Effect of JAK Inhibitors on Release of CXCL9, CXCL10 and CXCL11 from Human Airway Epithelial Cells

Abstract

BackgroundCD8+ T-cells are located in the small airways of COPD patients and may contribute to pathophysiology. CD8+ cells express the chemokine receptor, CXCR3 that binds CXCL9, CXCL10 and CXCL11, which are elevated in the airways of COPD patients. These chemokines are released from airway epithelial cells via activation of receptor associated Janus kinases (JAK). This study compared the efficacy of two structurally dissimilar pan-JAK inhibitors, PF956980 and PF1367550, and the glucocorticosteroid dexamethasone, in BEAS-2B and human primary airway epithelial cells from COPD patients and control subjects.MethodsCells were stimulated with either IFNγ alone or with TNFα, and release of CXCL9, CXCL10 and CXCL11 measured by ELISA and expression of CXCL9, CXCL10 and CXCL11 by qPCR. Activation of JAK signalling was assessed by STAT1 phosphorylation and DNA binding.ResultsThere were no differences in the levels of release of CXCL9, CXCL10 and CXCL11 from primary airway epithelial cells from any of the subjects or following stimulation with either IFNγ alone or with TNFα. Dexamethasone did not inhibit CXCR3 chemokine release from stimulated BEAS-2B or primary airway epithelial cells. However, both JAK inhibitors suppressed this response with PF1367550 being ~50-65-fold more potent than PF956980. The response of cells from COPD patients did not differ from controls with similar responses regardless of whether inhibitors were added prophylactically or concomitant with stimuli. These effects were mediated by JAK inhibition as both compounds suppressed STAT1 phosphorylation and DNA-binding of STAT1 and gene transcription.ConclusionsThese data suggest that the novel JAK inhibitor, PF1367550, is more potent than PF956980 and that JAK pathway inhibition in airway epithelium could provide an alternative anti-inflammatory approach for glucocorticosteroid-resistant diseases including COPD.

Isorhapontigenin, a bioavailable dietary polyphenol, suppresses airway epithelial cell inflammation through a corticosteroid‐independent mechanism

“…v ‐1 ) collagen‐coated flasks. Cells were extracted from the lung tissues of healthy non‐smokers and COPD patients using methods described previously (Fenwick et al, ). COPD patients fulfilled the Global Initiative for Chronic Obstructive Lung Disease criteria (GOLD, ).…”

“…cDNA was diluted to 5 ng·μL −1 using RNase‐free water to be used for subsequent assays. Gene expression was measured using the Taqman real‐time PCR on a 7500 Real Time PCR system (Applied Biosystems, Paisley, UK) as described previously (Fenwick et al, ). mRNA expression levels of the samples were calculated using their C t values by interpolation from the standard curve constructed.…”

Kinases as Novel Therapeutic Targets in Asthma and Chronic Obstructive Pulmonary Disease

“…These chemokines are induced by IFNg through the activation of JAKs and phosphorylation of predominantly STAT1 (Tudhope et al, 2007). Pan-JAK inhibitors are effective in reducing IFNg-induced secretion of CXCR3-activating chemokines from human airway epithelial cells, demonstrating their potential as antiinflammatory treatments in COPD (Fenwick et al, 2015). B. Jakinibs.…”

Role of epithelial chemokines in the pathogenesis of airway inflammation in asthma (Review)

“…On this basis, these associated signaling molecules have also been used as potential targets of anti-inflammatory treatment. It has been identified that the inhibition of JAK pathway in the airway epithelium may provide an alternative anti-inflammatory approach to glucocorticosteroid-resistant diseases in vitro (36). It has also been observed that the long-acting β2 agonists downregulate poly I:C-induced CXCL10 expression in bronchial epithelial cells via the β2 adrenoreceptor-cyclic adenosine monophosphate and JNK pathways in vitro (37).…”