mRNA 表达分析证实系统性红斑狼疮患者中 CD44 剪接受损。

mRNA expression analysis confirms CD44 splicing impairment in systemic lupus erythematosus patients.

机构信息

Department of Biomedicine & Prevention, Genetics Section, University of Rome "Tor Vergata", Rome, Italy.

Lupus Clinic, Dipartimento di Scienze cliniche internistiche, anestesiologiche e cardiovascolari, Sapienza University of Rome, Rome, Italy.

出版信息

Lupus. 2021 Jun;30(7):1086-1093. doi: 10.1177/09612033211004725. Epub 2021 Apr 1.

DOI:10.1177/09612033211004725

PMID:33794704

Abstract

BACKGROUND

Systemic Lupus Erythematosus (SLE) is a complex chronic autoimmune disease characterized by several immunological alterations. T cells have a peculiar role in SLE pathogenesis, moving from the bloodstream to the peripheral tissues, causing organ damage. This process is possible for their increased adherence and migration capacity mediated by adhesion molecules, such as CD44. Ten different variant isoforms of this molecule have been described, and two of them, CD44v3 and CD44v6 have been found to be increased on SLE T cells compared to healthy controls, being proposed as biomarkers of disease and disease activity. The process of alternative splicing of transcripts is not fully understood. We investigated the mRNA expression of and and also analyzed possible splicing regulators (ESRP1 molecule and rs9666607 polymorphism) in a cohort of SLE patients compared to healthy controls.

METHODS

This study involved 18 SLE patients and 18 healthy controls. Total RNA and DNA were extracted by peripheral blood mononuclear cells. The expression study was conducted by quantitative RT-polymerase chain reaction, using SYBR Green protocol. Genotyping of rs9666607 SNP was performed by direct sequencing.

RESULTS

mRNA expression was higher in SLE patients compared to healthy controls (p = 0.028). mRNA ratio in healthy controls was strongly unbalanced towards isoform v3 compared to SLE patients (p = 0.002) and decreased progressively from healthy controls to the SLE patients in remission and those with active disease (p = 0.015). The expression levels of and mRNA correlated with the disease duration (p = 0.038, Pearson r = 0.493 and p = 0.038, Pearson r = 0.495, respectively). Splicing regulator expression positively correlated with CD44v6 expression in healthy controls (p = 0.02, Pearson r = 0.532) but not in SLE patients. The variant A allele of rs9666607 of was associated with higher level of global mRNA (p = 0.04) but not with the variant isoforms.

CONCLUSIONS

In SLE patients, the increase in CD44v6 protein correlates with a higher transcript level of this isoform, confirming an impairment of splicing in the disease, whose regulatory mechanisms require further investigation.

摘要

背景

系统性红斑狼疮（SLE）是一种复杂的慢性自身免疫性疾病，其特征为多种免疫学改变。T 细胞在 SLE 的发病机制中具有特殊作用，它们可以从血液迁移到外周组织，导致器官损伤。这种迁移能力的增强可能是由黏附分子（如 CD44）介导的。该分子已被描述具有 10 种不同的变异异构体，其中两种，即 CD44v3 和 CD44v6，在 SLE T 细胞中较健康对照组升高，被提议作为疾病和疾病活动的生物标志物。然而，关于转录本的可变剪接过程尚未完全阐明。我们在一组 SLE 患者中研究了和的 mRNA 表达情况，并与健康对照组进行了比较，同时分析了可能的剪接调节因子（ESRP1 分子和 rs9666607 多态性）。

方法

本研究共纳入 18 例 SLE 患者和 18 例健康对照者。采用外周血单个核细胞提取总 RNA 和 DNA。采用 SYBR Green 法进行定量 RT-PCR 检测。采用直接测序法对 rs9666607SNP 进行基因分型。

结果

SLE 患者的 mRNA 表达高于健康对照组（p=0.028）。健康对照组中，与 SLE 患者相比，v3 异构体的 mRNA 比例明显失衡（p=0.002），并且从健康对照组到缓解期 SLE 患者和活动期 SLE 患者逐渐降低（p=0.015）。和 mRNA 的表达水平与疾病持续时间相关（p=0.038，Pearson r=0.493 和 p=0.038，Pearson r=0.495）。健康对照组中，ESRP1 表达与 CD44v6 表达呈正相关（p=0.02，Pearson r=0.532），而在 SLE 患者中则无相关性。基因 rs9666607 的 A 等位基因与的总 mRNA 水平升高相关（p=0.04），但与变异异构体无关。