TNF-α and IL-1β-activated human mesenchymal stromal cells increase airway epithelial wound healing in vitro via activation of the epidermal growth factor receptor

Abstract

BackgroundMesenchymal stromal cells (MSCs) are investigated for their potential to reduce inflammation and to repair damaged tissue. Inflammation and tissue damage are hallmarks of chronic obstructive pulmonary disease (COPD) and MSC infusion is a promising new treatment for COPD. Inflammatory mediators attract MSCs to sites of inflammation and affect their immune-modulatory properties, but little is known about their effect on regenerative properties of MSCs. This study investigates the effect of the pro-inflammatory cytokines TNF-α and IL-1β on the regenerative potential of MSCs, using an in vitro wound healing model of airway epithelial cells.MethodsStandardized circular wounds were created by scraping cultures of the airway epithelial cell line NCI-H292 and primary bronchial epithelial cells cultured at the air-liquid interface (ALI-PBEC), and subsequently incubated with MSC conditioned medium (MSC-CM) that was generated in presence or absence of TNF-α/IL-1β. Remaining wound size was measured up to 72 h. Phosphorylation of ERK1/2 by MSC-CM was assessed using Western blot. Inhibitors for EGFR and c-Met signaling were used to investigate the contribution of these receptors to wound closure and to ERK1/2 phosphorylation. Transactivation of EGFR by MSC-CM was investigated using a TACE inhibitor, and RT-PCR was used to quantify mRNA expression of several growth factors in MSCs and NCI-H292.ResultsStimulation of MSCs with the pro-inflammatory cytokines TNF-α and IL-1β increased the mRNA expression of various growth factors by MCSs and enhanced the regenerative potential of MSCs in an in vitro model of airway epithelial injury using NCI-H292 airway epithelial cells. Conditioned medium from cytokine stimulated MSCs induced ERK1/2 phosphorylation in NCI-H292, predominantly via EGFR; it induced ADAM-mediated transactivation of EGFR, and it induced airway epithelial expression of several EGFR ligands. The contribution of activation of c-Met via HGF to increased repair could not be confirmed by inhibitor experiments.ConclusionOur data imply that at sites of tissue damage, when inflammatory mediators are present, for example in lungs of COPD patients, MSCs become more potent inducers of repair, in addition to their well-known immune-modulatory properties.

Canine adipose tissue-derived mesenchymal stem cells pre-treated with TNF-alpha enhance immunomodulatory effects in inflammatory bowel disease in mice

“…Previous studies have revealed that human MSCs stimulated with pro-inflammatory cytokines (such as TNF-α and IL-1β) can improve the secretory effects of immunomodulatory soluble factors (Broekman et al, 2016;Heo et al, 2011). Here, we also showed that TNF-α-stimulated cAT-MSCs released significantly higher concentrations of TSG-6 (2.5-fold) and PGE2 (3-fold), relative to naive cAT-MSCs.…”

“…Fan et al also revealed that human MSCs stimulated with interleukin (IL)-1β showed enhanced efficacy in mice with colitis (Fan et al, 2012). In addition, recent studies have demonstrated that MSCs pre-treated with pro-inflammatory cytokines showed enhanced secretory abilities (Broekman et al, 2016;Heo et al, 2011). However, few studies have assessed the therapeutic effects of pro-inflammatory cytokine-stimulated canine MSCs.…”

IL‐33 induces production of autoantibody against autologous respiratory epithelial cells: a potential mechanism for the pathogenesis of COPD

“…In addition, this phenomenon was accompanied by elevated, local expression of a range of pro‐inflammatory cytokines, including IL‐6 and TNF‐ α , although not IL‐1 β , which have also been implicated in causing respiratory epithelial cell damage and promoting autoimmunity in patients with COPD …”

Human Mesenchymal Stem Cell–Educated Macrophages Are a Distinct High IL-6–Producing Subset that Confer Protection in Graft-versus-Host-Disease and Radiation Injury Models

“…Classically IL-1β is described as an acute phase reactant early during inflammation, but in certain immunologic contexts plays a major role in tissue repair. IL-1β levels may contribute to early bone healing 53 and prime MSCs to promote healing of airway epithelial cells 54 . TGF-β classically is involved in fibrosis and protects cell survival during radiation 55 , structural changes of tissues via EMT and inhibition of cell proliferation 56 , and has a protective role in acute GVHD 57 .…”