DataSheet1_Crosstalk between TBK1/IKKε and the type I interferon pathway contributes to tubulointerstitial inflammation and kidney tubular injury.PDF

The type I interferon (TI-IFN) pathway regulates innate immunity, inflammation, and apoptosis during infection. However, the contribution of the TI-IFN pathway or upstream signaling pathways to tubular injury in kidney disease is poorly understood. Upon observing evidence of activation of upstream regulators of the TI-IFN pathway in a transcriptomics analysis of murine kidney tubulointerstitial injury, we have now addressed the impact of the TI-IFN and upstream signaling pathways on kidney tubulointerstitial injury. In cultured tubular cells and kidney tissue, IFNα/β binding to IFNAR activated the TI-IFN pathway and recruited antiviral interferon-stimulated genes (ISG) and NF-κB-associated proinflammatory responses. TWEAK and lipopolysaccharide (LPS) signaled through TBK1/IKKε and IRF3 to activate both ISGs and NF-κB. In addition, TWEAK recruited TLR4 to stimulate TBK1/IKKε-dependent ISG and inflammatory responses. Dual pharmacological inhibition of TBK1/IKKε with amlexanox decreased TWEAK- or LPS-induced ISG and cytokine responses, as well as cell death induced by a complex inflammatory milieu that included TWEAK. TBK1 or IRF3 siRNA prevented the TWEAK-induced ISG and inflammatory gene expression while IKKε siRNA did not. In vivo, kidney IFNAR and IFNβ were increased in murine LPS and folic acid nephrotoxicity while IFNAR was increased in human kidney biopsies with tubulointerstitial damage. Inhibition of TBK1/IKKε with amlexanox or IFNAR neutralization decreased TI-IFN pathway activation and protected from kidney injury induced by folic acid or LPS. In conclusion, TI-IFNs, TWEAK, and LPS engage interrelated proinflammatory and antiviral responses in tubular cells. Moreover, inhibition of TBK1/IKKε with amlexanox, and IFNAR targeting, may protect from tubulointerstitial kidney injury.

I型干扰素（TI-IFN）通路调控感染期间的固有免疫、炎症和凋亡。然而，TI-IFN通路或上游信号通路在肾脏疾病中导致肾小管损伤的贡献理解尚不充分。通过对小鼠肾小管间质性损伤转录组学分析的观察，我们发现TI-IFN和上游信号通路对肾小管间质性损伤的影响。在培养的肾小管细胞和肾脏组织中，IFNα/β与IFNAR结合激活了TI-IFN通路，并招募了抗病毒干扰素刺激基因（ISG）以及与NF-κB相关的促炎反应。TWEAK和脂多糖（LPS）通过TBK1/IKKε和IRF3信号，激活了ISGs和NF-κB。此外，TWEAK招募TLR4以刺激TBK1/IKKε依赖的ISG和炎症反应。使用氨甲环酸双重药理抑制TBK1/IKKε可减少TWEAK或LPS诱导的ISG和细胞因子反应，以及由包括TWEAK在内的复杂炎症环境诱导的细胞死亡。TBK1或IRF3 siRNA可防止TWEAK诱导的ISG和炎症基因表达，而IKKε siRNA则不能。在体内，小鼠脂多糖和叶酸肾毒性模型中肾脏IFNAR和IFNβ增加，而在人类肾脏活检中，IFNAR在具有肾小管间质性损伤的活检样本中增加。使用氨甲环酸或IFNAR中和抑制TBK1/IKKε可减少TI-IFN通路激活，并保护免受叶酸或LPS诱导的肾脏损伤。综上所述，TI-IFNs、TWEAK和LPS在肾小管细胞中引发相互关联的促炎和抗病毒反应。此外，使用氨甲环酸抑制TBK1/IKKε和针对IFNAR的治疗可能保护免受肾小管间质性肾脏损伤。