良性成人家族性肌陣攣性癲癇的遺傳學(xué)特征及致病基因的定位
發(fā)布時(shí)間：2018-08-03 19:01
【摘要】：良性成人家族性肌陣攣癲癇(benign adult familial myoclonic epilepsy, BAFME)是一種常染色體顯性遺傳的，主要以四肢遠(yuǎn)端震顫、肌陣攣伴或不伴全面強(qiáng)直-陣攣發(fā)作的癲癇綜合征。至今，全世界約有100個(gè)家系在日本、意大利、荷蘭、法國(guó)、土耳其及中國(guó)等地被相繼發(fā)現(xiàn)并報(bào)道。但本病尚未被納入到2001年國(guó)際抗癲癇聯(lián)盟的癲癇綜合征的分類(lèi)當(dāng)中。 本病早期報(bào)道源于日本，日本學(xué)者Wakeno在1975年報(bào)道了一個(gè)以家族性震顫和癲癇發(fā)作為表現(xiàn)的家系，而后陸續(xù)報(bào)道了多個(gè)類(lèi)似家系，，并經(jīng)電生理研究確定其震顫均來(lái)源于大腦皮質(zhì)，且均為常染色體顯性遺傳。1991年Yasuda在報(bào)道兩個(gè)家系的基礎(chǔ)上，總結(jié)了本病具有良性病程和常染色體顯性遺傳的特點(diǎn)，首次提出了“良性成人家族性肌陣攣癲癇”的概念。在1999年，Mikami使用連鎖分析的方法將患有該病的一個(gè)3代27人的家系致病基因定位于8q23.3-24.1，從而發(fā)現(xiàn)了本病的第一個(gè)致病基因位點(diǎn)。2003年，Strianon對(duì)一個(gè)意大利的BAFME家系進(jìn)行基因連鎖分析后發(fā)現(xiàn)，其致病基因與2p11-q12.2具有連鎖關(guān)系，又發(fā)現(xiàn)了一個(gè)新的致病基因位點(diǎn)，從而說(shuō)明了該病的遺傳異質(zhì)性。2006年，我國(guó)鄧飛燕等人通過(guò)對(duì)一BAFME大家系的遺傳學(xué)分析，將致病基因定位于10p15；2010年Depienne等人將一法國(guó)FCMTE家系致病基因定位于5p15.31-p15；2012年Yeetong將一泰國(guó)BAFME家系致病基因定位在3q26.32-3q28。然而，目前為止尚未克隆出該病的致病基因。僅有日本學(xué)者Atsushi等人在2003年提出了CSMD3可能為BFAME的致病候選基因。CSMD3是一種編碼跨膜蛋白的較大的基因，位于人類(lèi)染色體8q23.3-q24.1區(qū)段，即BAFME被定位的區(qū)段。該基因包含73個(gè)外顯子，長(zhǎng)度跨越1.2Mb。主要在成人及胎兒的腦組織中表達(dá)。 自1991年Yasuda首次提出了BAFME的概念以來(lái)，BAFME綜合征的研究不超過(guò)20余年歷史，其中大多為病例報(bào)道，僅有屈指可數(shù)的幾個(gè)遺傳學(xué)研究報(bào)道，雖取得了一些研究成果，但是尚缺乏系統(tǒng)的對(duì)BAFME致病基因研究報(bào)道。尤其我國(guó)對(duì)BAFME家系很少有研究報(bào)道，對(duì)多代多人發(fā)病的大家系報(bào)道更是少見(jiàn)。我國(guó)作為人口大國(guó)，家系患病人群數(shù)量也相對(duì)較多，而B(niǎo)AFME的基因突變特點(diǎn)卻不明確，因此對(duì)于中國(guó)BAFME家系的調(diào)查研究有很大意義。而且積極開(kāi)展本病的臨床及分子遺傳學(xué)研究有助于提高臨床醫(yī)生對(duì)本病的認(rèn)識(shí)、了解本病及其他特發(fā)性癲癇的分子發(fā)病機(jī)制，并為本病的診斷、治療及遺傳咨詢提供基礎(chǔ)。 課題組調(diào)查的為遼寧省沈陽(yáng)市的一個(gè)家系，對(duì)家系調(diào)查分析后，確診為良性成人家族性肌陣攣性癲癇。課題組與該家系成員進(jìn)行溝通并講述實(shí)驗(yàn)研究的目的以及意義，在其簽署知情同意書(shū)后，對(duì)其進(jìn)行遺傳學(xué)調(diào)查、病史采集、臨床資料搜集、輔助檢查及采集外周血。該家系共4代43人，患者9人，其中男患2人，女患7人�，F(xiàn)存4代40人，男女患病機(jī)會(huì)均等，除第4代尚未到發(fā)病年齡外，其余每代均有發(fā)病患者，符合常染色體顯性遺傳特點(diǎn)�；颊甙l(fā)病年齡多在30～40歲左右，呈良性病程。所有發(fā)病患者的臨床表現(xiàn)都比較相似，大多以四肢遠(yuǎn)端震顫為首發(fā)癥狀，隨后或幾年后出現(xiàn)全身性強(qiáng)直-陣攣發(fā)作，服用抗癲癇藥物對(duì)緩解癥狀有效，β受體阻滯劑或飲酒無(wú)效，可除外原發(fā)性震顫。該家系是一個(gè)遺傳關(guān)系很明確，發(fā)病人數(shù)較多的呈常染色體顯性遺傳的BAFME大家系。 采集該家系成員及部分成員配偶的外周血，提取白細(xì)胞DNA進(jìn)行致病基因定位研究。實(shí)驗(yàn)思路為首先采用聚合酶鏈?zhǔn)椒磻?yīng)（PCR）及PCR產(chǎn)物測(cè)序法對(duì)家系中先證者進(jìn)行BAFME可疑致病基因CSMD3的突變檢測(cè)；如果結(jié)果為陰性，則對(duì)已知的BAFME基因進(jìn)行STR熒光標(biāo)記物連鎖分析，明確是否與目前已知的幾個(gè)染色體區(qū)段連鎖，盡可能將該家系的致病基因定位到某個(gè)染色體區(qū)段；如果結(jié)果仍為陰性，與現(xiàn)已知的基因位點(diǎn)均不連鎖，致病基因不在幾個(gè)候選染色體區(qū)段，則可以選擇全基因組掃描定位法。將致病基因定位在某染色體的特定區(qū)域后，進(jìn)行候選基因篩選，選擇該區(qū)域內(nèi)與癲癇有關(guān)的基因進(jìn)行突變檢測(cè)，從而明確該家系的致病基因。 首先，應(yīng)用PCR產(chǎn)物測(cè)序分析法對(duì)先證者CSMD3基因的73個(gè)外顯子進(jìn)行PCR擴(kuò)增產(chǎn)物測(cè)序，測(cè)序結(jié)果與GenBank人類(lèi)CSMD3gDNA序列進(jìn)行比較，未發(fā)現(xiàn)任何DNA序列變異，既沒(méi)有發(fā)現(xiàn)多態(tài)也沒(méi)有發(fā)現(xiàn)與疾病相關(guān)的突變，說(shuō)明本家系不存在CSMD3基因突變。而后，對(duì)目前已報(bào)道的5個(gè)染色體區(qū)段進(jìn)行連鎖分析，結(jié)果顯示染色體8q23.3-q24.1、2p11.1-q12.2、3q26.32-3q28、10p15區(qū)段的多個(gè)STR標(biāo)記連鎖分析結(jié)果顯示不支持連鎖，因此考慮本家系致病基因并不在上述4個(gè)染色體區(qū)段。而針對(duì)5p15.31-p15首先選擇了4個(gè)STR標(biāo)記，連鎖分析結(jié)果表明現(xiàn)不能完全肯定與否定致病基因是否位于該區(qū)段，之后，為進(jìn)一步明確結(jié)果，我們?cè)谠搮^(qū)段增加了8個(gè)STR位點(diǎn)，結(jié)果顯示，D5S486在θ=0.0時(shí)，LOD值為2.8，實(shí)驗(yàn)結(jié)果支持該家系致病基因與該位點(diǎn)的連鎖,考慮致病基因在5p15.31-p15染色體區(qū)段。 本實(shí)驗(yàn)研究了一良性成人家族性肌陣攣性癲癇的臨床癥狀、遺傳學(xué)特點(diǎn)并定位了致病基因位點(diǎn)。為良性成人家族性肌陣攣性癲癇的研究提供了經(jīng)驗(yàn)，也為該病的遺傳學(xué)研究提供了思路,并首次將國(guó)內(nèi)的良性成人家族性肌陣攣性癲癇家系致病基因定位在染色體5p15.31-p15區(qū)段。
[Abstract]:Benign adult familial myoclonus epilepsy (benign adult familial myoclonic epilepsy, BAFME) is a kind of autosomal dominant hereditary, mainly with distal tremor, myoclonus, or no total tonic clonic seizure syndrome. Up to now, about 100 families in the world are in Japan, Italy, Holland, France, Turkey and China. The disease has not been included in the classification of epilepsy syndrome of the International Antiepileptic Union in 2001.
The early report of this disease originated in Japan. In 1975, Japanese scholar Wakeno reported a family with familial tremor and epileptic hair. After that, several similar families were reported, and their tremors were all derived from the cerebral cortex by electrophysiological study, and all two families were reported to be the dominant hereditary.1991 year Yasuda. On the basis of this, the concept of "benign adult familial myoclonus epilepsy" was first proposed. In 1999, Mikami used the method of linkage analysis to locate the pathogenic gene of a family of 3 generation and 27 people with the disease in 8q23.3-24.1, and found the first one of the disease. The genetic linkage analysis of a Italy BAFME family in.2003 was found by Strianon. The pathogenic gene was linked to 2p11-q12.2, and a new gene locus was found. The genetic heterogeneity of the disease was explained in.2006 years. Deng Feiyan and others in our country passed the genetics of a BAFME family. Analysis, the pathogenic gene was located in 10p15; in 2010, Depienne et al. Located a French FCMTE family gene in 5p15.31-p15; in 2012 Yeetong, a BAFME family in Thailand was located in 3q26.32-3q28., but so far, the pathogeny gene of the disease has not been cloned. Only Japanese scholar Atsushi et al. In 2003 3 the possible pathogenetic candidate gene for BFAME,.CSMD3, is a large gene encoding the transmembrane protein, located in the 8q23.3-q24.1 section of the human chromosome, the section of the BAFME that is located. The gene contains 73 exons, and the length of the 1.2Mb. is mainly expressed in the brain tissue of the adult and the fetus.
Since Yasuda first proposed the concept of BAFME in 1991, the study of BAFME syndrome is not more than 20 years old, most of which are cases reported, only a few of the few genetic studies have been reported. Although some research results have been obtained, there is no systematic report on the study of BAFME pathogenic genes. Especially in China, there are few BAFME families in China. It is more rare to report on the incidence of the onset of multiple generation of people. As a country with a large population, the number of families with families is relatively large, but the genetic mutation characteristics of BAFME are not clear. Therefore, it is of great significance for the investigation and study of the Chinese BAFME family. To improve the understanding of the disease by clinicians and to understand the molecular pathogenesis of the disease and other idiopathic epilepsy, and to provide the basis for the diagnosis, treatment and genetic counseling of this disease.
The subject group investigated a family in Shenyang city of Liaoning province. After the investigation and analysis of the family, the group was diagnosed as familial myoclonic epilepsy of benign adults. The group communicated with the members of the family and explained the purpose and significance of the experimental study. After signing the informed consent book, the group had a genetic investigation, a history collection, and a clinical data search. There were 4 generations and 43 patients and 9 patients, including 2 men and 7 women, 4 generation 40, equal opportunities for both men and women, except for fourth generations that had not reached the age of onset, and the rest of the generation had the characteristics of autosomal dominant inheritance. The age of the patients was about 30~40 years old and had a benign course. The clinical manifestations of all the patients were similar, mostly with distal tremor of the extremities as the first symptom, followed by a generalized tonic clonic seizure later or a few years later, taking antiepileptic drugs to relieve the symptoms, the beta blocker or alcohol ineffective, except for the primary tremor. The family is a clear genetic relationship and the number of patients. A large number of autosomal dominant BAFME families.
The peripheral blood of the spouses of the family members and some members of the family was collected to extract the leukocyte DNA for the location of the pathogenic gene. The experiment was to detect the mutation of the suspected BAFME gene CSMD3 by polymerase chain reaction (PCR) and the sequencing of PCR products. If the result was negative, the known BAFME gene was found. A linkage analysis of STR fluorescent markers is carried out to determine whether it is possible to locate the family's pathogenic gene to a certain chromosome segment as far as possible, and if the result is still negative, it is not linked to the known gene loci, and the pathogenic gene is not in several candidate chromosome segments, then the whole gene can be selected. After locating the pathogenic gene in a specific region of a chromosome, the candidate gene was screened and the genes related to epilepsy in the region were selected for mutation detection so as to identify the pathogenic genes of the family.
First, the PCR product sequencing analysis was used to sequence the PCR amplification products of the 73 exons of the CSMD3 gene of the precursor. The sequencing results were compared with the CSMD3gDNA sequence of the GenBank human, and no mutation of the DNA sequence was found. Neither the polymorphism nor the disease related mutations were found, which showed that the family did not have the CSMD3 gene mutation. After the linkage analysis of the 5 reported chromosomal segments, the results showed that the results of multiple STR marker linkage analysis in the chromosome 8q23.3-q24.1,2p11.1-q12.2,3q26.32-3q28,10p15 section showed that the linkage was not supported. Therefore, the pathogenic genes in the family line were not in the 4 chromophore segments, and 4 were selected for 5p15.31-p15. STR markers, the result of linkage analysis showed that it was not fully affirmed and denying that the pathogenic gene was located in the area, and then, to further clarify the results, we added 8 STR loci to the section. The results showed that the LOD value was 2.8 when D5S486 was at theta =0.0. The experimental results supported the linkage of the pathogenic gene of the family with the site, considering the pathogenic gene in 5. P15.31-p15 chromosome section.
This study studied the clinical symptoms, genetic characteristics and loci of a benign adult familial myoclonic epilepsy, providing experience for the study of familial myoclonic epilepsy in benign adults, and for the genetic study of the disease, and for the first time the family myoclonic epilepsy home in China. The pathogenic gene was located in chromosome 5p15.31-p15 region.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R742.1
本文編號(hào)：2162705
[Abstract]:Benign adult familial myoclonus epilepsy (benign adult familial myoclonic epilepsy, BAFME) is a kind of autosomal dominant hereditary, mainly with distal tremor, myoclonus, or no total tonic clonic seizure syndrome. Up to now, about 100 families in the world are in Japan, Italy, Holland, France, Turkey and China. The disease has not been included in the classification of epilepsy syndrome of the International Antiepileptic Union in 2001.
The early report of this disease originated in Japan. In 1975, Japanese scholar Wakeno reported a family with familial tremor and epileptic hair. After that, several similar families were reported, and their tremors were all derived from the cerebral cortex by electrophysiological study, and all two families were reported to be the dominant hereditary.1991 year Yasuda. On the basis of this, the concept of "benign adult familial myoclonus epilepsy" was first proposed. In 1999, Mikami used the method of linkage analysis to locate the pathogenic gene of a family of 3 generation and 27 people with the disease in 8q23.3-24.1, and found the first one of the disease. The genetic linkage analysis of a Italy BAFME family in.2003 was found by Strianon. The pathogenic gene was linked to 2p11-q12.2, and a new gene locus was found. The genetic heterogeneity of the disease was explained in.2006 years. Deng Feiyan and others in our country passed the genetics of a BAFME family. Analysis, the pathogenic gene was located in 10p15; in 2010, Depienne et al. Located a French FCMTE family gene in 5p15.31-p15; in 2012 Yeetong, a BAFME family in Thailand was located in 3q26.32-3q28., but so far, the pathogeny gene of the disease has not been cloned. Only Japanese scholar Atsushi et al. In 2003 3 the possible pathogenetic candidate gene for BFAME,.CSMD3, is a large gene encoding the transmembrane protein, located in the 8q23.3-q24.1 section of the human chromosome, the section of the BAFME that is located. The gene contains 73 exons, and the length of the 1.2Mb. is mainly expressed in the brain tissue of the adult and the fetus.
Since Yasuda first proposed the concept of BAFME in 1991, the study of BAFME syndrome is not more than 20 years old, most of which are cases reported, only a few of the few genetic studies have been reported. Although some research results have been obtained, there is no systematic report on the study of BAFME pathogenic genes. Especially in China, there are few BAFME families in China. It is more rare to report on the incidence of the onset of multiple generation of people. As a country with a large population, the number of families with families is relatively large, but the genetic mutation characteristics of BAFME are not clear. Therefore, it is of great significance for the investigation and study of the Chinese BAFME family. To improve the understanding of the disease by clinicians and to understand the molecular pathogenesis of the disease and other idiopathic epilepsy, and to provide the basis for the diagnosis, treatment and genetic counseling of this disease.
The subject group investigated a family in Shenyang city of Liaoning province. After the investigation and analysis of the family, the group was diagnosed as familial myoclonic epilepsy of benign adults. The group communicated with the members of the family and explained the purpose and significance of the experimental study. After signing the informed consent book, the group had a genetic investigation, a history collection, and a clinical data search. There were 4 generations and 43 patients and 9 patients, including 2 men and 7 women, 4 generation 40, equal opportunities for both men and women, except for fourth generations that had not reached the age of onset, and the rest of the generation had the characteristics of autosomal dominant inheritance. The age of the patients was about 30~40 years old and had a benign course. The clinical manifestations of all the patients were similar, mostly with distal tremor of the extremities as the first symptom, followed by a generalized tonic clonic seizure later or a few years later, taking antiepileptic drugs to relieve the symptoms, the beta blocker or alcohol ineffective, except for the primary tremor. The family is a clear genetic relationship and the number of patients. A large number of autosomal dominant BAFME families.
The peripheral blood of the spouses of the family members and some members of the family was collected to extract the leukocyte DNA for the location of the pathogenic gene. The experiment was to detect the mutation of the suspected BAFME gene CSMD3 by polymerase chain reaction (PCR) and the sequencing of PCR products. If the result was negative, the known BAFME gene was found. A linkage analysis of STR fluorescent markers is carried out to determine whether it is possible to locate the family's pathogenic gene to a certain chromosome segment as far as possible, and if the result is still negative, it is not linked to the known gene loci, and the pathogenic gene is not in several candidate chromosome segments, then the whole gene can be selected. After locating the pathogenic gene in a specific region of a chromosome, the candidate gene was screened and the genes related to epilepsy in the region were selected for mutation detection so as to identify the pathogenic genes of the family.
First, the PCR product sequencing analysis was used to sequence the PCR amplification products of the 73 exons of the CSMD3 gene of the precursor. The sequencing results were compared with the CSMD3gDNA sequence of the GenBank human, and no mutation of the DNA sequence was found. Neither the polymorphism nor the disease related mutations were found, which showed that the family did not have the CSMD3 gene mutation. After the linkage analysis of the 5 reported chromosomal segments, the results showed that the results of multiple STR marker linkage analysis in the chromosome 8q23.3-q24.1,2p11.1-q12.2,3q26.32-3q28,10p15 section showed that the linkage was not supported. Therefore, the pathogenic genes in the family line were not in the 4 chromophore segments, and 4 were selected for 5p15.31-p15. STR markers, the result of linkage analysis showed that it was not fully affirmed and denying that the pathogenic gene was located in the area, and then, to further clarify the results, we added 8 STR loci to the section. The results showed that the LOD value was 2.8 when D5S486 was at theta =0.0. The experimental results supported the linkage of the pathogenic gene of the family with the site, considering the pathogenic gene in 5. P15.31-p15 chromosome section.
This study studied the clinical symptoms, genetic characteristics and loci of a benign adult familial myoclonic epilepsy, providing experience for the study of familial myoclonic epilepsy in benign adults, and for the genetic study of the disease, and for the first time the family myoclonic epilepsy home in China. The pathogenic gene was located in chromosome 5p15.31-p15 region.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R742.1
【參考文獻(xiàn)】
相關(guān)期刊論文 前7條
1 顧鳴敏,陳仁彪;多基因遺傳病遺傳分析的研究進(jìn)展[J];國(guó)外醫(yī)學(xué).遺傳學(xué)分冊(cè);1998年02期
2 劉書(shū)迎，武曉青，姚建民;24小時(shí)遙測(cè)腦電圖對(duì)癲癇的診斷價(jià)值[J];臨床腦電學(xué)雜志;1994年01期
3 陳竺,李偉俞,曼熊慧,陳賽娟;人類(lèi)基因組計(jì)劃的機(jī)遇和挑戰(zhàn):Ⅱ.對(duì)我國(guó)人類(lèi)基因組研究的若干思考[J];生命的化學(xué);1998年05期
4 廖成,趙慕鈞,李載平;定位候選克隆[J];生命的化學(xué);1999年02期
5 潘映輻;癲癇診斷中病史和腦電圖的臨床意義[J];中國(guó)醫(yī)刊;2002年04期
6 向燕群,唐北沙,沈璐,施小六,江泓,嚴(yán)新翔,郭伍英,吳志國(guó),劉月娥;良性家族性嬰兒驚厥疾病基因定位的初步研究[J];中華神經(jīng)科雜志;2000年05期
7 顧衛(wèi)紅;王國(guó)相;肖晶;王慕一;周永興;;常染色體顯性遺傳成人癲沲、震顫伴共濟(jì)失調(diào)臨床及致病基因定位研究[J];中華神經(jīng)科雜志;2006年06期
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