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The serine protease inhibitor SerpinA3N attenuates neuropathic pain by inhibiting T cell–derived leukocyte elastase

Abstract

Neuropathic pain is a major, intractable clinical problem and its pathophysiology is not well understood. Although recent gene expression profiling studies have enabled the identification of novel targets for pain therapy1,2,3,4, classical study designs provide unclear results owing to the differential expression of hundreds of genes across sham and nerve-injured groups, which can be difficult to validate, particularly with respect to the specificity of pain modulation5. To circumvent this, we used two outbred lines of rats6, which are genetically similar except for being genetically segregated as a result of selective breeding for differences in neuropathic pain hypersensitivity7. SerpinA3N, a serine protease inhibitor, was upregulated in the dorsal root ganglia (DRG) after nerve injury, which was further validated for its mouse homolog. Mice lacking SerpinA3N developed more neuropathic mechanical allodynia than wild-type (WT) mice, and exogenous delivery of SerpinA3N attenuated mechanical allodynia in WT mice. T lymphocytes infiltrate the DRG after nerve injury and release leukocyte elastase (LE), which was inhibited by SerpinA3N derived from DRG neurons. Genetic loss of LE or exogenous application of a LE inhibitor (Sivelastat) in WT mice attenuated neuropathic mechanical allodynia. Overall, we reveal a novel and clinically relevant role for a member of the serpin superfamily and a leukocyte elastase and crosstalk between neurons and T cells in the modulation of neuropathic pain.

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Figure 1: Serpina3n is upregulated in rats with low neuropathic pain sensitivity.
Figure 2: Serpina3n is upregulated in mouse lumbar DRGs after spared nerve injury, a model of neuropathic pain.
Figure 3: SerpinA3N attenuates mechanical allodynia after nerve injury.
Figure 4: Leukocyte elastase (LE) is a substrate for SerpinA3N, and it promotes neuropathic allodynia.

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Acknowledgements

The authors are grateful to R. LeFaucheur for secretarial assistance and to D. Baumgartl-Ahlert for technical assistance. We acknowledge support from the Interdisciplinary Neurobehavioral Core, Heidelberg, for the behavioral experiments performed here. We are grateful to M. Meister for his help in the analysis of FACS data. This work was supported by a European Research Council (ERC) Advanced Investigator Grant (PAINPLASTICITY; project no. 294293) to R.K., by grants from the Deutsche Forschungsgemeinschaft (DFG; MA 2081/4-1) to M.A.M., by grants from the Deutsche Forschungsgemeinschaft (SFB 938) to B.A., by grants from the US National Institutes of Health (5 R01 NS038253 and 2R37NS039518 to C.W., and R01 NS074430 to M.C.), and by a grant from the Israel Science Foundation to M.D. M.A.M. and R.K. are members of the Molecular Medicine Partnership Unit, Heidelberg. R.K. is a principal investigator in the Excellence Cluster 'CellNetworks' of Heidelberg University. M.A.M. is a member of the German Center for Lung Research. L.V. was partially supported by a PhD fellowship from CellNetworks and by the Hartmut-Hoffmann Berling International Graduate School for Cellular and Molecular Biology. M.S. was partially supported by a post-doctoral fellowship from CellNetworks. A.L. was partially supported by the RO1DE022912 grant from the US National Institutes of Health.

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L.V., D.E.S., A.L., K.K.B., M.S., D.H., S.P., P.R., R.S.G., C.N., S.G. and M.C. performed experiments and analyzed data; R.K., M.C., C.J.W., S.D.L., M.D., B.A., M.A.M. designed and supervised experiments; R.K., L.V. and M.C. primarily wrote the manuscript.

Corresponding author

Correspondence to Rohini Kuner.

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The authors have a patent application pending based on the data herein on the use of leukocyte elastase inhibitors against pain of neuropathic origin (patent application number: EP14200012.4 at the European Patent Office).

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Vicuña, L., Strochlic, D., Latremoliere, A. et al. The serine protease inhibitor SerpinA3N attenuates neuropathic pain by inhibiting T cell–derived leukocyte elastase. Nat Med 21, 518–523 (2015). https://doi.org/10.1038/nm.3852

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