Anna Michalak-Stoma

Tryptophan (TRP) and its metabolites are involved in numerous physiological and pathological processes within the skin. They may influence melanoma biology, but their role remains insufficiently understood. This study investigated the effects of TRP, anthranilic acid (AA), and cinnabarinic acid (CBA) in melanoma cells in vitro and their potential interaction with the B-Raf proto-oncogene, serine/threonine kinase (BRAF) inhibitor vemurafenib. A375, G361, and SK-MEL-3 melanoma cell lines and a THP-1-derived macrophage model were used. Cell proliferation and metabolic activity were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, migration using a scratch assay, and cell-cycle distribution by flow cytometry. Western blotting was used to analyze selected signaling proteins. Reactive oxygen species (ROS) and nitric oxide (NO) production were evaluated. Bioinformatic analyses investigated associations between BRAF and genes related to tryptophan metabolism and the kynurenine pathway. All compounds showed dose-dependent antiproliferative effects, with prolonged exposure increasing efficacy. TRP and AA increased G0/G1 cell-cycle arrest, whereas CBA increased the G2/M population. TRP also reduced A375 cell migration. All three compounds enhanced the antiproliferative effect of vemurafenib, with CBA showing the strongest effect (approximately 20%). Western blot analysis indicated modulation of p21 expression and ERK1/2 phosphorylation, suggesting implications for cell cycle and stress signaling pathways. Moreover, conditioned medium from A375 cells exposed to tested compounds affected cell viability, ROS, and NO production in THP-1-derived macrophages. TRP and its metabolites may be considered potential supportive agents for melanoma BRAF-targeted therapy. However, further in vivo studies are needed to clarify their therapeutic value and to elucidate the underlying molecular mechanisms.

CD (cluster of differentiation) 69 and CD25 are considered early and late markers of the activation of lymphocytes, respectively. CD25 is a part of the IL-2 receptor and is present on the surface of immune and non-immune cells, with high amounts on activated lymphocytes and regulatory T cells. CD69 is expressed on various types of white blood cells, including newly activated lymphocytes, lymphocytes infiltrating tissues isolated from subjects with chronic auto-inflammatory diseases, several subtypes of memory T cells and regulatory T cells. Primarily, CD69 was considered to be an early marker of the activation of lymphocytes, but, right now, data derived from in vitro and in vivo studies have revealed the immunomodulatory role of this surface antigen. In 84 patients with psoriasis, of whom 28 were treated with different biologic drugs, as well as in 29 healthy control subjects, the expression of CD25 and CD69 on different subtypes of peripheral blood mononuclear cells (PBMCs) was studied with the use of flow cytometry. Significantly higher levels of CD3/CD69-, CD8/CD69- and CD19/CD69-positive PBMCs as well as within CD3+ cells were present in subjects suffering from psoriasis when compared to healthy controls. In patients with psoriasis who were treated with biologic drugs, the levels of CD3/CD69-, CD4/CD69- and CD19/CD69-positive PBMCs, and CD3/CD69 within CD3+ cells, CD4/CD69 within CD4+ cells, CD4/CD25 within CD4+ cells and CD19/CD69 within CD19+ cells were significantly higher than before therapy. Our results support a role for activation markers, especially CD69, in psoriasis. Further research is warranted to fully clarify their significance in this common dermatosis, especially during biologic treatment.

Psoriasis is nowadays recognized as a multifactorial systemic disease with complex and not fully understood pathogenesis. In psoriatic patients, the increased cardiovascular disease (CVD) risk and frequent comorbidities like obesity are observed. The aim of this study was to investigate differences in miRNA (miR-22-3p, miR-133a-3p, miR-146a-5p, miR-369-3p, and Let-7b-5p) involved in CVD risk among psoriatic patients with overweight/obesity and with normal weight. The study comprised 28 male psoriatic patients and 16 male healthy controls. miRNA isolated from peripheral blood mononuclear cells was reverse-transcribed and RT-qPCR was performed. We have found decreased levels of miR-22, miR-133a, miR-146a, and miR-369 among the psoriatic patients. There was a statistically significant difference in miR-22 and miR-146a levels between psoriatic patients with overweight/obesity and with normal weight. There were positive correlations between miR-22 and miR-146a levels and psoriatic arthritis (PsA) in psoriatic patients with normal weight and between the miR-133a level and PsA in the overweight/obese patients. The decreased levels of selected miRNA are consistent with the levels observed in CVD indicating their impact on the CVD risk in psoriatic patients. miR-22 and miR-146 may be recognized as one of the contributing factors in the obesity-CVD-psoriasis network.

Psoriasis is nowadays recognized as a multifactorial systemic disease with complex and still not fully understood pathogenesis. In psoriatic patients the increased cardiovascular disease (CVD) risk and frequent comorbidities like obesity are observed. The aim of this study was to investigate differences in miRNA (miR-22-3p, miR-133a-3p, miR-146a-5p, miR-369-3p and Let-7b-5p) involved in CVD risk among psoriatic patients with overweight/obesity and with normal weight. The study comprised 28 male psoriatic patients and 16 male healthy controls. miRNA isolated from peripheral blood mononuclear cells was reverse-transcribed and RT-qPCR was performed. We have found the decreased levels of miR-22, miR-133a, miR-146a, and miR-369 among the psoriatic patients. There was a statistically significant difference in miR-22 and miR-146a levels between psoriatic patients with overweight/obesity and with normal weight. There were positive correlations between miR-22 and miR-146a levels and psoriatic arthritis (PsA) in psoriatic patients with normal weight and between miR-133a level and PsA in the overweight/obese patients. The decreased levels of selected miRNA are consistent with their levels observed in CVD indicating their impact in the CVD risk in psoriatic patients. miR-22 and miR-146 may be recognized as one of the contributing factors in the obesity-CVD-psoriasis network.

The programmed death-1 (PD-1) receptor plays a major physiological role in the maintenance of immune tolerance and, by interaction with its ligands (PD-L1 and PD-L2), prevents the development of multiple immune-mediated diseases. There is growing evidence of the PD-1/PD-L1 pathway playing an important role in the pathogenesis of psoriasis. In total, 84 subjects with psoriasis were included in this study, together with 29 healthy subjects as a control group. Twenty-eight of the psoriatic patients were treated with biologic therapy (TNF-alpha, interleukin (IL)-12/23, or IL-17 inhibitors). The amounts of PD-1- and PD-L1-positive T-cells in peripheral blood were evaluated using flow cytometry. Significantly lower levels of peripheral blood mononuclear cells (PBMCs) with the expression of PD-1 and PD-L1 were found in psoriatic patients compared to healthy individuals, i.e., CD3/PD-1-, CD3/PD-L1-, CD4/PD-1-, CD4/PD-L1-, CD8/PD-L1-, CD19/PD-1-, and CD19/PD-L1-positive cells. Biologic treatment resulted in the elevation of CD3/PD-L1- and CD8/PD-L1- and a decrease in CD8/PD-1-positive PBMCs. Our results confirm previous observations of the PD-1/PD-L1 pathway being disrupted in psoriasis, and that these disturbances may play an important role in development of the disease. Biologic drugs may reverse several abnormalities observed within this pathway, which may explain their excellent efficacy in the treatment of psoriasis. Further research should be conducted to fully explain the results obtained.

Programmed-death-1 (PD-1) receptor plays a major physiological role in maintainance of im-mune tolerance, and, by interaction with its ligands (PD-L1 and PD-L2), prevents the develop-ment of multiple immune-mediated diseases. There is growing evidence of an important role of PD-1/PD-L1 pathway in pathogenesis of psoriasis. 84 subjects with psoriasis were included into the study, together with 29 healthy subjects as a control group. 28 of psoriatic patients were treated with biologice therapy (TNF-alpha, interleukin (IL)-12/23 or IL-17 inhibitors). Amounts of PD1 and PD-L1 positive T-cells in peripheral blood were evaluated using flow cytometry. Significantly lower levels of peripheral blood mononuclear cells (PBMCs) with the expression of PD-1 and PD-L1 were found in psoriatic patients when compared to healthy individuals, i.e. CD3/PD1, CD3/PDL1, CD4/PD1, CD4/PDL1, CD8/PDL1, CD19/PD1, CD19/PDL1 positive cells. Biologic treatment resulted in elevation of CD3/PDL1, CD8/PDL1 and decrease of CD8/PD1 posi-tive PBMCs. Our results confirm previous observations of disrupted PD-1/PD-L1 pathway in psoriasis, and that this disturbances may play an important role in development of the disease. Biologic drugs may reverse several abnormalities observed within this pathway, which may ex-plain their excellent efficacy in the treatment of psoriasis. Further research should be conducted to fully explain the results obtained.

Psoriasis is an autoimmune and autoinflammatory disorder that has a significant impact on patient quality of life. The aim of the study was to assess the immune profiles of patients with psoriasis with multiple cytokine analysis. Fifty-two male psoriatic patients and 24 healthy male volunteers were recruited. Granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-gamma), interleukin (IL)-1 beta, IL-2, Il-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-13, IL-17A, IL-18, IL-21, IL-22, IL-23, IL-27, and tumor necrosis factor (TNF)-alpha were measured in patients’ serum with a Th1/Th2/Th9/Th17/Th22/Treg Cytokine 18-Plex Human ProcartaPlex Panel, based on Luminex xMAP technology. The median fluorescence intensities of serum GM-CSF, IL-2, IL-5, IL-10, IL-13, IL-17A, IL-21, and IL-22 were not intensive enough to calculate the cytokine concentration. We observed elevated levels of IL-6 (P=0.001) and IL-9 (P=0.003) in patients, compared with the control group. The levels of IL-1beta (P=0.008) and IL-27 (P=0.006) were decreased. In patients with psoriatic arthritis, we noticed a decreased level of IL-9 compared with that in patients without arthritis (P=0.034). The levels of IL-12 (P<0.05) and IL-18 (P<0.05) correlated positively with the Psoriasis Area and Severity Index. We found negative correlations of IL-9 (P<0.05), IL-12 (P<0.05), and IL-23 (P<0.05) with the age of psoriatic patients; IL-12 (P<0.05) and IL-23 (P<0.05) with psoriasis duration; and IL-6 (P<0.05) and IL-9 (P<0.05) with the Nail Psoriasis Severity Index. Multiple cytokine analysis seems to be an important form of individual immune profile assessment before treatment selection.

Morphea, also known as localized scleroderma (LoS), is a chronic autoimmune disease of the connective tissue. The clinical picture of LoS distinguishes between active and inactive lesions. Sometimes the clinical findings are challenging to identify, and therefore, the need for additional methods is emphasized. Our study aimed to demonstrate the characteristic dermoscopic features in morphea skin lesions, focusing on demonstrating features in active and inactive lesions. In our patients (n = 31) with histopathologically proven LoS, we performed clinical evaluation of lesions (n = 162): active/inactive and according to both disease activity (modified localized scleroderma severity index, mLoSSI) and damage (localized scleroderma skin damage index, LoSDI) parameters. In addition, we took into account compression locations to determine whether skin trauma, a known etiopathogenetic factor in LoS, affects the dermoscopic pattern of the lesions. We performed a dermoscopy of the lesions, categorizing the images according to the severity within the observed field. We showed that within the active lesions (clinically and with high mLoSSI), white clouds and linear branching vessels had the highest severity. These features decreased within the observed field in inactive lesions and with high LoSDI. Brownish structureless areas were most intense in inactive lesions with high LoSDI. Erythematous areas, linear branching vessels, dotted vessels, and crystalline structures were statistically significant for pressure locations. We have shown dermoscopy is a valuable tool to assess the activity or inactivity of lesions, which translates into appropriate therapeutic decisions and the possibility of monitoring the patient during and after therapy for possible relapse.

Advances in genotypic technologies enable identification of possible associations between genetic variants of certain genes and increased risk of developing plaque psoriasis or psoriatic arthritis. The aim of the study was to analyze the NOTCH3 (6746T>C) (rs1044009) and PSMA6 (-8C>G) (rs1048990) polymorphisms and their role in genetic susceptibility to psoriasis. The study included 158 psoriatic patients and 100 healthy controls. The frequencies of the NOTCH3 genotypes differed between the psoriatic patients and healthy controls (p = 0.050). No differences were found in the distribution of PSMA6 genotypes and alleles between the psoriatic patients and healthy controls. The studied psoriatic patients presented a higher frequency of the CC genotype of PSMA6 compared to the healthy controls (8.8% vs. 2%, respectively). Psoriatic arthritis was more frequent among patients with the CC genotype of PSMA6 (p = 0.059). CC homozygosity of NOTCH3 was more commonly observed in the studied psoriatic patients than in the healthy controls (OR = 4.76, p= 0.032). The obtained data suggest that genetic variants of NOTCH3 (6746T>C) and PSMA6 (-8C>G) genes may play significant roles in psoriatic patients. Further studies are necessary to unequivocally determine their role as genetic risk factors of psoriasis development.

Seborrheic keratosis (SK) is the most common benign tumour of epidermal origin. In most cases, it is simple to recognize in the clinical examination. However, sometimes SK can be a problematic lesion. We present the cases of two patients with seborrheic keratosis in whom we diagnosed the skin cancer through dermoscopic and histopathological examinations. The article aims to draw attention to the need for dermoscopic examinations to be included for an accurate assessment of the nevi not only by dermatologists but also not-specialized doctors. We would like to underline that many skin cancers share the similar features of malignancy, and competence and capability to interpret the dermoscopic pictures correctly are important for early recognition of malignant lesion. Very often malignant skin cancers can be hidden among benign lesions like seborrheic keratosis or they can be imitators of benign lesions. Amongst all cases of imposing SK, basal cell carcinoma, squamous cell carcinoma, melanoma is the most important differential diagnosis, of which their dermoscopic features will be discussed in this article.