Ruxolitinib inhibits poly(I:C) and type 2 cytokines‐induced CCL5 production in bronchial epithelial cells: A potential therapeutic agent for severe eosinophilic asthma

Abstract

Rationale Severe eosinophilic asthma is characterized by airway eosinophilia and corticosteroid‐resistance, commonly overlapping with type 2 inflammation. It has been reported that chemokine (C‐C motif) ligand 5 (CCL5) is involved in the exacerbation of asthma by RNA virus infections. Indeed, treatment with a virus‐associated ligand and a T helper type 2 cell (Th2) cytokine can synergistically stimulate CCL5 production in bronchial epithelial cells. We aimed to evaluate the mechanisms underlying CCL5 production in this in vitro model and to assess the potential of Janus kinase 1 (JAK1) as a novel therapeutic target via the use of ruxolitinib. Methods We stimulated primary normal human bronchial epithelial (NHBE) cells and BEAS‐2B cells with poly(I:C) along with interleukin‐13 (IL‐13) or IL‐4, and assessed CCL5 production. We also evaluated the signals involved in virus‐ and Th2‐cytokine‐induced CCL5 production and explored a therapeutic agent that attenuates the CCL5 production. Results Poly(I:C) stimulated NHBE and BEAS‐2B cells to produce CCL5. Poly(I:C) and IL‐13 increased CCL5 production. Poly(I:C)‐induced CCL5 production occurred via the TLR3–IRF3 and IFNAR/JAK1–phosphoinositide 3‐kinase (PI3K) pathways, but not the IFNAR/JAK1–STATs pathway. In addition, IL‐13 did not augment poly(I:C)‐induced CCL5 production via the canonical IL‐13R/IL‐4R/JAK1–STAT6 pathway but likely via subsequent TLR3‐IRF3‐IFNAR/JAK1‐PI3K pathways. JAK1 was identified to be a potential therapeutic target for severe eosinophilic asthma. The JAK1/2 inhibitor, ruxolitinib, was demonstrated to more effectively decrease CCL5 production in BEAS‐2B cells than fluticasone propionate. Conclusion We have demonstrated that JAK1 is a possible therapeutic target for severe corticosteroid‐resistant asthma with airway eosinophilia and persistent Th2‐type inflammation, and that ruxolitinib has potential as an alternative pharmacotherapy.

Disrupting TSLP–TSLP receptor interactions via putative small molecule inhibitors yields a novel and efficient treatment option for atopic diseases

“…TSLP activates JAK1 and JAK2 and subsequently phosphorylates downstream signaling proteins such as STAT3, STAT5 and STAT6. To identify the secondary messengers that stimulate Th2 cytokine secretion in primary human CD4 + T cells, TSLP-activated T cells were treated with inhibitors of JAK1/2 (ruxolinitib (Sada et al, 2021 ) and AZD1870 (Hedvat et al, 2009 )), STAT3/5 (SH-4-54) (Haftchenary et al, 2013 ), STAT5 (573108) (Müller et al, 2008 ), and STAT6 (AS1517499) (Chiba et al, 2009 ) inhibitors showing that targeting the JAK-STAT pathway impairs IL-4 and IL-13 secretion (Appendix Fig. S3 ).…”

Distinct Effects of Respiratory Viral Infection Models on miR-149-5p, IL-6 and p63 Expression in BEAS-2B and A549 Epithelial Cells

“…There are different reports on the concentrations of poly (I:C) used to stimulate epithelial cells [ 20 , 42 , 43 ]. Thus, we initially investigated the dose-dependent effect of poly (I:C) (0.01–10 μg/mL) on the viability of BEAS-2B or A549 cells.…”

Viral interference between severe acute respiratory syndrome coronavirus 2 and influenza A viruses

“…As we observed no severe damage in HAEs infected with SARS-CoV-2, we could confirm that ruxolitinib effectively inhibited the ISG expression. Although we did not evaluate the cytotoxic concentrations of both inhibitors in HAEs, concentrations of at least 5 µM of ruxolitinib and 6 µM of BX795 were not shown to cause any cytotoxicity in various cell lines, including human airway cells [51–56]. For instance, nasal HAEs have been exposed to 10 µM of ruxolitinib [50] and 6 µM of BX795 [4, 22, 23] without any cytotoxicity being noted.…”