北方汉族人群中发作性运动诱发性运动障碍的临床及分子遗传学研究

发布时间：2018-04-20 03:06

本文选题：发作性运动诱发性运动障碍 + 基因突变　；参考：《天津医科大学》2015年硕士论文

【摘要】：目的发作性运动诱发性运动障碍(paroxysmal kinesigenic dyskinesia,PKD)是一种罕见的、突然发作的运动障碍性疾病。目前认为位于16号染色体上的富脯氨酸跨膜蛋白2(proline-rich transmembrane protein 2,PRRT2)基因可能是PKD的致病基因。因此,为了明确PRRT2基因突变与汉族人群PKD发生间的关系以及PRRT2基因突变与PKD患者临床表型的关联,我们开展了如下研究,旨在:1.了解PRRT2基因相关的生物学信息,包括基因结构、编码蛋白序列的结构以及该基因编码区单核苷酸多态性(single nucleotide polymorphisms,SNPs),为后续研究做准备;2.明确汉族PKD患者PRRT2基因突变频率以及PRRT2基因突变位点信息;3.探讨PKD患者中PRRT2基因突变与临床表型的关联。方法1.通过生物网络数据库查找PRRT2基因,并进行相关生物信息学分析,包括:查找该基因的基因组定位、结构信息以及该基因编码蛋白质序列的跨膜结构信息等。2.收集3个中国北方汉族PKD家系共12例患者及其26例无症状家属、10例散发PKD患者和30例正常对照,由两名神经内科医生对纳入本研究的所有患者进行详细检查,然后记录临床资料,并抽取其外周血。3.对纳入本研究的所有患者行PRRT2基因全外显子测序,来明确PRRT2基因突变是否为北方汉族人群PKD的致病基因,随后在正常对照中对相应位点进行验证。4.应用SWISS-MODEL对PKD患者和正常对照PRRT2基因编码的氨基酸序列进行二级结构的预测分析。5.对纳入本研究的患者随访半年,观察对给予卡马西平等抗癫痫药物治疗的效果,并采用统计学的方法比较携带PRRT2基因突变的患者(阳性组)和未携带PRRT2基因突变的患者(阴性组)临床表型的差异。结果1.PRRT2基因定位于16p11.2,基因全长3794bp,含有4个外显子,m RNA全长2606bp,CDS在m RNA的302~1324bp区域,编码341个氨基酸。PRRT2蛋白存在两个跨膜区域,分别在第269~289和318~338氨基酸之间。PRRT2基因编码区共存在42个非同义SNPs和19个同义SNPs。2.全外显子测序的结果发现,在6例家族性PKD患者和3例散发PKD患者中发现PRRT2基因第2号外显子发生杂合点突变412 CG(Pro138Ala),使编码的脯氨酸(Proine)变成丙氨酸(Valine);在6例家族性PKD患者和2例散发PKD患者中发现PRRT2基因第3号外显子发生杂合突变917 CA(Ala306Asp),使编码的丙氨酸(Valine)变成天冬氨酸(Aspartic acid),但在所有正常对照者中均未发现这两个基因突变位点。3.蛋白质结构预测分析结果显示PRRT2基因发生917 CA(Ala306Asp)突变后,其蛋白质二级结构未发生变化,但由于PRRT2基因编码的氨基酸序列相似度较低,未能给出412 CG(Pro138Ala)位点突变的预测结果。4.将纳入本研究的22例PKD患者分为PRRT2基因突变阳性组(发现突变)和基因突变阴性组(未发现突变),PRRT2基因突变阳性组PKD患者较基因突变阴性组PKD患者发病年龄更早(平均发病年龄:10.31 vs 16.44,P=0.027);临床症状是否对称和发病频率在两组中的差异具有统计学意义(P=0.007,P=0.027);在发作先兆方面,两组患者差异无统计学意义(P=0.054)。经过6个月随访后发现:卡马西平治疗对携带PRRT2基因突变的患者全部有效(11/11,100%),但对未携带PRRT2基因突变的患者仅部分有效(4/9,44.4%)。5.通过家系内比较,发现3个中国北方汉族PKD家系均有子代较父代发病年龄提前,病情加重的现象。结论1.在中国北方汉族人群中,PRRT2基因在家族性PKD患者中的突变频率相对较高。PRRT2基因的2个错义突变在纳入本研究的患者中被检测出来,因此412 CG(Pro138Ala)和917 CA(Ala306Asp)这两个错义突变可能是PKD的致病原因。2.携带PRRT2基因突变的患者较不携带PRRT2基因突变的患者发病年龄更早,发作时的症状更为对称,发作更为频繁。同时研究显示携带PRRT2基因突变的患者对卡马西平等抗癫痫药物治疗的反应性更好,因此PRRT2基因检测可能对PKD患者的临床治疗具有潜在的指导意义。3.中国北方汉族PKD家系中可能存在遗传早现现象。
[Abstract]:Objective paroxysmal kinesigenic dyskinesia (PKD) is a rare, sudden onset of dyskinesia. It is now considered that the proline rich transmembrane protein 2 (proline-rich transmembrane protein 2, PRRT2) gene located on chromosome 16 may be a pathogenic gene of PKD. Therefore, in order to clear PR The relationship between RT2 gene mutation and the occurrence of PKD in the Han population, and the association of PRRT2 gene mutations with the clinical phenotype of PKD patients. We have conducted the following studies: 1. to understand the biological information related to the PRRT2 gene, including the structure of the gene, the structure of the encoded protein sequence and the single nucleotide Po (single nucleotide PO). Lymorphisms, SNPs), prepare for the follow-up study; 2. clarify the frequency of PRRT2 mutation and the mutation site information of PRRT2 gene in the PKD patients of Han nationality; 3. explore the association between the mutation of the PRRT2 gene and the clinical phenotype in the PKD patients. Method 1. find the PRRT2 gene by the biological network database and carry out the related bioinformatics analysis, including the search of the gene. The genomic location, structural information and the transmembrane structure information of the gene encoding protein sequence were collected from 3 Chinese northern Han PKD families and 26 cases of asymptomatic families, 10 cases of PKD patients and 30 normal controls. Two neurosurgeon examined all the patients in this study, and then examined all the patients in this study. The clinical data were recorded and the peripheral blood.3. was extracted from all the patients in the study. The PRRT2 gene exon sequencing was performed to determine whether the PRRT2 gene mutation was the pathogenic gene of PKD in the northern Han population. Subsequently, the corresponding loci were verified in normal controls by the.4. application of SWISS-MODEL to the PKD patients and the normal control PRRT2 genes. The predictive analysis of the amino acid sequence of the two stage structure.5. was followed up for six months in the patients who were included in the study. The results were observed for the treatment of C Masi Bing and other antiepileptic drugs. Statistical methods were used to compare the differences in the clinical phenotype of patients (positive group) with PRRT2 gene mutation (positive group) and those who did not carry the PRRT2 gene mutation (negative group). Results the 1.PRRT2 gene was located in 16p11.2, with a full length of 3794bp, 4 exons, m RNA full length 2606bp, CDS in 302~1324bp region of M RNA, and two transmembrane regions encoding 341 amino acid.PRRT2 proteins. There were 42 unsynonymous and 19 synonyms in the gene coding region between the first 269~289 and the amino acids. Exon sequencing results found that the heterozygous mutation 412 CG (Pro138Ala) of the PRRT2 gene exon second was found in 6 familial PKD patients and 3 patients with PKD, and the encoded proline (Proine) was transformed into alanine (Valine), and the heterozygous exons of the PRRT2 gene were found in 6 familial PKD patients and 2 sporadic PKD patients. The mutation of 917 CA (Ala306Asp) changed the encoded alanine (Valine) into aspartic acid (Aspartic acid), but none of the two gene mutations found in all the normal controls showed that the protein structure of the PRRT2 gene was 917 CA (Ala306Asp) mutation, and the protein two structure did not change, but it was due to PRRT2. The gene encoded amino acid sequence was less similar and failed to predict the mutation of 412 CG (Pro138Ala) site..4. would be included in 22 PKD patients in this study into PRRT2 gene mutation positive group (discovery mutation) and gene mutation negative group (no mutation), and PRRT2 gene mutation positive group PKD patients were compared with PKD patients with gene mutation negative group. The age of the disease was earlier (average age of onset: 10.31 vs 16.44, P=0.027); the differences in the symmetry and frequency of clinical symptoms in the two groups were statistically significant (P=0.007, P=0.027); there was no statistical difference between the two groups (P=0.054) in the onset omen (P=0.054). After 6 months of follow-up, it was found that the C Masi Bing therapy had a mutation of the PRRT2 gene. The patients were all effective (11/11100%), but only partially effective (4/9,44.4%).5. of the patients who did not carry the PRRT2 gene mutation (4/9,44.4%) through family comparison, it was found that all of the PKD families of the Han nationality in the north of China had the earlier age of the parent generation and the aggravation of the disease. Conclusion 1. in the Han population of northern China, the PRRT2 gene is in the familial PKD patients. The 2 missense mutations of a relatively high mutation frequency of the.PRRT2 gene were detected in the patients enrolled in this study, so 412 CG (Pro138Ala) and 917 CA (Ala306Asp), two missense mutations, may be the cause of PKD in the pathogenesis of.2. with the PRRT2 gene mutation in the patients who are not carrying the PRRT2 gene mutation earlier, at the time of the onset of the attack. The symptoms are more symmetrical and more frequent. At the same time, the study shows that patients with PRRT2 gene mutations have better reactivity to C Masi Bing and other antiepileptic drugs, so PRRT2 gene detection may have a potential guiding significance for the clinical treatment of PKD patients.3. may exist in the PKD family of the Han nationality in Northern China.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R741

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