Asbestos-induced Disruption of Calcium Homeostasis Induces Endoplasmic Reticulum Stress in Macrophages [Cell Biology]

October 16th, 2014 by Ryan, A. J., Larson-Casey, J. L., He, C., Murthy, S., Carter, A. B.

Though the mechanisms for fibrosis development remain largely unknown, recent evidence indicates that endoplasmic reticulum (ER) stress and activation of the unfolded protein response (UPR) may act as an important fibrotic stimulus in diseased lungs. ER stress is observed in lungs of patients with idiopathic pulmonary fibrosis. In this study we evaluated if ER stress and the UPR was present in macrophages exposed to chrysotile asbestos and if ER stress in macrophages was associated with asbestos-induced pulmonary fibrosis. Macrophages exposed to chrysotile had elevated transcript levels of several ER stress genes. Macrophages loaded with the Ca2+-sensitive dye Fura-2-AM showed that cytosolic Ca2+ increased significantly within minutes after chrysotile exposure and remained elevated for a prolonged time. Chrysotile-induced increases in cytosolic Ca2+ were partially inhibited by either anisomycin, an inhibitor of passive Ca2+ leak from the ER, or BAPTA-AM, an intracellular Ca2+ chelator known to deplete ER Ca2+ stores. Anisomycin inhibited X-box binding protein 1 (XBP1) mRNA splicing and reduced immunoglobulin binding protein (BiP) levels, whereas BAPTA-AM increased XBP1 splicing and BiP expression, suggesting that ER calcium depletion may be one factor contributing to ER stress in cells exposed to chrysotile. To evaluate ER stress in vivo, asbestos-exposed mice showed fibrosis development, and alveolar macrophages from fibrotic mice showed increased expression of BiP. Bronchoalveolar macrophages from asbestosis patients showed increased expression of several ER stress genes compared to normal subjects. These findings suggest that alveolar macrophages undergo ER stress, which is associated with fibrosis development.