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Slit3 secreted from M2-like macrophages increases sympathetic activity and thermogenesis in adipose tissue

Nature Metabolism volume 3, pages 1536–1551 (2021)Cite this article

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Abstract

Beiging of white adipose tissue (WAT) is associated with an increase of anti-inflammatory M2-like macrophages in WAT. However, mechanisms through which M2-like macrophages affect beiging are incompletely understood. Here, we show that the macrophage cytokine Slit3 is secreted by adipose tissue macrophages and promotes cold adaptation by stimulating sympathetic innervation and thermogenesis in mice. Analysing the transcriptome of M2-like macrophages in murine inguinal WAT (iWAT) after cold exposure, we identify Slit3 as a secreted cytokine. Slit3 binds to the ROBO1 receptor on sympathetic neurons to stimulate Ca2+/calmodulin-dependent protein kinase II signalling and norepinephrine release, which enhances adipocyte thermogenesis. Adoptive transfer of Slit3-overexpressing M2 macrophages to iWAT promotes beiging and thermogenesis, whereas mice that lack Slit3 in myeloid cells are cold-intolerant and gain more weight. Our findings shed new light on the integral role of M2-like macrophages for adipose tissue homeostasis and uncover the macrophage–Slit3–sympathetic neuron–adipocyte signalling axis as a regulator of long-term cold adaptation.

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Fig. 1: Slit3 is cold sensitive and secreted by M2 macrophages.
Fig. 2: Slit3-overexpressed M2 cells increase thermogenesis.
Fig. 3: Slit3 activates TH activity in sympathetic nerves.
Fig. 4: SNS denervation in iWAT blocked Slit3-induced thermogenesis.
Fig. 5: Monocytic deletion of Slit3 impairs thermogenesis.
Fig. 6: Slit3 stimulates the phosphorylation of TH via PKA/CaMKII.
Fig. 7: ROBO1 is the receptor for Slit3 to stimulate NE production.
Fig. 8: Knockdown of ROBO1 in iWAT blocks thermogenesis.

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Data availability statement

The raw RNA-sequence data used to generate Fig. 1b,c are available at the Sequence Read Archive (accession no. PRJNA744369). The data used to generate the main results shown in Figs. 1–8 are available as source data. Source data including uncropped western blots and raw microscopy images are provided with this paper or upon request from the corresponding authors: S.Q., e-mail shuwenqian2013@163.com or shuwenqian@fudan.edu.cn and Q.-Q.T., e-mail qqtang@shmu.edu.cn.

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Acknowledgements

We thank GemPharmatech Co., Ltd, Nanjing, China for producing the Slit3f/f mice. This work was supported by the National Key R&D Program of the Ministry of Science and Technology of China (grant no. 2018YFA0800401) to Q.-Q.T., and National Natural Science Foundation grant nos. 31670787 to S.Q., 81730021 to Q.-Q.T. and 81970754 to Y.T.

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Authors and Affiliations

  1. Key Laboratory of Metabolism and Molecular Medicine of the Ministry of Education, Department of Biochemistry and Molecular Biology of School of Basic Medical Sciences and Department of Endocrinology and Metabolism of Zhongshan Hospital, Fudan University, Shanghai, China

    Yi-Na Wang, Yan Tang, Zhihui He, Linyuan Wang, Yang Liu, Qiqi Yang, Dongning Pan, Shuwen Qian & Qi-Qun Tang

  2. Key Laboratory of Metabolism and Molecular Medicine of the Ministry of Education, Institutes of Biomedical Sciences, Fudan University, Shanghai, China

    Hong Ma

  3. State Key Laboratory of Medical Neurobiology and MOE Frontier Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai, China

    Cuiqing Zhu

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Contributions

S.Q., Q.-Q.T. and Y.-N.W. conceived and designed the experiments. S.Q. and Y.-N.W. wrote the paper. S.Q. and Q.-Q.T. administrated the project. Q.-Q.T., S.Q. and Y.T. acquired the funding. Y.-N.W., S.Q., Y.T., Z.H., H.M., L.W., Y.L., Q.Y. and C.Z. performed the experiments. Y.-N.W., D.P., C.Z. and S.Q. reviewed and edited the paper.

Corresponding authors

Correspondence to Shuwen Qian or Qi-Qun Tang.

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The authors declare no competing interests.

Additional information

Peer review information Nature Metabolism thanks Sheng Bi and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Primary Handling Editor: Isabella Samuelson.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–8 and Table 1.

Reporting Summary

Supplementary Data 1

RNA-sequencing data of all room temperature versus cold differentially expressed genes.

Supplementary Data 2

All data presented in the graphs in the figures and supplementary figures.

Supplementary Data 3

Raw microscopic images for the main figures.

Supplementary Data 4

Uncropped western blot images for the supplementary figures.

Supplementary Data 5

Statistic results of exact P values for the supplementary figures.

Supplementary Data 6

Raw microscopic images for all supplementary figures.

Source data

Source Data Fig. 1

Uncropped western blot images.

Source Data Fig. 1

Statistic results of exact P values.

Source Data Fig. 2

Uncropped western blot images.

Source Data Fig. 2

Statistic results of exact P values.

Source Data Fig. 3

Uncropped western blot images.

Source Data Fig. 3

Statistic results of exact P values.

Source Data Fig. 4

Uncropped western blot images.

Source Data Fig. 4

Statistic results of exact P values.

Source Data Fig. 5

Uncropped western blot images.

Source Data Fig. 5

Statistic results of exact P values.

Source Data Fig. 6

Uncropped western blot images.

Source Data Fig. 6

Statistic results of exact P values.

Source Data Fig. 7

Uncropped western blot images.

Source Data Fig. 7

Statistic results of exact P values.

Source Data Fig. 8

Uncropped western blot images.

Source Data Fig. 8

Statistic results of exact P values.

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Wang, YN., Tang, Y., He, Z. et al. Slit3 secreted from M2-like macrophages increases sympathetic activity and thermogenesis in adipose tissue. Nat Metab 3, 1536–1551 (2021). https://doi.org/10.1038/s42255-021-00482-9

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