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A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia

Abstract

Neuronal death in cerebral ischemia is largely due to excitotoxic mechanisms, which are known to activate the c-Jun N-terminal kinase (JNK) pathway. We have evaluated the neuroprotective power of a cell-penetrating, protease-resistant peptide that blocks the access of JNK to many of its targets. We obtained strong protection in two models of middle cerebral artery occlusion (MCAO): transient occlusion in adult mice and permanent occlusion in 14-d-old rat pups. In the first model, intraventricular administration as late as 6 h after occlusion reduced the lesion volume by more than 90% for at least 14 d and prevented behavioral consequences. In the second model, systemic delivery reduced the lesion by 78% and 49% at 6 and 12 h after ischemia, respectively. Protection correlated with prevention of an increase in c-Jun activation and c-Fos transcription. In view of its potency and long therapeutic window, this protease-resistant peptide is a promising neuroprotective agent for stroke.

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Figure 1: In vitro kinase assays showing sensitivity and specificity of JNK-inhibitory peptides.
Figure 2: Effects of NMDA on cortical neurons.
Figure 3: Time course of NMDA neurotoxicity and neuroprotection by L-JNKI-1, D-JNKI-1 and two control peptides, 'Tat-empty' (Tat sequence without JBD20) and 'L-JNKI-1 mut' (L-JNKI-1 with six amino acids mutated to alanine20).
Figure 4: Transient ischemia in mice.
Figure 5: Protection by D-JNKI-1 against permanent focal ischemia in P14 rats.

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Acknowledgements

This work was supported by grants 31-50598.97, 32-54119.98, 31-61736.00, 32-65139.01 and 3200-68306.02 from the Swiss National Science Foundation, the San Paolo Bank (Italy) and the Alzheimer Project (Italian Ministry of Health). We are particularly grateful to P. Nicod and the Botnar Foundation for human and financial support. We thank E. Bernardi, I. Favre, V. Mottier and A. Oberson for assistance; R. Kraftsik for help with computation and statistics; and J.-Y. Chatton and A. Volterra for critical comments on the manuscript.

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Correspondence to Tiziana Borsello.

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C.B. receives partial remuneration from the vendor, Alexis, for the sale of D-JNKI-1 and L-JNKI-1.

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Borsello, T., Clarke, P., Hirt, L. et al. A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nat Med 9, 1180–1186 (2003). https://doi.org/10.1038/nm911

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