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A Staphylococcus aureus pore-forming toxin subverts the activity of ADAM10 to cause lethal infection in mice

Abstract

Staphylococcus aureus is a major cause of human disease, responsible for half a million infections and approximately 20,000 deaths per year in the United States alone1,2. This pathogen secretes α-hemolysin, a pore-forming cytotoxin that contributes to the pathogenesis of pneumonia3,4,5. α-hemolysin injures epithelial cells in vitro by interacting with its receptor, the zinc-dependent metalloprotease ADAM10 (ref. 6). We show here that mice harboring a conditional disruption of the Adam10 gene in lung epithelium are resistant to lethal pneumonia. Investigation of the molecular mechanism of toxin-receptor function revealed that α-hemolysin upregulates ADAM10 metalloprotease activity in alveolar epithelial cells, resulting in cleavage of the adherens junction protein E-cadherin. Cleavage is associated with disruption of epithelial barrier function, contributing to the pathogenesis of lethal acute lung injury. A metalloprotease inhibitor of ADAM10 prevents E-cadherin cleavage in response to Hla; similarly, toxin-dependent E-cadherin proteolysis and barrier disruption is attenuated in ADAM10-knockout mice. Together, these data attest to the function of ADAM10 as the cellular receptor for α-hemolysin. The observation that α-hemolysin can usurp the metalloprotease activity of its receptor reveals a previously unknown mechanism of pore-forming cytotoxin action in which pathologic insults are not solely the result of irreversible membrane injury and defines ADAM10 inhibition as a strategy to attenuate α-hemolysin-induced disease.

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Figure 1: ADAM10 contributes to lethal S. aureus pneumonia.
Figure 2: Hla induces ADAM10-dependent epithelial barrier disruption and E-cadherin cleavage.
Figure 3: Hla is required for E-cadherin cleavage and disruption of epithelial barrier function in S. aureus pneumonia.
Figure 4: An ADAM10-specific metalloprotease inhibitor prevents Hla-mediated injury.

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Acknowledgements

We thank T. Golovkina for assistance with mouse breeding strategies, A. Crofts for pilot studies of E-cadherin cleavage in vitro, D. Gruenert (University of California San Francisco) for 16HBE14o- cells, R. Tweten (The University of Oklahoma Health Sciences Center) for the provision of recombinant PLY, J. Whitsett (Cincinnati Children's Medical Center) for SP-C-rtTA-(tetO)7CMV-Cre mice, O. Schneewind for discussions and comments on the manuscript, C. Labno for microscopy support, T. Li for immunohistochemistry support, and the Integrated Microscopy and Immunohistochemistry Facilities at the University of Chicago. This work was supported by the Departments of Pediatrics and Microbiology at the University of Chicago. M.E.P. was partially supported by US National Institutes of Health grant T32 GM007183. The authors acknowledge membership in and support from the Region V 'Great Lakes' Regional Center for Excellence (US National Institutes of Health award 2-U54-AI-057153).

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Contributions

I.I. performed mouse infection modeling, in vivo E-cadherin cleavage studies and ECIS studies. N.I. performed mouse breeding and genetic analysis and assisted with infection modeling. G.A.W. performed siRNA transfections, analyzed ADAM10 expression on 16HBE14o- cells and performed cellular assays of metalloprotease activity. M.E.P. examined the effects of GI254023X on toxin binding and performed ECIS experiments. K.M.F. generated the HlaPPL mutant. Y.W. performed ECIS experiments. J.B.W. performed cellular assays of metalloprotease activity, E-cadherin cleavage and immunofluorescence microscopy and wrote the paper. All authors discussed the results and commented on the manuscript.

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Correspondence to Juliane Bubeck Wardenburg.

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Inoshima, I., Inoshima, N., Wilke, G. et al. A Staphylococcus aureus pore-forming toxin subverts the activity of ADAM10 to cause lethal infection in mice. Nat Med 17, 1310–1314 (2011). https://doi.org/10.1038/nm.2451

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