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MicroRNA-99a mimics inhibit M1 macrophage phenotype and adipose tissue inflammation by targeting TNFα

Cellular & Molecular Immunology volume 16, pages 495–507 (2019)Cite this article

Abstract

In human adipose tissue and obesity, miR-99a expression is negatively correlated with inflammation. Therefore, the present study investigated the role of miR-99a in macrophage phenotype activation and adipose tissue inflammation. M2 BMDMs showed a significant increase in miR-99a expression when compared to the M0 and M1 phenotypes. Phenotype-switching experiments established an association between upregulated miR-99a expression and the M2 phenotype. Overexpression of miR-99a prevented M1 phenotype activation and attenuated bactericidal activity. Likewise, knockdown of miR-99a abolished M2 phenotype activation. By means of in silico target prediction tools and a luciferase reporter assay, TNFα was identified as a direct target of miR-99a. Knockdown of TNFα recapitulated the effect of miR-99a overexpression in M1 BMDMs. In a db/db mice model, miR-99a expression was reduced in eWAT and F4/80+ ATMs. Systemic overexpression of miR-99a in db/db mice attenuated adipocyte hypertrophy with increased CD301 and reduced CD86 immunostaining. Flow cytometry analysis also showed an increased M2 and a reduced M1 macrophage population. Mimics of miR-99a also improved the diabetic dyslipidemia and insulin signaling in eWAT and liver, with an attenuated expression of gluconeogenesis and cholesterol metabolism genes in the liver. Furthermore, adoptive transfer of miR-99a-overexpressing macrophages in the db/db mice recapitulated in vivo miR-99a mimic effects with increased M2 and reduced M1 macrophage populations and improved systemic glucose, insulin sensitivity, and insulin signaling in the eWAT and liver. The present study demonstrates that miR-99a mimics can regulate macrophage M1 phenotype activation by targeting TNFα. miR-99a therapeutics in diabetic mice reduces the adipose tissue inflammation and improves insulin sensitivity.

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Acknowledgements

The authors gratefully acknowledge the technical help provided by Mr. A.L. Vishwakarma and Mr. C.P. Pandey. This work was supported by the CSIR-Network project: “Towards a holistic understanding of complex diseases: Unraveling the threads of complex disease (THUNDER)”. A.J. and M.M. are supported by UGC, New Delhi, P.M. is supported by CSIR, New Delhi, and S.S.R. is supported by THUNDER. CDRI Communication number: 9683

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  1. Division of Pharmacology, Council of Scientific and Industrial Research-Central Drug Research Institute (CSIR-CDRI), Lucknow, 226031, U.P., India

    Anant Jaiswal, Mohita Maurya & Manoj Kumar Barthwal

  2. Academy of Scientific and Innovative Research, New Delhi, 110025, India

    Sukka Santosh Reddy & Preeti Maurya

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Correspondence to Manoj Kumar Barthwal.

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Jaiswal, A., Reddy, S.S., Maurya, M. et al. MicroRNA-99a mimics inhibit M1 macrophage phenotype and adipose tissue inflammation by targeting TNFα. Cell Mol Immunol 16, 495–507 (2019). https://doi.org/10.1038/s41423-018-0038-7

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