眼科遺傳疾病的分子遺傳學(xué)分析
發(fā)布時(shí)間：2018-10-21 07:53
【摘要】： 眼遺傳病是當(dāng)前兒童和青少年的主要致盲性眼病之一。根據(jù)遺傳方式可分為四種主要類(lèi)型:常染色體顯性遺傳病、常染色體隱性遺傳病,性染色體連鎖遺傳病,和線(xiàn)粒體遺傳病。在眾多眼遺傳病中,先天性白內(nèi)障和視神經(jīng)萎縮占有重要位置,本論文對(duì)這兩種眼科遺傳病進(jìn)行分子遺傳研究,開(kāi)展遺傳連鎖分析和致病突變檢測(cè)。 白內(nèi)障是指晶狀體出現(xiàn)渾濁的病理狀態(tài),按發(fā)病時(shí)間可被分為先天性和年齡相關(guān)性白內(nèi)障。先天性白內(nèi)障通常在患者出生時(shí)或不久就出現(xiàn),該病呈現(xiàn)出高度的遺傳以及臨床異質(zhì)性。 (1)在一全白內(nèi)障中國(guó)家系中,我們發(fā)現(xiàn)致病基因與微衛(wèi)星多態(tài)標(biāo)記D16S3043連鎖,進(jìn)一步單倍型分析將其定位于D16S515和D16S415標(biāo)記之間,對(duì)該區(qū)域內(nèi)HSF4基因測(cè)序發(fā)現(xiàn)第74位密碼子由精氨酸突變?yōu)榻M氨酸(p.R74H),該突變?cè)诩蚁抵信c疾病共分離,同時(shí)對(duì)150個(gè)正常對(duì)照的RFLP分析也沒(méi)有發(fā)現(xiàn)該突變。該結(jié)果豐富了HSF4基因突變導(dǎo)致的白內(nèi)障的臨床表現(xiàn)譜。 (2)在另一后極性白內(nèi)障家系中,連鎖和單倍型分析將該家系致病基因定位在11q22-22.3區(qū)域,最大LOD值為4.52,對(duì)CRYAB基因測(cè)序發(fā)現(xiàn)第20位密碼子由脯氨酸突變?yōu)榻z氨酸(p.P20S),該突變?cè)诖思蚁抵信c白內(nèi)障共分離,在200個(gè)正常人中,用SSCP分析未檢測(cè)到這一突變。該結(jié)果有力地證實(shí)了CRYAB基因突變可以導(dǎo)致白內(nèi)障。 視神經(jīng)纖維退化和相關(guān)血管系統(tǒng)病變會(huì)導(dǎo)致視力受損,這種病理學(xué)狀態(tài)被稱(chēng)為視神經(jīng)萎縮(OA)。遺傳性O(shè)A可分為兩大類(lèi):原發(fā)性O(shè)A和繼發(fā)性O(shè)A,原發(fā)性O(shè)A包括常染色體視神經(jīng)萎縮(ADOA)和Leber’s視神經(jīng)萎縮(LHON)等類(lèi)型。由于OA遺傳和臨床表型的高度異質(zhì)性,因此其診斷較為困難。 (3)通過(guò)一個(gè)ADOA家系的連鎖分析以及測(cè)序和RFLP,鑒定出OPA1基因的一個(gè)錯(cuò)義突變(p.G401D),經(jīng)RFLP分析證明該突變只存在于該家系患者,不存在于家系中健康人和150個(gè)正常人中,而且病人呈現(xiàn)出視神經(jīng)萎縮以及聽(tīng)力受損綜合征狀。該結(jié)果支持OPA1突變導(dǎo)致的視神經(jīng)萎縮可伴隨有聽(tīng)力受損。 (4)我們對(duì)一個(gè)LHON家系進(jìn)行臨床分析,發(fā)現(xiàn)家系內(nèi)患者整體預(yù)后良好,實(shí)屬罕見(jiàn);對(duì)家系成員進(jìn)行了線(xiàn)粒體DNA的3個(gè)原發(fā)突變位點(diǎn)檢測(cè),發(fā)現(xiàn)病人攜帶mtG11778A位點(diǎn)突變,而且多名病人和突變攜帶者呈現(xiàn)出異質(zhì)性突變;對(duì)相關(guān)繼發(fā)突變位點(diǎn)的檢測(cè)沒(méi)有發(fā)現(xiàn)次級(jí)突變的存在。該結(jié)果表明可能存在其它因子對(duì)該家系的表型起調(diào)節(jié)作用。 所有這些分子遺傳學(xué)研究為闡明先天性白內(nèi)障和視神經(jīng)疾病的病理學(xué),以及提供更好的遺傳咨詢(xún)和最終發(fā)展有效疾病治療策略提供了有益的幫助。
[Abstract]:Eye-hereditary disease is one of the leading causes of blindness in children and adolescents. According to the genetic pattern, there are four main types: autosomal dominant disease, autosomal recessive hereditary disease, sex chromosome linked genetic disease, and mitochondrial genetic disease. Congenital cataract and optic atrophy play an important role in many ophthalmic hereditary diseases. In this paper, molecular genetic analysis, genetic linkage analysis and pathogenic mutation detection of these two kinds of ophthalmic hereditary diseases were carried out. Cataract refers to the pathological state of lens opacity, which can be divided into congenital cataract and age-related cataract according to the onset time. Congenital cataract usually occurs at birth or shortly after birth, showing a high degree of genetic and clinical heterogeneity. We found that the pathogenicity gene was linked to microsatellite polymorphic marker D16S3043, and further haplotype analysis mapped it between D16S515 and D16S415 markers. The HSF4 gene in this region was sequenced from arginine to histidine (p.R74H) at codon 74. The mutation was isolated from the disease at home, and the mutation was not detected by RFLP analysis of 150 normal controls. The results enriched the clinical manifestations of cataract caused by mutation of HSF4 gene. (2) in another pedigree with post-polar cataract, linkage and haplotype analysis located the pathogenic gene in 11q22-22.3 region. The maximum LOD value was 4.52. The CRYAB gene was sequenced from proline to serine (p.P20S). The mutation was separated from cataract in this pedigree and was not detected by SSCP analysis in 200 normal subjects. The results strongly confirm that mutations in the CRYAB gene can lead to cataracts. Optic nerve fiber degeneration and associated vascular system lesions can lead to visual impairment, a pathological condition known as optic nerve atrophy (OA).) Hereditary OA can be divided into two categories: primary OA and secondary OA, primary OA including autosomal optic atrophy (ADOA) and Leber's optic atrophy (LHON). Due to the high heterogeneity of genetic and clinical phenotypes of OA, Therefore, it is difficult to diagnose OPA1 gene. (3) A missense mutation (p.G401D) of OPA1 gene was identified by linkage analysis, sequencing and RFLP, analysis of a ADOA pedigree. It does not exist in healthy people and 150 normal people, and the patient presents optic atrophy and hearing loss syndrome. The results support that the optic atrophy caused by OPA1 mutation may be accompanied by hearing impairment. (4) A clinical analysis of a LHON family showed that the overall prognosis of the patients in the family was good, which was rare. Three primary mutation sites of mitochondrial DNA were detected among family members. It was found that the patients carried mtG11778A mutation, and many patients and mutants showed heterogeneity. No secondary mutation was found in the detection of related secondary mutation sites. These results suggest that there may be other factors regulating the phenotype of the family. All of these molecular genetic studies are helpful to clarify the pathology of congenital cataract and optic nerve diseases, to provide better genetic counseling and to develop effective treatment strategies.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R77;R394
本文編號(hào)：2284451
[Abstract]:Eye-hereditary disease is one of the leading causes of blindness in children and adolescents. According to the genetic pattern, there are four main types: autosomal dominant disease, autosomal recessive hereditary disease, sex chromosome linked genetic disease, and mitochondrial genetic disease. Congenital cataract and optic atrophy play an important role in many ophthalmic hereditary diseases. In this paper, molecular genetic analysis, genetic linkage analysis and pathogenic mutation detection of these two kinds of ophthalmic hereditary diseases were carried out. Cataract refers to the pathological state of lens opacity, which can be divided into congenital cataract and age-related cataract according to the onset time. Congenital cataract usually occurs at birth or shortly after birth, showing a high degree of genetic and clinical heterogeneity. We found that the pathogenicity gene was linked to microsatellite polymorphic marker D16S3043, and further haplotype analysis mapped it between D16S515 and D16S415 markers. The HSF4 gene in this region was sequenced from arginine to histidine (p.R74H) at codon 74. The mutation was isolated from the disease at home, and the mutation was not detected by RFLP analysis of 150 normal controls. The results enriched the clinical manifestations of cataract caused by mutation of HSF4 gene. (2) in another pedigree with post-polar cataract, linkage and haplotype analysis located the pathogenic gene in 11q22-22.3 region. The maximum LOD value was 4.52. The CRYAB gene was sequenced from proline to serine (p.P20S). The mutation was separated from cataract in this pedigree and was not detected by SSCP analysis in 200 normal subjects. The results strongly confirm that mutations in the CRYAB gene can lead to cataracts. Optic nerve fiber degeneration and associated vascular system lesions can lead to visual impairment, a pathological condition known as optic nerve atrophy (OA).) Hereditary OA can be divided into two categories: primary OA and secondary OA, primary OA including autosomal optic atrophy (ADOA) and Leber's optic atrophy (LHON). Due to the high heterogeneity of genetic and clinical phenotypes of OA, Therefore, it is difficult to diagnose OPA1 gene. (3) A missense mutation (p.G401D) of OPA1 gene was identified by linkage analysis, sequencing and RFLP, analysis of a ADOA pedigree. It does not exist in healthy people and 150 normal people, and the patient presents optic atrophy and hearing loss syndrome. The results support that the optic atrophy caused by OPA1 mutation may be accompanied by hearing impairment. (4) A clinical analysis of a LHON family showed that the overall prognosis of the patients in the family was good, which was rare. Three primary mutation sites of mitochondrial DNA were detected among family members. It was found that the patients carried mtG11778A mutation, and many patients and mutants showed heterogeneity. No secondary mutation was found in the detection of related secondary mutation sites. These results suggest that there may be other factors regulating the phenotype of the family. All of these molecular genetic studies are helpful to clarify the pathology of congenital cataract and optic nerve diseases, to provide better genetic counseling and to develop effective treatment strategies.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R77;R394
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
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5 賈小云;王燕;黎仕強(qiáng);肖學(xué)珊;張清炯;郭向明;;中國(guó)人LHON患者mtDNA繼發(fā)突變的篩查[J];中國(guó)優(yōu)生與遺傳雜志;2005年11期
，本文編號(hào)：2284451
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