A new approach for the study of lung smooth muscle phenotypes and its application in a murine model of allergic airway inflammation

Jesus Paez-Cortez, Boston University School of Medicine
Ramaswamy Krishnan, Beth Israel Deaconess Medical Center
Anneliese Arno, Boston University School of Medicine
Linh Aven, Boston University School of Medicine
Sumati Ram-Mohan, Beth Israel Deaconess Medical Center
Kruti R. Patel, Boston University School of Medicine
Jining Lu, Boston University School of Medicine
Oliver D. King, University of Massachusetts Medical School
Xingbin Ai, Boston University School of Medicine
Alan Fine, Boston University School of Medicine

Copyright 2013 Paez-Cortez et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Phenotypes of lung smooth muscle cells in health and disease are poorly characterized. This is due, in part, to a lack of methodologies that allow for the independent and direct isolation of bronchial smooth muscle cells (BSMCs) and vascular smooth muscle cells (VSMCs) from the lung. In this paper, we describe the development of a bi-fluorescent mouse that permits purification of these two cell populations by cell sorting. By subjecting this mouse to an acute allergen based-model of airway inflammation that exhibits many features of asthma, we utilized this tool to characterize the phenotype of so-called asthmatic BSMCs. First, we examined the biophysical properties of single BSMCs from allergen sensitized mice and found increases in basal tone and cell size that were sustained ex vivo. We then generated for the first time, a comprehensive characterization of the global gene expression changes in BSMCs isolated from the bi-fluorescent mice with allergic airway inflammation. Using statistical methods and pathway analysis, we identified a number of differentially expressed mRNAs in BSMCs from allergen sensitized mice that code for key candidate proteins underlying changes in matrix formation, contractility, and immune responses. Ultimately, this tool will provide direction and guidance for the logical development of new markers and approaches for studying human lung smooth muscle.