Institutional Repository - Research Portal Dépôt Institutionnel - Portail de la Recherche University of Namurresearchportal.unamur.be RESEARCH OUTPUTS / RÉSULTATS DE RECHERCHE Characterization of CYP26B1-selective inhibitor, DX314, as a potential therapeutic for keratinization disorders Veit, Joachim; DE GLAS, VALERIE; Balau, Benoit; Liu, Haoming; Bourlond, Florence; Paller, Amy; POUMAY, Yves; Diaz, Philippe Published in: The journal of investigative dermatology Author(s)DOI: - Auteur(s) : 10.1016/j.jid.2020.05.090 Publication date: 2021 PublicationDocument date Version - Date de publication : Peer reviewed version Link to publication Citation for pulished version (HARVARD): Veit, J, DE GLAS, VALERIE, Balau, B, Liu, H, Bourlond, F, Paller, A, POUMAY, Y & Diaz, P 2021, Permanent'Characterization link - Permalien of CYP26B1-selective : inhibitor, DX314, as a potential therapeutic for keratinization disorders', The journal of investigative dermatology, vol. 141, pp. 72-83. https://doi.org/10.1016/j.jid.2020.05.090 Rights / License - Licence de droit d’auteur : General rights Copyright and moral rights for the publications made accessible in the public portal 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. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal ? 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BibliothèqueDownload date: Universitaire 07. oct.. 2021 Moretus Plantin Journal of Investigative Dermatology Characterization of CYP26B1-selective Inhibitor, DX314, as a Potential Therapeutic for Keratinization Disorders Journal: Journal of Investigative Dermatology Manuscript ID JID-2020-0105.R2 Article Type:ForOriginal Review Article Only Date Submitted by the 29-Apr-2020 Author: Complete List of Authors: Veit, Joachim; University of Montana Missoula, Biomedical and Pharmaceutical Sciences De Glas, Valerie; University of Namur, URPHYM-NARLIS Balau, Benoît; University of Namur, URPHYM-NARLIS Liu, Haoming; Northwestern University Feinberg School of Medicine, Departments of Dermatology and Pediatrics Bourlond, Florence; Hôpital Erasme, Université Libre de Bruxelles, Service de Dermatologie Paller, Amy; Northwestern University Feinberg School of Medicine, Departments of Dermatology and Pediatrics Poumay, Yves; University of Namur, URPHYM-NARLIS Diaz, Philippe; University of Montana Missoula, Biomedical and Pharmaceutical Sciences; DermaXon LLC, R&D Drug Development, Keratinization Disorders, Barrier Function, Keywords: Pharmacology, Ichthyosis Page 1 of 51 Journal of Investigative Dermatology 1 2 3 Editor comments: 4 Section/Deputy Editor: 1 5 6 Comments to the Author: 7 Revision is responsive to reviewer comments and improved by revision. 8 9 We would like to thank the editors for their time and feedback regarding our submission. 10 11 The only change we are submitting in this revision is to a minor figure reference issue we found 12 in the Figure 4 legend. The reference to parts "(a,c,d)" in the last sentence was changed to 13 14 correctly read "(a,b,d)". This correction was highlighted and underlined in yellow in the 15 manuscript text. 16 17 18 Reviewer comments: 19 For Review Only 20 Reviewer: 1 21 Comments to the Author 22 This is a very careful Revision of an excellent paper. My concern on the experimental design used, namely 23 a simultaneous co-tratment with all trans retinoic acid in the keratinocyte cultures is well and fully 24 25 adressed and authors Point out that in the rhino mice model no cotratment was used, but similar effects 26 were achieved. Moreover, they clarify that in their in vitro conditions no RA precursors and no 27 appreciable levels of all trans retinoic acid will be present which justifies their Approach and they 28 provide several prominent references like that of Giltaire et al 2009 on this culture model. All specific 29 30 points have been fully and carefully adressed. No further comment. 31 32 33 Reviewer: 2 34 Comments to the Author 35 I believe that the authors have responded adequately to my questions. 36 37 We would like to sincerely thank reviewer #1 and reviewer #2 for their time, effort and 38 comments regarding our submission. 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Journal of Investigative Dermatology Page 2 of 51 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 For Review Only 20 21 22 23 24 25 26 27 28 29 30 527x355mm (96 x 96 DPI) 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Page 3 of 51 Journal of Investigative Dermatology 1 2 3 Informative Title: 4 5 Characterization of CYP26B1-selective Inhibitor, DX314, as a Potential Therapeutic for 6 7 Keratinization Disorders 8 9 10 Short Title: 11 12 DX314: Keratinization Disorder Therapeutic 13 14 15 16 Joachim G.S. Veit1, Valérie De Glas2, Benoît Balau2, Haoming Liu4, Florence Bourlond5, 17 4 2 1,3 18 Amy S. Paller , Yves Poumay , Philippe Diaz 19 For Review Only 20 21 1Department of Biomedical and Pharmaceutical Sciences, University of Montana, Missoula, MT 22 2 23 URPHYM-NARLIS, University of Namur, Namur, Belgium 24 3DermaXon LLC, Missoula, MT 25 26 4Departments of Dermatology and Pediatrics, Northwestern University Feinberg School of 27 28 Medicine, Chicago, IL 29 5 30 Service de Dermatologie, Hôpital Erasme, Université Libre de Bruxelles, Belgique 31 32 33 Correspondence: Philippe Diaz, Department of Biomedical and Pharmaceutical Sciences, 34 35 University of Montana, 32 Campus Dr., Missoula, MT, 59812. Email: 36 37 [email protected] 38 39 40 ORCIDs: 41 42 Joachim G.S. Veit: https://orcid.org/0000-0001-5908-0648 43 Valérie De Glas: https://orcid.org/0000-0001-7366-2601 44 45 Benoît Balau: https://orcid.org/0000-0002-7314-860X 46 47 Haoming Liu: https://orcid.org/0000-0002-3190-5422 48 49 Florence Bourlond: https://orcid.org/0000-0003-1071-3306 50 Amy S. Paller: https://orcid.org/0000-0001-6187-6549 51 52 Yves Poumay: http://orcid.org/0000-0001-5200-3367 53 54 Philippe Diaz: https://orcid.org/0000-0002-2339-8803 55 56 57 58 1 59 60 Journal of Investigative Dermatology Page 4 of 51 1 2 3 Abbreviations: RAMBA, retinoic acid metabolism blocking agents; atRA, all-trans-retinoic acid; 4 5 RHE, reconstructed human epidermis; TEER, transepithelial electrical resistance; TEWL, 6 7 transepidermal water loss; CYP26, cytochrome p450 family 26; DD, Darier disease; LI, lamellar 8 ichthyosis; RXLI, Recessive x-linked ichthyosis. See supplementary materials for additional 9 10 abbreviations (Table S4) 11 12 13 14 Funding: This work was supported by the by the National Institute of Arthritis and 15 Musculoskeletal and Skin Disease of the National Institutes of Health: R44AR069416 (PD), 16 17 P20GM103546 (PD, JV). The content of this paper is solely the responsibility of the authors and 18 19 does not necessarily reflectFor the official Review views of the National Only Institutes of Health. 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 2 59 60 Page 5 of 51 Journal of Investigative Dermatology 1 2 3 ABSTRACT 4 5 6 Inhibition of cytochrome P450 (CYP)-mediated retinoic acid (RA) metabolism by RA 7 8 metabolism blocking agents (RAMBAs) increases endogenous retinoids and is an alternative to 9 10 retinoid therapy. Currently available RAMBAs (i.e. liarozole and talarozole) tend to have fewer 11 12 adverse effects than traditional retinoids but lack target specificity. Substrate-based inhibitor 13 14 15 DX314 has enhanced selectivity for RA-metabolizing enzyme CYP26B1 and may offer an 16 17 improved treatment option for keratinization disorders such as congenital ichthyosis and Darier 18 19 disease. In this study we useFor RT-qPCR, Review RNA sequencing, Only pathway, upstream regulator, and 20 21 22 histological analyses to demonstrate that DX314 can potentiate the effects of all-trans-RA 23 24 (atRA) in healthy and diseased reconstructed human epidermis (RHE). We unexpectedly 25 26 discovered that DX314, but not atRA or previous RAMBAs, appears to protect epidermal barrier 27 28 29 integrity. Additionally, DX314-induced keratinization and epidermal proliferation effects are 30 31 observed in a rhino mice model. Altogether, results indicate that DX314 inhibits atRA 32 33 metabolism with minimal off-target activity and shows therapeutic similarity to topical retinoids 34 35 in vitro and in vivo. Findings of a unique barrier-protecting effect require further mechanistic 36 37 38 study but may lead to a novel strategy in barrier-reinforcing therapies. DX314 is a unique and 39 40 promising candidate compound for further study and development in the context of 41 42 keratinization disorders. 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 3 59 60 Journal of Investigative Dermatology Page 6 of 51 1 2 3 INTRODUCTION 4 5 6 Therapeutics targeting retinoid biopathways have been implemented in the clinical 7 8 treatment of keratinization disorders such as the congenital ichthyoses (Vahlquist et al. 2008), 9 10 Darier disease (DD) (Casals et al. 2009; Cooper and Burge 2003; Dicken et al. 1982; Steijlen et 11 12 al. 1993), and other skin disorders (e.g.