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: 10525231
• 负责人: Darrell N. Kotton
• 金额: $ 65.39万
• 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/10525231
• 关键词: ABCA3 gene; Acceleration; Aging; Basal Cell; Biological Assay; Biological Models; Cells; Collaborations; Data; Development; Disease; Engineering; Epithelial Cells; Epithelium; Event; Fibroblasts; Fibrosis; Functional disorder; Genes; Genetic Polymorphism; Genetic Recombination; Goals; Human; Hybrid Cells; In Vitro; Individual; Interstitial Lung Diseases; Interstitial Pneumonia; Literature; Lung; MUC5B gene; Mesenchymal; Mesenchyme; Modeling; Morbidity - disease rate; Mutation; Myofibroblast; Organoids; Pathogenesis; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Publishing; Pulmonary Fibrosis; Recombinants; Reporter; Sampling; Secretory Cell; Source; System; TP53 gene; Telomerase; Telomere Shortening; Testing; Tissues; alveolar epithelium; 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/文摘