Article | Published:

An epithelial circadian clock controls pulmonary inflammation and glucocorticoid action

Nature Medicine volume 20, pages 919926 (2014) | Download Citation

Abstract

The circadian system is an important regulator of immune function. Human inflammatory lung diseases frequently show time-of-day variation in symptom severity and lung function, but the mechanisms and cell types underlying these effects remain unclear. We show that pulmonary antibacterial responses are modulated by a circadian clock within epithelial club (Clara) cells. These drive circadian neutrophil recruitment to the lung via the chemokine CXCL5. Genetic ablation of the clock gene Bmal1 (also called Arntl or MOP3) in bronchiolar cells disrupts rhythmic Cxcl5 expression, resulting in exaggerated inflammatory responses to lipopolysaccharide and an impaired host response to Streptococcus pneumoniae infection. Adrenalectomy blocks rhythmic inflammatory responses and the circadian regulation of CXCL5, suggesting a key role for the adrenal axis in driving CXCL5 expression and pulmonary neutrophil recruitment. Glucocorticoid receptor occupancy at the Cxcl5 locus shows circadian oscillations, but this is disrupted in mice with bronchiole-specific ablation of Bmal1, leading to enhanced CXCL5 expression despite normal corticosteroid secretion. The therapeutic effects of the synthetic glucocorticoid dexamethasone depend on intact clock function in the airway. We now define a regulatory mechanism that links the circadian clock and glucocorticoid hormones to control both time-of-day variation and the magnitude of pulmonary inflammation and responses to bacterial infection.

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Acknowledgements

We thank J.S. Takahashi, Southwestern University, for the gift of the Per2-luc mice, F. Scott and A. Hughes for help with imaging lung tissue slices and J. Woodburn for technical help. We thank M.H. Hastings for comments on the manuscript. The work was supported by research grants to A.L. from GlaxoSmithKline and the Biotechnology and Biological Sciences Research Council, UK (BB/D004357/1, BB/K003119/1 and BB/K003097/1), to D.R. from the Wellcome Trust and the UK National Institute for Health Research Musculoskeletal Biomedical Research Unit Manchester, as well as the US National Institute of Diabetes and Digestive and Kidney Diseases grant P01 DK59820 to F.D. and US National Institutes of Health grants 1RO1HL105834 and 1RO1AI099479 to G.S.W.

Author information

Affiliations

  1. Faculty of Medical and Human Sciences, University of Manchester, Manchester Academic Health Sciences Centre, Manchester, UK.

    • Julie Gibbs
    • , Laura Matthews
    • , Nan Yang
    • , Toryn Poolman
    • , Marie Pariollaud
    • , Stuart Farrow
    •  & David Ray
  2. Faculty of Life Sciences, University of Manchester, Manchester, UK.

    • Louise Ince
    • , Ben Saer
    • , Nicola Begley
    •  & Andrew Loudon
  3. Division of Neonatology, Children's Hospital of Philadelphia, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

    • Junjie Mei
    •  & G Scott Worthen
  4. Manchester Collaborative Centre for Inflammation Research, Core Technology Facility, University of Manchester, Manchester, UK.

    • Thomas Bell
    •  & Tracy Hussell
  5. Respiratory Therapy Area, GlaxoSmithKline, Stevenage, UK.

    • Stuart Farrow
  6. Dan L. Duncan Cancer Center, Baylor College of Medicine, Houston, Texas, USA.

    • Francesco DeMayo

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Contributions

J.G., D.R., S.F. and A.L. conceived the project. J.G., D.R., G.S.W. and A.L. designed the experiments, and J.G., D.R. and A.L. wrote the paper. J.G. performed all in vivo experiments, oversaw all breeding programs, as well as measures of cellular infiltrates, qPCR gene and cytokine protein expression studies and all statistical analyses; L.I. carried out all adrenalectomies and the associated in vivo studies and downstream analyses; L.M. performed transfection studies; M.P. undertook studies of human bronchiolar epithelial cells; J.M. and G.S.W. undertook all studies of CXCL5-null mice; N.Y. performed ChIP studies; T.P. analyzed DEX-responsive genes in the lung; T.B. and T.H. led pulmonary S. pneumoniae infection studies; F.D. developed the Ccsp-icre mouse model; B.S. and N.B. managed the transgenic animal colony and genotyping; and B.S. undertook the radioactive in situ hybridization studies.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to David Ray or Andrew Loudon.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–14 and Supplementary Tables 1–4

Videos

  1. 1.

    CCSP-bmal–/– lung video.

    Video showing bioluminescence over time in a lung slice from a CCSP-bmal–/– mouse on a PER2::luc background. The slice was recorded for 3 d.

  2. 2.

    Wild-type lung video.

    Video showing bioluminescence over time in a lung slice from a wild-type mouse on a PER2::luc background. The slice was recorded for 3 d.

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DOI

https://doi.org/10.1038/nm.3599

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