与家族性系统性红斑狼疮有关的PLD2基因新突变及其功能分析  

Identification and functional analysis of a novel phospholipase D2 gene mutation associated with familial systemic lupus erythematosus

作　　者：彭琳 袁新科 陈李笑 陈斯佳 陈科[2] PENG Lin;YUAN Xinke;CHEN Lixiao;CHEN Sijia;CHEN Ke(Department of Nephrology,First Hospital of Changsha,Changsha 410005;Department of Endocrinology,Third Xiangya Hospital,Central South University,Changsha 410013,China)

机构地区：[1]长沙市第一医院肾内科,长沙410005 [2]中南大学湘雅三医院内分泌科,长沙410013

出　　处：《中南大学学报（医学版）》2021年第3期234-239,共6页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(81870589);湖南省卫生健康委员会科研课题(B2017030,B2019135)。

摘　　要：目的:系统性红斑狼疮(systemic lupus erythematosus,SLE)是严重危害人类健康的自身免疫性结缔组织病,遗传因素在SLE的发病中起关键作用。本研究探讨与家族性SLE有关的磷脂酶D2(phospholipase D2,PLD2)基因新突变,并进一步探讨该突变致SLE的可能机制。方法:收集SLE患者、患者父母及147例正常对照的静脉血,抽提全血DNA。对患者及其父母行全基因组高通量测序,并对测序结果采用多种生物信息学方法进行分析。进一步构建野生型(wild type,wt)、突变型(mutant type,mu)及阴性对照PLD2质粒并转染293细胞,采用蛋白质印迹法检测各组293细胞中调控PLD2-Ras信号通路的关键基因HRAS的蛋白质表达水平。结果:在该SLE家系中,女性SLE患者及其母亲、Ⅱ及Ⅲ代各1例有典型的SLE临床表现,且均较早出现狼疮性肾炎,其遗传特点符合常染色体显性遗传。发现患者及其母亲新的PLD2杂合突变(c.2722C>T),且患者父亲及其他正常对照中未发现该突变。与转染wtPLD2质粒和阴性对照PLD2质粒比较,转染muPLD2质粒的293细胞中HRAS的蛋白质表达水平显著上调(均P<0.05)。结论:PLD2 c.2722C>T突变可能是导致该SLE家系发病的原因之一。Objective: Systemic lupus erythematosus(SLE) is a kind of autoimmune inflammatory connective tissue disease which seriously endangers human health. Genetic factors play a key role in the pathogenesis of SLE. This study aims to investigate a novel phospholipase D2(PLD2) mutation associated with familial SLE, and further explore the underlying mechanism of the mutation in SLE.Methods: The blood samples from a SLE patient, the patient’s parents, and 147 normal controls were collected and DNA was extracted. Whole genome high-throughput sequencing was performed in the patient and her parents and the results were further analyzed by various bioinformatics methods. The wild type(wt), mutant type(mu), and negative control PLD2 plasmids were further constructed and transfected into 293 cells.The expression level of HRAS protein in 293 cells was detected by Western blotting.Results: In this SLE family, the female SLE patient and her mother, 1 in generation Ⅱ and 1 in generation Ⅲ had typical clinical manifestations of SLE, and all of them had lupus nephritis at early stage. The genetic characteristics are consistent with autosomal dominant inheritance. A novel PLD2 heterozygous mutation(c. 2722 C>T) was found in the patient and her mother, but not in her father and other normal controls. Compared with wt PLD2 plasmid and negative control PLD2 plasmid, the expression of HRAS in 293 cells transfected with mu PLD2 plasmid was significantly up-regulated(both P<0.05).Conclusion: PLD2 c. 2722 C>T mutation may be one of the pathogeny of SLE in this family.

关 键 词：系统性红斑狼疮 PLD2基因 HRAS 全基因组高通量测序 PLD2-Ras信号通路 

分 类 号：R593.241[医药卫生—内科学]