The pathology of Pulmonary Hypoplasia (PH) includes reduced lung mass, insufficient ability of the lungs to stay expanded due to a deficiency in surfactant protein production, a poorly differentiated lining of the air sac (alveolar epithelium), and a reduction of gas exchange. Our long term goal is to alleviate neonatal PH using stem cell therapy to augment premature lung tissue with in vitro stem cell derived distal airway cells. The specific hypothesis is that recapitulation of key developmental events by providing transcription and growth factors will optimize derivation of distal airway cells. This includes a 2 step process with derivation of definitive endoderm followed by differentiation into distal airway cells. Furthermore, providing a 3-dimensional scaffold will enhance our yield of distal airway cells by providing important spatial cues. In the lung, distal airway development and integrity is essential for proper gas exchange, surfactant production and survival. The focus of this grant proposal is to explore methods of deriving distal airway cells from embryonic stem cells.
Our specific aims i nclude: 1: Optimize derivation of definitive endoderm cells from murine embryonic stem cells, specifically by manipulating the Wnt and nodal pathway or by providing small molecules. 2: Optimize derivation of distal airway cells through growth factor supplementation, specifically FGF2, 7,10. 3: Optimize spatial cues utilizing 3-dimensional hydrogel scaffolds with potential transplantation in vivo, time permitting.

Public Health Relevance

Preterm delivery with resultant immature lungs (pulmonary hypoplasia) is a major problem in obstetrics and accounts for more than 70% of perinatal mortality. In 2004, 12.5% of births in the United States were preterm. These immature lungs have reduced numbers of mature epithelial cells which results in insufficient surfactant production (a protein used to keep the distal airways open) as well as decreased vasculature. We propose to utilize murine embryonic stem cells to derive distal airway cells for possible utilization in pulmonary hypoplasia. This exploratory study may help identify optimal methods of deriving distal airway cells that can potentially be utilized for treatment of pulmonary diseases such as pulmonary hypoplasia.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL104258-04
Application #
8486333
Study Section
Lung Injury, Repair, and Remodeling Study Section (LIRR)
Program Officer
Blaisdell, Carol J
Project Start
2010-07-01
Project End
2015-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
4
Fiscal Year
2013
Total Cost
$327,249
Indirect Cost
$27,089
Name
University of Connecticut
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
022254226
City
Farmington
State
CT
Country
United States
Zip Code
06030
Wagner, Darcy E; Bonvillain, Ryan W; Jensen, Todd et al. (2013) Can stem cells be used to generate new lungs? Ex vivo lung bioengineering with decellularized whole lung scaffolds. Respirology 18:895-911
Girard, Eric D; Jensen, Todd J; Vadasz, Stephanie D et al. (2013) Automated procedure for biomimetic de-cellularized lung scaffold supporting alveolar epithelial transdifferentiation. Biomaterials 34:10043-55