The University of Manchester Research IL-36 Promotes Systemic IFN-I Responses in Severe Forms of Psoriasis DOI: 10.1016/j.jid.2019.08.444 Document Version Accepted author manuscript Link to publication record in Manchester Research Explorer Citation for published version (APA): Catapano, M., Vergnano, M., Romano, M., Mahil, S. K., Choon, S., Burden, A. D., Young, H. S., Carr, I. M., Lachmann, H. J., Lombardi, G., Smith, C. H., Ciccarelli, F. D., Barker, J. N., & Capon, F. (2019). IL-36 Promotes Systemic IFN-I Responses in Severe Forms of Psoriasis. Journal of Investigative Dermatology. https://doi.org/10.1016/j.jid.2019.08.444 Published in: Journal of Investigative Dermatology Citing this paper Please note that where the full-text provided on Manchester Research Explorer is the Author Accepted Manuscript or Proof version this may differ from the final Published version. If citing, it is advised that you check and use the publisher's definitive version. General rights Copyright and moral rights for the publications made accessible in the Research Explorer are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. Takedown policy If you believe that this document breaches copyright please refer to the University of Manchester’s Takedown Procedures [http://man.ac.uk/04Y6Bo] or contact [email protected] providing relevant details, so we can investigate your claim. Download date:03. Oct. 2021 Journal of Investigative Dermatology Interleukin-36 promotes systemic Type-I IFN responses in severe forms of psoriasis Journal: Journal of Investigative Dermatology Manuscript ID JID-2019-0205.R1 Article Type:ForOriginal Review Article Only Date Submitted by the 14-Jun-2019 Author: Complete List of Authors: Catapano, Marika; King's College London, Division of Genetics and Molecular Medicine Vergnano, Marta; King's College London, Division of Genetics and Molecular Medicine Romano, Marco; King's College London, Department of Immunobiology, School of Immunology & Microbial Sciences Mahil, Satveer; King’s College London, Department of Medical and Molecular Genetics Choon, Siew-Eng; Hospital Sultanah Aminah, Department of Dermatology Burden, David; University of Glasgow, Department of Dermatology Young, Helen; The University of Manchester, Dermatology Research Centre Carr, Ian; University of Leeds, School of Medicine Lachmann, Helen; University College London, National Amyloidosis Centre and Centre for Acute Phase Proteins, Division of Medicine Lombardi, Giovanna; King's College London, MRC Centre for Transplantation Smith, Catherine; Kings College London, St John's Institute of Dermatology Ciccarelli, Francesca; Francis Crick Institute, Cancer Systems Biology Laboratory Barker, Jonathan; St John's Institute of Dermatology, St Thomas' Hospital, Division of Genetics and Moilecular Medicine Capon, Francesca; King's College London, Division of Genetics and Molecular Medicine Autoinflammatory Diseases, Cytokines, Inflammatory Skin Diseases, Keywords: Neutrophils, Psoriasis Page 1 of 62 Journal of Investigative Dermatology Head of School Department of 1 King’s College London Faculty of Life Sciences Professor Mathias Gautel Medical & 2 Molecular Genetics 3 & Medicine Head of Department Floor 8 Tower Wing 4 Professor Tim J P Hubbard Guy’s Hospital 5 School of Basic and London SE1 9RT 6 Medical Biosciences 7 8 http://www.kcl.ac.uk/gm 9 Please reply to: Francesca Capon 10 mReader in Inflammation Genetics 11 e-mail: [email protected] 12 13 14 15 16 17 14th June 2019 18 19 20 21 Prof Mark C Udey, For Review Only 22 Editor, Journal of Investigative Dermatology 23 24 25 Dear Prof Udey, 26 Re: MS# JID-2019-0205 27 28 We are submitting revised version of the above manuscript entitled “Interleukin-36 promotes 29 30 systemic Type-I IFN responses in severe psoriasis “. 31 32 To address the reviewers’ comments, we have undertaken the analysis of additional datasets, 33 provided more information on our patient resources and included further experimental data on IL- 34 36R expression. We believe that the manuscript, which has also been edited for clarity, is much 35 36 improved and will now meet the criteria for publication on the Journal of Investigative Dermatology. 37 38 39 I look forward to hearing from you and remain 40 41 Yours sincerely 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 www.kcl.ac.uk Journal of Investigative Dermatology Page 2 of 62 1 2 3 RESPONSE TO REVIEWERS 4 5 Reviewer 1 6 The data is reliable… however, there are too many miss spelling in the text. 7 8 We have carefully proofread the text and eliminated typos as well as instances of inconsistent spelling 9 One concern is as if the author checked IL-36RN mutation or IL-1RN mutation in GPP patients or not. 10 Based on the diagnosis of GPP, we have screened all patients for IL36RN mutations. This has now been 11 st 12 clarified in the Methods (p.11, 1 paragraph). 13 Add the comment that treatment with IL-17 antibody or IL-17R inhibitor will decrease skin 14 15 inflammation and production of IL-36 from keratinocytes, leading to the down regulation of Type-I IFN. 16 The comment has been added to the Discussion (p.10, 3rd paragraph). 17 18 Reviewer 2 19 1) It is questionable if any of the findings described is specific for IL-36 as compared to other pro- 20 21 inflammatory cytokines andFor in particular Review members of the Only IL-1 family. Importantly Swindell et al (Front 22 Immunol. 2018 Jan 29; 9:80) have demonstrated that IL1 and IL-36 signalling is essentially the same. 23 It is likely that all effects shown here would be equally inducible by other IL-1 family members some of 24 which are also upregulated in psoriatic inflammation. 25 We have revised the Discussion (p.8, 2nd paragraph), to quote the important work of Swindell et al and 26 27 acknowledge the similarities between IL-1 and IL-36 signalling. At the same time, we note that Swindell 28 did not identify any of our interferon signature genes among the transcripts induced by IL-1. In keeping 29 with this observation, we find that no IFN signature genes are up-regulated in the whole-blood of 30 patients with CAPS, a condition caused by excessive IL-1 signalling. This pattern was observed in two 31 independent datasets, the second of which was analysed as part of the revision process. The result of 32 33 this new analysis are reported in Supplementary Figure S1. 34 2) A significant body of literature highlights the role of LL-37 in psoriatic inflammation being an 35 36 important contributor to type I IFN responses also in pustular phenotype development. LL-37 is neither 37 mentioned nor included in experimental setups in this paper and should be considered to gain a more 38 balanced information on which factors contribute to clinically relevant IFN expression. Conrad et 39 al (Nat Commun. 2018 Jan 2;9(1):25) and other publications by Gilliet group. 40 The reviewer makes a very interesting point, which we have initially addressed by querying the 41 42 expression of CAMP (the gene encoding LL-37) in our transcriptomic datasets. This revealed that CAMP 43 is up-regulated in the skin of psoriatic patients (fold change >2.0, FDR <0.05), but not in their blood. 44 In keeping with the latter observation, real-time PCR showed that IL-36 treatment does not increase 45 CAMP expression in PBMCs. 46 47 LL37 48 PBMCs were stimulated with IL-36 49 3 50 for the indicated time points. Data 51 2 represent the mean (+/- SEM) of 52 triplicate stimulations 53 1 54 0 55 0h 3h 6h 8h 24h 56 57 Finally, we observed that CAMP whole-blood expression does not correlate with the up-regulation of 58 Type-I IFN genes in either GPP or PsV samples (r<0.1 in both datasets). This suggests that LL-37 is 59 unlikely to be driving the Type-I IFN responses we observed at the systemic level. 60 1 Page 3 of 62 Journal of Investigative Dermatology 1 2 3 We have now edited the Discussion (p.9, final paragraph) to mention these negative findings and 4 quote the work of the Gilliet group, which provides the underlying context. 5 6 3) The clinical relevance of an increase IFN signature (which clearly overlaps with IFNg signature – the 7 latter is known to be highly expressed in psoriasis) remains to be demonstrated. To further explore the 8 9 significance of the induced IFN response a comparison to a disease recognised for its type I IFN pathway 10 activation such as systemic lupus should be included. Some comparison with other diseases is based 11 only on literature data. Figure 1F as it stands gives too little information to demonstrated convincing 12 evidence. Has the same cell population been analysed, was treatment given to the patients, are patient 13 groups regarding to basic demographics comparable etc. 14 15 First of all, we have edited the Methods (RNA-seq data analysis, p.11) to clarify that we did not base 16 the comparisons with other diseases on gene lists published in the literature. On the contrary, we 17 retrieved raw sequence files for every dataset and processed them with the same computational 18 pipeline that was used for the QC and differential expression analysis of our own data. Thus, the up- 19 regulated genes reported in Figures 1f and S1 were identified through a robust, standardised process. 20 21 To fully document these analyses,For we Review have also added a Onlysupplementary Table S4, which summarises 22 the key features of the publicly available datasets and shows that they were all generated in whole- 23 blood samples obtained from European individuals.