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Toll-like receptor 4 and high-mobility group box-1 are involved in ictogenesis and can be targeted to reduce seizures

Nature Medicine volume 16, pages 413–419 (2010)Cite this article

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Abstract

Brain inflammation is a major factor in epilepsy, but the impact of specific inflammatory mediators on neuronal excitability is incompletely understood. Using models of acute and chronic seizures in C57BL/6 mice, we discovered a proconvulsant pathway involving high-mobility group box-1 (HMGB1) release from neurons and glia and its interaction with Toll-like receptor 4 (TLR4), a key receptor of innate immunity. Antagonists of HMGB1 and TLR4 retard seizure precipitation and decrease acute and chronic seizure recurrence. TLR4-defective C3H/HeJ mice are resistant to kainate-induced seizures. The proconvulsant effects of HMGB1, like those of interleukin-1β (IL-1β), are partly mediated by ifenprodil-sensitive N-methyl-d-aspartate (NMDA) receptors. Increased expression of HMGB1 and TLR4 in human epileptogenic tissue, like that observed in the mouse model of chronic seizures, suggests a role for the HMGB1-TLR4 axis in human epilepsy. Thus, HMGB1-TLR4 signaling may contribute to generating and perpetuating seizures in humans and might be targeted to attain anticonvulsant effects in epilepsies that are currently resistant to drugs.

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Figure 1: HMGB1 and TLR4 immunoreactivity in the CA1 pyramidal layer of hippocampi of kainic acid–injected C57BL/6 mice.
Figure 2: HMGB1 and TLR4 immunoreactivity in the hippocampi of control subjects and subjects with TLE-HS.
Figure 3: Dose-dependent proconvulsant effect of HMGB1 in wild-type mice and absence of HMGB1 effect in Tlr4Lps-d mice.
Figure 4: Anticonvulsant effects of BoxA, Lps-Rs and Cyp in acute seizure models.
Figure 5: Anticonvulsant effects of BoxA and Lps-Rs in the chronic seizure model.
Figure 6: Effect of ifenprodil on acute and chronic seizures.

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Acknowledgements

This work was supported by a CARIPLO Foundation grant (A.V., M.E.B. and C.R.), by Parents Against Childhood Epilepsy (A.V.), Associazione Italiana Contro l'Epilessia (T.R.), EU FP6 project EPICURE (LSH-CT-2006-037315) (A.V.), EU FP7 project NeuroGlia (No. 202167) and National Epilepsy Fund (NEF 09-05) (E.A.) and a grant from Regione Lombardia to HMGBiotech. J.L. is a recipient of an EU fellowship in the International Graduate Programme in Molecular Medicine training program. We acknowledge F. De Ceglie for technical support in photograph preparation.

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Authors and Affiliations

  1. Department of Neuroscience, Mario Negri Institute for Pharmacological Research, Milano, Italy

    Mattia Maroso, Silvia Balosso, Teresa Ravizza & Annamaria Vezzani

  2. Department of Genetics and Cell Biology, San Raffaele University and San Raffaele Research Institute, Milano, Italy

    Jaron Liu & Marco E Bianchi

  3. Department (Neuro) Pathology, Academisch Medisch Centrum, Amsterdam, The Netherlands

    Eleonora Aronica & Anand M Iyer

  4. The Netherlands Foundation (Stichting Epilepsie Instellingen Nederland), Heemstede, The Netherlands

    Eleonora Aronica

  5. Department of Biotechnology and Molecular Sciences, University of Insubria, Varese, Italy

    Carlo Rossetti

  6. Department of Environmental Health Sciences, Mario Negri Institute for Pharmacological Research, Milano, Italy

    Carlo Rossetti & Monica Molteni

  7. HMGBiotech srl, Milano, Italy

    Maura Casalgrandi

  8. Department of Regenerative Medicine, San Raffaele University and San Raffaele Research Institute, Milano, Italy

    Angelo A Manfredi

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Contributions

M. Maroso and S.B. conducted the pharmacological studies in the experimental models of seizures and performed the analysis of data; T.R. conducted the immunohistochemical analysis in the experimental models and analyzed the data; J.L. performed the western blot analyses and in vitro experiments; E.A. and A.M.I. conducted the immunohistochemical studies in human tissues and data analysis; C.R. and M. Molteni characterized and provided the TLR4 antagonists used in the pharmacological experiments; M.C. produced HMGB1 and BoxA; A.A.M. contributed experimental suggestions and participated in manuscript editing; M.E.B. together with A.V. supervised all phases of the project and wrote the manuscript.

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Correspondence to Annamaria Vezzani.

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Declaration: M.M. is employed by and has personal financial interests in Bluegreen Biotech. M.E.B. is founder and part-owner of HMGBiotech, a biotech that sells products and idea based on the functions of HMGB proteins. M.C. is an employee of HMGBiotech.

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Maroso, M., Balosso, S., Ravizza, T. et al. Toll-like receptor 4 and high-mobility group box-1 are involved in ictogenesis and can be targeted to reduce seizures. Nat Med 16, 413–419 (2010). https://doi.org/10.1038/nm.2127

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