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

基本信息

批准号：
10318560
负责人：
Darrell N. Kotton
金额：
$ 67.57万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
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/10318560
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

项目摘要

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.

项目概要/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），需要治疗由于对启动和延续机制的不完全理解而限制了选择疾病。现在越来越多的文献表明肺上皮功能障碍在这些事件中发挥了作用导致下游成纤维细胞激活，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 中肺上皮细胞都表现出端粒缩短，表明肺上皮功能障碍可能引发 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制。然而，无法访问患者特定的人类上皮间质模型系统，测试选择有限基因多态性或端粒酶扰动如何诱导上皮细胞变化的假设可能在机制上有助于 IPF。在这里，我们建议开发一种基于人类类器官的体外 IPF研究模型系统。我们建立了诱导多能干细胞生物样本库由患有散发性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 iPSC 的体外分化来应用此存储库突变（与正常 iPSC 相比）到各种气道和肺泡肺上皮细胞中，目的是生成内在上皮功能障碍的还原论、仅上皮细胞 3D 培养模型我们认为可能引发肺纤维化。在目标 2 中，我们通过以下方式增强了该模型的复杂性：引入由 iPSC 衍生的肺上皮与人类并列组成的人类类器官间充质谱系，以模拟假设的上皮-间质相互作用使 IPF 永久化。最后，在目标 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。