iPSC-Derived Airway Basal Cells to Model Human Airway Development and Disease

iPSC 衍生的气道基底细胞用于模拟人类气道发育和疾病

基本信息

批准号：
9769850
负责人：
Finn Hawkins
金额：
$ 41.17万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9769850
关键词：
Activities of Daily Living Address Adopted Adult Airway Disease Alveolar Autologous Basal Cell Cell Fate Control Cell Line Cell Therapy Cells Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Data Derivation procedure Development Disease Disease model Engineering Epithelial Epithelial Cells Exhibits FGF10 gene FGFR2 gene Fibroblast Growth Factor Genes Genetic Genetic Models Genetic Transcription Goals Human Human Development In Vitro Individual Lead Lung Lung diseases Maintenance Modeling Morbidity - disease rate Mus Mutation Pathway interactions Pharmaceutical Preparations Play Predictive Value Property Protocols documentation Public Health Reporter Research Role Seminal Signal Pathway Signal Transduction Somatic Cell Stem cells Technology Testing Time Tissues Translating United States airway epithelium base bone cell type drug development human data human disease human model improved in vivo induced pluripotent stem cell injured airway lung development mortality mouse model novel organ growth progenitor programs reconstitution regenerative response self-renewal single-cell RNA sequencing stem stem cell technology tool

项目摘要

Project Summary: Our long-term objective is to improve our understanding of human lung development and disease using iPSC directed differentiations to access cells from the same human being, at various stages of lung development. Basal cells are epithelial stem/progenitor cells of the airway. They exhibit self-renewal and multi-lineage differentiation in vitro and in vivo and can reconstitute the injured airway epithelium. Deriving functional basal cells from induced pluripotent stem cells (iPSCs) would have broad ranging applications for disease modeling, drug development, and potential cell-based therapies. The goal of this proposal is to (1) identify for the first time the signaling pathways that control human airway BC specification, (2) determine the in vitro conditions that support self-renewal and expansion of iPSC-derived BCs, (3) compare the functional and transcriptional profile of these cells to bone-fide adult BCs, and (4) employ these engineered cells to model the monogenic airway disease cystic fibrosis (CF). The central hypothesis is that precise step-wise modulation of developmental pathways can be used to dissect the mechanisms of human basal cell specification, maturation and differentiation and to model airway diseases. We present preliminary data demonstrating the feasibility of deriving BCs from human iPSCs, including functional and single-cell RNA- Seq (scRNA-Seq) characterizations. We introduce for the first time a novel human iPSC reporter line to track and purify putative BCs. Based on mouse studies and our preliminary human data we hypothesize that FGF and Hippo signaling play key roles in human BC specification and subsequent maintenance/amplification of the progenitor pool. We propose a detailed comparison of iPSC-derived stem cells to their in-vivo counterparts to establish their similarity but also harness the information to improve the differentiation protocols for iPSCs.

项目摘要：我们的长期目标是提高我们对人类肺发育和疾病的理解 IPSC指示分化以访问来自同一人的细胞，在肺的各个阶段发展。基底细胞是气道的上皮干/祖细胞。他们表现出自我更新，体外和体内的多条不通分化，可以重建受伤的气道上皮。衍生来自诱导多能干细胞（IPSC）的功能基础细胞将具有广泛的应用程序疾病建模，药物开发和潜在的基于细胞的疗法。该提议的目的是（1）首次确定控制人类气道BC规范的信号通路，（2）确定支持IPSC衍生BC的自我更新和扩展的体外条件，（3）比较这些细胞对骨易于的成年BC的功能和转录曲线，（4）采用这些工程的细胞对单基因气道疾病囊性纤维化（CF）进行建模。中心假设是那样精确开发途径的逐步调制可用于剖析人基底的机制细胞规范，成熟和分化，并模拟气道疾病。我们提出初步数据证明从人IPSC中得出BC的可行性，包括功能性和单细胞RNA- SEQ（SCRNA-SEQ）表征。我们首次介绍了新颖的人类IPSC记者系列跟踪和净化推定的BCS。根据小鼠研究和我们的初步人类数据，我们假设 FGF和HIPPO信号在人类BC规范中起关键作用以及随后的关键作用祖细胞池的维护/扩增。我们提出了IPSC衍生的STEM的详细比较细胞到其体内对应物，以建立它们的相似性，但也利用信息来改善 IPSC的分化协议。