A novel ex-vivo approach to study lung injury and repair

Abstract

Degenerative lung diseases such as chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis are incurable. The genesis of these diseases is either an inadequate or aberrant tissue repair processes yet current treatments cannot actively repair injured tissue. A promising approach to drive tissue repair is the stimulation of Wnt signalling, which has a crucial role in lung epithelial development and alveolar epithelial cell function in lung disease (Uhl FE, <i>et al.</i> Eur Respir J. 2015, 46(4):1150). However, the precise molecular mechanisms that target the pathway to drive repair are not well established. Pharmacological modification of the Wnt pathway is a promising approach to stimulate lung repair. To investigate the efficacy of novel pro-repair molecules, an acid drop injury is made on a spatially restricted area of mouse precision cut lung slices (PCLS). Key aspects of tissue repair are assessed at 24, 48, 72 h by automated quantification of cell proliferation and differentiation (Ki67 and Hopx) and pro-surfactant protein C (proSp-C, a marker for ATII cells/an early repair response, Fig. 1A). Post imaging, additional analysis is conducted on injured and non-injured areas of PCLS (Fig. 1B) by qPCR&nbsp;to analyse&nbsp;pro-repair genes (Fig. 1C). This study reports the establishment of a novel ex-vivo model for studies of lung injury and repair. The techniques developed in this work are likely to provide a versatile new tool&nbsp;in respiratory research.