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.

Public Health Relevance

The proposed project is of relevance to public health because lung diseases are a major cause of morbidity and mortality in the United States. Upon conclusion of this project we will understand the mechanism of basal cell specification and differentiation and have developed a tool, an iPSC-derived lung stem cell, with broad reaching implications for our understanding of human lung disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL139799-01A1
Application #
9596528
Study Section
Lung Injury, Repair, and Remodeling Study Section (LIRR)
Program Officer
Lin, Sara
Project Start
2018-09-01
Project End
2023-08-31
Budget Start
2018-09-01
Budget End
2019-08-31
Support Year
1
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Boston University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
Hawkins, Finn J; Kotton, Darrell N (2018) Pulmonary Ionocytes Challenge the Paradigm in Cystic Fibrosis. Trends Pharmacol Sci 39:852-854