中，我们验证了不同端粒酶途径突变 导致共同的肺上皮扰动，包括短端粒和p53激活，然后 导致下游间充质活化。