Aquaporin 3 inhibition Attenuates Imiquimod-Induced Psoriatic Symptoms in a Murine Model

Aquaporin 3 inhibition Attenuates Imiquimod-Induced Psoriatic Symptoms in a Murine Model | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Short Report Aquaporin 3 inhibition Attenuates Imiquimod-Induced Psoriatic Symptoms in a Murine Model Ryosuke Okubo, Manami Tanaka, Masato Yasui, Mariko Hara-Chikuma This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5145495/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 01 Apr, 2025 Read the published version in Molecular Biology Reports → Version 1 posted 9 You are reading this latest preprint version Abstract Background: Aquaporin 3 (AQP3) is highly expressed in both keratinocytes and T cells within psoriatic skin. Previous studies have demonstrated that AQP3 knockout mice show reduced development of psoriatic symptoms in murine models. This study aims to evaluate the effect of AQP3 inhibition on psoriasis progression. Methods and results: AQP3 conditional knockout mice were generated to assess the role of AQP3 expression in keratinocytes and T cells in psoriasis pathogenesis. In an imiquimod (IMQ)-induced psoriasis model, psoriatic symptoms were mitigated only in mice with keratinocyte-specific AQP3 deletion. Additionally, treatment with an anti-AQP3 monoclonal antibody (mAb) significantly reduced IMQ-induced psoriasis symptoms in wild-type mice. Conclusions: AQP3 inhibition presents a promising approach for the treatment of psoriasis. Aquaporin 3 psoriasis antibody therapy Figures Figure 1 Figure 2 Figure 3 Introduction Psoriasis is a common chronic inflammatory skin disease affecting 2 to 3% of the global population [1]. It is characterized by keratinocyte hyperproliferation, abnormal differentiation, and infiltration of inflammatory cells into the skin and epidermis [2]. The IL-23/Th17 axis plays a pivotal role in the pathogenesis of psoriasis, with both keratinocyte activation and T-cell-mediated immune responses contributing to disease progression [3–5]. While current treatments, including topical agents, phototherapy, biologics targeting TNF-α, IL-17, and IL-23, as well as small-molecule inhibitors, have improved disease management, there remains a substantial unmet need for more effective and safer therapeutic options. Aquaporin-3 (AQP3) is a membrane protein that transports water and small molecules, including glycerol and hydrogen peroxide (H 2 O 2 ). AQP3-mediated transport has been shown to contribute to cell proliferation, migration, and inflammation [6, 7]. In the skin, AQP3 is expressed in keratinocytes and immune cells, such as T cells [8, 9]. Our previous study demonstrated that AQP3 knockout mice exhibited reduced pathology in a psoriasis model induced by imiquimod (IMQ) or IL-23 application [10]. Then, we proposed that AQP3-mediated H 2 O 2 transport is essential for NF-kB activation and the inflammatory response in keratinocytes during psoriasis development. Building on these findings, we developed a monoclonal antibody (mAb) against AQP3 that specifically binds to an extracellular epitope on both human and mouse AQP3, inhibiting the transport of water, glycerol, and H 2 O 2 [11]. Our previous research demonstrated that administration of this anti-AQP3 mAb effectively suppressed liver fibrosis and cancer progression in experimental mouse models [11, 12]. In this study, we aimed to evaluate the therapeutic potential of AQP3 inhibition via anti-AQP3 mAb in a murine model of psoriasis, assessing its effects on disease development and progression. Materials and methods Mice C57BL/6 mice were purchased from Japan SLC, Inc. Mice with loxP-flanked AQP3 alleles (AQP3 fl/fl ) were generated by Cyagen (USA) using ES genome engineering. T cell-specific AQP3-deficient mice were created by crossing AQP3 fl/fl mice with CD4-Cre Tg mice, originally provided by Dr. Wilson [13]. For keratinocyte-specific AQP3 conditional knockout mice (AQP3 fl/fl K14-CreERT +/− ), AQP3 fl/fl mice were crossed with K14-CreERT mice [14], which was administered tamoxifen (Nacalai, 50 mg/Kg body weight, dissolved in corn oil) intraperitoneally for five consecutive days to induce AQP3 deletion. All animal experiments were approved by the President of Keio University, following review and approval by the Institutional Animal Care and Use Committee and the Genetic Modification Safety Committee of Keio University School of Medicine. Imiquimod-induced psoriasis model Imiquimod cream (5% IMQ, BESELNA Cream, Mochida Pharmaceutical Co., Ltd.) was applied daily to the back (62.5 mg, 2 × 2 cm²) or ear (10 mg) for 6–7 consecutive days. Anti-AQP3 mAb [11] or a matched isotype control (mouse IgG2a isotype control, clone #C1.18.4, BioXcell) was administered intraperitoneally (12.5 mg/Kg body weight) on days 1 and 4. The Psoriasis Area and Severity Index (PASI) scoring system was used to assess the severity of erythema and scaling, with scores ranging from 0–3 [5]. Skin samples were collected 24 hours after the final IMQ application. Histopathological examination Skin tissue samples were fixed in 10% neutral buffered formalin, then embedded in paraffin or O.C.T. compound (Sakura Finetek JAPAN). Sections were stained with hematoxylin and eosin (H&E), and epidermal thickness was measured using Fiji software [15]. RNA extraction and real-time quantitative RT-PCR Total RNA was extracted using TRIzol (ThermoFisher, Waltham, MA, USA) and reverse-transcribed into cDNA using the PrimeScript RT reagent kit (Takara Bio, Mountain View, CA, USA). Quantitative reverse transcription PCR (RT-PCR) was performed with SYBR Green I (Takara Bio) on a CFX Opus Real-Time PCR System (Bio-Rad). Flow cytometry analysis Single-cell suspensions from the spleen were stained with antibodies against CD3 and CD4 (eBioscience), followed by anti-AQP3 mAb [11] and anti-mouse IgG conjugated with Alexa Fluor 488 (Thermo Fisher). Stained cells were analyzed using a CytoFLEX flow cytometer (Beckman Coulter Life Sciences, Brea, CA) and FlowJo software (Treestar). Immunoblotting The epidermis of the mouse was separated from the dermis by incubating in phosphate-buffered saline (PBS) at 60°C for 10 seconds. The epidermis was then lysed with an extraction buffer containing 250 mM sucrose, 1 mM EDTA, and 1% protein inhibitor cocktail (Sigma-Aldrich). For immunoblot analysis, a polyclonal AQP3 antibody (Millipore) and horseradish peroxidase-conjugated secondary antirabbit IgG antibody (Cell Signaling Technology) were used for detection via ECL (GE Healthcare, Piscataway, NJ). Statistical analysis Statistical analysis was performed using the two-tailed t-test, one-way ANOVA or two-way ANOVA with GraphPad Prism 8 (San Diego, CA, USA). A p-value of less than 0.05 was considered statistically significant for all tests. Results AQP3 deficiency in T cells had little effect on IMQ-induced psoriasis development To investigate which cell type’s expression of AQP3 contributes more significantly to the pathogenesis of psoriasis, we established AQP3 conditional knockout (cKO) mice in T cells or keratinocytes (Fig. 1 A). Successful deletion of AQP3 in T cells was confirmed by quantitative RT-PCR and fluorescence-activated cell sorting ( FACS) analysis compared to control mice (AQP3 fl/fl CD4-Cre −/− ) (Fig. 1 B, C). We topically applied IMQ to the ear skin to induce a psoriasis-like phenotype, characterized by noticeable ear thickening. No significant difference in ear thickness was observed between control and T cell-specific AQP3 cKO mice (Fig. 1 D). AQP3 deficiency in keratinocytes attenuates IMQ-induced psoriasis-like symptoms Next, we examined the role of keratinocyte AQP3 in psoriasis development using keratinocyte-specific AQP3 cKO mice. AQP3 deletion was induced by tamoxifen injection before initiating the psoriasis model (Fig. 2 A). The deletion of AQP3 in the epidermis was confirmed at both the protein and mRNA levels through Western blotting and quantitative RT-PCR, respectively (Fig. 2 B, C). Topical application of IMQ resulted in ear swelling, which was significantly reduced in keratinocyte-specific AQP3 cKO mice compared to control mice (Fig. 2 D). These results indicate that AQP3 expression in keratinocytes, rather than T cells, contributes to the development of IMQ-induced psoriasis. Administration of AQP3 mAb suppressed IMQ-induced psoriatic symptoms To evaluate the therapeutic potential of targeting AQP3 in psoriasis, mice were administered either AQP3 mAb or an IgG control during the development of IMQ-induced psoriasis (Fig. 3 A). Psoriasis-like symptoms were quantified using the PASI scores [5]. Administration of AQP3 mAb significantly reduced visible erythema and scaling compared to control IgG treatment, suggesting a reducing in inflammation and keratinocyte proliferation (Fig. 3 B, C). H&E staining revealed that IMQ-induced epidermal hyperplasia was suppressed in mice treated with AQP3 mAb compared to those receiving control IgG (Fig. 3 D). Additionally, quantitative RT-PCR analysis demonstrated that the IMQ-induced expression levels of S100A8 and S100A7—key proteins involved in inflammation and keratinocyte proliferation—were significantly decreased in the epidermis of AQP3 antibody-treated mice. Treatment with AQP3 mAb also reduced mRNA expression of Ki67, a marker of cell proliferation, consistent with the observation that epidermal thickening was suppressed (Fig. 3 E). Collectively, these data indicate that anti-AQP3 mAb treatment can effectively reduce the severity of IMQ-induced psoriasis-like symptoms, potentially by reducing keratinocyte hyperproliferation. Discussion Our study demonstrated that AQP3 inhibition can effectively suppress psoriasis, highlighting the crucial role of AQP3 in the pathophysiology of the disease. This finding not only corroborates previous research identifying AQP3 as a key factor in various skin conditions [10, 16, 17] but also reinforces its potential as a therapeutic target specifically for psoriasis. Given the growing evidence linking AQP3 to inflammatory diseases, our study contributes to the understanding that AQP3 inhibition may be a promising new approach for treating inflammatory skin disorders, particularly psoriasis. Using AQP3 cKO mice, we found that AQP3 in keratinocytes is more critical for psoriasis development than AQP3 expression in T cells. This suggests a more direct and pivotal role for keratinocyte AQP3 in psoriasis than previously recognized. Our findings are consistent with reports indicating that AQP3 overexpression in keratinocytes leads to excessive cell proliferation and epidermal hyperplasia, which are characteristics typical of psoriatic lesions [9, 10]. Therefore, it is important to focus on pathways specific to keratinocytes, as well as T cells, when considering therapeutic options for psoriasis. Abnormal interactions between keratinocytes and immune cells are key factors driving psoriasis progression [18, 19]. The inflammatory response causes keratinocyte hyperproliferation and perpetuates the chronic inflammatory cycle underlying psoriatic pathology. Although the pathogenesis of psoriasis is multifaceted and not fully understood, targeting AQP3 in keratinocytes may provide a strategy to interrupt this cycle and better regulate the disease. Additionally, we previously demonstrated that AQP3-mediated cellular H 2 O 2 functions as a second messenger, regulating NF-κB activation and inflammatory responses during psoriatic pathology [10]. Future studies will explore the mechanisms by which AQP3 inhibition through the AQP3 antibody effectively prevents psoriasis development. Given the challenges associated with current psoriasis treatments, including the side effects of biologics [20] and JAK inhibitors [21], our findings suggest the potential for more selective and targeted therapies. In this study, we used AQP3 mAb to investigate the efficacy of AQP3 inhibition in psoriasis pathology, as there are currently no available AQP3 inhibitors for human or mouse administration. The results from the AQP3 cKO experiments are expected to encourage the development of small-molecule inhibitors targeting AQP3 for topical application. This approach may not only enhance symptom management for psoriasis patients but also deepen our understanding of the underlying mechanisms of the disease, paving the way for future therapeutic developments. However, an important limitation of our study is the lack of long-term data regarding the safety and efficacy of AQP3 inhibition. While our findings highlight its short-term benefits, further research is needed to assess the potential risks and therapeutic durability of prolonged AQP3 inhibition. Addressing these questions is crucial for translating AQP3 inhibition into a viable treatment option for chronic psoriasis. Abbreviations AQP3, aquaporin 3 H 2 O 2 , hydrogen peroxide AQP3 mAb, anti-AQP3 monoclonal antibody cKO, conditional knockout TMX, tamoxifen RT-PCR, reverse transcription PCR FACS, fluorescence-activated cell sorting PBS, phosphate-buffered saline Declarations Competing interests The authors declare no competing interests. Ethical Statements: All animal experiments were approved by the President of Keio University, following the consideration by the Institutional Animal Care and Use Committee of Keio University (Approval no: 16075) and by Genetic Modification Safety Committee, Keio University School of Medicine (approval no. 28–029). Funding: This work was supported by Research Ministry of Education, Culture, Sports, Science (21K06974, M.H-C). Author Contribution M.H-C. conceived the study. R.O., M.T., and M.H-C., performed the experiments and analyzed the data. R.O. and M.H-C. wrote the manuscript. All authors reviewed the manuscript Acknowledgements. This work was supported by Research Ministry of Education, Culture, Sports, Science (21K06974, M.H-C). References Nestle FO, Kaplan DH, Barker J (2009) Psoriasis. New England Journal of Medicine 361:496–509. https://doi.org/10.1056/NEJMra0804595 Boehncke W-H, Schön MP (2015) Psoriasis. The Lancet 386:983–994. https://doi.org/10.1016/S0140-6736(14)61909-7 Hawkes JE, Chan TC, Krueger JG (2017) Psoriasis Pathogenesis and the Development of Novel, Targeted Immune Therapies. J Allergy Clin Immunol 140:645–653. https://doi.org/10.1016/j.jaci.2017.07.004 Zeichner JA, Armstrong A (2016) The Role of IL-17 in the Pathogenesis and Treatment of Psoriasis. J Clin Aesthet Dermatol 9:S3–S6 van der Fits L, Mourits S, Voerman JSA, et al (2009) Imiquimod-Induced Psoriasis-Like Skin Inflammation in Mice Is Mediated via the IL-23/IL-17 Axis1. The Journal of Immunology 182:5836–5845. https://doi.org/10.4049/jimmunol.0802999 Hara-Chikuma M, Verkman AS (2008) Prevention of Skin Tumorigenesis and Impairment of Epidermal Cell Proliferation by Targeted Aquaporin-3 Gene Disruption. Molecular and Cellular Biology 28:326–332. https://doi.org/10.1128/MCB.01482-07 Hara-Chikuma M, Chikuma S, Sugiyama Y, et al (2012) Chemokine-dependent T cell migration requires aquaporin-3–mediated hydrogen peroxide uptake. Journal of Experimental Medicine 209:1743–1752. https://doi.org/10.1084/jem.20112398 Verkman AS, Anderson MO, Papadopoulos MC (2014) Aquaporins: important but elusive drug targets. Nat Rev Drug Discov 13:259–277. https://doi.org/10.1038/nrd4226 Soler DC, Young AE, Griffith AD, et al (2017) Overexpression of AQP3 and AQP10 in the skin exacerbates psoriasiform acanthosis. Exp Dermatol 26:949–951. https://doi.org/10.1111/exd.13307 Hara-Chikuma M, Satooka H, Watanabe S, et al (2015) Aquaporin-3-mediated hydrogen peroxide transport is required for NF-κB signalling in keratinocytes and development of psoriasis. Nat Commun 6:7454. https://doi.org/10.1038/ncomms8454 Hara-Chikuma M, Tanaka M, Verkman AS, Yasui M (2020) Inhibition of aquaporin-3 in macrophages by a monoclonal antibody as potential therapy for liver injury. Nat Commun 11:5666. https://doi.org/10.1038/s41467-020-19491-5 Tanaka M, Ito A, Shiozawa S, Hara-Chikuma M (2022) Anti-tumor effect of aquaporin 3 monoclonal antibody on syngeneic mouse tumor model. Transl Oncol 24:101498. https://doi.org/10.1016/j.tranon.2022.101498 Makar KW, Pérez-Melgosa M, Shnyreva M, et al (2003) Active recruitment of DNA methyltransferases regulates interleukin 4 in thymocytes and T cells. Nat Immunol 4:1183–1190. https://doi.org/10.1038/ni1004 Vasioukhin V, Degenstein L, Wise B, Fuchs E (1999) The magical touch: Genome targeting in epidermal stem cells induced by tamoxifen application to mouse skin. Proceedings of the National Academy of Sciences 96:8551–8556. https://doi.org/10.1073/pnas.96.15.8551 Schindelin J, Arganda-Carreras I, Frise E, et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682. https://doi.org/10.1038/nmeth.2019 Tricarico PM, Mentino D, De Marco A, et al (2022) Aquaporins Are One of the Critical Factors in the Disruption of the Skin Barrier in Inflammatory Skin Diseases. Int J Mol Sci 23:4020. https://doi.org/10.3390/ijms23074020 Chen M, Peng Q, Tan Z, et al (2023) Targeting Aquaporin-3 Attenuates Skin Inflammation in Rosacea. Int J Biol Sci 19:5160–5173. https://doi.org/10.7150/ijbs.86207 Lowes MA, Russell CB, Martin DA, et al (2013) The IL-23/T17 pathogenic axis in psoriasis is amplified by keratinocyte responses. Trends in Immunology 34:174–181. https://doi.org/10.1016/j.it.2012.11.005 Tonel G, Conrad C (2009) Interplay between keratinocytes and immune cells—Recent insights into psoriasis pathogenesis. The International Journal of Biochemistry & Cell Biology 41:963–968. https://doi.org/10.1016/j.biocel.2008.10.022 Jiang Y, Chen Y, Yu Q, Shi Y (2023) Biologic and Small-Molecule Therapies for Moderate-to-Severe Psoriasis: Focus on Psoriasis Comorbidities. BioDrugs 37:35–55. https://doi.org/10.1007/s40259-022-00569-z Miot HA, Criado PR, de Castro CCS, et al (2023) JAK-STAT pathway inhibitors in dermatology. Anais Brasileiros de Dermatologia 98:656–677. https://doi.org/10.1016/j.abd.2023.03.001 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 01 Apr, 2025 Read the published version in Molecular Biology Reports → Version 1 posted Editorial decision: Revision requested 26 Nov, 2024 Reviews received at journal 26 Nov, 2024 Reviewers agreed at journal 29 Oct, 2024 Reviews received at journal 04 Oct, 2024 Reviewers agreed at journal 27 Sep, 2024 Reviewers invited by journal 25 Sep, 2024 Editor assigned by journal 24 Sep, 2024 Submission checks completed at journal 24 Sep, 2024 First submitted to journal 24 Sep, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5145495","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":382777615,"identity":"604555ee-3026-442b-a958-49b785b06711","order_by":0,"name":"Ryosuke Okubo","email":"","orcid":"","institution":"Keio University","correspondingAuthor":false,"prefix":"","firstName":"Ryosuke","middleName":"","lastName":"Okubo","suffix":""},{"id":382777616,"identity":"a451050b-5713-41bc-83e1-ce91b0af8d47","order_by":1,"name":"Manami Tanaka","email":"","orcid":"","institution":"Keio University","correspondingAuthor":false,"prefix":"","firstName":"Manami","middleName":"","lastName":"Tanaka","suffix":""},{"id":382777617,"identity":"d5ba5938-fdff-4cef-b402-255e0b97f318","order_by":2,"name":"Masato Yasui","email":"","orcid":"","institution":"Keio University","correspondingAuthor":false,"prefix":"","firstName":"Masato","middleName":"","lastName":"Yasui","suffix":""},{"id":382777618,"identity":"9fc259d0-891d-46b0-b285-e0d169eec86c","order_by":3,"name":"Mariko Hara-Chikuma","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA6UlEQVRIiWNgGAWjYJCCgw1AwoCZDcS2MWBgBlISJGhJI04LI1gLA1jLYQOCjtJtP/vw4IwKBnlzdrY0qRs154352ZkPMFjuwK3F7Ey6wcENZxgMdzazHZPOOXbbTLKZLYFB8gweLQfSGA4+bGNIMDjM3iadw3bbxuAwjwGDZBseLeefIWv5d87GnqCWG0BbNoK1AB2W23bAzICZoBagLTPOSBhuOMyWbJ3bl2wscZgt4QBev5xPY/7YU2Ejb3D+mOHtnG92hv39hw8+lsQTYlCAFnWHJRsIakEDjB9J1jIKRsEoGAXDGAAAbqFPv53fL8UAAAAASUVORK5CYII=","orcid":"","institution":"Keio University","correspondingAuthor":true,"prefix":"","firstName":"Mariko","middleName":"","lastName":"Hara-Chikuma","suffix":""}],"badges":[],"createdAt":"2024-09-24 13:38:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5145495/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5145495/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11033-025-10444-z","type":"published","date":"2025-04-01T15:57:41+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":70539554,"identity":"74464013-21c8-4bb9-a1bd-818c0556babf","added_by":"auto","created_at":"2024-12-04 07:48:33","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":70651,"visible":true,"origin":"","legend":"\u003cp\u003e(A) Schematic diagram of the generation of conditional knockout (cKO) mice targeting AQP3 expression in T cells and keratinocytes. (B) mRNA expression of AQP3 in isolated CD4\u003csup\u003e+ \u003c/sup\u003eT cells as measured by real-time RT-PCR, with 18S rRNA as the internal reference (mean ± SE, n = 3, **p \u0026lt; 0.01 by unpaired t-test). (C) Flow cytometric analysis of AQP3 expression in gated CD3\u003csup\u003e+\u003c/sup\u003e CD4\u003csup\u003e+\u003c/sup\u003e T cells; representative FACS analysis is shown. (D) IMQ was applied to the ears of control and cKO mice for six consecutive days, and ear thickness was measured on the seventh day (mean ± SE, n = 5, ****p \u0026lt; 0.0001 by two-way ANOVA).\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-5145495/v1/f0aaf5701b2055711643fca8.png"},{"id":70539553,"identity":"198fc74e-2fa9-4acd-bb19-6ac11dadd8f4","added_by":"auto","created_at":"2024-12-04 07:48:33","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":47894,"visible":true,"origin":"","legend":"\u003cp\u003e(A) Schedule for the IMQ-induced psoriatic model in AQP3 keratinocyte-specific conditional knockout (cKO) mice. TMX, tamoxifen injection. (B) mRNA expression of AQP3 in isolated keratinocytes as measured by real-time RT-PCR, with 18S rRNA as the internal reference (mean ± SE, n = 3, **p \u0026lt; 0.01 by unpaired t-test). (C) AQP3 expression in isolated epidermis determined by Western blotting. (D) IMQ was applied to the ears of control and cKO mice for six consecutive days, and ear thickness was measured (mean ± SE, n = 6, **p \u0026lt; 0.01, ****p \u0026lt; 0.0001 by two-way ANOVA).\u003c/p\u003e","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-5145495/v1/45c05cbd708e77cbb65e9493.png"},{"id":70539887,"identity":"7a98641b-0478-48a3-9787-ab16180c83d6","added_by":"auto","created_at":"2024-12-04 07:56:33","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":145311,"visible":true,"origin":"","legend":"\u003cp\u003e(A) Diagram depicting the experimental design of IMQ application. (B) Gross appearance of mouse back skin on day 7. (C) Erythema and scaling were scored from days 1 to 7 based on the PASI (mean ± SE, n = 5, *p \u0026lt; 0.05, **p \u0026lt; 0.01 by two-tailed unpaired t-test). (D) (Left) H\u0026amp;E staining of dorsal skin. Scale bar; 200 mm. (Right) Epidermal thickness (mean ± SE,n = 5, *p \u0026lt; 0.05, ****p \u0026lt; 0.0001 by one-way ANOVA). (E) mRNAexpressions of S100A8, S100A7, and Ki67 in ear skin, with 18S rRNA as the internal reference (mean ± SE, n = 4, *p \u0026lt; 0.05, ***p \u0026lt; 0.001 by one-way ANOVA).\u003c/p\u003e","description":"","filename":"Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-5145495/v1/4a8acb08811d8797f44c27f3.png"},{"id":80082132,"identity":"4dd36185-c80a-4b5e-8aa4-84de79d67a96","added_by":"auto","created_at":"2025-04-07 16:07:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":782773,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5145495/v1/277c125b-2403-481f-8246-547d7cdfc45d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Aquaporin 3 inhibition Attenuates Imiquimod-Induced Psoriatic Symptoms in a Murine Model","fulltext":[{"header":"Introduction","content":"\u003cp\u003ePsoriasis is a common chronic inflammatory skin disease affecting 2 to 3% of the global population [1]. It is characterized by keratinocyte hyperproliferation, abnormal differentiation, and infiltration of inflammatory cells into the skin and epidermis [2]. The IL-23/Th17 axis plays a pivotal role in the pathogenesis of psoriasis, with both keratinocyte activation and T-cell-mediated immune responses contributing to disease progression [3\u0026ndash;5]. While current treatments, including topical agents, phototherapy, biologics targeting TNF-α, IL-17, and IL-23, as well as small-molecule inhibitors, have improved disease management, there remains a substantial unmet need for more effective and safer therapeutic options.\u003c/p\u003e \u003cp\u003eAquaporin-3 (AQP3) is a membrane protein that transports water and small molecules, including glycerol and hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e). AQP3-mediated transport has been shown to contribute to cell proliferation, migration, and inflammation [6, 7]. In the skin, AQP3 is expressed in keratinocytes and immune cells, such as T cells [8, 9]. Our previous study demonstrated that AQP3 knockout mice exhibited reduced pathology in a psoriasis model induced by imiquimod (IMQ) or IL-23 application [10]. Then, we proposed that AQP3-mediated H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e transport is essential for NF-kB activation and the inflammatory response in keratinocytes during psoriasis development.\u003c/p\u003e \u003cp\u003eBuilding on these findings, we developed a monoclonal antibody (mAb) against AQP3 that specifically binds to an extracellular epitope on both human and mouse AQP3, inhibiting the transport of water, glycerol, and H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e [11]. Our previous research demonstrated that administration of this anti-AQP3 mAb effectively suppressed liver fibrosis and cancer progression in experimental mouse models [11, 12]. In this study, we aimed to evaluate the therapeutic potential of AQP3 inhibition via anti-AQP3 mAb in a murine model of psoriasis, assessing its effects on disease development and progression.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMice\u003c/h2\u003e \u003cp\u003eC57BL/6 mice were purchased from Japan SLC, Inc. Mice with loxP-flanked AQP3 alleles (AQP3\u003csup\u003efl/fl\u003c/sup\u003e) were generated by Cyagen (USA) using ES genome engineering. T cell-specific AQP3-deficient mice were created by crossing AQP3\u003csup\u003efl/fl\u003c/sup\u003e mice with CD4-Cre Tg mice, originally provided by Dr. Wilson [13]. For keratinocyte-specific AQP3 conditional knockout mice (AQP3\u003csup\u003efl/fl\u003c/sup\u003e K14-CreERT\u003csup\u003e+/\u0026minus;\u003c/sup\u003e), AQP3\u003csup\u003efl/fl\u003c/sup\u003e mice were crossed with K14-CreERT mice [14], which was administered tamoxifen (Nacalai, 50 mg/Kg body weight, dissolved in corn oil) intraperitoneally for five consecutive days to induce AQP3 deletion. All animal experiments were approved by the President of Keio University, following review and approval by the Institutional Animal Care and Use Committee and the Genetic Modification Safety Committee of Keio University School of Medicine.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eImiquimod-induced psoriasis model\u003c/h3\u003e\n\u003cp\u003eImiquimod cream (5% IMQ, BESELNA Cream, Mochida Pharmaceutical Co., Ltd.) was applied daily to the back (62.5 mg, 2 \u0026times; 2 cm\u0026sup2;) or ear (10 mg) for 6\u0026ndash;7 consecutive days. Anti-AQP3 mAb [11] or a matched isotype control (mouse IgG2a isotype control, clone #C1.18.4, BioXcell) was administered intraperitoneally (12.5 mg/Kg body weight) on days 1 and 4. The Psoriasis Area and Severity Index (PASI) scoring system was used to assess the severity of erythema and scaling, with scores ranging from 0\u0026ndash;3 [5]. Skin samples were collected 24 hours after the final IMQ application.\u003c/p\u003e\n\u003ch3\u003eHistopathological examination\u003c/h3\u003e\n\u003cp\u003eSkin tissue samples were fixed in 10% neutral buffered formalin, then embedded in paraffin or O.C.T. compound (Sakura Finetek JAPAN). Sections were stained with hematoxylin and eosin (H\u0026amp;E), and epidermal thickness was measured using Fiji software [15].\u003c/p\u003e\n\u003ch3\u003eRNA extraction and real-time quantitative RT-PCR\u003c/h3\u003e\n\u003cp\u003eTotal RNA was extracted using TRIzol (ThermoFisher, Waltham, MA, USA) and reverse-transcribed into cDNA using the PrimeScript RT reagent kit (Takara Bio, Mountain View, CA, USA). Quantitative reverse transcription PCR (RT-PCR) was performed with SYBR Green I (Takara Bio) on a CFX Opus Real-Time PCR System (Bio-Rad).\u003c/p\u003e\n\u003ch3\u003eFlow cytometry analysis\u003c/h3\u003e\n\u003cp\u003eSingle-cell suspensions from the spleen were stained with antibodies against CD3 and CD4 (eBioscience), followed by anti-AQP3 mAb [11] and anti-mouse IgG conjugated with Alexa Fluor 488 (Thermo Fisher). Stained cells were analyzed using a CytoFLEX flow cytometer (Beckman Coulter Life Sciences, Brea, CA) and FlowJo software (Treestar).\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eImmunoblotting\u003c/h2\u003e \u003cp\u003eThe epidermis of the mouse was separated from the dermis by incubating in phosphate-buffered saline (PBS) at 60\u0026deg;C for 10 seconds. The epidermis was then lysed with an extraction buffer containing 250 mM sucrose, 1 mM EDTA, and 1% protein inhibitor cocktail (Sigma-Aldrich). For immunoblot analysis, a polyclonal AQP3 antibody (Millipore) and horseradish peroxidase-conjugated secondary antirabbit IgG antibody (Cell Signaling Technology) were used for detection via ECL (GE Healthcare, Piscataway, NJ).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eStatistical analysis was performed using the two-tailed t-test, one-way ANOVA or two-way ANOVA with GraphPad Prism 8 (San Diego, CA, USA). A p-value of less than 0.05 was considered statistically significant for all tests.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eAQP3 deficiency in T cells had little effect on IMQ-induced psoriasis development\u003c/h2\u003e \u003cp\u003eTo investigate which cell type\u0026rsquo;s expression of AQP3 contributes more significantly to the pathogenesis of psoriasis, we established AQP3 conditional knockout (cKO) mice in T cells or keratinocytes (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). Successful deletion of AQP3 in T cells was confirmed by quantitative RT-PCR and fluorescence-activated cell sorting \u003cb\u003e(\u003c/b\u003eFACS) analysis compared to control mice (AQP3 \u003csup\u003efl/fl\u003c/sup\u003e CD4-Cre\u003csup\u003e\u0026minus;/\u0026minus;\u003c/sup\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB, C). We topically applied IMQ to the ear skin to induce a psoriasis-like phenotype, characterized by noticeable ear thickening. No significant difference in ear thickness was observed between control and T cell-specific AQP3 cKO mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eD).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eAQP3 deficiency in keratinocytes attenuates IMQ-induced psoriasis-like symptoms\u003c/h2\u003e \u003cp\u003eNext, we examined the role of keratinocyte AQP3 in psoriasis development using keratinocyte-specific AQP3 cKO mice. AQP3 deletion was induced by tamoxifen injection before initiating the psoriasis model (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). The deletion of AQP3 in the epidermis was confirmed at both the protein and mRNA levels through Western blotting and quantitative RT-PCR, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB, C). Topical application of IMQ resulted in ear swelling, which was significantly reduced in keratinocyte-specific AQP3 cKO mice compared to control mice (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eD). These results indicate that AQP3 expression in keratinocytes, rather than T cells, contributes to the development of IMQ-induced psoriasis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eAdministration of AQP3 mAb suppressed IMQ-induced psoriatic symptoms\u003c/h2\u003e \u003cp\u003eTo evaluate the therapeutic potential of targeting AQP3 in psoriasis, mice were administered either AQP3 mAb or an IgG control during the development of IMQ-induced psoriasis (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). Psoriasis-like symptoms were quantified using the PASI scores [5]. Administration of AQP3 mAb significantly reduced visible erythema and scaling compared to control IgG treatment, suggesting a reducing in inflammation and keratinocyte proliferation (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB, C). H\u0026amp;E staining revealed that IMQ-induced epidermal hyperplasia was suppressed in mice treated with AQP3 mAb compared to those receiving control IgG (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAdditionally, quantitative RT-PCR analysis demonstrated that the IMQ-induced expression levels of S100A8 and S100A7\u0026mdash;key proteins involved in inflammation and keratinocyte proliferation\u0026mdash;were significantly decreased in the epidermis of AQP3 antibody-treated mice. Treatment with AQP3 mAb also reduced mRNA expression of Ki67, a marker of cell proliferation, consistent with the observation that epidermal thickening was suppressed (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE). Collectively, these data indicate that anti-AQP3 mAb treatment can effectively reduce the severity of IMQ-induced psoriasis-like symptoms, potentially by reducing keratinocyte hyperproliferation.\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eOur study demonstrated that AQP3 inhibition can effectively suppress psoriasis, highlighting the crucial role of AQP3 in the pathophysiology of the disease. This finding not only corroborates previous research identifying AQP3 as a key factor in various skin conditions [10, 16, 17] but also reinforces its potential as a therapeutic target specifically for psoriasis. Given the growing evidence linking AQP3 to inflammatory diseases, our study contributes to the understanding that AQP3 inhibition may be a promising new approach for treating inflammatory skin disorders, particularly psoriasis.\u003c/p\u003e \u003cp\u003eUsing AQP3 cKO mice, we found that AQP3 in keratinocytes is more critical for psoriasis development than AQP3 expression in T cells. This suggests a more direct and pivotal role for keratinocyte AQP3 in psoriasis than previously recognized. Our findings are consistent with reports indicating that AQP3 overexpression in keratinocytes leads to excessive cell proliferation and epidermal hyperplasia, which are characteristics typical of psoriatic lesions [9, 10]. Therefore, it is important to focus on pathways specific to keratinocytes, as well as T cells, when considering therapeutic options for psoriasis.\u003c/p\u003e \u003cp\u003eAbnormal interactions between keratinocytes and immune cells are key factors driving psoriasis progression [18, 19]. The inflammatory response causes keratinocyte hyperproliferation and perpetuates the chronic inflammatory cycle underlying psoriatic pathology. Although the pathogenesis of psoriasis is multifaceted and not fully understood, targeting AQP3 in keratinocytes may provide a strategy to interrupt this cycle and better regulate the disease. Additionally, we previously demonstrated that AQP3-mediated cellular H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e functions as a second messenger, regulating NF-κB activation and inflammatory responses during psoriatic pathology [10]. Future studies will explore the mechanisms by which AQP3 inhibition through the AQP3 antibody effectively prevents psoriasis development.\u003c/p\u003e \u003cp\u003eGiven the challenges associated with current psoriasis treatments, including the side effects of biologics [20] and JAK inhibitors [21], our findings suggest the potential for more selective and targeted therapies. In this study, we used AQP3 mAb to investigate the efficacy of AQP3 inhibition in psoriasis pathology, as there are currently no available AQP3 inhibitors for human or mouse administration. The results from the AQP3 cKO experiments are expected to encourage the development of small-molecule inhibitors targeting AQP3 for topical application. This approach may not only enhance symptom management for psoriasis patients but also deepen our understanding of the underlying mechanisms of the disease, paving the way for future therapeutic developments. However, an important limitation of our study is the lack of long-term data regarding the safety and efficacy of AQP3 inhibition. While our findings highlight its short-term benefits, further research is needed to assess the potential risks and therapeutic durability of prolonged AQP3 inhibition. Addressing these questions is crucial for translating AQP3 inhibition into a viable treatment option for chronic psoriasis.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eAQP3, aquaporin 3\u003c/p\u003e\n\u003cp\u003eH\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, hydrogen peroxide\u003c/p\u003e\n\u003cp\u003eAQP3 mAb, anti-AQP3 monoclonal antibody\u003c/p\u003e\n\u003cp\u003ecKO, conditional knockout\u003c/p\u003e\n\u003cp\u003eTMX, tamoxifen\u003c/p\u003e\n\u003cp\u003eRT-PCR, reverse transcription PCR\u003c/p\u003e\n\u003cp\u003eFACS, fluorescence-activated cell sorting\u003c/p\u003e\n\u003cp\u003ePBS, phosphate-buffered saline\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical Statements:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll animal experiments were approved by the President of Keio University, following the consideration by the Institutional Animal Care and Use Committee of Keio University (Approval no: 16075) and by Genetic Modification Safety Committee, Keio University School of Medicine (approval no. 28\u0026ndash;029).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by Research Ministry of Education, Culture, Sports, Science (21K06974, M.H-C).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eM.H-C. conceived the study. R.O., M.T., and M.H-C., performed the experiments and analyzed the data. R.O. and M.H-C. wrote the manuscript. All authors reviewed the manuscript\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by Research Ministry of Education, Culture, Sports, Science (21K06974, M.H-C).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eNestle FO, Kaplan DH, Barker J (2009) Psoriasis. New England Journal of Medicine 361:496\u0026ndash;509. https://doi.org/10.1056/NEJMra0804595\u003c/li\u003e\n\u003cli\u003eBoehncke W-H, Sch\u0026ouml;n MP (2015) Psoriasis. The Lancet 386:983\u0026ndash;994. https://doi.org/10.1016/S0140-6736(14)61909-7\u003c/li\u003e\n\u003cli\u003eHawkes JE, Chan TC, Krueger JG (2017) Psoriasis Pathogenesis and the Development of Novel, Targeted Immune Therapies. J Allergy Clin Immunol 140:645\u0026ndash;653. https://doi.org/10.1016/j.jaci.2017.07.004\u003c/li\u003e\n\u003cli\u003eZeichner JA, Armstrong A (2016) The Role of IL-17 in the Pathogenesis and Treatment of Psoriasis. J Clin Aesthet Dermatol 9:S3\u0026ndash;S6\u003c/li\u003e\n\u003cli\u003evan der Fits L, Mourits S, Voerman JSA, et al (2009) Imiquimod-Induced Psoriasis-Like Skin Inflammation in Mice Is Mediated via the IL-23/IL-17 Axis1. The Journal of Immunology 182:5836\u0026ndash;5845. https://doi.org/10.4049/jimmunol.0802999\u003c/li\u003e\n\u003cli\u003eHara-Chikuma M, Verkman AS (2008) Prevention of Skin Tumorigenesis and Impairment of Epidermal Cell Proliferation by Targeted Aquaporin-3 Gene Disruption. Molecular and Cellular Biology 28:326\u0026ndash;332. https://doi.org/10.1128/MCB.01482-07\u003c/li\u003e\n\u003cli\u003eHara-Chikuma M, Chikuma S, Sugiyama Y, et al (2012) Chemokine-dependent T cell migration requires aquaporin-3\u0026ndash;mediated hydrogen peroxide uptake. Journal of Experimental Medicine 209:1743\u0026ndash;1752. https://doi.org/10.1084/jem.20112398\u003c/li\u003e\n\u003cli\u003eVerkman AS, Anderson MO, Papadopoulos MC (2014) Aquaporins: important but elusive drug targets. Nat Rev Drug Discov 13:259\u0026ndash;277. https://doi.org/10.1038/nrd4226\u003c/li\u003e\n\u003cli\u003eSoler DC, Young AE, Griffith AD, et al (2017) Overexpression of AQP3 and AQP10 in the skin exacerbates psoriasiform acanthosis. Exp Dermatol 26:949\u0026ndash;951. https://doi.org/10.1111/exd.13307\u003c/li\u003e\n\u003cli\u003eHara-Chikuma M, Satooka H, Watanabe S, et al (2015) Aquaporin-3-mediated hydrogen peroxide transport is required for NF-\u0026kappa;B signalling in keratinocytes and development of psoriasis. Nat Commun 6:7454. https://doi.org/10.1038/ncomms8454\u003c/li\u003e\n\u003cli\u003eHara-Chikuma M, Tanaka M, Verkman AS, Yasui M (2020) Inhibition of aquaporin-3 in macrophages by a monoclonal antibody as potential therapy for liver injury. Nat Commun 11:5666. https://doi.org/10.1038/s41467-020-19491-5\u003c/li\u003e\n\u003cli\u003eTanaka M, Ito A, Shiozawa S, Hara-Chikuma M (2022) Anti-tumor effect of aquaporin 3 monoclonal antibody on syngeneic mouse tumor model. Transl Oncol 24:101498. https://doi.org/10.1016/j.tranon.2022.101498\u003c/li\u003e\n\u003cli\u003eMakar KW, P\u0026eacute;rez-Melgosa M, Shnyreva M, et al (2003) Active recruitment of DNA methyltransferases regulates interleukin 4 in thymocytes and T cells. Nat Immunol 4:1183\u0026ndash;1190. https://doi.org/10.1038/ni1004\u003c/li\u003e\n\u003cli\u003eVasioukhin V, Degenstein L, Wise B, Fuchs E (1999) The magical touch: Genome targeting in epidermal stem cells induced by tamoxifen application to mouse skin. Proceedings of the National Academy of Sciences 96:8551\u0026ndash;8556. https://doi.org/10.1073/pnas.96.15.8551\u003c/li\u003e\n\u003cli\u003eSchindelin J, Arganda-Carreras I, Frise E, et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676\u0026ndash;682. https://doi.org/10.1038/nmeth.2019\u003c/li\u003e\n\u003cli\u003eTricarico PM, Mentino D, De Marco A, et al (2022) Aquaporins Are One of the Critical Factors in the Disruption of the Skin Barrier in Inflammatory Skin Diseases. Int J Mol Sci 23:4020. https://doi.org/10.3390/ijms23074020\u003c/li\u003e\n\u003cli\u003eChen M, Peng Q, Tan Z, et al (2023) Targeting Aquaporin-3 Attenuates Skin Inflammation in Rosacea. Int J Biol Sci 19:5160\u0026ndash;5173. https://doi.org/10.7150/ijbs.86207\u003c/li\u003e\n\u003cli\u003eLowes MA, Russell CB, Martin DA, et al (2013) The IL-23/T17 pathogenic axis in psoriasis is amplified by keratinocyte responses. Trends in Immunology 34:174\u0026ndash;181. https://doi.org/10.1016/j.it.2012.11.005\u003c/li\u003e\n\u003cli\u003eTonel G, Conrad C (2009) Interplay between keratinocytes and immune cells\u0026mdash;Recent insights into psoriasis pathogenesis. The International Journal of Biochemistry \u0026amp; Cell Biology 41:963\u0026ndash;968. https://doi.org/10.1016/j.biocel.2008.10.022\u003c/li\u003e\n\u003cli\u003eJiang Y, Chen Y, Yu Q, Shi Y (2023) Biologic and Small-Molecule Therapies for Moderate-to-Severe Psoriasis: Focus on Psoriasis Comorbidities. BioDrugs 37:35\u0026ndash;55. https://doi.org/10.1007/s40259-022-00569-z\u003c/li\u003e\n\u003cli\u003eMiot HA, Criado PR, de Castro CCS, et al (2023) JAK-STAT pathway inhibitors in dermatology. Anais Brasileiros de Dermatologia 98:656\u0026ndash;677. https://doi.org/10.1016/j.abd.2023.03.001\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"molecular-biology-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mole","sideBox":"Learn more about [Molecular Biology Reports](https://www.springer.com/journal/11033)","snPcode":"11033","submissionUrl":"https://submission.nature.com/new-submission/11033/3","title":"Molecular Biology Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Aquaporin 3, psoriasis, antibody therapy","lastPublishedDoi":"10.21203/rs.3.rs-5145495/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5145495/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground:\u003c/h2\u003e \u003cp\u003eAquaporin 3 (AQP3) is highly expressed in both keratinocytes and T cells within psoriatic skin. Previous studies have demonstrated that AQP3 knockout mice show reduced development of psoriatic symptoms in murine models. This study aims to evaluate the effect of AQP3 inhibition on psoriasis progression.\u003c/p\u003e\u003ch2\u003eMethods and results:\u003c/h2\u003e \u003cp\u003eAQP3 conditional knockout mice were generated to assess the role of AQP3 expression in keratinocytes and T cells in psoriasis pathogenesis. In an imiquimod (IMQ)-induced psoriasis model, psoriatic symptoms were mitigated only in mice with keratinocyte-specific AQP3 deletion. Additionally, treatment with an anti-AQP3 monoclonal antibody (mAb) significantly reduced IMQ-induced psoriasis symptoms in wild-type mice.\u003c/p\u003e\u003ch2\u003eConclusions:\u003c/h2\u003e \u003cp\u003eAQP3 inhibition presents a promising approach for the treatment of psoriasis.\u003c/p\u003e","manuscriptTitle":"Aquaporin 3 inhibition Attenuates Imiquimod-Induced Psoriatic Symptoms in a Murine Model","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-12-04 07:48:29","doi":"10.21203/rs.3.rs-5145495/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-11-26T09:19:42+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-11-26T06:47:43+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"48413116901427652657834647763677083298","date":"2024-10-29T16:02:32+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-10-05T01:36:01+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"289589455540695769920827826168248046488","date":"2024-09-27T15:12:38+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-09-25T13:44:57+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-09-25T03:57:26+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-09-25T03:56:38+00:00","index":"","fulltext":""},{"type":"submitted","content":"Molecular Biology Reports","date":"2024-09-24T13:22:39+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"molecular-biology-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mole","sideBox":"Learn more about [Molecular Biology Reports](https://www.springer.com/journal/11033)","snPcode":"11033","submissionUrl":"https://submission.nature.com/new-submission/11033/3","title":"Molecular Biology Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"47b5c15c-9ef7-4cdd-93ce-9315598b4b38","owner":[],"postedDate":"December 4th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-04-07T16:04:20+00:00","versionOfRecord":{"articleIdentity":"rs-5145495","link":"https://doi.org/10.1007/s11033-025-10444-z","journal":{"identity":"molecular-biology-reports","isVorOnly":false,"title":"Molecular Biology Reports"},"publishedOn":"2025-04-01 15:57:41","publishedOnDateReadable":"April 1st, 2025"},"versionCreatedAt":"2024-12-04 07:48:29","video":"","vorDoi":"10.1007/s11033-025-10444-z","vorDoiUrl":"https://doi.org/10.1007/s11033-025-10444-z","workflowStages":[]},"version":"v1","identity":"rs-5145495","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5145495","identity":"rs-5145495","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}