Abstract
Netherton syndrome (NS) is a rare, severe type of ichthyosis, often lethal in neonates, for which there is no therapy. Spink5−/− mice recapitulate major NS hallmarks and die homogeneously within 5 h from birth due to severe epidermal barrier defect leading to dehydration. Spink5−/−Klk5−/− mice survive neonatal lethality, indicating that KLK5 could be a drug target for NS. Nevertheless, after a week, these mice developed epidermal inflammation and signs of barrier defect leading to lethality. Here we tested whether anti-TNFα strategy in combination with anti-KLK5 could provide a long-term effective therapy for NS. Deletion of Tnfa in Spink5−/− suppressed the inflammatory phenotype but did not rescue neonatal lethality of Spink5−/− indicating that anti-TNFα therapy alone would not be sufficient to treat NS. Interestingly, in Spink5−/−Klk5−/−Tnfa−/− mice, NS features were rescued, and mice lived normally for 16–18 months. For the first time, evidence is provided that a combination of anti-TNFα and anti-KLK5 therapeutics represents an effective therapeutic strategy for NS. Notably, anti-TNFα factors are marketed and used widely, while LMW KLK5 inhibitors are being developed.
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Acknowledgements
We would like to thank Prof George Kollias (BSRC Alexander Fleming, Athens, Greece) and Prof Andriew McKenzie (MRC Laboratory of Molecular Biology, Cambridge, UK) for kindly providing the Tnfα−/− and the Spink5−/− mice, respectively. We also thank Dr. Andreas Seferlis (Laboratory of Electron Microscopy and Microanalysis, University of Patras, Greece) for technical assistance with scanning electron microscopy, and finally, Dr. Dalila Darmoul for kindly offering the quenched PAR2 peptide.
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This study is funded, in part, by the research project (Skink5 LS4-2139) implemented within the framework of the Action “Supporting Postdoctoral Researchers” of the Operational Program “Education and Lifelong Learning” (Action’s Beneficiary: General Secretariat for Research and Technology) and is co-financed by the European Social Fund (ESF) and the Greek State.
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EZ investigation, methodology, data curation, validation; GP methodology, validation; GS conceptualization, supervision, funding acquisition, resources, validation; all authors participated in writing the original draft.
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All experiments with animals were carried out in accordance with the EU and national legal and ethical regulations following governmental approval of the experimental protocol (Veterinary Department of Regional Units of Achaia, Region of Western Greece).
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Fig. S1
Deletion of Tnfa in Spink5-/-Klk5-/- mice is associated with attenuated expression of proinflammatory cytokines. Relative expression of proinflammatory cytokines and quantification of mast cells infiltrating the dermis of newborn mice (A) and 4-month old mice (B). The mRNA levels of proinflammatory cytokines were determined by RT-qPCR. Expression is normalized against Hprt1 and shown as fold difference to wt expression. Values represent mean ± S.E.M. (PNG 153 kb)
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Zingkou, E., Pampalakis, G. & Sotiropoulou, G. Cocktails of KLK5 Protease Inhibitors and Anti-TNFα Therapeutics: an Effective Treatment for Netherton Syndrome. J Clin Immunol 42, 597–605 (2022). https://doi.org/10.1007/s10875-021-01195-0
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DOI: https://doi.org/10.1007/s10875-021-01195-0