149例中国脑性瘫痪儿童的致病基因研究

发布时间：2018-07-04 20:14

本文选题：脑性瘫痪 + KANK1　；参考：《佳木斯大学》2017年硕士论文

【摘要】：目的:检测我国先天性脑性瘫痪(简称脑瘫)儿童KANK1、GAD1和AP4复合物(包括AP4E1、AP4B1、AP4M1和AP4S1)基因突变的状况,初步探讨其基因型与临床表型的关系,探寻先天性脑瘫的病因,为脑瘫的预防及早期诊断探索新的方法。方法:选择在黑龙江省小儿脑性瘫痪防治疗育中心、河南中医药大学第一附属医院、安徽医科大学第一附属医院、深圳市儿童医院、西安中医脑病医院等院就诊的1-12岁先天性脑瘫儿童149例。对脑瘫儿童进行全面临床评估,详细收集病史等临床资料,采集患儿、父母和(或)其同胞的外周静脉血,提取DNA,运用高通量测序技术对KANK1、GAD1和AP4复合物基因进行DNA检测,并对发现的变异进行Sanger测序验证。结果:149例脑瘫男女比例为1.87:1(男97例,女52例),均为足月出生,无明显高危因素。脑瘫类型分布由高到低依次为痉挛型双瘫48例(32.21%),混合型27例(18.12%),痉挛型四肢瘫26例(17.45%),痉挛型偏瘫25例(16.78%),不随意运动型20例(13.42%),共济失调型3例(2.01%);其中痉挛型最多,共99例(66.44%)。149例脑瘫合并症前五位为:智力落后、语言障碍、小头畸形、视觉功能障碍和癫痫。经二代测序及Sanger验证,共发现6个致病性基因突变,涉及6例患儿(4%),受累基因为KANK1和AP4S1。携带基因突变的脑瘫儿童均为男性,均无脑瘫家族史。其中,2例合并智力落后、语言功能障碍、小头畸形的痉挛型双瘫的同胞患儿,携带KANK1基因的插入突变(c.1274_1275ins AGCTGT,p.427_428ins Ala Val);1例合并智力落后、双眼内斜视、语言障碍、小头畸形、行为异常和孤独症谱系障碍的痉挛型四肢瘫患儿,携带KANK1基因的错义突变(c.2167CT、p.Arg723Cys);1例无合并症的痉挛型偏瘫患儿,携带KANK1基因的错义突变(c.3482AG,p.Asn1161Ser);1例合并智力落后、语言发育迟缓和小头畸形的痉挛型四肢瘫患儿,携带KANK1基因的错义突变(c.2110GA,p.Asp704Asn);1例合并智力落后、语言障碍、小头畸形和癫痫的痉挛型四肢瘫患儿,携带KANK1基因的缺失突变(c.3103_3105del,p.1039del)和AP4S1基因的无义突变(c.289CT,p.Arg97Ter)。以上突变均来源于患儿父亲,AP4S1基因为常染色体隐性遗传,KANK1基因的遗传方式尚不明确。除AP4S1基因的无义突变有报道外,其它突变均为首次报道。携带致病基因突变的患者中伴发小头畸形(83%)的比例较未携带突变的患者(16%)高,差异具有统计学意义(P0.05)。结论:1.KANK1和AP4S1基因突变是中国脑瘫儿童的致病因素之一;2.KANK1和AP4S1基因突变可导致痉挛型脑瘫;3.KANK1和AP4S1基因突变所致脑瘫更易伴有小头畸形;4.脑瘫儿童的KANK1基因突变均来源于其父亲;5.KANK1基因突变的临床表型存在变异。
[Abstract]:Objective: to detect the gene mutations of KANK1T GAD1 and AP4 complex (including AP4E1AP-4B1A1AP4M1 and AP4S1) in children with congenital cerebral palsy (CP) in China, and to explore the relationship between genotypes and clinical phenotypes, and to explore the etiology of congenital cerebral palsy. To explore a new method for the prevention and early diagnosis of cerebral palsy. Methods: the first affiliated Hospital of Henan University of traditional Chinese Medicine, the first affiliated Hospital of Anhui Medical University, and the Children's Hospital of Shenzhen City were selected. 149 cases of congenital cerebral palsy children aged 1-12 years in Xi'an Encephalopathy Hospital. The clinical data of children with cerebral palsy were evaluated, the clinical data such as history were collected in detail, the peripheral venous blood of children, parents and / or their sibling were collected, the DNA was extracted, and the DNA of KANK1 + GAD1 and AP4 complex genes were detected by high-throughput sequencing technique. The mutation was sequenced by Sanger. Results the ratio of male to female in 149 cases of cerebral palsy was 1.87: 1 (97 males and 52 females). The distribution of cerebral palsy from high to low were spastic diplegia in 48 cases (32.21%), mixed type in 27 cases (18.12%), spastic tetraplegia in 26 cases (17.45%), spastic hemiplegia in 25 cases (16.78%), involuntary motor type in 20 cases (13.42%), ataxia type in 3 cases (2.01%). A total of 99 cases (66.44%). 149 cases of cerebral palsy complicated by the first five: mental retardation, language disorders, microcephaly, visual dysfunction and epilepsy. Six pathogenicity gene mutations were identified by second generation sequencing and Sanger analysis, involving 6 children (4%), involving KANK1 and AP4S1. All the children with cerebral palsy with gene mutation were male and had no family history of cerebral palsy. Among them, 2 children with mental retardation, language dysfunction, spastic diplegia with microcephaly, 1 case with mental retardation, binocular esotropia, language disorder, microcephaly, and microcephaly were accompanied by c.1274_1275ins AGCT GTP.427428ins Ala Val. In children with spastic quadriplegia with abnormal behavior and autism spectrum disorder, the missense mutation of KANK1 gene (c. 2167CTP. Arg723Cys) and the missense mutation of KANK1 gene (c. 3482AGp.Asn1161Ser) combined with mental retardation were found in one child with spastic hemiplegia without complication. Children with spastic quadriplegia with language retardation and microcephaly, one patient with kANK1 gene missense mutation (c.2110GAp.Asp704Asn), one child with mental retardation, language disorder, microcephaly and epilepsy spastic quadriplegia. The deletion mutation of KANK1 gene (c. 3105 delp. 1039del) and the nonsense mutation of AP4S1 gene (c. 289 CTP. Arg97Ter). All of the above mutations originated from the father of the child, AP4S1 because the genetic pattern of the autosomal recessive gene KANK1 was not clear. Except for the reported nonsense mutation of AP4 S1 gene, all other mutations were reported for the first time. The proportion of microcephaly with microcephaly (83%) was higher in patients with pathogenic gene mutation than that in patients without mutation (16%), the difference was statistically significant (P0.05). Conclusion 1. The mutation of KANK1 and AP4S1 gene is one of the pathogenic factors of cerebral palsy in Chinese children. Secondly, the mutation of KANK1 and AP4S1 gene may lead to spastic cerebral palsy. 3. The mutation of KANK1 and AP4S1 gene is more likely to cause cerebral palsy associated with microcephaly. KANK1 gene mutations in children with cerebral palsy all originated from the clinical phenotypic variation of KANK1 gene mutations.
【学位授予单位】：佳木斯大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742.3

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