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The E3 ubiquitin ligase midline 1 promotes allergen and rhinovirus-induced asthma by inhibiting protein phosphatase 2A activity

Nature Medicine volume 19, pages 232–237 (2013)Cite this article

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Abstract

Allergic airway inflammation is associated with activation of innate immune pathways by allergens. Acute exacerbations of asthma are commonly associated with rhinovirus infection. Here we show that, after exposure to house dust mite (HDM) or rhinovirus infection, the E3 ubiquitin ligase midline 1 (MID1) is upregulated in mouse bronchial epithelium. HDM regulates MID1 expression in a Toll-like receptor 4 (TLR4)– and tumor necrosis factor–related apoptosis-inducing ligand (TRAIL)-dependent manner. MID1 decreases protein phosphatase 2A (PP2A) activity through association with its catalytic subunit PP2Ac. siRNA-mediated knockdown of MID1 or pharmacological activation of PP2A using a nonphosphorylatable FTY720 analog in mice exposed to HDM reduces airway hyperreactivity and inflammation, including the expression of interleukin-25 (IL-25), IL-33 and CCL20, IL-5 and IL-13 release, nuclear factor (NF)κB activity, p38 mitogen-activated protein kinase (MAPK) phosphorylation, accumulation of eosinophils, T lymphocytes and myeloid dendritic cells, and the number of mucus-producing cells. MID1 inhibition also limited rhinovirus-induced exacerbation of allergic airway disease. We found that MID1 was upregulated in primary human bronchial epithelial cells upon HDM or rhinovirus exposure, and this correlated with TRAIL and CCL20 expression. Together, these findings identify a key role of MID1 in allergic airway inflammation and links innate immune pathway activation to the development and exacerbation of asthma.

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Figure 1: MID1 silencing reduces allergen-induced airway hyperreactivity, allergic inflammation and increases PP2A activity.
Figure 2: Pharmacological activation of PP2A decreases HDM-induced airway hyperreactivity and allergic airway inflammation.
Figure 3: MID1 inhibition attenuates rhinovirus-induced airway inflammation and asthma exacerbations.
Figure 4: MID1 in allergen and rhinovirus exposed human airway epithelial cells.

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Acknowledgements

This study was supported by the National Health and Medical Research Council (NH&MRC 631075 and 1011153) (J.M., P.S.F., N.V.), the Hunter Medical Research Institute (J.M., P.A.B.W., P.S.F.), the Hunter Children's Research Foundation (J.M., P.S.F., A.P.d.S.) and an NH&MRC Health Practitioner Research Fellowship to J.M. (455623). N.W.B. was supported by a project grant from Asthma UK (06-050) and S.L.J. by a Chair from Asthma UK (CH1155). This work was supported in part by MRC Centre grant G1000758 and ERC FP7 Advanced grant 233015 (to S.L.J.). We would like to thank M. Smyth, Peter MacCallum Cancer Centre, and J. Peschon, Amgen, for providing Tnfsf10−/− mice and S. Akira, Osaka University, for providing Tlr4−/− and Myd88−/− mice. We appreciate technical assistance from C. Cesar de Souza Alves, F. Eyers, J. Girkin, J. Grehan, H. MacDonald, M. Morten, K. Parsons, S. Reeves, L. Sokulsky and the staff from the animal care facilities of the contributing institutes.

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Author notes

  1. Adam Collison and Luke Hatchwell: These authors contributed equally to this work.

Authors and Affiliations

  1. Experimental and Translational Respiratory Group, University of Newcastle and Hunter Medical Research Institute, Newcastle, New South Wales, Australia

    Adam Collison, Luke Hatchwell, Ana Pereira de Siqueira & Joerg Mattes

  2. Priority Research Centre for Asthma and Respiratory Diseases, University of Newcastle and Hunter Medical Research Institute, Newcastle, New South Wales, Australia

    Adam Collison, Luke Hatchwell, Peter A B Wark, Ana Pereira de Siqueira, Melinda Tooze, Paul S Foster & Joerg Mattes

  3. Discipline of Medical Biochemistry, School of Biomedical Science and Pharmacy, University of Newcastle, Newcastle, New South Wales, Australia

    Nicole Verrills & Helen Carpenter

  4. Department of Respiratory and Sleep Medicine, John Hunter Hospital, Newcastle, New South Wales, Australia

    Peter A B Wark & Melinda Tooze

  5. Lowy Cancer Research Centre, Prince of Wales Clinical School, University of New South Wales, Sydney, New South Wales, Australia

    Anthony S Don

  6. School of Chemistry, University of New South Wales, Sydney, New South Wales, Australia

    Jonathan C Morris

  7. Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital Medical Center, University of Cincinnati, Cincinnati, Ohio, USA

    Nives Zimmermann & Marc E Rothenberg

  8. Section of Airway Disease Infection, National Heart and Lung Institute, Medical Research Council and Asthma UK Centre in Allergic Mechanisms of Asthma, Imperial College London, Norfolk Place, London, UK

    Nathan W Bartlett & Sebastian L Johnston

  9. Paediatric Respiratory and Sleep Medicine Unit, Newcastle Children's Hospital, Kaleidoscope, Newcastle, New South Wales, Australia

    Joerg Mattes

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Contributions

A.C. and L.H. performed and designed mouse and cell culture experiments, analyzed data, generated figures and edited the manuscript. P.A.B.W. and M.T. performed and supervised studies on healthy subjects and subjects with asthma and performed cell culture experiments. N.V. and H.C. performed and analyzed PP2Ac measurements and immunoprecipitation and designed experiments. N.V. edited the manuscript. A.S.D. and J.C.M. synthesized AAL(S) for use as an activator of PP2A and developed the dosing regiment. N.Z. and M.E.R. coordinated and assisted in microarray array analysis. N.W.B. and S.L.J. assisted in design of experiments, provided RV1B for further propagation and cDNAs and edited the manuscript. A.P.d.S. coordinated and supervised mouse and human studies. P.S.F. supervised mouse studies, interpreted data and edited the manuscript. J.M. conceptualized, coordinated, designed and supervised mouse and human studies, interpreted and analyzed data, and drafted and edited the manuscript. All authors contributed to data discussion and revised the manuscript.

Corresponding author

Correspondence to Joerg Mattes.

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Competing interests

J.M., A.C, L.H. and P.S.F. have filed a patent application under the Patent Cooperation Treaty that relates to the described data on MID1 function in this manuscript.

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Collison, A., Hatchwell, L., Verrills, N. et al. The E3 ubiquitin ligase midline 1 promotes allergen and rhinovirus-induced asthma by inhibiting protein phosphatase 2A activity. Nat Med 19, 232–237 (2013). https://doi.org/10.1038/nm.3049

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