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
View: Text | PDF | Corrigendum
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

×

Figure 5

NPFF increases the proliferation of ATMs.

Options: View larger image (or click on image) Download as PowerPoint
NPFF increases the proliferation of ATMs. (A) Mice were injected with Br...
(A) Mice were injected with BrdU, and ATMs were isolated after BrdU administration as shown. FACS analysis of BrdU+ ATMs from eWAT. BrdU incorporation was also assessed in blood cells after 4 hours. SSC-A, side scatter area. (B) SVF cells pooled from eWAT from 6 lean mice were cultured in vitro (Control). SVF cells were incubated with clodronate liposomes for 1 hour (Depletion). Apoptotic cells were removed by washing, and surviving cells were allowed to recover in fresh medium for 1 hour and 4 hours. Scale bar: 100 μm. (C) FACS analysis of cells from the experiment shown in B (left). Relative transcription of Ccne and Emr1 in SVF cells treated with clodronate liposomes (blue bars) and allowed to recover for 2 hours (green bars) (right). Dashed lines indicate gene transcription level in control cells. (D) Overview of ATM cultures treated with vehicle or 0.5 nM NPFF (18 hours). Scale bars: 50 μm. (E) Number of ATMs after treatment with vehicle or 0.5 nM NPFF for 18 hours. (F) Proliferation of mouse ATMs cultured in vitro and treated for 18 hours with 0.5 nM NPFF. ATMs were pooled from 3 to 5 mice and treated in triplicate in D–F. (G) Proliferation of human ATMs in response to treatment with 0.5 nM NPFF for 18 hours. ATMs were isolated and cultured in triplicate from each donor. (H) HFD-fed mice were treated with vehicle or NPFF as shown in Figure 2H. Relative transcription of the macrophage markers Emr1 and Cd64 (also known as Fcgr1a) was measured in SVF. Each data point represents pooled ATMs from eWAT of 2 mice. The experiment was conducted 2 times. Quantity of ATMs in eWAT after treatment with vehicle or NPFF (n = 6). The values for iWAT are shown in Supplemental Figure 16G. (I) Proliferation of in vitro cultured WT and Npffr2-KO ATMs treated with 0.5 nM NPFF for 4 hours. ATMs were pooled from 6 mice and treated in triplicate. The experiment was conducted 6 times. *P < 0.05, **P < 0.01, and ***P < 0.001, by unpaired, 2-tailed Student’s t test.