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Dynamic regulation of cardiolipin by the lipid pump Atp8b1 determines the severity of lung injury in experimental pneumonia

Nature Medicine volume 16, pages 1120–1127 (2010)Cite this article

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Abstract

Pneumonia remains the leading cause of death from infection in the US, yet fundamentally new conceptual models underlying its pathogenesis have not emerged. We show that humans and mice with bacterial pneumonia have markedly elevated amounts of cardiolipin, a rare, mitochondrial-specific phospholipid, in lung fluid and find that it potently disrupts surfactant function. Intratracheal cardiolipin administration in mice recapitulates the clinical phenotype of pneumonia, including impaired lung mechanics, modulation of cell survival and cytokine networks and lung consolidation. We have identified and characterized the activity of a unique cardiolipin transporter, the P-type ATPase transmembrane lipid pump Atp8b1, a mutant version of which is associated with severe pneumonia in humans and mice. Atp8b1 bound and internalized cardiolipin from extracellular fluid via a basic residue–enriched motif. Administration of a peptide encompassing the cardiolipin binding motif or Atp8b1 gene transfer in mice lessened bacteria-induced lung injury and improved survival. The results unveil a new paradigm whereby Atp8b1 is a cardiolipin importer whose capacity to remove cardiolipin from lung fluid is exceeded during inflammation or when Atp8b1 is defective. This discovery opens the door for new therapeutic strategies directed at modulating the abundance or molecular interactions of cardiolipin in pneumonia.

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Figure 1: Quantification of cardiolipin in subjects with pneumonia.
Figure 2: Biophysical effects of cardiolipin.
Figure 3: Effect of cardiolipin on lung structure and epithelial cell viability.
Figure 4: Bacteria-induced lung injury after Atp8b1 overexpression.
Figure 5: Bacteria-induced lung injury in Atp8b1-mutant mice.
Figure 6: Bacterial lung injury after CBD peptide administration.

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Acknowledgements

We thank M.E. Anderson, M.J. Welsh, J. Zabner and M. Gladwin for critical review of the manuscript and helpful suggestions. The Atp8b1G308V/G308V 129S1/SvlmJ mutant mice14 were a generous gift from L. Bull (University of California–San Francisco). Nontypable H. influenza strain 12 bacteria were kindly provided by D. Look (University of Iowa)29. Antibodies to ATP8b1 were generous gifts from D. Ortiz (Tufts University)30 and M. Ananthanarayanan (Mount Sinai School of Medicine)31. This material is based upon work supported, in part, by the US Department of Veterans Affairs, Veterans Health Administration, Office of Research and Development, Biomedical Laboratory Research and Development. This work was supported by a Merit Review Award from the US Department of Veterans Affairs and US National Institutes of Health R01 grants HL068135, HL080229, HL081784, HL096376, HL097376 and HL098174 (to R.K.M.), HL70755, HL094488 and NIOSH OH008282 (to V.E.K.) and K23 HL075402 and U01 HL102288 (to L.D.). The contents do not represent the views of the Department of Veterans Affairs or the US government.

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Author notes

  1. Nancy B Ray and Lakshmi Durairaj: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Internal Medicine, University of Iowa, Iowa City, Iowa, USA

    Nancy B Ray, Lakshmi Durairaj, Alan J Ryan, Florita C Henderson, Christopher A Etscheidt, Diann M McCoy, Marianna Agassandian, Satya N Mathur, Jessica C Sieren & Geoffrey McLennan

  2. Department of Internal Medicine, Acute Lung Injury Center of Excellence, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    Bill B Chen, Bryan J McVerry, Michael Donahoe, Alisa K Waltenbaugh, Christopher P O'Donnell, Emily C Hayes-Rowan, Tiffany A Coon & Rama K Mallampalli

  3. Department of Biochemistry, University of Iowa, Iowa City, Iowa, USA

    Phillip L Butler & Lokesh Gakhar

  4. Department of Environmental and Occupational Health, Center for Free Radical and Antioxidant Health, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    Yulia Y Tyurina & Valerian E Kagan

  5. Department of Radiology, University of Iowa, Iowa City, Iowa, USA

    Geoffrey McLennan

  6. Department of Biomedical Engineering, University of Iowa, Iowa City, Iowa, USA

    Geoffrey McLennan

  7. Medical Specialty Service Line, Veteran's Affairs Pittsburgh Healthcare System, Pittsburgh, Pennsylvania, USA

    Rama K Mallampalli

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Contributions

N.B.R. designed and executed cardiolipin-ATP8b1 binding, in vitro imaging and immunological studies and wrote the manuscript. L.D. edited the manuscript and conducted the human studies. B.B.C. performed in vitro (cardiolipin uptake, biochemical and molecular) experiments and all mouse studies. B.J.M. and M.D. contributed to human studies and statistical analyses. A.K.W., T.A.C., M.A., P.L.B., F.C.H. and S.N.M. assisted with in vitro studies. A.J.R. and C.P.O. assisted with mouse studies. D.M.M., E.C.H.-R. and C.A.E. conducted cardiolipin analysis. L.G. conducted surfactant studies. J.C.S. and G.M. designed and conducted in vivo imaging. V.E.K. designed and executed mass spectrometry of cardiolipin, with assistance from Y.Y.T., and provided editorial suggestions. R.K.M. revised the manuscript and directed the study.

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Correspondence to Rama K Mallampalli.

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Ray, N., Durairaj, L., Chen, B. et al. Dynamic regulation of cardiolipin by the lipid pump Atp8b1 determines the severity of lung injury in experimental pneumonia. Nat Med 16, 1120–1127 (2010). https://doi.org/10.1038/nm.2213

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