Role of LncRNA NEAT-1 in pathogenesis of psoriasis in Egyptian patients

Abstract


Introduction
Psoriasis is a chronic, painful, and disabling disease with no known cure that has a substantial adverse effect on people's quality of life [1].Psoriasis has a wide variation in reported frequency among nations, from 0.09 to 11.4 percent [2], making psoriasis a major issue barrier breakdown, and immune system malfunction are all proposed as major contributors to this complex disease [3].
. RNA transcripts longer than 200 nucleotides that are not translated into proteins are called long non-coding RNAs.RNA polymerase II (RNA Pol II) is the primary transcriber of LncRNAs [4].LncRNAs regulate gene expression at several stages, involving histone modification, transcription, posttranscription, translation, & post-translation.
Dysregulation of LncRNAs participates in the pathogenesis of multiple autoimmune diseases [5].
NEAT1 is a new nuclear long noncoding RNA that clocks in at ~3.2 kb.It regulates gene expression through diverse mechanisms, including mRNA retention, mRNA breakage, A-to-I editing, and protein capture.Although NEAT-1's role in psoriasis remains unclear, it is possible that it contributes to the disease's development in a variety of ways.Overexpression of NEAT1 in SLE individuals' monocytes was found to elevate IL-6, CCL2, & CXCL10 production [6].They participate in the development of psoriasis.
Moreover, NEAT-1 acts through the MAPKrelated axis, which plays a role in psoriasis pathogenesis [7].
The current study aimed to determine the expression levels of LncRNA NEAT-1 in the serum of psoriasis patients.Also, to investigate its function in the pathogenesis of the disease and whether it can be used as a potential diagnostic marker for psoriasis.

Subjects
This study was carried out on a total of sixty participants; group I consisted of forty people diagnosed with psoriasis who had not been taking any therapy for at least one month prior to the start of the trial.They received care at the dermatology outpatient clinic located inside the Faculty of Medicine at Fayoum

University
Hospital.Psoriasis, chronic dermatological conditions, or systemic disorders involving renal or liver diseases, as well as malnutrition, did not develop in any of the twenty healthy participants who served as controls in Group II.These individuals were age-and gender-matched.

Exclusion criteria
• Age below 20 and above 60 years.
• Patients with other autoimmune diseases.
• Pregnant or lactating females.
• cases with hematological or solid malignancies.
• cases with concurrent or present history of systemic or cutaneous infections.
All study participants gave their written

Sample collection and storage
• A complete medical history and physical examination were performed on all subjects.
• Each participant's antecubital vein was venipuncture for 10 ml of blood after they fasted overnight and under sterile circumstances.

Results
This
Figure 1 shows that there was a significant increase in lncRNA NEAT-1 levels in psoriasis cases compared with controls (P<0.001).Table 2 shows no significant distinction in NEAT-1 levels among male and female psoriasis patients (P = 0.76), and between different courses of the disease (P =0.4).Table 3 shows that there was a statistically significant positive association between PASI score and age (P = 0.007) and platelet/lymphocyte ratio (P = 0.027) in psoriasis patients.Also, there was a statistically significant positive correlation between age of onset and each of age (P = 0.001) and triglycerides (P = 0.04), while there was a significant negative correlation between age of onset and HDL (P = 0.025).Also, there was a significant positive correlation among the ratios of platelets to lymphocytes and neutrophils to lymphocytes (P = 0.003).The ROC analysis showed that the diagnostic value of lncRNA NEAT-1 as a predictor in differentiating between cases of psoriasis and controls (p < 0.001, AUC = 1) at a cutoff value of 1.47 with sensitivity of 97%, specificity of 99%, & accuracy of 98 (Table 4, Figure 2).

Discussion
Psoriasis is a chronic, stressful, and disabling disease for which there is no cure and In another study about psoriasis, they found that CXCL10 was the most highly ranked gene and indicated positive relationships with the other six hub genes, suggesting it may be the most important gene.The CXCL10 RT-qPCR data further confirmed this forecast.Psoriatic skin lesions and serum CXCL10 levels have been found to be elevated in numerous investigations [13].
In SLE, NEAT1 increases the level of CXCL-10.The latter plays a role in the pathogenesis of psoriasis [14], so this might be leading to an immunological response in inflammatory and immune disorders [18].For example, upregulation of NEAT1 was reported in bronchial asthma [19] and SLE [20].
Overexpression of NEAT1 has also been demonstrated to increase levels of reactive oxygen species, which are known to aggravate immunological responses, herpes simplex infection, and inflammatory responses [21].We hypothesized that NEAT-1 has a role in the immunological and inflammatory responses observed in psoriasis based on the available information.
Consistent with previous research, we found a link between NEAT1 expression and disease activity in SLE [20].In addition, Wang Galectin-7 [22].Also, in another study performed to decipher the role of lncRNAs in psoriasis utilizing RT-qPCR, they found that Meg9 and NEAT-1 illustrated a trend towards reduced expression, but the results were not statistically significant [23].
Serum triglyceride (TG) and cholesterol levels were found to be significantly higher in psoriasis patients compared with controls (p = 0.02, 0.001 correspondingly), whereas serum high-density lipoprotein (HDL) levels were found to be significantly lower (p = 0.01).This was consistent with another study performed to estimate the lipid profile in psoriasis patients in comparison with controls, where the results were as follows: Individuals with psoriasis had considerably higher TG levels and significantly lower high-density lipoprotein (HDL) levels in contrast to people without psoriasis (p = 0.001 and 0.013, respectively) [24].
Our study also showed that there was an increase in neutrophil/lymphocyte ratio and platelet/lymphocyte ratio in psoriasis patients compared with controls, and this was consistent with another study performed to estimate these parameters in psoriasis patients, and they found that they were all significantly higher in moderate to severe psoriasis patients [25].Also, there was no significant variation among the two groups concerning AST and ALT; this was consistent with another study that found the activity and concentration of liver function markers (ALT and AST) did not differ significantly between healthy individuals and cases with psoriasis [26].Our study showed that there was no significant distinction amongst the two groups concerning serum creatinine levels, and this is consistent with another study that found the median levels of serum creatinine were not significantly different among psoriatic cases and healthy controls [27].
informed permission.The Ethics Committee of Fayoum University's School of Medicine has given the green light to all research including human subjects.All procedures involving human subjects complied with the Helsinki Declaration, an ethical norm established by the World Medical Association.

1 )
being extracted employing a miRNeasy mini-Kit (Qiagen, Valenica, CA, USA) extraction kit.B. Reverse transcription reactions A total of eleven microliters of RNA, two microliters of genomic DNA elimination (GE), and seven microliters of reverse transcription mix were combined into a final volume of twenty microliters to execute reverse transcription on total RNA.The RNAs were reverse transcribed into cDNAs with an RT-PCR kit utilizing RT2 first strand KIT Cat.No. 330404 (Qiagen, Valencia, CA, USA), in accordance with the instructions provided by the manufacturer.C. Quantitative Real-time PCR (qPCR) for Detection of Lnc RNA NEAT-1 RT-qPCR was performed using the Rotor-gene Q real-time PCR system (Qiagen, USA).We used RT2 SYBR Green ROX qPCR and the housekeeping gene (GAPDH; Catalog no: 330701 LPH31725A, Accession no: ENST00000496049.0) in a total volume of 25 μL.The PCR cycling procedure for quantifying lncRNA NEAT-1 begins with a 10-minute incubation at 95°C, followed by 40 cycles at 95°C for 15 seconds and 60°C for 60 seconds.2−ΔΔCt was utilized to calculate the serum fold changes of NEAT-1.The values for FC controls were set to one.
Package for the Social Sciences (SPSS) version 22 was employed for data collection, codification & analysis.Categorical data were compared using the Chisquare (χ2) test.The mean and standard deviation were employed to summarize numerical data.Both the independent-t test and the Mann-Whitney U test were used to assess the degree of similarity between the two sets of information.When comparing courses, the Kruskal-Wallis test was utilized.ROC analysis was done to detect the diagnostic value of serum NEAT-1 for individuals with psoriasis.Pearson's correlation coefficient was utilized to establish associations amongst quantitative variables [10].

Figure 2 :
Figure 2: ROC curve analysis of lncRNA NEAT-1 for the differentiation between psoriasis patients and controls.

FUMJ
has a great negative influence on individuals' quality of life.Genetic factors play a significant role in the pathogenesis of the disease.Immunity also plays a role in the pathogenesis of psoriasis through T cells, dendritic cells, natural killer cells, Langerhans cells, and neutrophils[11].In agreement with the findings of investigations of LncRNAs in immune cells (dendritic cells, neutrophils, T cells, monocytes, macrophages, & B cells), it was discovered that the degree of expression of LncRNA was connected with the differentiation, development, and activation of immune cells[12].NEAT1 is a conserved, long, noncoding RNA whose significance in immunity is still largely unknown.The purpose of this study was to examine the expression levels of LncRNA NEAT1 in the serum of psoriasis patients, its role in the pathogenesis of the disease, and whether it could be used as a potential marker for the diagnosis of psoriasis.To the best of our knowledge, this was the first study to detect the level of NEAT-1 in the serum of psoriatic patients.In the current research, the expression of LncRNA NEAT-1 was detected by RT-PCR in psoriatic patients and contrasted with controls.The serum expression of LncRNA NEAT-1 was significantly higher in patients than controls.According to the ROC test, NEAT1 showed a highly significant value in psoriasis (p value < 0.001, AUC = 1) at a cutoff value of 1.47 with sensitivity of 97%, specificity of 99%, & , which could be useful to establish a new diagnostic marker.There were many studies that showed that many other autoimmune disorders involving SLE had previously implicated NEAT1.It had been established that NEAT1 was overexpressed in SLE patients' monocytes.In order to increase the expression of IL-6, CCL2, and CXCL10, this lncRNA phosphorylates JNK & ERK.Thus, it is obvious that NEAT-1 increases the level of IL-6.The latter plays a role in the development of psoriasis.This might be considered a mechanism for the action of NEAT1 in psoriasis development.
considered another mechanism for the role of NEAT-1 in the development of psoriasis.Cell proliferation, differentiation, gene expression, and apoptosis are all regulated by the MAPK kinases, which together form an important collection of signaling pathways [15].MAPKs are activated by phosphorylation, and once activated, they phosphorylate other intracellular kinases and transcription factors.One of the targets of the p38 MAPK signaling cascade is a protein kinase called MAPKactivated protein kinase 2 (MK2).Psoriatic lesions were shown to have larger quantities of activated MK2 than other tissues [16].These findings could support our result, as P38MAPK participates in the development of psoriasis and NEAT-1 acts through the MAPK-related axis.NEAT-1 knockdown inhibited the Ras-MAPK pathway [17].This might be considered an explanation for the role played by NEAT1 in psoriasis development.Multiple inflammasomes (NLRP3, NLRC4, and AIM2) & proinflammatory cytokines (CXCL10, IL-6, & IL-1β) are released when NEAT-1 is upregulated, et al. (2017) discovered that blocking NEAT1 results in better skin lesion healing [21].In contrast with our results, another study was performed to clarify the mechanism underlying the influence of paeoniflorin (PF) on the proliferation and emigration of psoriatic keratinocytes.The expressions of NEAT-1 and Galectin-7 in skin tissues from psoriatic patients and healthy subjects were evaluated, and there was a reduced expression of NEAT-1 and Galectin-7.PF suppressed the proliferation and emigration of psoriatic HaCat cells by increasing the expressions of NEAT-1 and

Table 1 :
Demographic data of control subjects and psoriasis patients.

Table 2 :
Risk factors among the studied groups.

Table 3 :
Correlations between various studied parameters in psoriasis patients.

Table 4 : Sensitivity and specificity of lncRNA NEAT-1 in diagnosis of psoriasis.
AUC: area under the curve, P value: probability value, CI: confidence interval.