Interleukin-1α inhibits transforming growth factor-β1 and β2-induced extracellular matrix production, remodeling and signaling in human lung fibroblasts: Master regulator in lung mucosal repair

Background: Lung fibroblasts play a central role in maintaining lung homeostasis and facilitating repair through the synthesis and organization of the extracellular matrix (ECM). This study investigated the cross-talk between interleukin-1 alpha (IL-1 alpha) and transforming growth factor-beta (TGF-beta) signaling, two key regulators in tissue repair and fibrosis, in the context of lung fibroblast repair in the healthy lung. Results: Stimulation of lung fibroblasts with TGF-beta 1 and TGF-beta 2 induced collagen-I and fibronectin protein expression (p < 0.05), a response inhibited with co-treatment with IL-1 alpha (p < 0.05). Additionally, TGF-beta 1 and TGF-beta 2 induced myofibroblast differentiation, and collagen-I gel contraction, which were both suppressed by IL1 alpha (p < 0.05). In contrast, interleukin (IL)-6, IL-8 and thymic stromal lymphopoietin induced by IL-1 alpha, were unaffected by TGF-beta 1 or TGF-beta 2. Mechanistically, IL-1 alpha administration led to the suppression of TGF-beta 1 and TGF beta 2 signaling, through downregulation of mRNA and protein for TGF-beta receptor II and the downstream adaptor protein TRAF6, but not through miR-146a that is known to be induced by IL-1 alpha. Discussion: IL-1 alpha acts as a master regulator, modulating TGF-beta 1 and TGF-beta 2-induced ECM production, remodeling, and myofibroblast differentiation in human lung fibroblasts, playing a vital role in balancing tissue repair versus fibrosis. Further research is required to understand the dysregulated cross-talk between IL-1 alpha and TGF-beta signaling in chronic lung diseases and the exploration of therapeutic opportunities. Methods: Primary human lung fibroblasts (PHLF) were treated with media control, or 1 ng/ml IL-1 alpha with or without 50 ng/ml TGF-beta 1 or TGF-beta 2 for 1, 6 and 72 h. Cell lysates were assessed for the expression of ECM proteins and signaling molecules by western blot, miRNA by qPCR, mRNA by RNA sequencing and cell supernatants for cytokine production by ELISA. PHLFs were also seeded in non-tethered collagen-I gels to measure contraction, and myofibroblast differentiation using confocal microscopy.