Gordon综合征患者WNK基因突变研究  

Mutations of WNK kinases in patients with Gordon syndrome

作　　者：张翀[1] 秦岭[1] 王朝晖[1] 王伟铭[1] 任红[1] 张文[1] 严富洪[1] 谢静远[1] 陈楠[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院肾内科,上海200025

出　　处：《肾脏病与透析肾移植杂志》2009年第3期226-231,290,共7页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：上海市重点学科(T0201);上海市卫生局重点学科基金(05III001);国家自然科学基金(30670972)

摘　　要：目的:通过对Gordon综合征(PHA2)患者的WNK4和WNK1基因外显子扩增测序,研究我国PHA2患者WNK基因的突变特点。方法:通过查阅住院病史,收集不明原因高血压伴高血钾而肾功能正常的患者,进行临床和实验室检查,确诊PHA2。采集患病者及患病/非患病亲属的外周血,抽提DNA,设计17对引物扩增WNK4的19个外显子,27对引物扩增WNK1的28个外显子,纯化后送测序,测序结果进行序列比对。结果:发现PHA2患者4例,其中2例表现为家族遗传性,另2例为散发。先证者及所有患病家系成员共8人,年龄6～79岁,均表现为不明原因的高血压、高血钾,肾功能正常。8例中的5例伴有肾小管酸中毒,6例伴有高氯血症。DNA扩增后测序比对发现,其中1例散发患者为WNK4基因的杂合错义突变,位于17号外显子,造成氨基酸序列的改变,赖氨酸突变为谷氨酸(K1169E)。对其2个非患病女儿以及200个正常对照的WNK4基因检测均未发现该突变。其它2个患病家系及1个散发患者未发现WNK4基因突变,所有患者和健康者均未发现WNK1外显子突变。结论:PHA2是一种少见的单基因遗传疾病,漏误诊率很高。已报道的WNK4致病位点均位于螺旋-螺旋结构域附近。我们发现的K1169E为HGMD未收录的新突变,也位于螺旋-螺旋结构域,该位点氨基酸在不同物种高度保守,而且该位点氨基酸的变化伴有电荷的改变,可能导致蛋白质构象的改变。Objective：To investigate mutations of WNK kinases in patients with Gordon syndrome （pseudohypoaldosteronism type 2,PHA 2）. Methodology：Four pedigrees with PHA 2 were included in this study. The diagnose criteria of PHA 2 are one or more individuals in a family had an unknown origin of hypertension,elevated serum potassium levels,and normal glomerular filtration rate,with or without renal tubule acidosis and hyperchloraemia. We designed 17 and 27 pairs of primers respectively,for PCR-amplify 19 exons of WNK 4 gene and 28 exons of WNK1 gene in genomic DNA and then performed direct sequencing to analyze the PCR products. Results：Four patients were diagnoised as PHA 2 after searching the medical history records of hospitalized patients in the past 18 years and subsequent physical and biochemistry examination. 2 of them had a family history of hypertension and hyperpotassaemia. We found a new heterozygote missense mutation （3505A→G） K1169E in exon 17 in one sporadic patient with PHA 2. His father,who died many years ago,also had a doubtful history of hypertension. His 2 daughters haven＇t any clinical manifestations of PHA 2. This patient,but none of the unaffected members of this family and 200 unrelated unaffected control subjects,has a single base substitution that produces a K1169E substitution in WNK 4. Conclusion：We found a novel heterozygote missense mutation of WNK 4 gene in a Chinese patient with PHA 2. This charge-changing mutation lies in the coiled-coil domain of WNK 4,and this residue is also highly conserved among its known orthologs in many species,similar to those mutation loci ever reported.

关 键 词：Gordon综合征 突变 多聚酶链反应 测序 WNK4 WNK1 

分 类 号：R596[医药卫生—内科学]