Gene Expression Profiling of Lung Myofibroblasts Reveals the Anti-Fibrotic Effects of Cyclosporine

Abstract

Cyclosporine, a calcineurin inhibitor, is a potent immunosuppressive agent that acts chiefly through the inactivation of T-lymphocytes. Several clinical studies have demonstrated the effectiveness of cyclosporine for treating fibrotic lung disease, but the underlying mechanism remains elusive. We hypothesized that cyclosporine exerts direct effects against fibrogenesis of lung myofibroblasts, and aimed to elucidate the mechanism of this anti-fibrotic effect through gene-expression profiling using DNA microarray analysis. We found that cyclosporine suppressed the expression of alpha-smooth muscle actin and collagen type I in myofibroblasts that had been differentiated from a fetal human lung fibroblast cell line by induction with transforming growth factor (TGF)-β. Furthermore, microarray analysis revealed that cyclosporine downregulated 57 genes whose expression levels were increased by TGF-β, and up-regulated 73 genes, whose expression was decreased by TGF-β. Classifying these 57 down-regulated and 73 up-regulated genes with the Database for Annotation, Visualization and Integrated Discovery (DAVID) web tool, we have identified the involvement of several functional categories, including innate immunity, cytokine interaction, growth factor, and cancer pathway. Of the identified genes, we selected three fibrosis-related genes, insulin-like growth factor binding protein 2 (IGFBP2), inhibitor of DNA binding 1 (ID1) and peroxisome proliferator-activated receptor gamma (PPARG), and validated their expression patterns by quantitative reverse transcription-polymerase chain reaction. Cyclosporine treatment decreased the expression levels of IGFBP2 and ID1, but increased PPARG expression. These results suggest that cyclosporine is a potent anti-fibrotic agent acting on myofibroblasts. Therefore, cyclosporine shows potential as a novel remedy for fibrotic lung disease.

Raised serum levels of IGFBP-1 and IGFBP-2 in idiopathic pulmonary fibrosis

“…Besides TGF-β, there has been recently a growing interest for the axis IGFs/IGFBPs in fibrosing process. A recently published study has shown a link between the production of TGF-β and the production of insulin-like growth factor binding protein-2 (IGFBP-2) [ 10 ] known to be related to IPF [ 11 ] within myofibroblasts cells derived from the lung. Moreover, IGFBP-2 was found at significantly higher level in the bronchoalveolar lavage of children with interstitial lung disease [ 12 ] relative to healthy subjects.…”

Scleral remodelling in myopia and its manipulation: a review of recent advances in scleral strengthening and myopia control

“…Cyclosporine A, an immunomodulatory drug, has been shown to increase TGF-β, α-SMA, and type I collagen expression in fibroblasts from some tissues (119,120), although in other tissue types it has inhibitory effects (121,122). Interestingly, cyclosporine A has recently been shown to reduce experimental myopia progression, although whether this was effected through changes in α-SMA expression was not investigated (123).…”

Chronic Ocular Complications of Stevens–Johnson Syndrome and Toxic Epidermal Necrolysis: The Role of Systemic Immunomodulatory Therapy

“…Hirota et al reported that CSA down-regulates the expression of a variety of genes that are increased by TGF-b, a fibrogenic cytokine, in lung myofibroblasts. 80 Furthermore, excessive TGF-b activity may lead to increased ocular surface inflammation, neovascularization, and cicatrization. 80,81 De Rojas et al described the use of cyclosporine either as monotherapy or in combination with another IMT in five patients with chronic ocular SJS/TEN.…”

“…80 Furthermore, excessive TGF-b activity may lead to increased ocular surface inflammation, neovascularization, and cicatrization. 80,81 De Rojas et al described the use of cyclosporine either as monotherapy or in combination with another IMT in five patients with chronic ocular SJS/TEN. 1 Of note, OCP patients tend to respond poorly to calcineurin inhibitors; in one study, 490% failed this therapy.…”