Wcisło-Dziadecka Dominika, Simka Klaudia, Kaźmierczak Agata, Kruszniewska-Rajs Celina, Gola Joanna, Grabarek Beniamin, Hybiak Jolanta, Grillon Catherine, Mazurek Urszula, Łos Marek J
Department of Skin Structural Studies, Chair of Cosmetology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Katowice, Poland.
Department of Internal Medicine, School of Public Health in Bytom, Medical University of Silesia, Katowice, Poland.
Cell Physiol Biochem. 2018;50(2):525-537. doi: 10.1159/000494166. Epub 2018 Oct 11.
BACKGROUND/AIMS: Psoriasis, an autoimmune diseases of the skin, characterized by patches of abnormal/inflammed skin, although not usually life-threatening, it causes severe discomfort, esthetic impairments, and may lead to impaired social functions and social withdrawal. Besides UV-phototherapy, various anti-inflammatory treatments are applied, depending on the severity of symptoms. In 2008, adalimumab (fully humanized human anti-TNF antibody) was launched for the treatment of psoriasis. In the quest to better understand the pathomechanism of adalimumab's therapeutic effects, and the acquired resistance to the drug, we have investigated how its administration affect the regulation of the expression of selected caspases, including those activated by inflammosome.
The research was initially carried out on normal human dermal fibroblasts (NHDF) treated with adalimumab for 2, 8 and 24 hours in vitro. Then, expression profile of genes encoding caspases and their regulatory micro-RNAs was determined with the use of oligonucleotide microarray. The validation of the microarray results was carried out by qRT-PCR. The in vitro study was followed by ex-vivo investigation of adalimumab's effects on the expression of caspase-6 in blood of the psoriatic patients. The samples were collected before, and 2 hours after adalimumab's administration and the analysis was determined by qRT-PCR.
The result of the analysis indicated that introduction of adalimumab to the NHDF culture resulted in the change of the transcription activity of genes encoding caspases and genes encoding miRNAs. The analysis revealed 5 different miRNA molecules regulating the expression of: CASP2, CASP3 and CASP6. There were no statistically significant differences in the expression of gene encoding caspase-6 in the patients' blood before and 2 hours after the anti-TNF drug administration.
We have found that adalimumab administration affects caspases expression, thus they may be used as molecular markers for monitoring the therapy with the use of an anti-TNF drugs, including adalimumab. It is likely that the mechanisms responsible for changed expression profiles of genes encoding caspase-2,-3, and -6, may be caused by the upregulation of the respective microRNA molecules. Increased expression of genes encoding specific caspases may induce inflammatory processes, as well as trigger apoptosis. Furthermore, the proapoptotic activity of caspases may be enhanced by miRNA molecules, which exhibit proapoptotic function. The overexpression of such miRNAs was observed in our study.
背景/目的：银屑病是一种皮肤自身免疫性疾病，其特征为皮肤出现异常/炎症斑块，虽通常不危及生命，但会导致严重不适、美观受损，并可能导致社交功能受损和社交退缩。除紫外线光疗外，还会根据症状严重程度采用各种抗炎治疗方法。2008年，阿达木单抗（完全人源化抗TNF抗体）被批准用于治疗银屑病。为了更好地理解阿达木单抗的治疗作用病理机制以及对该药物的获得性耐药性，我们研究了其给药方式如何影响所选半胱天冬酶（包括由炎性小体激活的半胱天冬酶）表达的调节。
该研究最初在体外将阿达木单抗处理2、8和24小时的正常人皮肤成纤维细胞（NHDF）上进行。然后，使用寡核苷酸微阵列确定编码半胱天冬酶及其调节性微小RNA的基因的表达谱。通过qRT-PCR对微阵列结果进行验证。体外研究之后，对阿达木单抗对银屑病患者血液中半胱天冬酶-6表达的影响进行了离体研究。在给药前及给药后2小时采集样本，并通过qRT-PCR进行分析。
分析结果表明，将阿达木单抗引入NHDF培养物会导致编码半胱天冬酶的基因和编码微小RNA的基因的转录活性发生变化。分析发现5种不同的微小RNA分子调节CASP2、CASP3和CASP6的表达。抗TNF药物给药前和给药后2小时患者血液中编码半胱天冬酶-6的基因表达无统计学显著差异。
我们发现阿达木单抗给药会影响半胱天冬酶的表达，因此它们可用作监测包括阿达木单抗在内的抗TNF药物治疗的分子标志物。编码半胱天冬酶-2、-3和-6的基因表达谱变化的机制可能是由各自微小RNA分子的上调引起的。编码特定半胱天冬酶的基因表达增加可能会诱导炎症过程以及引发细胞凋亡。此外，微小RNA分子可能会增强半胱天冬酶的促凋亡活性，这些分子具有促凋亡功能。在我们的研究中观察到了此类微小RNA的过表达。
