Supplemental oxygen (hyperoxia) with/without continuous nasal positive airway pressure (CPAP) to preterm infants is associated with airway hyperreactivity (AHR) proceeding to wheezing and asthma. Understanding mechanisms by which hyperoxia and CPAP induce sustained AHR represents our long-term goal, and an unmet clinical need. We propose the initial stretch imposed by CPAP on more compliant bronchial airways in premature infants, particularly with added hyperoxia, is contributory. AHR involves greater [Ca2+]i and contractility of airway smooth muscle (ASM), and remodeling mediated partly by ASM proliferation. Our published studies and preliminary work using human fetal airway cells and neonatal mouse models show moderate hyperoxia (50% O2) and mechanical stretch not only enhance ASM contractility and proliferation, but also bronchial epithelial arginase, raising the question of whether and how epithelial arginase and neonatal AHR are linked. We propose the novel and intriguing idea that arginase-derived polyamines (e.g. spermine) have downstream effects on a novel ASM target: extracellular Ca2+ sensing receptor (CaSR). Although well-known for regulating body Ca2+, there is limited information on CaSR in lung, and none in postnatal airways. Preliminary data in developing human airways show high expression of ASM CaSR that responds to extracellular Ca2+ ([Ca2+]o) or spermine, and enhances [Ca2+]I, contractility and proliferation. Hyperoxia and stretch each increase CaSR expression and function. In neonatal mice exposed to 50% O2 and/or CPAP which show sustained AHR, epithelial arginase and ASM CaSR are increased, while inhibitors of arginase (nor-NOHA) and CaSR (calcilytic NPS2143) blunt AHR. Our overall hypothesis is that the epithelial arginase-ASM CaSR axis contributes to AHR in the context of hyperoxia and CPAP exposure in prematurity. We will examine this idea via 3 Aims.
Aim 1 : In developing bronchial epithelium, determine the effect of hyperoxia and/or stretch on the arginase pathway;
Aim 2 : In developing ASM, determine the role of CaSR in enhanced [Ca2+]i/contractility and proliferation induced by hyperoxia and/or stretch;
Aim 3 : In neonatal mouse models of hyperoxia and/or CPAP exposure, determine the role of the arginase-CaSR axis in AHR and airway remodeling. 18-22 week human fetal bronchial epithelial cells (fBECs) and ASM cells (fASM) are exposed to hyperoxia (<60% O2) +/- 0-15% stretch (representing 0 to high CPAP) with continuous 5% oscillations, mimicking clinical hyperoxia +/- CPAP in spontaneously breathing premies. fBEC arginase pathway and downstream effects (Aim 1), and role of fASM CaSR in [Ca2+]i/contractility and proliferation following hyperoxia +/- stretch (Aim 2) are examined, with signaling pathways such as RhoA/Rho kinase and MAPKs.
In Aim 3, airway structural, functional and molecular changes are assessed in neonatal WT and smooth muscle CaSR KO mice exposed to 50% O2 +/- intermittent CPAP (3, 6, 9 cmH2O) for 7 days, with 14 days recovery (mimicking human conditions). Effects of inhibiting arginase (nor-NOHA) vs. CaSR (NPS2143) are tested towards establishing clinical significance.

Public Health Relevance

Babies born prematurely often require substantial respiratory support to overcome the critical period while their immature lungs continue to develop. Unfortunately, clinical interventions including provision of supplemental oxygen (hyperoxia) and even non-invasive continuous positive pressure (CPAP) to an immature respiratory system following preterm birth can predispose to chronic lung diseases such as wheezing and asthma which remain major health problems throughout childhood and beyond. The primary goal of this proposal is to understand how the initial exposure to hyperoxia and CPAP affects structure and function of the developing airway even after these interventions are stopped. We are focusing on the novel idea that the mechanical stretch imposed by CPAP on the airway (on top of the forces of breathing by the baby and supplemental oxygen) result in activation of mechanisms that promote airway hypercontractility and excessive growth of airway cells, all resulting in a thicker, more reactive airway while the baby and the lungs are growing. Our investigations will not only further our understanding of the role of such factors in early airway development, but also open up new therapeutic strategies to interfere with the detrimental effects of CPAP and oxygen exposure.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Natarajan, Aruna R
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Mayo Clinic, Rochester
United States
Zip Code
Cheon, In Su; Son, Young Min; Jiang, Li et al. (2018) Neonatal hyperoxia promotes asthma-like features through IL-33-dependent ILC2 responses. J Allergy Clin Immunol 142:1100-1112
Han, MeiLan K; Arteaga-Solis, Emilio; Blenis, John et al. (2018) Female Sex and Gender in Lung/Sleep Health and Disease. Increased Understanding of Basic Biological, Pathophysiological, and Behavioral Mechanisms Leading to Better Health for Female Patients with Lung Disease. Am J Respir Crit Care Med 198:850-858
Di Fiore, Juliann M; Dylag, Andrew M; Honomichl, Ryan D et al. (2018) Early inspired oxygen and intermittent hypoxemic events in extremely premature infants are associated with asthma medication use at 2 years of age. J Perinatol :
Martin, Richard J (2018) The unfortunate tale of immature respiratory control superimposed on an immature lung. Pediatr Res 84:153-154
Vogel, Elizabeth R; Britt Jr, Rodney D; Faksh, Arij et al. (2017) Moderate hyperoxia induces extracellular matrix remodeling by human fetal airway smooth muscle cells. Pediatr Res 81:376-383
Prakash, Y S; Pabelick, Christina M; Sieck, Gary C (2017) Mitochondrial Dysfunction in Airway Disease. Chest 152:618-626
Aravamudan, Bharathi; Goorhouse, Katelyn J; Unnikrishnan, Ghanashyam et al. (2017) Differential Expression of Estrogen Receptor Variants in Response to Inflammation Signals in Human Airway Smooth Muscle. J Cell Physiol 232:1754-1760
Freeman, Michelle R; Sathish, Venkatachalem; Manlove, Logan et al. (2017) Brain-derived neurotrophic factor and airway fibrosis in asthma. Am J Physiol Lung Cell Mol Physiol 313:L360-L370
Britt Jr, Rodney; Prakash, Y S (2017) The -Omic Approach to Understanding Glucocorticoid Effects in Smooth Muscle: Diving for Pearls. Am J Respir Cell Mol Biol 57:147-148
Aravamudan, Bharathi; Thompson, Michael; Sieck, Gary C et al. (2017) Functional Effects of Cigarette Smoke-Induced Changes in Airway Smooth Muscle Mitochondrial Morphology. J Cell Physiol 232:1053-1068

Showing the most recent 10 out of 99 publications