Full-length IL-33 regulates Smad3 phosphorylation and gene transcription in a distinctive AP2-dependent manner.
Full-length IL-33 regulates Smad3 phosphorylation and gene transcription in a distinctive AP2-dependent manner.
复制标题
DOI：
10.1016/j.cellimm.2020.104203
复制
发表时间：
2020-11
影响因子：
4.3
通讯作者：
Atamas SP
中科院分区：
文献类型：
--
作者：
Luzina IG;Fishelevich R;Hampton BS;Courneya JP;Parisella FR;Lugkey KN;Baleno FX;Choi D;Kopach P;Lockatell V;Todd NW;Atamas SP
IL-33已成为免疫、炎症和纤维化反应的中心介质。许多研究都集中在成熟的IL-33上，但前体全长IL-33（FLIL 33）的表达升高也涉及一系列疾病，包括组织纤维化。我们先前报道并且现在证实，在来自健康供体和特发性肺纤维化患者的原代人肺成纤维细胞中，FLIL 33的过表达诱导TGF-β的关键促纤维化信号传导介质Smad 3的磷酸化。目前，我们证明FLIL 33诱导的Smad 3磷酸化不被抗TGF-β抗体消除，但被ALK 5/TGFBR 1特异性和Smad 3特异性抑制消除，表明FLIL 33作用不依赖于TGF-β，但依赖于其受体TGFBR。蛋白质印迹分析显示，FLIL 33过表达增加了衔接蛋白复合物2（AP 2）的AP 2A 1和AP 2B 1亚基的水平，但不影响亚细胞分布，AP 2是一种已知的TGFBR结合伴侣。siRNA介导的抑制这些亚基阻断FLIL 33诱导的Smad 3磷酸化，而AP 2亚基过表达诱导Smad 3磷酸化，即使在FLIL 33的情况下。RNA-Seq转录组学分析显示，用TGF-β刺激成纤维细胞诱导了许多基因表达水平的重大变化，而FLIL 33的过表达诱导了少量基因的适度表达变化。此外，qRT-PCR测试表明，尽管诱导Smad 3磷酸化，但FLIL 33不诱导胶原基因转录，甚至轻度减弱TGF-β诱导的胶原I和III mRNA水平。我们得出结论，FLIL 33通过TGF-β非依赖性但TGF-β受体和AP 2依赖性机制诱导Smad 3磷酸化，并且具有有限的下游转录组学后果。
IL-33 has emerged as a central mediator of immune, inflammatory, and fibrotic responses. Many studies have focused on mature IL-33, but elevated expression of the precursor, full-length IL-33 (FLIL33), has also been implicated in a spectrum of diseases, including tissue fibrosis. We previously reported and now confirmed that overexpression of FLIL33 induced phosphorylation of the key profibrotic signaling mediator of TGF-β, Smad3, in primary human lung fibroblasts from healthy donors and idiopathic pulmonary fibrosis patients. Presently, we demonstrate that FLIL33-induced Smad3 phosphorylation was not abrogated by anti-TGF-β antibody but was abrogated by ALK5/TGFBR1-specific and Smad3-specific inhibition, indicating that FLIL33 effect was independent of TGF-β but dependent on its receptor, TGFBR. Western blotting analyses revealed that FLIL33 overexpression increased levels, but did not affect subcellular distribution, of the AP2A1 and AP2B1 subunits of the adaptor protein complex 2 (AP2), a known TGFBR binding partner. siRNA-mediated inhibition of these subunits blocked FLIL33-induced Smad3 phosphorylation, whereas AP2 subunit overexpression induced Smad3 phosphorylation even in the absence of FLIL33. RNA-Seq transcriptomic analyses revealed that fibroblast stimulation with TGF-β induced major changes in expression levels of numerous genes, whereas overexpression of FLIL33 induced modest expression changes in a small number of genes. Furthermore, qRT-PCR tests demonstrated that despite inducing Smad3 phosphorylation, FLIL33 did not induce collagen gene transcription and even mildly attenuated TGF-β-induced levels of collagen I and III mRNAs. We conclude that FLIL33 induces Smad3 phosphorylation through a TGF-β-independent but TGF-β receptor- and AP2- dependent mechanism and has limited downstream transcriptomic consequences.
登录
查看更多内容
影响因子：
--
作者：
Luzina, Irina G.;Todd, Nevins W.;Nacu, Natalia;Lockatell, Virginia;Choi, Jung;Hummers, Laura K.;Atamas, Sergei P.
通讯作者：
Atamas, Sergei P.
影响因子：
2.9
作者：
ABE, M;HARPEL, JG;RIFKIN, DB
通讯作者：
RIFKIN, DB
DOI：
10.4049/jimmunol.1200259
发表时间：
2012-07-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Luzina IG;Pickering EM;Kopach P;Kang PH;Lockatell V;Todd NW;Papadimitriou JC;McKenzie AN;Atamas SP
通讯作者：
Atamas SP
影响因子：
4.8
作者：
Chung, Hyunjae;Ramachandran, Rithwik;Muruve, Daniel A.
通讯作者：
Muruve, Daniel A.
影响因子：
3.3
作者：
Mitchell, H;Choudhury, A;Leof, EB
通讯作者：
Leof, EB