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.

项目摘要/摘要 特发性肺纤维化（IPF）仍然是一种致命的间质性肺疾病（ILD） 选项受到对启动和永久机制的不完全理解的限制 疾病。现在越来越多的文献暗示肺上皮功能障碍是在事件中发挥作用 这导致下游成纤维细胞激活，最终导致无端纤维化。这些研究， 以及以下观察结果，即各种形式的IPF肺上皮细胞显示出缩短的端粒， 表明肺上皮功能障碍可能启动IPF，并加速衰老表型或 端粒酶途径异常可能导致这种发病机理。但是，没有访问 特定于患者的人类上皮 - 间质模型系统，测试选择有限 假设基因多态性或端粒酶扰动引起的上皮上皮变化如何 可能在机械上有助于IPF。在这里，我们建议开发基于人体器官的体外 IPF研究的模型系统。我们已经建立了诱导多能茎的生物座 来自零星或家族性肺纤维化的个体产生的细胞（IPSC）。在目标1中我们 通过指导携带端粒酶的库IPF IPF的体外差异来应用此存储库 突变（与正常的IPSC）进入各种气道和肺泡上皮细胞 产生还原性的，仅上皮的3D培养模型的内在上皮功能障碍 我们认为可能启动肺纤维化。在AIM 2中，我们通过 引入由IPSC衍生的肺上皮组成的人类器官与人并列 间充质谱系为了模拟假设的上皮 - 间质相互作用 永久性IPF。最后，在AIM 3中，我们检验了不同端粒酶途径突变的假设 导致共同的肺上皮扰动，包括短端粒和p53激活，然后 导致下游间充质激活。