Four novel ATP2C1 mutations in Chinese patients with Hailey–Hailey disease

Hailey–Hailey disease (HHD) is a kind of autosomal dominant dermatosis. The ATP2C1 gene has been identified as the pathogenic gene of HHD since 2000. In this study, direct DNA sequencing was used to identify ATP2C1 gene mutations in four Chinese families and two sporadic cases with HHD. The entire coding and flanking intronic sequences of ATP2C1 were screened for mutations and five heterozygous mutations of the ATP2C1 gene were detected in the four pedigrees and two sporadic cases with HHD. Four of them were novel, including three frame‐shift mutations (c.1330delC, c.888_889insT, c.478_479insA) and one nonsense mutation (c.1720C>T). These data added new variants to the database of ATP2C1 mutations associated with HHD.     

Hailey-Hailey病四个家系的ATP2C1基因突变分析

目的检测Hailey-Hailey病(HHD)4个家系的致病基因ATP2C1,鉴定其突变位点和突变类型。方法采集4个HHD家系成员共9例患者和6名正常人,与100名无关健康对照者外周静脉血各2 ml,提取全基因组DNA。运用聚合酶链反应(PCR)扩增ATP2C1基因的全部28个外显子及其侧翼内含子序列,扩增产物纯化后进行DNA直接测序,BLAST比对分析其突变位点和突变方式。结果在9例HHD患者中共检出了3个ATP2C1基因致病突变:c.888_889ins T(p.296Tfs X2)、c.1330del C(p.443Qfs X33)和c.2416CT(p.Arg806X)。在4个HHD家系的6名正常者和100名健康对照者中均未发现上述突变。结论在9个HHD家系患者中存在2个移码突变(c.888_889ins T和c.1330del C)及1个无义突变(c.2416CT),其中2个移码突变为首次报道。这些突变的发现有助于HHD的诊断,并丰富了HHD相关ATP2C1突变数据库。     

Four novel mutations in ATP2C1 found in Chinese patients with Hailey-Hailey disease

BACKGROUND: Familial benign chronic pemphigus or Hailey-Hailey disease (HHD; OMIM 169600) is an autosomal dominant blistering disease. Pathogenic mutations in ATP2C1 encoding a novel Ca2+ pump have recently been identified. OBJECTIVES: To identify mutations in ATP2C1 in Chinese patients with HHD. METHODS: Eleven unrelated Chinese patients with HHD were subjected to mutation detection in ATP2C1. Eight of them had a family history of HHD. The 27 coding exons and their flanking sequences were amplified and sequenced. RESULTS: Five of the 11 patients were identified to have heterozygous mutations including three nonsense mutations and two splicing mutations in ATP2C1. CONCLUSIONS: Four novel mutations, nonsense mutations S887X and W795X and splicing mutations 118-1 g-->a and 1890+1del(gtgag)ins53, were found in this series of Chinese patients with HHD.     

隐性营养不良型大疱性表皮松解症4例COL7A1基因突变分析

【摘要】 目的 分析4例隐性营养不良型大疱性表皮松解症（RDEB）患儿基因突变与临床表型情况。方法 收集4例RDEB患儿临床资料,提取患儿及其父母外周血DNA,使用先天性大疱性表皮松解症多基因芯片进行高通量测序,确定致病基因位点后,采用Sanger测序法进行双向验证。结果 4例患儿基因检测均显示COL7A1基因复合杂合突变,共8个突变位点。例1为中度泛发型RDEB,存在父源c.7828C>T和母源c.448G>A突变;例2为中度泛发型,存在父源c.3625_3635del和母源 c.2726_2728del突变;例3为局限型,存在父源突变c.682+1G>A和等位基因突变c.5532+1G>A,其父母外周血DNA未发现c.5532+1G>A突变;例4为重度泛发型,存在父源c.5196delC和母源c.500_501insAGGG突变。其中c.2726_2728del和c.5196delC突变既往未见报道。结论 4例RDEB患儿均存在COL7A1基因复合杂合突变,可能为其致病原因。其中,双等位基因移码突变携带者的表型重于双等位基因剪切位点、错义和无义、移码和氨基酸缺失及插入组成的复合杂合突变携带者表型。     

Heterogeneous mutations of the ATP2C1 gene causing Hailey–Hailey disease in Hong Kong Chinese

Background Hailey–Hailey disease (HHD) is a rare autosomal dominant dermatosis. It causes suprabasilar acantholysis leading to vesicular and crusted erosions affecting the flexures. Mutation of ATP2C1 gene encoding the human secretory pathway Ca²⁺/Mn²⁺‐ATPase (hSPCA1) was identified to be the cause of this entity. Objective The aim of this study was to study the mutational profile of the ATP2C1 gene in Hong Kong Chinese patients with HHD. Methods Patients with the clinical diagnosis of HHD proven by skin biopsy were included in this study. Mutation analysis was performed in 17 Hong Kong Chinese patients with HHD. Results Ten mutations in the ATP2C1 gene were found. Six of these were novel mutations. The novel mutations included a donor splice site mutation (IVS22+1G>A); a missense mutation (c.1049A>T); two deletion mutations (c.185_188delAGTT and c.923_925delAAG); an acceptor splice site mutation (IVS21‐1G>C) and an insertion mutation (c.2454dupT). Conclusion The six novel mutations provide additions to the HHD mutation database. No hot‐spot mutation was found and high allelic heterogeneity was demonstrated in the Hong Kong Chinese patients.     

Mutation analysis of ATP2C1 gene in Taiwanese patients with Hailey-Hailey disease

BACKGROUND: Hailey-Hailey disease (HHD) is an autosomal dominant disorder with recurrent eruption of vesicles and bullae involving predominantly the neck, groin and axillary regions. Histopathology shows suprabasal cleavage in epidermal cells. Recent studies have revealed that HHD is caused by mutations in the ATP2C1 gene encoding a novel Ca2+ pump. OBJECTIVES: To analyse the mutations of the ATP2C1 gene in Taiwanese patients with HHD. METHODS: In total, five familial and two sporadic cases of HHD were retrieved from the medical records. The diagnosis of HHD was made based on the characteristic clinical features and histopathological evidence. All 27 exons and flanking intron boundaries were amplified by polymerase chain reaction and products analysed by direct sequencing. RESULTS: We identified six novel mutations and one reported mutation: three deletion mutations (nt884-904del, 1459delCTCA, 1975delA), two non-sense mutations (R39X, R783X), one mis-sense mutation (A730T) and one splicing mutation (483 + 2T-->A). The non-sense mutation R39X had been reported previously; the other six mutations are novel mutations. CONCLUSIONS: These results demonstrate that a spectrum of ATP2C1 gene mutations is present in Taiwanese HHD patients.     

Mutations in the ATP2C1 gene in Chinese patients with Hailey-Hailey disease

Hailey-Hailey disease (HHD; MIM 16960) is a rare autosomal dominant hereditary disorder characterized by recurrent eruption of vesicles and bullae, predominantly involving the body folds. It is caused by heterozygous mutations in the ATP2C1 gene, encoding the human secretory pathway Ca2+/Mn2+-ATPase protein 1 (hSPCA1). When we studied Chinese patients with HHD, we found two different heterozygous mutations, Q506X and G353V, the former previously reported in a Hungarian patient, and the latter being a novel mutation. In a 38-year-old patient from a four-generation pedigree with a 3-year history of severe recurrent blisters, we identified a C-->T transition at nucleotide 1696, c(1696C-->T), in exon 17 of ATP2C1, resulting in a nonsenes mutation, Gln506X, which resulted in a premature termination codon. In the second patient, who represented a occurrence of sporadic Hailey-Hailey disease, a G-->T transversion of nucleotide, c(G1238T), in exon 13 of ATP2C1 was detected, which resulted in a Gly353-->Val amino acid substitution (G353V). Our molecular findings further demonstrate that the mutational events in the human ATP2C1 gene encoding the hSPCA1 pump play an important role in the pathogenesis of HHD.     

毛囊角化病一家系及三例散发患者ATP2A2基因突变研究

目的：对毛囊角化病（Darier's disease, DD）一家系及三例散发患者进行ATP2A2基因的突变分析。方法：收集先证者及其家系成员、散发病例的临床资料和外周血,采用PCR技术扩增ATP2A2基因所有编码区及侧翼序列,用Sanger法测序检测潜在的突变,选取与患者无亲缘关系的100例健康人作为对照,同时对已报道的ATP2A2基因突变进行文献回顾。结果：家系中三例患者均检测出ATP2A2基因第5号外显子c.380 G>A(p.G127D)新发错义突变;散发患者S1检测出第13号外显子C.1676G>A(p.R559Q)错义突变,散发患者S2检测出第14号外显子c. 2001C>T(p.D667D)同义突变,散发患者S3未检测出突变。结论：本研究中共发现三个突变,其中c.380G>A(p.G127D)在中国人群中首次报道,拓展了ATP2A2的基因突变谱。     

Detection and comparison of two types of ATP2C1 gene mutations in Chinese patients with Hailey–Hailey disease

The gene ATP2C1 is identified as the defective gene in Hailey–Hailey disease (HHD). The nonsense and missense are two common types of mutations and have ,respectively, been detected in many HHD patients. The aims of our study were to identify the pathogenic ATP2C1 abnormality in Chinese HHD patients, and to compare nonsense and missense mutations in vivo to provide further understanding of the molecular and the physiological basis of HHD. The nucleotide sequencing of the ATP2C1 gene was performed in HHD patients, unaffected family members and 100 unrelated individuals. Meanwhile, we detected and analyzed the clinical manifestations, the expression of ATP2C1 mRNA and hSPCA1 protein in the two types of mutations. Three heterozygous mutations were identified, including a previously reported nonsense mutation (R799X), two novel missense mutations (D644G) and (R417K). The results of comparisons between two types of mutations showed that the common clinical features, the similarly low-level expressions of ATP2C1 mRNA and hSPCA1 protein, but the ATP2C1 mRNA expression of nonsense mutation was lower than missense mutation and even less than half the level of normal people. Our findings expand the known spectrum of ATP2C1 mutations in HHD. We supported the haploinsufficiency theory as prevalent mechanism in both types of mutations, and believed that the differences of ATP2C1 mRNA expressions in peripheral blood may relate with the type of mutation and reflect the state of illness of patients.     

慢性家族性良性天疱疮五家系ATP2C1基因突变分析

目的：检测中国汉族慢性家族性良性天疱疮5家系ATP2C1基因突变情况。方法：提取5家系中12例患者、13名表型正常及100名正常对照外周血基因组DNA，经PCR扩增后进行DNA测序，并使用Chromas软件解析。结果：家系1中3例患者存在ATP2C1基因第18号外显子c.1738A>G（p.I580V）突变，家系2 中2例患者存在ATP2C1基因第25号外显子c.2416C>T(p.R806*)突变，家系3中3例患者存在ATP2C1基因第15号外显子c.1250G>A（p.R417K）突变，家系4和家系5 中ATP2C1基因未发现突变。上述家系内表型正常个体及100名正常对照中均未检测到相应突变。结论：ATP2C1基因突变可能在3例汉族HHD家系内发挥致病作用。     

Two novel mutations of the ATP2C1 gene in Chinese patients with Hailey–Hailey disease

Hailey–Hailey disease (HHD; OMIM 169600) is an autosomal dominant blistering disease. Pathogenic mutations in ATP2C1 encoding the human secretory pathway Ca²⁺/Mn²⁺-ATPase protein 1 (hSPCA1) have been identified since 2000. The aim of this study was to report a Chinese pedigree and a sporadic case of HHD and to explore the genetic mutations. The Chinese pedigree and the sporadic case of typical HHD were subjected to mutation detection of ATP2C1. The 27 coding exons and their flanking sequences were amplified and sequenced. The heterozygous C to T transition at nucleotide 2753 in exon 26 and G to T transition at nucleotide 2090 in exon 21 of the ATP2C1 gene were identified in a pedigree and a sporadic case of HHD, respectively. The C2753T transition resulted in a novel nonsense mutation of glutamine codon (CAG) to a stop codon (TAG) at amino acid residue 865 (Q865X) and the G2090T transition resulted in a novel missense mutation of glycine condon (GGA) to Valine (GUA) at amino acid residue 645 (G645V) in hSPCA1. This study should be useful for genetic counseling and prenatal diagnosis for affected families and in expanding the repertoire of ATP2C1 mutations underlying HHD. This work was supported by grant from the Natural Science Foundation of China (30471564).     

Mutation analysis of the ATP2C1 gene in Taiwanese patients with Hailey-Hailey disease

BackgroundHailey-Hailey disease (HHD) is an autosomal dominant disorder with recurrent pruritic vesicles and erosions, and scaly erythematous plaques, particularly involving intertriginous areas such as the neck, axillae, groins and perineum. Histopathology shows intraepidermal vesiculation with acantholysis in the suprabasal layer. It is caused by heterozygous mutations in the ATP2C1 gene, which encodes for the human secretory pathway Ca²⁺/Mn²⁺ ATPase 1. In this study, we analyze the mutations of the ATP2C1 gene in 26 Taiwanese patients with HHD.MethodsIn total, 21 familial cases from seven families and 5 sporadic cases (including 7 previously reported) were retrieved from the medical records. The diagnosis of HHD was made based on the characteristic clinical features and histopathological evidence. All 27 exons and flanking intron boundaries were amplified by polymerase chain reaction and the products were analyzed by direct sequencing.ResultsWe identified three nonsense mutations (R39X, R468X, R783X), two splice-site mutations (483 + 2t→a, 832G→A), four deletion mutations (nt884-904del, 1459delCTCA, 1874delA, 1975delA) and one missense mutation (A730T). Two unrelated families with nonsense mutation R783X had the comorbidity of chronic schizophrenia since the third decade.ConclusionsWe report two novel mutations (832G→A and 1874delA) of ATP2C1 involved in HHD. The nonsense mutation R783X might represent a mutational “hotspot” in the ATP2C1 gene. The present study demonstrates that a spectrum of ATP2C1 gene mutations is present in Taiwanese HHD patients.     

家族性良性天疱疮患者ATP2C1基因突变研究

目的 探讨3个家族性良性天疱疮家系和1例散发患者的ATP2C1基因突变。方法 采取家系中患病成员外周血,应用外周血细胞DNA抽提、PCR扩增和DNA直接测序等方法检测ATP2C1基因突变情况,用反向测序验证突变,用100例无血缘关系个体作正常人对照。结果 在2个家族性良性天疱疮家系和1例散发患者中发现3个未曾报道的错义突变。家系1第20外显子2048位碱基G→A,导致错义突变R619K;家系2第8外显子853位碱基A→C,导致错义突变T221P;散发患者第23外显子2323位碱基T→C,导致错义突变Y711H。家系中非患病成员和100例无血缘关系正常人均未发现这些改变。在1个家族性良性天疱疮家系未检测到基因突变。结论 发现家族性良性天疱疮3种新的ATP2C1基因突变位点。     

家族性慢性良性天疱疮2家系的基因突变分析

目的检测两个中国家族性慢性良性天疱疮(Hailey-Hailey disease,HHD)家系ATP2C1基因的致病性突变。方法收集两个中国HHD家系的临床资料和外周血标本,用基因组抽提试剂盒提取外周血DNA,用PCR反应扩增ATP2C1基因的所有外显子编码区及其侧翼序列,然后对扩增产物进行直接测序,并与100例正常对照组进行比较。结果发现两个新的杂合性突变,包括一个无义突变(p.Q633X)和一个移码突变(c.2164insACAT),这两个突变在家系正常成员和100例正常对照中没有发现。结论该结果表明这两个ATP2C1基因新突变可能导致中国汉族人HHD的发病,增加了ATP2C1基因突变数据库新的突变位点。     

慢性家族性良性天疱疮两家系ATP2C1基因突变分析

目的对2例慢性家族性良性天疱疮(HHD)家系ATP2C1基因中可能存在的突变进行鉴别。方法收集2个HHD家系和100份无亲缘关系正常人外周血标本,采用聚合酶链反应方法扩增ATP2C1基因的全部外显子并测序,结果和Genbank中相应序列进行比对。结果家系1中所有患者ATP2C1基因检测到第17号外显子存在一个新的无义突变c.T1431A(p.C477X);家系2中所有患者第24外显子发现一个已报道的移码突变c.2374delTTTG(791LfsX9)。结论两个家系中存在ATP2C1基因的变异,导致编码蛋白的结构和功能发生改变。     

Three severe cases of EBS Dowling-Meara caused by missense and frameshift mutations in the keratin 14 gene

We report three unrelated patients affected at birth with an unusually severe form of epidermolysis bullosa simplex Dowling-Meara type (EBS-DM) because of mutations in KRT14 encoding keratin 14. Two patients were heterozygous for the previously described p.M119T mutation. The third patient was heterozygous for a novel c.1246delC mutation predicting the replacement of the helix termination peptide and the tail domain by a 25 amino-acid aberrant carboxyterminal sequence. At age 2 years, patients carrying the p.M119T mutation still suffered from severe EBS-DM, whereas the patient harboring the c.1246delC mutation has improved over time. These cases illustrate genotype-phenotype correlations and have implications for genetic counselling of EBS.     

慢性家族性良性天疱疮发病机制的研究进展

Hailey-Hailey病(HHD),又称为慢性家族性良性天疱疮,是一种少见的常染色体显性遗传病,以表皮棘层松解为特征,具体发病机制尚不清楚。HHD由钙依赖性ATP酶基因遗传缺陷引起,该基因编码人类分泌途径钙离子转运ATP酶1型(SPCA1),在角质形成细胞内高度表达。本文就近年来有关HHD发病机制的研究进展进行综述,为今后该病相关研究提供理论基础。     

Genetic diagnosis in a Chinese Hailey–Hailey disease pedigree with novel ATP2C1 gene mutation

Hailey–Hailey disease (HHD) is an autosomal dominant skin disorder characterized by recurrent eruption of vesicles and bullae at the sites of friction and in the intertriginous areas. Mutations in the ATP2C1 gene encoding the human secretory pathway calcium ATPase 1 (hSPCA1) have been identified as the causative mutations in HHD. In this study, we used direct sequencing and restriction endonuclease digestion to analyze mutations of the ATP2C1 gene in a Chinese three-generation pedigree. A heterozygous T-to-C transition at nucleotide 1004 in exon 12 of ATP2C1 gene was detected. After summarizing the reported cases with ATP2C1 mutation, we concluded that the T1004C transition resulted in a novel missense mutation of leucine condon (CTG) to proline (CCG) at amino acid residue 335(L335P) in hSPCA1. Here, a genetic diagnosis was made for the proband’s daughter before the clinical presentation. The study realized the molecular diagnosis in the HHD pedigree. Our findings should be useful for genetic counseling and prenatal diagnosis for the affected family and in demonstrating the critical role of the ATP2C1 gene in the pathogenesis of HHD further.     

Novel mutation in ATP2C1 gene in a Japanese patient with Hailey-Hailey disease

Hailey-Hailey disease (HHD) is an autosomal dominant disorder with recurrent eruption of vesicles and bullae involving predominantly the neck, groin and axillary regions. Histopathology shows suprabasal cleavage in epidermal cells. Recent studies have revealed that HHD is caused by mutations in the ATP2C1 gene encoding a novel Ca(2+) pump. We analyzed mutations of the ATP2C1 gene in 2 Japanese patients with HHD. The diagnosis of HHD was made based on the characteristic clinical features and histopathological evidence. All 27 exons and flanking intron boundaries were amplified by polymerase chain reaction and products analyzed by sequencing. As a result, we identified a novel missense mutation (A1087G) in exon 13 of the ATP2C1 gene in a patient. This mutation led the amino acid change from Thrto Ala in the phosphorylation protein domain. Another patient showed no mutation of the gene. These results demonstrate that a spectrum of ATP2C1 gene mutations is present in Japanese HHD patients.     

Genetic diagnosis of Hailey–Hailey disease in two Chinese families: novel mutations in the ATP2C1 gene

Hailey–Hailey disease (HHD; OMIM 169600), is an autosomal dominantly inherited disorder characterized by suprabasal cell separation of the epidermis. Mutations in ATP2C1, which encodes the human secretory pathway Ca²⁺/ Mn² ± ATPase protein 1 (hSPCA1), have been identified as the pathogenic gene of HHD without evidence of genetic heterogeneity. In this study, the ATP2C1 gene was screened in two typical Chinese pedigrees with HHD, and two specific novel mutations of the ATP2CL gene were identified. Family 1 had a 16‐base deletion mutation c.1068–1083del16 and family 2 had a substitution mutation c.1982T>G (p.Met661Arg). DNA sequencing of the three descendants of the probands revealed that they all had the normal genotype, indicating that there had been no transmission of the mutation.