Increased survival of very premature infants has been accompanied by an increased incidence of bronchopulmonary dysplasia (BPD). In the "new BPD," alveolar septa are thickened with collagen and 1-smooth muscle actin-positive, transforming growth factor (TGF)-?-positive myofibroblasts. We have isolated mesenchymal stromal cells from the tracheal aspirates of premature infants. These cells produce TGF-?1 and undergo TGF-?-induced myofibroblastic differentiation, suggesting that, in the absence of other signals, myofibroblastic differentiation represents the "default program" for mesenchymal stromal cell specialization. Mesenchymal stromal cells also appear to be potent biomarkers for the development of BPD;accounting for the potential influences of gender, birth weight and gestational age, isolation of mesenchymal stromal cells increased the adjusted odds ratio of BPD over 31-fold. Finally, our microarray data show that stromal cells express high levels of mRNAs encoding matricellular proteins, a subfamily of non-structural extracellular matrix proteins that regulate cell-matrix interactions and fiber deposition. Together, these data support our general hypothesis that mesenchymal stromal cell myofibroblastic differentiation and matricellular protein expression play critical roles in BPD pathogenesis. To test this hypothesis, we propose the following Specific Aims: 1. Correlate neonatal lung mesenchymal stromal cell matricellular protein expression with clinical outcomes. We hypothesize that: i) mesenchymal stromal cells maintain a stable pro-fibrotic phenotype of matricellular protein expression that is coupled with myofibroblastic differentiation;and (ii) expression of the matricellular proteins connective tissue growth factor (CTGF), SPARC and periostin predicts BPD development. 2. Examine the effects of matricellular protein expression on neonatal lung mesenchymal stromal cell proliferation and differentiation, as well as the roles of glycogen synthase kinase (GSK)-3? and ?-catenin in myofibroblastic differentiation in vitro. We hypothesize that (i) matricellular proteins induce mesenchymal stromal cell proliferation and are required for maximal TGF-?-induced myofibroblastic differentiation;and (ii) myofibroblastic differentiation is dependent on Nox4/GSK-3?/?-catenin signaling. 3. Examine the expression, localization, requirement and sufficiency of matricellular proteins and their downstream signaling intermediates for the BPD phenotype in vivo. We hypothesize that: i) fibroblast-specific expression of CTGF is sufficient for the BPD phenotype;(ii) SPARC and periostin are required for hyperoxia-induced hypoalveolarization and interstitial fibrosis;iii) mesenchyme-specific inhibition of GSK-3? is sufficient for the BPD phenotype;and iv) matricellular protein expression in increased in the lungs of infants with BPD. Completion of this work will provide insight into the pathogenesis of BPD, lead to the development of more accurate biomarkers, and provide new information on the role of matricellular proteins in lung fibrosis.

Public Health Relevance

Prematurity and bronchopulmonary dyplasia (BPD) are increasingly common causes of asthma in older children. This application seeks to understand the cellular and molecular mechanisms underlying the development of BPD, focusing on the role of mesenchymal stromal cells and their differentiation into myofibroblasts, the key effector cell in lung scarring. Insight provided from the proposed studies may lead to new methods to diagnose, prevent and treat BPD, a common precedent to childhood asthma.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL079339-08
Application #
8486304
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Blaisdell, Carol J
Project Start
2004-12-01
Project End
2016-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
8
Fiscal Year
2013
Total Cost
$370,090
Indirect Cost
$132,090
Name
University of Michigan Ann Arbor
Department
Pediatrics
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Whiteman, Eileen L; Fan, Shuling; Harder, Jennifer L et al. (2014) Crumbs3 is essential for proper epithelial development and viability. Mol Cell Biol 34:43-56
Anyanwu, Anuli C; Bentley, J Kelley; Popova, Antonia P et al. (2014) Suppression of inflammatory cell trafficking and alveolar simplification by the heme oxygenase-1 product carbon monoxide. Am J Physiol Lung Cell Mol Physiol 306:L749-63
Popova, Antonia P; Bentley, J Kelley; Cui, Tracy X et al. (2014) Reduced platelet-derived growth factor receptor expression is a primary feature of human bronchopulmonary dysplasia. Am J Physiol Lung Cell Mol Physiol 307:L231-9
Bentley, J Kelley; Chen, Qiang; Hong, Jun Young et al. (2014) Periostin is required for maximal airways inflammation and hyperresponsiveness in mice. J Allergy Clin Immunol 134:1433-42
Bozyk, Paul D; Bentley, J Kelley; Popova, Antonia P et al. (2012) Neonatal periostin knockout mice are protected from hyperoxia-induced alveolar simplication. PLoS One 7:e31336
Deng, Huan; Hershenson, Marc B; Lei, Jing et al. (2010) p70 Ribosomal S6 kinase is required for airway smooth muscle cell size enlargement but not increased contractile protein expression. Am J Respir Cell Mol Biol 42:744-52
Bentley, J Kelley; Popova, Antonia P; Bozyk, Paul D et al. (2010) Ovalbumin sensitization and challenge increases the number of lung cells possessing a mesenchymal stromal cell phenotype. Respir Res 11:127
Bentley, J Kelley; Deng, Huan; Linn, Marisa J et al. (2009) Airway smooth muscle hyperplasia and hypertrophy correlate with glycogen synthase kinase-3(beta) phosphorylation in a mouse model of asthma. Am J Physiol Lung Cell Mol Physiol 296:L176-84
Deng, Huan; Dokshin, Gregoriy A; Lei, Jing et al. (2008) Inhibition of glycogen synthase kinase-3beta is sufficient for airway smooth muscle hypertrophy. J Biol Chem 283:10198-207
Hershenson, Marc B; Brown, Melanie; Camoretti-Mercado, Blanca et al. (2008) Airway smooth muscle in asthma. Annu Rev Pathol 3:523-55

Showing the most recent 10 out of 12 publications