PU.1 promotes development of rheumatoid arthritis via repressing FLT3 in macrophages and fibroblast-like synoviocytes.
PU.1 promotes development of rheumatoid arthritis via repressing FLT3 in macrophages and fibroblast-like synoviocytes.
复制标题
PU.1 通过抑制巨噬细胞和成纤维样滑膜细胞中的 FLT3 促进类风湿关节炎的发展
DOI：
10.1136/ard-2022-222708
复制
发表时间：
2023-02
影响因子：
27.4
通讯作者：
中科院分区：
文献类型：
--
作者：
目的探讨类风湿关节炎（RA）发病过程中一种重要的转录因子PU.1的功能及其作用机制。方法采用免疫印迹和免疫组化（IHC）法检测RA患者滑膜中PU.1及其潜在靶点FMS样酪氨酸激酶3（FLT 3）的表达和定位。使用UREΔ（PU.1敲低）和FLT 3-ITD（FLT 3激活）小鼠建立胶原抗体诱导的关节炎（CAIA）。在体外研究中，使用SiRNA研究了PU.1和FLT 3对原代巨噬细胞和成纤维细胞样滑膜细胞（FLS）的影响。在机制上，进行荧光素酶报告基因测定、蛋白质印迹、FACS和IHC以显示PU.1对巨噬细胞和FLS中FLT 3转录的直接调节。最后，使用PU.1的小分子抑制剂DB2313，使用两种体内模型CAIA和胶原诱导的关节炎（CIA）进一步说明DB2313对关节炎的治疗作用。结果与骨关节炎患者及正常对照组相比，RA患者滑膜组织中PU.1的表达明显增强。FLT 3和p-FLT 3在RA中的表达模式与PU.1相反。CAIA模型表明，PU.1是体内关节炎发展的激活剂，而FLT 3是抑制剂。此外，来自体外测定的结果与体内结果一致：PU.1促进巨噬细胞和FLS的过度活化和炎症状态，而FLT 3具有相反的作用。此外，PU.1通过直接结合FLT 3的启动子区域来抑制其转录。PU.1抑制剂DB2313明显减轻了对CAIA和CIA模型中关节炎发展的影响。结论这些结果支持PU.1在RA中的作用，并可能通过直接抑制FLT 3而具有治疗意义。因此，靶向PU.1可能是治疗RA的一种潜在方法。
Objectives To uncover the function and underlying mechanism of an essential transcriptional factor, PU.1, in the development of rheumatoid arthritis (RA). Methods The expression and localisation of PU.1 and its potential target, FMS-like tyrosine kinase 3 (FLT3), in the synovium of patients with RA were determined by western blot and immunohistochemical (IHC) staining. UREΔ (with PU.1 knockdown) and FLT3-ITD (with FLT3 activation) mice were used to establish collagen antibody-induced arthritis (CAIA). For the in vitro study, the effects of PU.1 and FLT3 on primary macrophages and fibroblast-like synoviocytes (FLS) were investigated using siRNAs. Mechanistically, luciferase reporter assays, western blotting, FACS and IHC were conducted to show the direct regulation of PU.1 on the transcription of FLT3 in macrophages and FLS. Finally, a small molecular inhibitor of PU.1, DB2313, was used to further illustrate the therapeutic effects of DB2313 on arthritis using two in vivo models, CAIA and collagen-induced arthritis (CIA). Results The expression of PU.1 was induced in the synovium of patients with RA when compared with that in osteoarthritis patients and normal controls. FLT3 and p-FLT3 showed opposite expression patterns compared with PU.1 in RA. The CAIA model showed that PU.1 was an activator, whereas FLT3 was a repressor, of the development of arthritis in vivo. Moreover, results from in vitro assays were consistent with the in vivo results: PU.1 promoted hyperactivation and inflammatory status of macrophages and FLS, whereas FLT3 had the opposite effects. In addition, PU.1 inhibited the transcription of FLT3 by directly binding to its promoter region. The PU.1 inhibitor DB2313 clearly alleviated the effects on arthritis development in the CAIA and CIA models. Conclusions These results support the role of PU.1 in RA and may have therapeutic implications by directly repressing FLT3. Therefore, targeting PU.1 might be a potential therapeutic approach for RA.
登录
查看更多内容
DOI：
10.1038/s41577-020-00426-6
发表时间：
2021-03
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
Hosokawa H;Rothenberg EV
通讯作者：
Rothenberg EV
影响因子：
7.3
作者：
Tu J;Huang W;Zhang W;Mei J;Zhu C
通讯作者：
Zhu C
芍药苷 CP-25 的修饰化合物可抑制胶原诱导的关节炎小鼠的免疫反应和滑膜炎症。
影响因子：
5.6
作者：
Chen J;Wang Y;Wu H;Yan S;Chang Y;Wei W
通讯作者：
Wei W
DOI：
10.1038/s41573-020-00092-2
发表时间：
2021-03
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
Lee DSW;Rojas OL;Gommerman JL
通讯作者：
Gommerman JL
影响因子：
4.9
作者：
Bustamante MF;Garcia-Carbonell R;Whisenant KD;Guma M
通讯作者：
Guma M