Veldkamp Saskia R, Reugebrink Maud, Evers Sanne W, Moreau Thomas R J, Bondet Vincent, Armbrust Wineke, van den Berg J Merlijn, Hissink Muller Petra C E, Kamphuis Sylvia, Schatorjé Ellen, Delemarre Eveline M, van der Kooi Anneke J, Bader-Meunier Brigitte, Duffy Darragh, Rodero Mathieu P, Raaphorst Joost, van Royen-Kerkhof Annet, Jansen Marc H A, van Wijk Femke
Center for Translational Immunology, University Medical Center Utrecht, Utrecht, the Netherlands.
Department of Neurology, Amsterdam University Medical Centre, Location AMC, Amsterdam, the Netherlands.
Rheumatology (Oxford). 2025 May 15. doi: 10.1093/rheumatology/keaf227.
For IFN-driven diseases, such as juvenile dermatomyositis (JDM), there is a critical need for targeted therapies. We aimed to develop an in vitro model, using Siglec-1 as read-out, to evaluate inhibition of IFN-mediated responses with different JAK inhibitors (JAKi).
Healthy donor (HD) PBMCs were cultured with type I and II IFNs, TLR agonists, and plasma or serum from patients (JDM, DM, juvenile SLE, COVID-19) and HDs. Siglec-1 expression on CD14+ monocytes was analyzed using flow cytometry. Inhibitory assays involved pre-incubation with JAKi (filgotinib, tofacitinib, baricitinib, ruxolitinib, deucravacitinib) and interferon-α/β receptor (IFNAR)-blocking antibody. Correlations between plasma-induced Siglec-1 levels and clinical disease activity were analyzed in JDM patients, as well as correlations with IFN-α and -β plasma levels.
Siglec-1 was induced after 18 h of stimulation with type I IFNs and TLR-3/7/9 agonists, with minimal induction by IFN-y. IFNAR blockade prevented type I IFN and TLR-mediated induction. JAKi inhibited Siglec-1 induction by IFN-α and -β in a dose-dependent manner. Co-culture with plasma or serum from patients with IFN-driven diseases induced Siglec-1 expression on healthy monocytes, which could be inhibited by JAKi and IFNAR blockade. Siglec-1 levels induced by JDM plasma correlated strongly with clinical disease activity and IFN-β plasma levels.
Siglec-1 is an easy and reliable in vitro marker for type I IFN responses. Its induction can be inhibited by JAKi. The type I IFN signature in JDM is likely predominantly driven by IFN-β. This assay holds promise for precision treatment strategies in JDM and other IFN-driven diseases.
对于由干扰素驱动的疾病，如幼年皮肌炎（JDM），迫切需要靶向治疗。我们旨在开发一种以唾液酸结合免疫球蛋白样凝集素-1（Siglec-1）为指标的体外模型，以评估不同的JAK抑制剂（JAKi）对干扰素介导反应的抑制作用。
将健康供体（HD）的外周血单核细胞（PBMC）与I型和II型干扰素、Toll样受体（TLR）激动剂以及患者（JDM、皮肌炎（DM）、青少年系统性红斑狼疮（SLE）、新型冠状病毒肺炎（COVID-19））和HD的血浆或血清一起培养。使用流式细胞术分析CD14+单核细胞上Siglec-1的表达。抑制试验包括与JAKi（非戈替尼、托法替布、巴瑞替尼、鲁索替尼、氘可来昔替尼）和干扰素-α/β受体（IFNAR）阻断抗体预孵育。分析JDM患者血浆诱导的Siglec-1水平与临床疾病活动度之间的相关性，以及与血浆中干扰素-α和-β水平的相关性。
用I型干扰素和TLR-3/7/9激动剂刺激18小时后可诱导Siglec-1表达，而干扰素-γ诱导作用最小。阻断IFNAR可阻止I型干扰素和TLR介导的诱导。JAKi以剂量依赖性方式抑制干扰素-α和-β诱导的Siglec-1表达。与干扰素驱动疾病患者的血浆或血清共培养可诱导健康单核细胞上Siglec-1表达，这可被JAKi和阻断IFNAR抑制。JDM血浆诱导的Siglec-1水平与临床疾病活动度和血浆干扰素-β水平密切相关。
Siglec-1是I型干扰素反应的一种简便可靠的体外标志物。其诱导可被JAKi抑制。JDM中的I型干扰素特征可能主要由干扰素-β驱动。该试验有望用于JDM和其他干扰素驱动疾病的精准治疗策略。
