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TIMAP repression by TGFβ and HDAC3-associated Smad signaling regulates macrophage M2 phenotypic phagocytosis

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Abstract

TIMAP (TGFβ-inhibited membrane-associated protein) is an endothelium-enriched TGFβ downstream protein and structurally belongs to the targeting subunit of myosin phosphatase; however, the mechanism of TGFβ repressing TIMAP and its functional relevance to TGFβ bioactivity remain largely unknown. Here, we report that TIMAP is reduced in TGFβ-elevated mouse fibrotic kidney and highly expressed in macrophages. TGFβ repression of TIMAP is associated with HDAC3 upregulation and its recruitment by Smad2/3 at the Smad binding element on TIMAP promoter, whereas specific HDAC3 inhibition reversed the TIMAP repression, suggesting that TGFβ transcriptionally downregulates TIMAP through HDAC3-associated Smad signaling. Further investigation showed that TIMAP over-expression interrupted TGFβ-associated Smad signaling and TIMAP repression by TGFβ correlated with TGFβ-induced macrophage M2 polarization markers, migration, and phagocytosis—the processes promoted by phosphorylation of the putative TIMAP substrate myosin light chain (MLC). Consistently, TIMAP dephosphorylated MLC in macrophages and TGFβ induced macrophage migration and phagocytosis in TIMAP- and MLC phosphorylation-dependent manners, suggesting that TIMAP dephosphorylation of MLC constitutes an essential regulatory loop mitigating TGFβ-associated macrophage M2 phenotypic activities. Given that hyperactive TGFβ often causes excessive macrophage phagocytic activities potentially leading to various chronic disorders, the strategies targeting HDAC3/TIMAP axis might improve TGFβ-associated pathological processes.

Key message

  • TIMAP is enriched in the endothelium and highly expressed in macrophages.

  • TIMAP is suppressed by TGFβ via HDAC3-associated Smad signaling.

  • TIMAP inhibits TGFβ signaling and TGFβ-associated macrophage M2 polarization.

  • TIMAP dephosphorylation of MLC counteracts TGFβ-induced macrophage phagocytosis.

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Acknowledgments

This work was supported by research grants from the National Nature Science Foundation of China (81271301, 81470940, and 81670672) to W. Cao.

Authors’ contributions

JY performed most experiments and participated in manuscript writing; SY, FB, LL, and QT assisted in animal work, data collection, and some experiments. HW provided critical advice; WC designed and conceived the project, analyzed and arranged results, and wrote the manuscript. All the authors reviewed the manuscript.

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Authors and Affiliations

  1. Nanjing University School of Medicine, Jiangsu Key Laboratory of Molecular Medicine, 22 Hankou Road, Room 209, Science & Technology Building, Nanjing, 210093, China

    Jun Yang, Shasha Yin, Fangfang Bi, Lin Liu, Tian Qin, Hongwei Wang & Wangsen Cao

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  1. Jun Yang
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  2. Shasha Yin
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Correspondence to Wangsen Cao.

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Yang, J., Yin, S., Bi, F. et al. TIMAP repression by TGFβ and HDAC3-associated Smad signaling regulates macrophage M2 phenotypic phagocytosis. J Mol Med 95, 273–285 (2017). https://doi.org/10.1007/s00109-016-1479-z

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  • DOI: https://doi.org/10.1007/s00109-016-1479-z

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