本文選題：發(fā)作性運動誘發(fā)性運動障礙　切入點：PKD基因突變　出處：《鄭州大學(xué)》2014年博士論文　論文類型：學(xué)位論文

【摘要】：背景：發(fā)作性運動誘發(fā)性運動障礙(Paroxysmal Kinesigenit Dyskinesia，PKD)又稱發(fā)作性運動誘發(fā)性舞蹈手足徐動癥，是以在靜止情況下，起始隨意動作如起立、轉(zhuǎn)身、邁步、舉手等誘發(fā)的短暫的運動異常為臨床特征的神經(jīng)系統(tǒng)發(fā)作性疾病。其診斷要點為：運動誘發(fā)；發(fā)作持續(xù)時間短（1分鐘）；發(fā)作不伴有意識喪失或疼痛；排除其他器質(zhì)性疾��；神經(jīng)系統(tǒng)檢查正常。最近研究顯示，PKD是一種遺傳因素為主要發(fā)病因素的神經(jīng)系統(tǒng)疾病，目前認為，有家族史的病人約占PKD發(fā)病總數(shù)的60%，通常為常染色體顯性遺傳，其他病人則為散發(fā)。長期以來PKD的致病基因不明，最新的分子遺傳學(xué)研究發(fā)現(xiàn)：PKD的主要致病基因是PRRT2基因。目前，關(guān)于PKD人群中PRRT2基因突變檢測的相關(guān)研究較少，PRRT2基因相關(guān)基因型與表型的關(guān)系尚不明確，為了明確本組PKD患者中PRRT2基因突變頻率，及明確攜帶PRRT2基因突變對PKD病人表型的影響，我們開展了下列研究。 目的: 1.明確本研究納入PKD患者的PRRT2基因突變頻率及相關(guān)PRRT2基因突變位點信息； 2.探討本組PKD患者中PRRT2基因相關(guān)基因型與表型的關(guān)系。 方法: 1.連續(xù)納入2011年12月至2013年1月在鄭州大學(xué)第一附屬醫(yī)院神經(jīng)內(nèi)科就診的運動誘發(fā)性發(fā)作性運動障礙病人,所有病人均完成相關(guān)檢查，并有兩名神經(jīng)內(nèi)科專科醫(yī)師詳細進行體格檢查，所有納入病人均同意登記臨床資料、抽血及定期隨訪。 2.所有患者行PRRT2外顯子區(qū)域基因檢測，明確本組PKD患者中PRRT2基因突變頻率和突變位點，并在正常對照中檢測發(fā)現(xiàn)的PRRT2基因突變是否存在。 3.對本研究納入患者隨訪半年，觀察其對抗癲癇藥物卡馬西平治療的反應(yīng)性，并比較PRRT2突變陽性患者和不攜帶PRRT基因突變PKD患者的表型差異。 結(jié)果: 1.本研究共納入5個家系共11例家族性PKD患者和19例無家族史的散發(fā)PKD患者，其中男性患者21例，女性患者9例，平均發(fā)病年齡14.33±3.92歲。 2.在本組患者中，我們發(fā)現(xiàn)在5個PKD家系中的4個家系，共9名患者中PRRT2基因的c.649dupC (p.P217fsX7)突變是這些家系的致病突變，在19例散發(fā)PKD患者中同樣發(fā)現(xiàn)PRRT2基因的c.649dupC (p.P217fsX7)突變導(dǎo)致了6例散發(fā)PKD患者致病，在1例散發(fā)患者中發(fā)現(xiàn)了一個PRRT2基因的同義突變c.1011C/T（p.Gly337Gly），在其余患者中未發(fā)現(xiàn)PRRT2基因突變。 3.在納入PRRT2基因型-表型關(guān)系研究的27例PKD患者中，攜帶PRRT2基因突變的PKD病人發(fā)病年齡更早(平均發(fā)病年齡:11.62vs.17.21,P=0.01)；發(fā)病癥狀多呈對稱性，但在PRRT2基因突變陰性組中全部為單側(cè)起�。≒＝0.001)；PRRT2基因突變組病人每日發(fā)作次數(shù)更多((P＝0.038)。在發(fā)作先兆方面，兩組患者無明顯差異。(P＝0.585)。6個月隨訪結(jié)束后，我們發(fā)現(xiàn)：卡馬西平治療對PRRT2基因突變陽性組患者全部有效（13/13,100%），但對PRRT2基因突變陰性組患者僅部分病人有效（5/14，35.7%）。 結(jié)論: 1. PRRT2基因突變是絕大多數(shù)家族性PKD患者及部分散發(fā)型PKD患者的致病原因，c.649dupC (p.P217fsX7)熱點突變占到所有突變的93.75%，報道了一種可能致病的PRRT2基因c.1011C/T（p.Gly337Gly）同義突變。 2. PRRT2基因突變陽性的PKD患者發(fā)病年齡較早，發(fā)作更為頻繁且發(fā)作時癥狀對稱性更高。本研究提示PRRT2基因突變陽性對抗癲癇藥物卡馬西平的反應(yīng)性具有預(yù)測作用，PRRT2基因檢測可能對PKD患者的臨床治療具有潛在指導(dǎo)意義。 背景：遺傳性共濟失調(diào)（Hereditary ataxia, HA）是一組以慢性進行性小腦萎縮及小腦性共濟失調(diào)為主要特征的神經(jīng)系統(tǒng)遺傳變性病，戈登-福爾摩斯氏綜合征（Gordon Holmes Syndrome）是遺傳性共濟失調(diào)類疾病的一種特殊亞型，遺傳方式為常染色隱性遺傳，臨床表現(xiàn)為青春期起病的，逐漸進展的步態(tài)不穩(wěn)，，眼震等共濟失調(diào)癥狀；病人同時合并性腺功能障礙，男性病人表現(xiàn)為外生殖器發(fā)育不良，性腺幼稚，不育，女性病人表現(xiàn)為青春期后仍無月經(jīng)來潮及第二性征發(fā)育不全，臨床上盡管在對Gordon Holmes綜合征已有近100年時間的認識，但世界上僅有20余個該疾病家系被報道，其致病基因長期以來未被鑒定和克隆。2013年，其第一、二種致病基因RNF216和OTUD4被報道。 目的: 我們在前期工作中，收集到中國地區(qū)的首個Gordon Holmes綜合征家系，本研究擬在前期收集本家系的基礎(chǔ)上，通過外顯子測序技術(shù)，探尋Gordon Holmes綜合征的致病基因。 方法: 1收集本研究家系內(nèi)所有成員的臨床資料，并有兩名神經(jīng)內(nèi)科專科醫(yī)師詳細進行體格檢查，所有家系成員均同意登記臨床資料、抽血及定期隨訪。 2應(yīng)用全外顯子組捕獲+高通量測序技術(shù)對本家系內(nèi)2例患者及家系內(nèi)1位正常對照者進行全外顯子組測序，對外顯子組測序結(jié)果進行后期數(shù)據(jù)分析，尋找候選致病基因位點。 3對外顯子組測序發(fā)現(xiàn)的候選基因致病位點進行Sanger測序驗證，并在相關(guān)病人及正常對照中進行驗證。 結(jié)果: 1本研究中的Gordon Holmes綜合征家系內(nèi)兩名女性病人表現(xiàn)為青春期出現(xiàn)的共濟失調(diào)伴小腦萎縮，性腺發(fā)育障礙，同時合并輕度的認知功能障礙。病人父母及一個弟弟表現(xiàn)正常，符合常染色體隱性遺傳規(guī)律。 2本研究中的三個全外顯子組測序樣本每個樣本均產(chǎn)生了8Gb左右的數(shù)據(jù)量，平均測序深度達到了90%以上，目標序列覆蓋度超過99%，通過對SNP及Indel的注釋，在本研究中的三個樣本中，每個樣本均發(fā)現(xiàn)了大約90000個SNP和2000個Indel。通過（1）數(shù)據(jù)庫過濾，（2）隱性遺傳模型分析，（3）IBD分析（，4）SNP致病性預(yù)測，我們發(fā)現(xiàn)STUB1基因的c.737C-T（p.T246M）突變是本家系最可能的致病位點。 3在正常人群、其他共濟失調(diào)及Gordon Holmes綜合征患者中進行驗證，未發(fā)現(xiàn)STUB1基因突變，提示Gordon Holmes綜合征的致病基因具有異質(zhì)性。 結(jié)論:STUB1基因的c.737C-T（p.T246M）突變是本家系的最可能的致病位點，結(jié)合后續(xù)開展的體外功能研究及轉(zhuǎn)基因動物研究結(jié)果，提示：STUB1基因是GordonHolmes綜合征的第三種致病基因
[Abstract]:Background: paroxysmal kinesigenic dyskinesia (Paroxysmal Kinesigenit, Dyskinesia, PKD) also known as paroxysmal kinesigenic choreoathetosis, which is in the stationary case, initial random actions such as stand up, turn around, move, nervous system movement induced by different short hands often clinical features of paroxysmal diseases. The main points of diagnosis: exercise induced; short duration episodes (1 minutes); the attack was not accompanied by loss of consciousness or pain; exclusion of other organic disease; neurological examination is normal. Recent studies show that PKD is a disease of the nervous system, as the main risk factors of a genetic factor is believed to have a family history of the PKD patients accounted for about 60% of the total incidence, usually autosomal dominant, other patients are sporadic. For a long time the pathogenic gene of PKD is unknown, molecular genetics, the new study found: mainly caused by PKD The disease gene is PRRT2 gene. At present, about PRRT2 in the population of PKD gene mutation detection research, the relationship between PRRT2 gene genotype and phenotype is not clear, in order to determine the PRRT2 of the patients with PKD gene mutation frequency, and clearly carrying PRRT2 gene mutation effect on the phenotype of PKD disease people, we carried out the following study.
Objective:
1. the PRRT2 gene mutation frequency and the related PRRT2 gene mutation sites of the PKD patients were included in this study.
2. to investigate the relationship between genotype and phenotype of PRRT2 gene in PKD patients.
Method:
1. consecutive December 2011 to January 2013 in the First Affiliated Hospital of Zhengzhou University Department of Neurology of motor evoked seizures in patients with dyskinesia, all patients completed related examination, and two neurological physicians with physical examination, all patients agreed to register the clinical data, blood and regular follow-up.
2. in all patients, the PRRT2 exon region gene detection was performed to identify the mutation frequency and mutation site of PRRT2 gene in PKD patients, and whether the PRRT2 gene mutation detected in normal controls existed.
3. the patients were included in the study for half a year to observe the reaction of C Masi Bing against epilepsy drugs. The phenotypic difference between PRRT2 mutation positive patients and PKD patients without PRRT gene mutation was observed.
Result:
1., a total of 5 families, 11 family PKD patients and 19 sporadic PKD patients without family history were included in the study, including 21 male patients and 9 female patients, with an average age of 14.33 + 3.92 years.
2. in this group of patients, we found 4 families in 5 PKD families, a total of 9 patients in the PRRT2 gene of c.649dupC (p.P217fsX7) mutations in these families in 19 sporadic mutation, PRRT2 gene c.649dupC was also found in PKD patients (p.P217fsX7) mutation in 6 cases patients with sporadic PKD disease, in 1 sporadic patients found a synonymous mutation in PRRT2 gene c.1011C/T (p.Gly337Gly), PRRT2 gene mutation was not found in the remaining patients.
3. in PRRT2 genotype - phenotype of 27 PKD patients, PKD patients with a younger age of PRRT2 gene mutation (average age: 11.62vs.17.21, P=0.01); the incidence of symptoms was symmetrical, but mutations in PRRT2 negative group were all unilateral onset (P = 0.001); PRRT2 gene mutation patients daily episodes more ((P = 0.038). At the onset of aura, no significant difference between the two groups. (P = 0.585) after.6 months follow-up, we found that C Masi Bing therapy on PRRT2 gene mutation positive group were all effective (13/13100%), but the PRRT2 gene mutation only some patients effective negative group (5/14,35.7%).
Conclusion:
1. PRRT2 gene mutation is the cause of most familial PKD patients and some sporadic PKD patients. C.649dupC (p.P217fsX7) hotspot mutation accounts for 93.75% of all mutations. A possible pathogenic PRRT2 gene c.1011C/T (p.Gly337Gly) synonymous mutation is reported.
The onset age of patients with PKD positive early 2. mutations in the PRRT2 gene, more frequent episodes and symptoms of higher symmetry. This study suggests a role in predicting the response of the PRRT2 mutation of antiepileptic drug C Masi Bing, detection of PRRT2 gene may be of clinical patients with PKD treatment has potential significance.
Background: hereditary ataxia (Hereditary ataxia HA) is a chronic progressive cerebellar atrophy and cerebellar ataxia as genetic neural degeneration disease system is the main feature of the Gordon - Holmes syndrome (Gordon Holmes Syndrome) is a kind of disease of hereditary ataxia is a special subtype, genetic mode autosomal recessive, clinical manifestations of adolescent onset, progressive ataxia, nystagmus and ataxia; patients with gonadal dysfunction, male patient presented with genital hypoplasia, gonadal immature, infertility, female patients showed no menstruation after puberty and sex characteristics hypoplasia, clinical although in the understanding of Gordon Holmes syndrome in nearly 100 years, but only 20 more than the world in the family have been reported. The pathogenic gene has not been identified And cloned.2013, first, second of its pathogenic genes, RNF216 and OTUD4, were reported.
Objective:
In our previous work, collected the first Gordon Holmes Chinese area syndrome family, this study based on the pedigree collected earlier on by exome sequencing technology, to explore the pathogenic gene Gordon Holmes syndrome.
Method:
1 the clinical data of all the members in the family were collected, and two neurologes were examined in detail. All the family members agreed to register clinical data, blood sampling and regular follow-up.
2 Application of whole exome capture and high-throughput sequencing of the family in 2 patients and families in 1 normal control subjects by whole exome sequencing, exon sequencing results of group data analysis, identify genetic loci of candidate pathogenicity.
3 the pathogenicity site of the candidate gene was confirmed by Sanger sequencing and verified in the related patients and normal controls.
Result:
1 in this study Gordon Holmes syndrome family in two female patients presenting to the onset of puberty ataxia with cerebellar atrophy, gonadal dysgenesis, and mild cognitive impairment patients. Parents and a brother showed normal, consistent with autosomal recessive inheritance.
2 in this study three whole exome sequencing samples each have about 8Gb the amount of data, the average sequencing depth reached more than 90%, the target sequence coverage of more than 99%, according to the SNP and Indel notes, three samples in this study in each sample were found. About 90000 SNP and 2000 Indel. (1) through the filtering database (2), analysis of the recessive genetic model, (3) IBD (4, SNP) prediction analysis of pathogenicity, we found that STUB1 c.737C-T gene (p.T246M) mutation is pathogenic sites most likely family.
3 in the normal population, other ataxia and Gordon Holmes syndrome patients were verified, and no STUB1 gene mutation was found, suggesting that the Gordon Holmes syndrome gene is heterogeneous.
Conclusion: STUB1 gene c.737C-T (p.T246M) is the most likely pathogenic mutation pedigree, combined with the results, to carry out follow-up and in vitro functional study of transgenic animal study showed that the STUB1 gene is third kinds of pathogenic gene of GordonHolmes syndrome

【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R741


本文編號：1581891