Details

Autor(en) / Beteiligte

• Luzina, Irina G.
• Fishelevich, Rita
• Hampton, Brian S.
• Courneya, Jean-Paul
• Parisella, Francesca R.
• Lugkey, Katerina N.
• Baleno, Frank X.
• Choi, Dohyun
• Kopach, Pavel
• Lockatell, Virginia
• Todd, Nevins W.
• Atamas, Sergei P.

Titel

Full-length IL-33 regulates Smad3 phosphorylation and gene transcription in a distinctive AP2-dependent manner

Ist Teil von

• Cellular immunology, 2020-11, Vol.357, p.104203-104203, Article 104203

Ort / Verlag

Netherlands: Elsevier Inc

Erscheinungsjahr

2020

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• •Elevated full-length IL-33 induces Smad3 phosphorylation in human lung fibroblasts.•This effect is TGF-β-independent but TGFBR- and adaptor protein complex 2-dependent.•Unlike the impact of TGF-β, elevated FLIL33 modestly affects the transcriptome.•FLIL33 alone or combined with TGF-β does not activate collagen gene transcription.•Previously reported fibrotic potentiation by FLIL33 is likely nontranscriptional. 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.