Granulocyte macrophage colony-stimulating factor induces CCL17 production via IRF4 to mediate inflammation
Adrian Achuthan, Andrew D. Cook, Ming-Chin Lee, Reem Saleh, Hsu-Wei Khiew, Melody W.N. Chang, Cynthia Louis, Andrew J. Fleetwood, Derek C. Lacey, Anne D. Christensen, Ashlee T. Frye, Pui Yeng Lam, Hitoshi Kusano, Koji Nomura, Nancy Steiner, Irmgard Förster, Stephen L. Nutt, Moshe Olshansky, Stephen J. Turner, John A. Hamilton
Adrian Achuthan, Andrew D. Cook, Ming-Chin Lee, Reem Saleh, Hsu-Wei Khiew, Melody W.N. Chang, Cynthia Louis, Andrew J. Fleetwood, Derek C. Lacey, Anne D. Christensen, Ashlee T. Frye, Pui Yeng Lam, Hitoshi Kusano, Koji Nomura, Nancy Steiner, Irmgard Förster, Stephen L. Nutt, Moshe Olshansky, Stephen J. Turner, John A. Hamilton
View: Text | PDF
Research Article Inflammation

Granulocyte macrophage colony-stimulating factor induces CCL17 production via IRF4 to mediate inflammation

Abstract

Data from preclinical and clinical studies have demonstrated that granulocyte macrophage colony-stimulating factor (GM-CSF) can function as a key proinflammatory cytokine. However, therapies that directly target GM-CSF function could lead to undesirable side effects, creating a need to delineate downstream pathways and mediators. In this work, we provide evidence that GM-CSF drives CCL17 production by acting through an IFN regulatory factor 4–dependent (IRF4-dependent) pathway in human monocytes, murine macrophages, and mice in vivo. In murine models of arthritis and pain, IRF4 regulated the formation of CCL17, which mediated the proinflammatory and algesic actions of GM-CSF. Mechanistically, GM-CSF upregulated IRF4 expression by enhancing JMJD3 demethylase activity. We also determined that CCL17 has chemokine-independent functions in inflammatory arthritis and pain. These findings indicate that GM-CSF can mediate inflammation and pain by regulating IRF4-induced CCL17 production, providing insights into a pathway with potential therapeutic avenues for the treatment of inflammatory diseases and their associated pain.

Authors

Adrian Achuthan, Andrew D. Cook, Ming-Chin Lee, Reem Saleh, Hsu-Wei Khiew, Melody W.N. Chang, Cynthia Louis, Andrew J. Fleetwood, Derek C. Lacey, Anne D. Christensen, Ashlee T. Frye, Pui Yeng Lam, Hitoshi Kusano, Koji Nomura, Nancy Steiner, Irmgard Förster, Stephen L. Nutt, Moshe Olshansky, Stephen J. Turner, John A. Hamilton

×

Figure 10

GM-CSF regulates IRF4 expression via JMJD3 demethylase in human monocytes, and inhibition of JMJD3 demethylase ameliorates GM-CSF–driven inflammatory pain.

Options: View larger image (or click on image) Download as PowerPoint
GM-CSF regulates IRF4 expression via JMJD3 demethylase in human monocyte...
(A and B) Human monocytes were treated with GM-CSF (10 ng/ml) for indicated periods of time. (A) KDM6B, IRF4, and CCL17 mRNA expression (qPCR) and (B) JMJD3 activity were determined (n = 5). (C and D) Human monocytes were pretreated with JMJD3 inhibitor GSK-J4 (10 nM) or DMSO for 30 minutes before treatment with GM-CSF (10 ng/ml) for 16 hours. (C) Whole cell lysates were subjected to Western blotting with anti-JMJD3, anti-IRF4, anti–β-actin, anti-H3K27me3, and anti-H3 antibodies and (D) secreted CCL17 measured by ELISA (n = 4). (E) Human monocytes were pretreated with JMJD3 inhibitor GSK-J4 (10 nM) or DMSO for 30 minutes before treatment with GM-CSF (10 ng/ml) for 1 hour. ChIP analysis of the association of RNA Pol II, H3K27me3, and total H3 with the IRF4 TSS is expressed as percentage of input DNA (n = 4). (F) i.pl. injection of GM-CSF (20 ng) or saline in WT mice treated with/without GSK-J4 (25 mg/kg i.p. at t = –30 minutes). Pain development was measured (n = 11 per group). Graphs are plotted as mean ± SEM. P values were obtained using a 1-way ANOVA test (A, B, D, and E) and a 2-way ANOVA test (F). *P < 0.05; **P < 0.01, vs. t = 0; ***P < 0.001, saline vs. GM-CSF + vehicle. ####P < 0.0001, vehicle vs. GSK-J4.