Neuropeptide FF increases M2 activation and self-renewal of adipose tissue macrophages
Syed F. Hassnain Waqas, … , Christine M. Seroogy, Tamás Röszer
Syed F. Hassnain Waqas, … , Christine M. Seroogy, Tamás Röszer
Published June 5, 2017
Citation Information: J Clin Invest. 2017;127(7):2842-2854. https://doi.org/10.1172/JCI90152.
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Research Article Endocrinology Inflammation Article has an altmetric score of 35

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 1

Human and mouse ATMs express NPFFR2, which increases after IL-4 exposure.

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Human and mouse ATMs express NPFFR2, which increases after IL-4 exposure...
(A) Diagram of human cell types of the omental fat depot analyzed in this study. ATMs, CD68+CD14+ ATMs; SCs, CD68–CD14– stromal cells. (B) NPFFR2 transcription in SVF and adipocytes of human omental fat (n = 4). (C) FACS analysis of NPFFR2 expression in human ATMs and SCs. (D) NPFFR2 immunostaining in human omental fat. Scale bar: 25 μm. (E) TEM image of an NPFFR2+ human ATM. Scale bar: 0.5 μm. (F) Diagram of the analyzed mouse fat depots. (G) Relative transcription of Npffr2 in SVF of lean mice; each data point shows pools of cells from 3 mice. (H) FACS analysis of NPFFR2 expression in mouse ATMs and iMCs. (I) Immunostaining of NPFFR2 in mouse eWAT. Arrowheads indicate the cell border. Scale bars for both images: 25 μm. (J) TEM image of an NPFFR2+ mouse ATM. Scale bar: 0.5 μm. (K) Western blot of NPFFR2 in mouse ATMs. Lanes 1–3: ATMs from eWAT; lanes 4–6: ATMs from iWAT. (L) FACS analysis of NPFFR2 expression in mouse macrophages. PMFs, peritoneal macrophages; KCs, Kupffer cells; BMDMs, nonstimulated bone marrow–derived macrophages (n = 3). (M) Npffr2 transcription in mouse ATMs after 100 ng/ml LPS or 2.5 ng/ml IFN-γ treatment for 4 hours in vitro or after 3 months on a NCD or HFD (n = 3 and 5, respectively). (N) NPFFR2+ ATMs in NCD- or HFD-fed mice. Each data points represent pooled ATMs from 2 mice. The experiment was conducted 2 times. (O and P) Npffr2 and NPFFR2 transcription after 10 ng/ml IL-4 treatment of mouse (O) and human (P) ATMs (n = 3). Human ATMs were treated for 4 hours. *P < 0.05, **P < 0.01, and ***P < 0.001, by unpaired, 2-tailed Student’s t test (B, C, G, M, N, and P) and 1-way ANOVA with Dunnett’s post-hoc test (O). ac, adipocyte; fp, filopodia; iMCs, immature myeloid cells; iso, isotype; lp, lamellipodia; MFI, mean fluorescence intensity; mt, mitochondria; nc, nucleus; vs, vesicle.