Neuropeptide FF increases M2 activation and self-renewal of adipose tissue macrophages
Syed F. Hassnain Waqas, Anh Cuong Hoang, Ya-Tin Lin, Grace Ampem, Hind Azegrouz, Lajos Balogh, Julianna Thuróczy, Jin-Chung Chen, Ivan C. Gerling, Sorim Nam, Jong-Seok Lim, Juncal Martinez-Ibañez, José T. Real, Stephan Paschke, Raphaëlle Quillet, Safia Ayachi, Frédéric Simonin, E. Marion Schneider, Jacqueline A. Brinkman, Dudley W. Lamming, Christine M. Seroogy, Tamás Röszer
Syed F. Hassnain Waqas, Anh Cuong Hoang, Ya-Tin Lin, Grace Ampem, Hind Azegrouz, Lajos Balogh, Julianna Thuróczy, Jin-Chung Chen, Ivan C. Gerling, Sorim Nam, Jong-Seok Lim, Juncal Martinez-Ibañez, José T. Real, Stephan Paschke, Raphaëlle Quillet, Safia Ayachi, Frédéric Simonin, E. Marion Schneider, Jacqueline A. Brinkman, Dudley W. Lamming, Christine M. Seroogy, Tamás Röszer
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Research Article Endocrinology Inflammation

Neuropeptide FF increases M2 activation and self-renewal of adipose tissue macrophages

Abstract

The quantity and activation state of adipose tissue macrophages (ATMs) impact the development of obesity-induced metabolic diseases. Appetite-controlling hormones play key roles in obesity; however, our understanding of their effects on ATMs is limited. Here, we have shown that human and mouse ATMs express NPFFR2, a receptor for the appetite-reducing neuropeptide FF (NPFF), and that NPFFR2 expression is upregulated by IL-4, an M2-polarizing cytokine. Plasma levels of NPFF decreased in obese patients and high-fat diet–fed mice and increased following caloric restriction. NPFF promoted M2 activation and increased the proliferation of murine and human ATMs. Both M2 activation and increased ATM proliferation were abolished in NPFFR2-deficient ATMs. Mechanistically, the effects of NPFF involved the suppression of E3 ubiquitin ligase RNF128 expression, resulting in enhanced stability of phosphorylated STAT6 and increased transcription of the M2 macrophage–associated genes IL-4 receptor α (Il4ra), arginase 1 (Arg1), IL-10 (Il10), and alkylglycerol monooxygenase (Agmo). NPFF induced ATM proliferation concomitantly with the increase in N-Myc downstream-regulated gene 2 (Ndrg2) expression and suppressed the transcription of Ifi200 cell-cycle inhibitor family members and MAF bZIP transcription factor B (Mafb), a negative regulator of macrophage proliferation. NPFF thus plays an important role in supporting healthy adipose tissue via the maintenance of metabolically beneficial ATMs.

Authors

Syed F. Hassnain Waqas, Anh Cuong Hoang, Ya-Tin Lin, Grace Ampem, Hind Azegrouz, Lajos Balogh, Julianna Thuróczy, Jin-Chung Chen, Ivan C. Gerling, Sorim Nam, Jong-Seok Lim, Juncal Martinez-Ibañez, José T. Real, Stephan Paschke, Raphaëlle Quillet, Safia Ayachi, Frédéric Simonin, E. Marion Schneider, Jacqueline A. Brinkman, Dudley W. Lamming, Christine M. Seroogy, Tamás Röszer

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Figure 4

NPFF inhibits p-STAT6 decay and triggers M2 activation of ATMs.

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NPFF inhibits p-STAT6 decay and triggers M2 activation of ATMs. (A) Effe...
(A) Effect of the STAT6 inhibitor AS1517499 (200 nM) on NPFF-induced Arg1 and Il4ra transcription. ATMs were treated with 0.5 nM NPFF for 1 hour. (B) Western blot of p-STAT6/STAT6 in ATMs treated with 100 ng/ml IL-4 or 0.5 nM NPFF. Each lane corresponds to pooled ATMs from 3 to 5 mice. Results are representative of 3 independent experiments. (C) p-STAT6 levels following 10 ng/ml IL-4 and 0.5 nM NPFF treatment, as measured by in-cell ELISA. (D) Effect of 10 ng/ml IL-4 and 0.5 nM NPFF on the transcription of Il4ra in ATMs. (E) Effect of 10 ng/ml IL-4 and 0.5 nM NPFF on the transcription of Rnf128 in ATMs. (F) RNF128 levels in lysates of ATMs treated with 10 ng/ml IL-4, 0.5 nM NPFF, or their combination. In C–F, the dotted lines represent the levels with vehicle treatment. (G and H) Relative transcription and MFI of M2 markers in control and RNF128-overexpressing (RNF128-OE) J774A.1 macrophages after a 4-hour treatment with vehicle or 0.5 nM NPFF. (I) MFI of M2 markers in ATMs from WT and Rnf128-KO mice. Data points represent pooled ATMs from 2 mice. The experiment was conducted 2 times. (J) Percentage of M2 ATMs and relative transcription of Il10 in ATMs from WT and NPFFR2-overexpressing (NPFFR2-OE) mice (n = 6). (K) Relative transcription of Rnf128 in ATMs from vehicle-treated and NPFF-treated HFD-fed mice (treatment scheme as in Figure 2H). Data points represent pooled ATMs from 2 mice. The experiment was conducted 2 times. In A and C–F, the data points represent pooled ATMs from 3 mice. The experiment was conducted 3 times. (L and M) Arginase 1 expression (L) and transcriptional changes (M) in WT and Npffr2-KO ATMs evoked by 0.5 nM NPFF within 1 hour. ATMs harvested from 6 mice and treated in triplicate. The experiment was conducted 6 times. *P < 0.05, **P < 0.01, and ***P < 0.001 by 1-way ANOVA with Dunnett’s post-hoc test (A and C–H) and unpaired, 2-tailed Student’s t test (I–M).