Archives of Dermatological Research (2019) 311:535–544 https://doi.org/10.1007/s00403-019-01931-y ORIGINAL PAPER Narrowband UVB treatment induces expression of WNT7B, WNT10B and TCF7L2 in psoriasis skin Malin Assarsson1 · Jan Söderman2 · Albert Duvetorp3 · Ulrich Mrowietz4 · Marita Skarstedt2 · Oliver Seifert1,5 Received: 14 December 2018 / Accepted: 2 May 2019 / Published online: 14 May 2019 © The Author(s) 2019 Abstract WNT/β-catenin signaling pathways play a pivotal role in the human immune defense against infections and in chronic infammatory conditions as psoriasis. Wnt gene alterations are linked to known comorbidities of psoriasis as obesity, dia- betes and Crohn’s disease. The objective of this study was to investigate WNT7B, WNT10B, WNT16 and TCF7L2 gene and protein expression in lesional and non-lesional skin and in the peripheral blood of patients with chronic plaque psoria- sis compared with healthy individuals. To investigate the efect of narrowband UVB radiation, expression of these genes were analyzed before and after narrowband UVB treatment. Associations between single nucleotide polymorphisms for WNT7B, WNT10B, WNT16 and TCF7L2 genes and psoriasis were tested. Our results show signifcantly decreased WNT7B, WNT10B and TCF7L2 gene expression in lesional skin compared with non-lesional skin and healthy controls. Narrowband UVB treatment signifcantly increased expression of these genes in lesional skin. Immunohistochemistry shows increased WNT16 expression in lesional skin. No signifcant diferences in allele or genotype frequencies for Wnt or TCF7L2 gene polymorphisms were found between patient and control group. This study shows for the frst time signifcant UVB induced upregulation of WNT7B, WNT10B and TCF7L2 in patients with psoriasis and suggests a potential role of these genes in psoriasis pathogenesis. Keywords WNT/β-catenin signaling · WNT proteins · Infammation · Gene expression · Single nucleotide polymorphisms Introduction WNT proteins are a family of secreted glycoproteins with 19 isoforms. Recent reports suggest a role of WNTs in infam- mation, psoriasis and the human immune defense against Electronic supplementary material The online version of this article (https ://doi.org/10.1007/s0040 3-019-01931 -y) contains infections [6, 22, 23, 37, 40, 42]. WNTs can activate two supplementary material, which is available to authorized users. distinct signaling pathways: the canonical WNT/β-catenin pathway and the β-catenin-independent, non-canonical path- * Oliver Seifert way [9]. [email protected] Psoriasis is a chronic systemic infammatory disease char- 1 Division of Dermatology and Venereology, Ryhov Hospital, acterized by activation of both innate and adaptive immunity Region Jönköping County, 55185 Jönköping, Sweden [34]. Dendritic cells play a key role in linking innate and 2 Division of Medical Diagnostics, Ryhov Hospital, Region adaptive immunity and in balancing infammatory and regu- Jönköping County, Jönköping, Sweden latory responses. Interestingly, a recent study showed that 3 Division of Dermatology and Venereology, Skånes activation of WNT/β-catenin signaling in dendritic cells is University Hospital, Malmö, Sweden critical for promoting tolerance and limiting infammation 4 Psoriasis-Center, Department of Dermatology, University [45]. Medical Center Schleswig-Holstein, Campus Kiel, Kiel, In spite of the important roles WNT proteins play in Germany cell proliferation and differentiation and their role in 5 Department of Clinical and Experimental Medicine, Faculty innate immunity, not much is known about the expres- of Medicine and Health Sciences, Linköping University, sion and the potential function of WNT isoforms under Linköping, Sweden Vol.:(0123456789)1 3 536 Archives of Dermatological Research (2019) 311:535–544 pathophysiological conditions, such as in psoriasis. Table 1 Study population’s demographic data, psoriasis area and WNT16 plays a role in mediating keratinocyte prolif- severity index (PASI), body mass index (BMI) and age shown as mean and standard deviation (SD) eration [46], and one study shows lower WNT7B gene expression in lesional skin compared to non-lesional Psoriasis Control skin while WNT16 expression was unchanged [43]. Two Genotyping reports describe increased expression of WNT5A mRNA Study subjects (n female, % 170 (61, 36%) 365 (201, 55%) and protein in lesional skin of patients with psoriasis [22, female) 43], and interestingly, recent data suggest WNT5a to be a Age 51.7 (15.8) 55.4 (10.5) link between psoriasis, obesity and metabolic complica- Skin gene expression tions [17, 33]. The transcription factor 7-like 2 (TCF7L2) Study subjects (n female, % 32 (12, 38%) 20 (13, 65%) gene encodes a transcription factor involved in the WNT female) signaling pathway, and numerous studies show a correla- Age 54.8 (13.9) 45.7 (14.3) tion between TCF7L2 single nucleotide polymorphisms PASI 7.2 (6.5) – (SNPs) and dyslipidemia, metabolic syndrome and type BMI 27.5 (3.5) 23.9 (1.7) 2 diabetes, [19, 41] which are known associated diseases Immunohistochemistry of psoriasis [20]. Study subjects (n female, % 4 (2, 50%) 4 (2, 50%) The aim of this study was to describe the expression of female) WNT7B, WNT10B, WNT16 and TCF7L2 in lesional and Age 62.0 (12.1) 43.3 (10.8) non-lesional skin and in whole blood of patients with pso- PASI 6.7 (3.7) – riasis compared to healthy individuals and to investigate if Blood gene expression n SNPs in these genes can be correlated to psoriasis. Previ- Study subjects ( female, % 34 (15, 44%) 16 (9, 57%) female) ous studies show that ultraviolet (UV) radiation upregu- Age 57.6 (14.8) 42.5 (12.0) lates members of the WNT/β-catenin pathway [16, 50]. To PASI 6.4 (5.5) – investigate the efect of UV radiation on WNT7B, WNT10B, BMI 27.8 (4.5) 24.4 (2.0) WNT16 and TCF7L2, gene expression levels were analyzed Gene expression nbUVB treatment before and after narrowband UVB (nbUVB) treatment. Study subjects (n female, % 27 (7, 21%) – female) Age 48.3 (15.1) – Materials and methods PASI before 7.9 (4.5) – PASI after 2.1 (1.8) – BMI 26.5 (4.1) – Study subjects Patients with chronic plaque psoriasis were recruited from the outpatient clinic at the Division of Dermatology, Sampling methods Ryhov Hospital, Jönköping, Sweden. Healthy controls were recruited from patients visiting the same clinic for evaluation Full-thickness punch biopsies for WNT7B, WNT10B, of benign nevi. Whole blood samples for genotyping were WNT16 and TCF7L2 gene expression and immunohisto- obtained from these controls and from blood donors from chemistry (IHC) were taken from non-lesional skin (4 mm Ryhov Hospital, Jönköping, Sweden. Participants’ demo- in diameter; at least 10 cm distance from any psoriatic graphic data are summarized in Table 1. The type of analysis lesion) and from the active margin of a psoriatic plaque performed and the number of individuals included for each (4 mm diameter) after application of local anesthetics. In analysis are illustrated in Fig. 1. Study subjects did not use healthy controls, biopsies were obtained from correspond- any systemic or topical anti-psoriatic treatments 2 weeks ing anatomical sites. Immediately upon removal, biopsies prior to study inclusion. This study was conducted in com- were stored in either formalin for IHC or RNAlater, RNA pliance with good clinical practice and according to the Dec- Stabilization Reagent (Qiagen, Hilden, Germany) for gene laration of Helsinki Principles. Written informed consent expression analysis and stored at − 150 °C. was obtained from all subjects under protocols approved by Whole blood samples were taken from patients with the Local Ethics Committee, Linköping University, Sweden. psoriasis and controls for WNT7b, WNT10b, WNT16 and Sex, age and body mass index (BMI) were recorded for all TCF7L2 gene expression and genotyping. For genotyp- individuals and an assessment of disease severity using the ing venous blood samples were collected in BD Vacu- Psoriasis Area and Severity Index (PASI) was recorded for tainer EDTA tubes (BD Biosciences). The tubes were subjects with psoriasis. 1 3 Archives of Dermatological Research (2019) 311:535–544 537 Fig. 1 Illustration of patient fow and number of patients with chronic plaque psoriasis (n) and healthy control individuals (c) included in the study and the diferent WNT7B, WNT10B, WNT16 and TCF7L2 analysis performed (nb narrowband, IHC immunohistochemistry) centrifuged at 2000×g for 10 min and the buffy coat was Immunohistochemistry frozen in − 80 °C for subsequent DNA extraction and genotyping. Tempus™ Blood RNA tubes (Thermo Fisher WNT7B, WNT10B, WNT16 and TCF7L2 staining was per- Scientific Inc) were frozen in − 80 °C for subsequent formed using a standard protocol on 4 µm sections from RNA extraction and gene expression. formalin-fxed parafn-embedded tissue blocks as previ- For analysis of WNT7B, WNT10B, WNT16 and ously described [12]. Sections were subsequently incubated TCF7L2 gene expression in response to nbUVB treat- for 30 min at a concentration of 1:1000 with a primary ment, full-thickness 2 mm punch biopsies were taken rabbit anti-human WNT7B antibody (1.0 mg/mL, Sigma after application of local anesthetics from a single pso- Aldrich, St Louis, USA), with primary mouse anti-human riatic plaque and from non-lesional skin at least 10 cm WNT10B anti-body at a concentration of 1:50 (0.5 mg/mL, distance from any psoriatic lesion of patients before and R&D, Minneapolis, USA), with primary rabbit anti-human after receiving a full nbUVB treatment series according to WNT16 anti-body at a concentration of 1:40 (0.1 mg/mL, standard clinical protocol at the Division of Dermatology, Sigma Aldrich, St Louis, USA) and with primary mouse Ryhov Hospital. The location of the biopsies before treat- anti-human TCF7L2 antibody at a concentration of 1:200 ment was recorded to ensure that biopsies after treatment (1.0 mg/mL, Sigma Aldrich, St Louis, USA). were taken from approximately the same location. Detection of primary antibodies was performed using the MACH4 Universal HRP-Polymer Detection System (Biocare Medical, Concord, USA) in the IntelliPATH FLX system Phototherapy protocol (Biocare Medical, Concord, USA) as previously described [32].