Human Lung Parenchyma but Not Proximal Bronchi Produces Fibroblasts with Enhanced TGF-β Signaling and α-SMA Expression

Given the contribution various fibroblast subsets make to wound healing and tissue remodeling, the concept of lung fibroblast heterogeneity is of great interest. However, the mechanisms contributing to this heterogeneity are unknown. To this aim, we compared molecular and biophysical characteristics of fibroblasts concurrently isolated from normal human proximal bronchi (B-FBR) and distal lung parenchyma (P-FBR). Using quantitative RT-PCR, spontaneous expression of more than 30 genes related to repair and remodeling was analyzed. All P-FBR lines demonstrated significantly increased basal alpha-smooth muscle actin (alpha-SMA) mRNA and protein expression levels when compared with donor-matched B-FBR. These differences were not associated with sex, age, or disease history of lung tissue donors. In contrast to B-FBR, P-FBR displayed enhanced transforming growth factor (TGF)-beta/Smad signaling at baseline, and inhibition of either ALK-5 or neutralization of endogenously produced and activated TGF-beta substantially decreased basal alpha-SMA protein in P-FBR. Both B-FBR and P-FBR up-regulated alpha-SMA after stimulation with TGF-beta 1, and basal expression levels of TGF-beta 1, TGF-beta RI, and TGF-beta RII were not significantly different between fibroblast pairs. Blockade of metalloproteinase-dependent activation of endogenous TGF-beta did not significantly modify alpha-SMA expression in P-FBR. However, resistance to mechanical tension of these cells was significantly higher in comparison with B-FBR, and added TGF-beta 1 significantly increased stiffness of both cell monolayers. Our data suggest that in contrast with human normal bronchial tissue explants, lung parenchyma produces mesenchymal cells with a myofibroblastic phenotype by intrinsic mechanisms of TGF-beta activation in feed-forward manner. These results also offer a new insight into mechanisms of human fibroblast heterogeneity and their function in the airway and lung tissue repair and remodeling.