Generation & Characterization of a Lung Stem Cell “Tool Kit” to Generate Functional Lung Epithelial Lineages

Project Title: Generation & Characterization of a Lung Stem Cell “Tool Kit” to Generate Functional Lung Epithelial Lineages

Ancillary Study Principal Investigator: Paul Gadue Ph.D. and Darrell Kotten, M.D.

Hub Site Principal Investigator: Edward Morrisey, Ph.D.

Awarded Organizations: Trustees of the University of Pennsylvania

Abstract:

The goal of this proposal is the development of methods that will allow the derivation of pure populations of specified proximal and distal lung epithelial lineages from hPSCs, including lung disease-specific iPSCs. We propose a collaboration between eminent labs focused on early lung endoderm differentiation (Kotton and Gadue) and lung development and regeneration (Morrisey) to exploit recent technologies to generate a lung stem cell “tool kit” that can be used by investigators in the PCBC as well as other labs interested in lung stem cell/regenerative biology. Extending our published history of generating reporter tools in mouse PSCs (Longmire et al., 2012), this proposed set of tools for human PSCs will consist of fluorescent reporters for both multipotent lung endoderm progenitor cells (NKX2.1, SOX2) and reporters for functionally important lineages of the proximal (SCGB1A1/SCGB3A2) and distal (SFTPC) lung epithelium. This new set of reporters will be used to optimize current methods for derivation of lung epithelial lineages from hPSCs and define the importance of WNT signaling in this process. The advantage of these markers is that excellent mouse models have been generated that allow for isolation and lineage fate mapping of cells expressing these genes (Morrisey and Hogan, 2010). This provides a useful and rigorous side-by-side comparison between mouse and human lung endoderm progenitor cells during differentiation, which is critical to determine if the cell populations generated in vitro mirror cells in vivo. Importantly, this group of investigators has extensive experience in lung endoderm development and regeneration as well as WNT signaling making this a unique opportunity to explore these questions. This ancillary grant partners with Dr. Morrisey’s PCBC project focused on the role of WNT signaling in cardiac progenitor expansion and regeneration.