EP4 receptor-associated protein in macrophages protects against bleomycin-induced pulmonary inflammation in mice

Sei Higuchi, Risako Fujikawa, Taichi Ikedo, Kosuke Hayashi, Mika Yasui, Manabu Nagata, Masato Nakatsuji, Masayuki Yokode, Manabu Minami

Research output: Contribution to journalArticle

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Abstract

Excessive activation of inflammatory macrophages drives the pathogenesis of many chronic diseases. EP4 receptor-associated protein (EPRAP) has been identified as a novel, anti-inflammatory molecule in macrophages. In this study, we investigated the role of EPRAP using a murine model of bleomycin (BLM)-induced pulmonary inflammation. When compared with wild-type mice, EPRAP-deficient mice exhibited significantly higher mortality, and increased accumulation of macrophages and proinflammatory molecules in the lung 7 d post-BLM administration. Accordingly, the levels of phosphorylated p105, MEK1/2, and ERK1/2 were elevated in EPRAP-deficient alveolar macrophages following BLM administration. In contrast, macrophage-specific EPRAP overexpression decreased the production of proinflammatory cytokines and chemokines, suggesting that EPRAP in macrophages plays a key role in attenuating BLM-induced pulmonary inflammation. As EPRAP is phosphorylated after translation, we examined the role of posttranslational modifications in cellular inflammatory activation using mouse embryo fibroblasts (MEFs) expressing mutant EPRAP proteins. Expression of mutant EPRAP, in which serine-108 and serine-608 were replaced with alanine (EPRAP S108A/S608A), markedly suppressed TNF-a production in LPS-treated MEFs. Conversely, the serine phosphatase 2A (PP2A) inhibitor, cantharidic acid, increased LPS-induced TNF-α production in MEFs expressing wild-type EPRAP, but not in MEFs expressing EPRAP S108A/S608A. Immunoprecipitation analyses demonstrated that EPRAP associated with PP2A in both MEFs and alveolar macrophages from BLM-treated mice. Our data suggest that PP2A dephosphorylates EPRAP, which may be a crucial step in exertion of its anti-inflammatory properties. For these reasons, we believe the EPRAP-PP2A axis in macrophages holds the key to treating chronic inflammatory disorders.

Original languageEnglish
Pages (from-to)4436-4443
Number of pages8
JournalJournal of Immunology
Volume197
Issue number11
DOIs
Publication statusPublished - Dec 1 2016

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Bleomycin
Pneumonia
Macrophages
Proteins
Embryonic Structures
Fibroblasts
Phosphoric Monoester Hydrolases
Serine
Alveolar Macrophages
Anti-Inflammatory Agents
Protein Phosphatase 2
Macrophage Activation
Post Translational Protein Processing
Chemokines
Immunoprecipitation
Alanine
Chronic Disease

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

EP4 receptor-associated protein in macrophages protects against bleomycin-induced pulmonary inflammation in mice. / Higuchi, Sei; Fujikawa, Risako; Ikedo, Taichi; Hayashi, Kosuke; Yasui, Mika; Nagata, Manabu; Nakatsuji, Masato; Yokode, Masayuki; Minami, Manabu.

In: Journal of Immunology, Vol. 197, No. 11, 01.12.2016, p. 4436-4443.

Research output: Contribution to journalArticle

Higuchi, S, Fujikawa, R, Ikedo, T, Hayashi, K, Yasui, M, Nagata, M, Nakatsuji, M, Yokode, M & Minami, M 2016, 'EP4 receptor-associated protein in macrophages protects against bleomycin-induced pulmonary inflammation in mice', Journal of Immunology, vol. 197, no. 11, pp. 4436-4443. https://doi.org/10.4049/jimmunol.1502618
Higuchi, Sei ; Fujikawa, Risako ; Ikedo, Taichi ; Hayashi, Kosuke ; Yasui, Mika ; Nagata, Manabu ; Nakatsuji, Masato ; Yokode, Masayuki ; Minami, Manabu. / EP4 receptor-associated protein in macrophages protects against bleomycin-induced pulmonary inflammation in mice. In: Journal of Immunology. 2016 ; Vol. 197, No. 11. pp. 4436-4443.
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