EIF2AK3基因相关Wolcott-Rallison综合征1例并文献复习

患儿，女，1个月29 d。因抽搐6 d、发现血糖增高5 d入院。血糖波动于正常或增高，糖化血红蛋白因过高无法检测，尿糖+~++++，空腹C肽0.19 ng/mL，胰岛素11.68 μIU/mL。遗传性内分泌疾病基因Panel（检测基因412个，包含已知糖尿病相关基因49个）高通量测序发现患儿EIF2AK3基因存在新发c.2731_2732delAG和c.2980G > A复合杂合突变，均位于基因的激酶结构域。该婴儿被确诊为Wolcott-Rallison综合征（WRS）。WRS是一种罕见的常染色体隐性遗传疾病，以新生儿糖尿病、多发性骨骺发育不良和肝脏疾病为特征，新生儿糖尿病是WRS诊断的必备条件，EIF2AK3基因是WRS的致病基因。     

先天性甲状腺功能减低症35例甲状腺过氧化物酶基因突变检测

目的 检测35例先天性甲状腺功能减低症(CH)患儿甲状腺过氧化物酶(TPO)基因突变.方法 抽取35例先天性甲状腺功能减低症患儿外周血并提取DNA,用PCR扩增患儿TPO基因所有17个外显子、外显子-内含子交界区以及3'端和5'端非翻译区,用DNA测序技术和限制性内切酶检测基因突变,并对发现突变的CH患儿父母进行对照分析.结果 5例CH患儿存在TPO基因突变:1例为c.961A＞G和c.2422delT突变复合杂合子,1例为c.2268insT和c.1477G＞A突变复合杂合子,3例为c.2268insT突变纯合子.其中c.961 A＞G[p.Thr321 Ala]为未见文献报道的突变.结论 在35例先天性甲状腺功能减低症检测到4种TPO基因突变.

Abstract：

Objective To identify thyroid peroxidase (TPO) gene mutations in 35 patients with congenital hypothyroidism. Method Genomic DNA was isolated from peripheral blood samples of 35 patients with congenital hypothyroidism. All of the 17 exons and flanking introns of TPO gene were amplified by PCR, then the PCR products were sequenced bi-directionally and were analyzed by restriction endonucleases. Result One patient had compound heterozygous mutations c. 961A ＞ G/c. 2422delT, one was c. 2268insT/c. 1477G ＞ A, and three was homozygous mutation c. 2268insT. The TPO gene mutation c.961A ＞ G [p. Thr321Ala] was one novel mutation. Conclusion High frequency mutation in TPO gene was detected in patients with congenital hypothyroidism.     

真核翻译始动因子2-a激酶3基因突变致Wolcott-Rallison综合征1例

目的 对Wolcott - Rallison综合征的临床特征及遗传发病机制进行研究.方法 选取临床诊断为Wolcott - Rallison综合征的患儿及其父母为研究对象,运用PCR技术扩增患儿家系真核翻译始动因子2-α激酶3(EIF2AK3)基因的17个外显子区,用DNA直接测序技术,对患儿家系EIF2AK3基因的17个外显子进行基因突变分析.结果 在患儿及其父亲EIF2AK3基因的第9外显子区发现了1个杂合突变(1798A/T),氨基酸序列分析显示这个突变是一种无义突变,可导致EIF2AK3第532位C氨基酸残基形成终止密码子( C532STOP),形成1个532氨基酸残基组成的截短蛋白.患儿母亲EIF2AK3基因未发现相应突变(1798T/T).结论 在中国儿童中,EIF2AK3基因突变可导致Wolcott - Rallison综合征发生.     

Whole exome sequencing analysis of 6 children with dilated cardiomyopathy北大核心CSCD

Objective To explore the mutational loci associated with the occurrence of dilated cardiomyopathy (DCM). Methods Six children with DCM and 3 healthy children were recruited through the " Children DCM Susceptibility Gene Research Project" for a prospective study from December 2019 to June 2021. Six patients were aged from 4 months to 14 years, including 5 girls and 1 boy. Three healthy children were aged between 3-13 years, including 2 girls and 1 boy. Whole exome sequencing was performed on the research subjects, and the pathogenic genes were identified by bioinformatics methods. At the same time, the venous blood of the first - degree relatives of the corresponding children was collected, and the region with gene mutations was subjected to next - generation sequencing. Results A total of 4 mutational loci that might be related to DCM were identified. Case 1 was found to have a c. 2011-3C > G mutation in the jounctophiilin - 2 (JPH2) gene. The c. 2011-3 C > G mutation was homozygous (GG) in the child, but heterozygous (CG) in the parents. This child also had a c. G49415A mutation in the titin (TTN) gene. This c. G49415A mutation was homozygous (AA) in the child, but heterozygous (GA) in the parents. Case 4 was found to have a c. G23033A mutation in the TTN gene. The c. G23033A mutation was heterozygous (GA) in both the subject and the father, but a wild type (GG) in the mother. Case 5 was found to have a c. 16975_16978del mutation in the TTN gene. The c. 16975_16978del mutation was heterozygous (TCTTC/T) in the child and the father, but a wild type (TCTTC/TCTTC) in the mother. Conclusions A total of 4 pathogenic gene loci related to the pathogenesis of DCM are identified in this study. The finding enriches the DCM disease gene spectrum and provides targets for the implementation of precision medicine. © 2022 ＣｈｉｎＪＡｐｐｌＣｌｉｎＰｅｄｉａｔ. All rights reserved.     

新生儿起病的鸟氨酸转氨甲酰基酶缺乏症一家系的临床与生化及基因分析

目的 通过对1例新生儿期急性起病的鸟氨酸转氨甲酰基酶缺乏症(OTCD)患儿家系从临床、生化和鸟氨酸转氨甲酰基酶基因(OTC)进行分析,初步探讨OTC在产前诊断中的意义.方法 分析患儿的临床、生化等特点,利用质谱技术检测患儿、患儿父母及第2胎新生儿血、尿氨基酸和有机酸代谢产物,同时应用聚合酶链反应与DNA测序的方法对患儿、患儿父母血样、第2胎胎儿羊水细胞及出生后血样进行OTC基因突变检测.结果 患儿出生后3 d起病.表现为反应差,喂养困难,新生儿感染,生化检测发现高血氨,轻度代谢性酸中毒,符合尿素循环代谢障碍的临床诊断标准.患儿于出生后第9天死亡.2次血质谱检测均显示瓜氨酸浓度明显下降,分别为0.86 μm和1.06μm,尿质谱有机酸检测发现乳清酸升高(124μm/M肌酐),并伴乳酸明显升高.父母及第2胎新生儿血尿瓜氨酸和乳清酸等均正常.基因分析显示患儿9号外显子存在无义突变(C.958 C＞T),使编码精氨酸的密码变成终止密码.其母亲为杂合携带者;在患儿和母亲的9号和5号内含子分别存在插入和杂合突变,分别为C.1005+132 InsT和C.542+134 G＞G/A;而父亲血样和第2胎胎儿羊水细胞及出生后血样DNA分析均正常.结论 OTC基因C.958 C＞T的突变可引起新生儿期急性起病.该突变如发生于男性患儿,出生后不久即可出现症状,起病迅速,如未及时抢救,极易导致死亡,而C.1005+132 InsT和C.542+134 G＞G/A是否与新生儿起病的OTCD有关,还需进一步的分析研究.OTC基因分析可应用于产前诊断.

Abstract：

Objective This study aimed at understanding clinical features, biochemistry and gene mutation in one Chinese pedigree which had a neonatal-onset ornithine transcarbamylase deficiency ( OTCD )boy, and exploring the significance of ornithine transcarbamylase analysis in prenatal diagnosis.Method The clinical and biochemical data of one case were analyzed.The amino acids in blood and organic acids in urine were analyzed by mass spectrum technology.The OTC gene mutation was detected using polymerase chain reaction (PCR) and DNA direct sequencing for the case, his parents and the fetus amniocyte and her blood after birth.Result The age of onset was 3 days after birth, he began to have poor reaction, difficulty to feed, high blood ammonia, infection, slight metabolic acidosis, which were consistent with the clinical diagnosis of urea cycle disorders.The boy died at the age of 9 days.Citrulline of blood was detected twice,and were 0.86 μm and 1.06 μm, respectively.The orotic acid was elevated ( 124 μm/M Creatinine), and urine lactic acid was significantly elevated.The citrulline and orotic acid in his parents and their second baby were normal in DBS and urine.One nonsense mutation in the OTC gene was found at the exon 9 ( C.958 C ＞ T) and his mother was the heterozygote, which caused an arginine to terminate the code at position 320 of the protein (R320X).Two other mutations were also detected at intron 9 ( C.1005 + 132 InsT) and intron 5 (C.542 + 134 G ＞ G/A).But the analysis of his father's DNA, the fetus amniocyte and her bloodwas normal.Conclusion The mutation of C.958 C ＞ T in OTC gene may occur during neonatal period.This mutation would result in a very severe symptom, even die suddenly several days after birth, if it was a boy.It needs more researches to discuss whether the C.1005 + 132 InsT in intron 9 and C.542 + 134 G ＞G/A in intron 5 were associated with the neonatal-onset OTCD.The DNA analysis of OTC gene could be utilized for the prenatal diagnosis.     

Rett综合征的临床特点及MECP2基因突变分析

目的 分析典型的Rett综合征患者的临床特点,并对患儿甲基化CpG结合蛋白-2（MECP2）基因进行突变分析。方法 使用PCR扩增和测序的方法 对近期诊断的9例RETT综合征患儿及其父母MECP2基因的3个外显子进行检测。结果 在9例患儿中发现5例存在MECP2基因的杂合突变,突变率超过50%,其中1例为碱基插入导致的移码突变（c.913insT）;其余4例为点突变,分别为位于外显子3的c.316C>T（R106W）,位于外显子4的c.502C>T（R168X）、c.808C>T（R270X）和c.1126C>T（P376S）,其中c.913insT为首次发现的突变,这些患儿父母均未检测到突变。2例患儿MECP2基因突变位于转录抑制区域,与其他患儿相比,这2个患儿语言功能几乎丧失,且发育明显落后。结论 该研究确诊的5例Rett综合征的患儿存在MECP2基因突变,且多数位于外显子4上;MECP2蛋白转录抑制区域突变可能影响患儿语言功能及发育明显落后。     

黏多糖贮积症Ⅰ型二例基因突变检测及产前诊断

目的 对黏多糖贮积症Ⅰ型病例进行基因检测,对再孕母亲进行产前诊断.方法 用酶学检测方法 确诊2例黏多糖贮积症Ⅰ型先证者;采用PCR扩增技术结合序列分析的方法,检测2例先证者血液及其再孕母亲羊水细胞中IDUA基因外显子及其两侧内含子.结果 在2例经酶学检测确诊的先证者中共检测出4种基因突变:p.L238R、c.883InsC、c.531InsT、p.L346R,2种为插入突变,2种为错义突变.在先证者1母亲羊水细胞IDUA基因中未发现致病基因突变,羊水细胞中IDUA酶活性为9.0 nmol/(h·mg蛋白);在先证者2母亲羊水细胞IDUA基因中检测到与先证者相同的两个致病突变,羊水细胞中IDUA酶活性为0.5 mol/(h·mg蛋白).结论 从黏多糖贮积症Ⅰ型先证者发现的4种突变中,p.L346R为已知突变,p.L238R,c.883InsC,c.531InsT为首次发现的新突变.胎儿1未获得与先证者相同致病基因,为正常胎儿.胎儿2获得与先证者相同致病基因,为受累胎儿.产前基因诊断结果 与酶学产前诊断结果 相符合.

Abstract：

Objectiye Mucopolysaccharidosis type Ⅰ(MPS Ⅰ; MIM# 252800)is an autosomal recessive disease that results from the deficiency in the lysosomal enzyme α-L-iduronidase(IDUA).IDUA is one of the enzymes involved in degradation of glycosaminoglycans heparan sulphate and dermatan sulphate.The deficiency of IDUA leads to widespread accumulation of partially degraded mucopolysaccharides inside lysosomes,resulting in progressive cellular and multiorgan dysfunction. Up to now there is no definitely effective treatment for this disorder,therefore it is important to provide an accurate genetic diagnosis and prenatal diagnosis for the MPS Ⅰ families.This study was conducted to detect IDUA gene mutation in patients with MPS Ⅰ and make a definite diagnosis of homozygote or heterozygote and make first trimester prenatal diagnosis.Method The 2 male probands included in this study were diagnosed as MPS Ⅰ patients in Peking Union Medical College Hospital,case 1 was 2 years old and case 2 was 5 years old.Genomic DNA was extracted from leucocytes in the 2 patients and 2 mothers' cultured amniocytes.IDUA gene DNA sequence was amplified by polymerase chain reaction(PCR)and the PCR products were sequenced directly.Novel mutations were analyzed in 100 normal chromosomes.Result The genotype of case 1 was p.L238R/c.883InsC,while of case 2 was c.531InsT/p.L346R.The fetal case 1 did not inherit the same pathogenic inherited the same pathogenic mutations with the proband,the genotype of fetal 2 was c.531InsT/p.L346R,in 2 MPS Ⅰ patients,p.L238R,c.883InsC,c.531InsT were novel.The fetal case 1 was diagnosed as normal fetus while the fetus 2 was diagnosed as affected.The results of the two kinds of prenatal diagnostic methods were correspondent with each other.     

X-连锁淋巴细胞异常增生症一例及其家系基因和蛋白表达研究

目的 探讨1例中国X-连锁淋巴细胞异常增生症(XLP)患儿及其家系的临床特征、基因突变和外周血单个核细胞(PBMC)SAP蛋白表达.方法 患儿男,6岁,于5岁时发现右腰部肿物,活检提示为伯基特淋巴瘤.其胞兄及表兄均于1岁左右因重症传染性单核细胞增多症夭折.据临床表现、家族史、免疫学特征拟诊为XLP.提取患儿及部分亲属基因组DNA,采用PCR法扩增SH2D1A基因,PCR产物直接进行双向序列测定,采用流式细胞仪检测PBMC中SAP蛋白表达.结果 患儿在缓解期EBV-DNA检测为536.9拷贝/ml(＞500拷贝/ml为EBV阳性),其SH2D1A基因第2外显子462位核苷酸发生无义突变,碱基C突变为T,形成TGA终止密码子(Arg55X),患儿母亲、姨母及外祖母为该突变携带者.患儿PBMC中SAP蛋白表达水平明显下降,而携带者SAP蛋白表达未见异常.结论 通过临床及实验室检查,确诊1例XLP患儿及家系.男性重症EBV感染,甚或无EB病毒感染证据,但具有家族史的淋巴瘤患儿应考虑XLP.SAP蛋白流式细胞仪检测为快速、准确的诊断手段.

Abstract：

Objective X-linked lymphopmliferative disease(XLP),a genetic disorder characterized by immunodeficiency to Epstein-Barr virus(EBV)infection,has been linked to mutations in the SH2D1 A gene.XLP patient displays EBV associated fulminant infectious mononucleosis or hemophagocytie lymphohistocytosis,hypogammaglobulinemia or malignant lymphoma.Here we report the clinical features.gene mutation and SAP expression on PBMCs of a Chinese patient with XLP and potential carriers.Method A 6 years old male patient and his maternal relatives were enrolled in this study.The patient was found to have with a renal Burkitt lymphoma on the right waist at 5 years of age by accident.His elder brother and a maternally related cousin botIl died of multiple systemic organ dysfunction syndrome (MODS)due to fulminant infectious mononucleosis(FIM)at the age of one year.The patient and his maternal relatives were subjected to detection of SAP expression on the PBMCs by flow cytometry and gene mutation analysis of SH2D1A by using PCR based on genomic DNA.Result The patient exhibited 536.9copy/ml level of circulating EBV-DNA during remission.Sequence analysis showed that the patient harbored a nonsense mutation in exon 2(C462T),resulting in a premature stop codon(Arg55X).His mother and some of the matemal relatives were proved to be carriers of this mutation.SAP expression from the patient was significantly reduced as compared to normal individual and the carriers.Conclusion We identified a Chinese XLP ease genetically.Assessment of SAP expression on PBMCs by flow cytometry seemed to be an effective rapid diagnostic method for this disease.Absence of EBV infeetion does not diminish the possibility of XLP.     

Wolcott-Rallison syndrome: a clinical and genetic study of three children, novel mutation in EIF2AK3 and a review of the literature

Background: Wolcott-Rallison syndrome is a rare autosomal recessive condition characterized by early infancy onset diabetes mellitus and multiple epiphyseal dysplasia. So far, 17 children have been described in the world literature. Recently, mutations in the gene encoding EIF2AK3 have been shown to segregate with the syndrome in three affected families. Aims: We aimed to describe the clinical characterization and mutation analysis of a further child, and full clinical and follow-up details on our first family including the longest surviving child. Methods: Retrospective case notes review of three children presenting to the diabetic unit at our institution; mutation analysis of the EIF2AK3 gene in our most recent patient; and review of the literature on Wolcott-Rallison syndrome. Results: Previously unreported phenotypic features in our patients included developmental regression after episodes of hepatic failure, and pachygyria on brain imaging. We have identified a novel 4-base pair deletion (nt 3021-3024 del GAGA) in exon 13, which results in a frameshift and premature stop codon (R908 F/S +22X), causing premature truncation of the protein and abolition of the carboxy- segment of the catalytic domain.

Conclusions: Wolcott-Rallison syndrome causes early-onset diabetes and acute hepatic failure, before epiphyseal dysplasia is manifest. We have identified a novel mutation in EIF2AK3, and prenatal diagnosis may now be offered to affected families.     

Primary hypothyroidism and nipple hypoplasia in a girl with Wolcott–Rallison syndrome

Wolcott–Rallison syndrome (WRS), caused by mutation in the EIF2AK3 gene encoding the PERK enzyme, is the most common cause of permanent neonatal diabetes mellitus (PNDM) in consanguineous families and isolated populations. Besides PNDM, it also includes skeletal abnormalities, liver and renal dysfunction, and other inconsistently present features. We present two siblings, who are WRS patients, and are Albanians from Kosovo born to unrelated parents. The older sister presented with PNDM, exocrine pancreatic insufficiency, short stature, microcephaly, normocytic anemia, delay in speech development, skeletal abnormalities, primary hypothyroidism, and hypoplastic nipples. Sequencing of the EIF2AK3 gene identified a homozygous mutation R902X in exon 13. The younger brother was diagnosed with PNDM and died from hepatic failure suggesting that he has been suffering from WRS as well. Including one previously reported patient from Kosovo carrying the same homozygous mutation, there are three WRS patients from this very small, ethnically homogenous region suggesting founder effect in this population. Conclusion: We postulate that thyroid hypoplasia with primary subclinical hypothyroidism already reported in two WRS patients and nipple hypoplasia could also be the phenotypic reflection of the mutation of pleiotropic EIF2AK3 gene in secretory cells.     

Wolcott–Rallison syndrome due to the same mutation (W522X) in EIF2AK3 in two unrelated families and review of the literature*

Ozbek MN, Senée V, Aydemir S, Kotan LD, Mungan NO, Yuksel B, Julier C, Topaloglu AK. Wolcott–Rallison syndrome due to the same mutation (W522X) in EIF2AK3 in two unrelated families and review of the literature. Wolcott‐Rallison syndrome (WRS) is a rare autosomal recessive disorder characterized by an early‐infancy‐onset diabetes mellitus associated with a variety of multisystemic clinical manifestations. Here, we present six patients with WRS, carrying the same homozygous mutation (EIF2AK3‐W522X), from two unrelated Turkish families. This is the largest series of patients with the same mutation for this rare syndrome. In this communication we compare clinical features of these six patients with the other 34 patients who have been reported to date, and review the clinical features of WRS. All WRS patients presented first with symptoms of insulin dependent diabetes mellitus, with a mean age at onset of 2 months. All patients had skeletal dysplasia or early signs of it, and growth retardation. Many of the patients with WRS have been reported to have developmental delay, mental retardation, and learning difficulties; in contrast, none of our patients showed abnormal development at age up to 30 months. Acute attacks of hepatic failure were reported in 23 cases out of 37 patients; in 15 of those 23 cases an acute attack of renal failure accompanied the liver failure. Exocrine pancreatic deficiency has been reported in only four cases other than our four patients. Central hypothyroidism was observed in six of 28 cases. We propose that central hypothyroidism is not a component of WRS, but rather a reflection of euthyroid sick syndrome. Four of our patients experienced severe neutropenia, compared to only five of the 27 other cases, suggesting that the W522X mutation may be specifically associated with neutropenia. Other than the consistent features of diabetes mellitus and epiphyseal dysplasia, WRS patients are otherwise characterized by extensive phenotypic variability that correlates poorly to genotype.     

Wolcott Rallison Syndrome: A Rare Inherited Diabetes Mellitus

A 9-y-old boy was referred to authors’ institute for the management of insulin dependent diabetes mellitus. He was the product of third degree consanguineous marriage and was delivered by full term vaginal delivery with a birth weight of 2.75 kg. At 3 mo of age, he had presented with diabetic ketoacidosis and was on insulin regimen. Patient had delayed milestones and short stature. On follow up, child developed limb deformity and was diagnosed to have skeletal dysplasia. At the age of 9 y, patient was diagnosed to have cirrhosis of liver. Genetic analysis revealed homozygous EIF2AK3 nonsense mutation. It confirmed the diagnosis of Wolcott-Rallison syndrome. Patient’s mother was heterozygous for the same mutation.     

Apert 综合征 1 例临床特征与基因类型分析

目的分析Apert综合征(AS)临床特征与基因类型。方法回顾1例AS患儿的临床资料及患儿和其父亲FGFR2基因测序结果,并复习相关文献。结果男性患儿,1岁1个月,扁头,突眼,眼距宽,耳位低,下颌小,高腭弓,无腭裂,双手五指并指并挛缩,双足五趾并趾。FGFR2基因外显子7 c.758CG,p.P253R杂合变异,父亲未检测到相关基因突变,支持Apert综合征诊断。文献检索到AS个案24例,22例明显颅面部畸形,1例轻微畸形,1例无畸形;均有手足并指/趾畸形。基因类型为S252W 13例,P253R 3例,Alu元件插入3例,基因缺失2例,杂合突变2例,序列变异1例。结论 AS患儿颅面部畸形及手足并指/趾明显,FGFR2基因以S252W、P253R突变为主。     

3日龄男婴呼吸困难合并内脏反位

患儿,男,生后3d,因气促2d、发绀1d入院.主要临床表现为生后早期出现的呼吸困难,合并内脏反位,最终确诊为Kartagener综合征.予以氧疗、抗感染、雾化等治疗后,患儿病情好转.基因检测结果提示患儿DNAH5基因存在1个大片段杂合缺失和1个半合子突变:exon 48_50杂合缺失,c.7915C>T(p.R2639...     

EIF2S3基因变异致MEHMO综合征2例报告并文献复习

目的 探讨MEHMO综合征的临床特征与遗传学特点.方法 回顾分析2例确诊MEHMO综合征患儿的临床资料,并复习相关文献.结果 两例患儿均为男性.例1于2岁3个月就诊,表现为癫痫发作、智力低下、进行性痉挛性四肢瘫、小头畸形、面部畸形、身材矮小和隐睾等.例2于3月龄就诊,4个月15天龄死亡,表现为反复低血糖、小头畸形和小阴...     

丙酮酸脱氢酶E1ɑ亚单位缺陷导致Leigh综合征

Leigh综合征是由于线粒体呼吸链能量代谢障碍所导致的遗传性疾病,丙酮酸脱氢酶E1ɑ亚单位（pyruvate dehydrogenase complex E1 alpha subunit, PDHA1）缺陷是导致Leigh综合征的常见原因之一。该研究通过PDHA1基因分析首次确诊了1例中国患者。该患儿1岁后运动发育落后,无力,肌张力低下,肌腱反射消失,发热、感冒时间歇性加重,智力发育正常。肌电图检查、腓肠肌病理活检结果提示神经性损害。3岁时脑MRI扫描未见异常,5岁时脑MRI呈现双侧苍白球对称性损害,符合Leigh综合征表现。经基因分析证实患者及其母亲PDHA1基因外显子3存在C214T突变,导致丙酮酸脱氢酶复合物活性下降。经过维生素B1、辅酶Q10、左旋肉碱及低碳水化合物饮食治疗后,患儿运动能力显著改善,现在8岁,正常就学。PDHA1缺陷为X连锁遗传性疾病,表型复杂,临床诊断困难,该患儿以无力为主要表现,经基因诊断确诊。［中国当代儿科杂志,2007,9（3）：216-219］