中国一个先天性无虹膜症家系的临床分析及其与PAX6基因的相关性研究

发布时间：2018-03-29 21:07

本文选题：先天性无虹膜　切入点：临床分析　出处：《中南大学》2012年博士论文

【摘要】：第一章中国一个先天性无虹膜症家系的经典遗传学及临床分析目的分析中国一个先天性无虹膜家系的遗传模式及其临床特点。方法将就诊于中南大学湘雅二医院门诊的一个先天性无虹膜家系(ANL)进行详细家系调查并绘制家系图谱。对自愿参加研究的9名患者进行详细病史采集及眼部检查。4名可得到清晰黄斑光学相干断层扫描(optical coherence tomography, OCT)图像的患者采用仪器自动按黄斑9格分区分析视网膜厚度,取黄斑中央区读数为中央区视网膜厚度,取上方内圈、鼻侧内圈、下方内圈、颞侧内圈视网膜厚度平均值为旁中心内圈视网膜厚度,取上方外圈、鼻侧外圈、下方外圈、颞侧外圈为旁中心外圈视网膜厚度,同时随机选取10名正常志愿者作为正常对照。结果先天性无虹膜家系(ANL)符合常染色显性遗传模式。ANL家系在相同的家庭条件下年龄大者较年龄小者最佳矫正视力(bestcorrected visual acuity, BCVA)差,而在相同年龄的条件下则农民家庭视力较工人家庭BCVA差。患者中央区视网膜厚度为290.38±12.49μm,旁中心凹内圈视网膜厚度为298.34±14.26μm,旁中心凹外圈视网膜厚度为267.22±11.27μm；与正常志愿者比较,前者P0.05,后两者P0.05。先证者Ⅲ：9右眼晶体颞下方呈刀削状,左眼行超声乳化白内障手术及折叠型人工晶体植入,术中体会前囊膜脆弱,术后眩光现象严重。患者Ⅲ：16同时患精神分裂症；患者Ⅳ：18同时患精神异常,但未分型,还同时患先天性青光眼。患者Ⅳ：16是随访9人中残存虹膜组织最多者,其左眼虹膜组织较右眼多,且在左眼瞳孔区鼻上方见瞳孔残膜。结论(1)先天性无虹膜症ANL家系遗传模式为常染色体显性遗传；(2)先天性无虹膜症BCVA随年龄下降可能与晶体混浊及视网膜的光损伤有关；(3)先天性无虹膜症黄斑中心凹发育不良主要表现为黄斑中央区明显增厚,中心凹处存在除神经纤维层以外的各层视网膜结构；(4)先天性无虹膜患者晶体形态可发育异常,其白内障可能在达到某个程度后即迅速发展；(5)先天性无虹膜患者白内障手术时需综合考虑前囊膜脆弱及术后视觉质量问题；(6)先天性无虹膜ANL家系内存在虹膜形态及精神疾病的表现型变异；(7)先天性无虹膜患者瞳孔残膜为发病机制的虹膜组织萎缩过多学说提供支持。第二章中国一个常染色体显性遗传先天性无虹膜症家系PAX6基因序列及外显子拷贝数测定目的确定中国一个常染色体显性遗传先天性无虹膜症家系PAX6基因启动子区、外显子、外显子与内含子交界处是否存在基因变异以及外显子拷贝数是否有变化。方法提取中国一个常染色体显性遗传先天性无虹膜家系ANL先证者Ⅲ：9基因组DNA,采用PCR扩增直接测序法测定其PAX6基因3个转录异构体的2个启动子区、5’端非编码区(5'-untranslated region,5'-UTR)、编码区(coding region)、3’端非编码区(3'-untranslated region,3'-UTR)以及剪接位点的DNA序列,采用实时定量PCR(real-time quantitative PCR, qPCR)方法测定3个转录异构体所有外显子拷贝数,qPCR采用ΔΔCt相对定量数据分析方法。结果先证者Ⅲ：9的PAX6基因在转录异构体3启动子区检测到4个已报道的单核苷酸多态性(Single nucleotide polymorphism, SNP)rs1806155、rs5790870、rs1806156、rs1806157和rs1806180,并检测到1个未报道为SNP但与疾病表型无共分离现象的碱基替换g.-1217CT；在第12号内含子检测到1个SNP rs3026393;在3’-UTR检测到1个SNP rs1506。转录异构体1和转录异构体2启动子区因GC含量过高导致PCR经反复优化条件仍扩增失败。PAX6基因3个转录异构体各外显子拷贝数均为双拷贝。结论先天性无虹膜家系ANL的致病突变与PAX6外显子及外显子内含子交界处序列及拷贝数变异无关,与转录异构体3启动子区无关,但不能排除是否与转录异构体1和转录异构体2启动子区有关。第三章中国一个常染色体显性遗传先天性无虹膜症家系的基因连锁定位目的先天性无虹膜家系ANL的致病基因是否与PAX6基因有关。方法提取中国一个常染色体显性遗传先天性无虹膜家系ANL的2个分支现存21人基因组DNA,并在PAX6基因附近选取6个微卫星标记D11S904、D11S1324、D11S914、D11S1776、D11S907和D11S935进行连锁分析,采用LOD值两点连锁分析,并构建单体型。结果6个微卫星标记中在PAX6基因下游微卫星标记D11S1324取得LOD最大值3.16(θ=0),且其附近标记物的LOD值均在θ=0时取得大于1的正数。单体型分析表明6个微卫星标记构成的区域在家系中呈现出与疾病表型共分离现象。结论先天性无虹膜症家系ANL的致病基因定位与PAX6基因附近,并且位于PAX6基因下游的可能性大。
[Abstract]:The classic genetics and clinical analysis of a Chinese family with congenital irinsis
Objective to analyze the genetic pattern and clinical characteristics of a congenital ininirinid family in China.
Methods a congenital treatment in Xiangya No.2 Hospital of Central South University clinic aniridia family (ANL) detailed pedigree investigation and family maps were drawn. The 9 patients volunteered for the study detailed history collection and eye examination.4 can obtain clear macular optical coherence tomography (optical coherence tomography, OCT) instrument the 9 grid partition automatically according to the macular retinal thickness analysis by image were taken as the central retinal thickness of macular central readings, the top side of the nose below the inner ring, inner ring, inner ring, the inner retinal thickness of the temporal average paracentral inner retinal thickness, the top side of the nose below the outer ring, outer ring, outer ring, the outer ring is adjacent to the temporal side the center of the outer retinal thickness, and randomly selected 10 healthy volunteers as normal control.
The results of congenital aniridia family (ANL) with autosomal dominant genetic model.ANL family age in the same family conditions than the younger, the best corrected visual acuity (bestcorrected visual, acuity, BCVA), and in the same age under the condition of peasant family worker family vision was BCVA. Central retinal thickness with 290.38 + 12.49 m, parafoveal inner retinal thickness was 298.34 + 14.26 m, parafoveal outer retinal thickness was 267.22 + 11.27 m; compared with normal volunteers after the former P0.05, two P0.05. proband III: 9 under the right eye was cut crystal temporal shape, the left eye of phacoemulsification cataract surgery and foldable IOL implantation. The intraoperative experience of anterior capsule fragility, postoperative glare phenomenon is serious. At the same time: 16 patients of schizophrenia patients; IV: 18 patients with mental disorders, but not type, also from the first Natural glaucoma. Patients IV: 16, the most remaining iris tissues were followed up in 9 patients. The iris tissue in the left eye was more than that in the right eye, and the pupillary remnant membrane was seen above the nasal area in the left eye pupil area.
Conclusion (1) congenital aniridia family ANL is an autosomal dominant genetic model; (2) congenital aniridia with age BCVA decline may be related to lens opacity and retinal light injury; (3) congenital aniridia and foveal hypoplasia mainly manifested as thickening of central macular. The retinal nerve fiber layer in the layer structure outside the fovea; (4) congenital aniridia patients with abnormal development of the crystal morphology, in cataract may reach a certain level after the rapid development; (5) considering the quality problems of visual anterior capsule fragile and postoperative cataract surgery in patients with congenital aniridia required time; (6) congenital absence of phenotypic variation in iris morphology and iris ANL mental illness within the family; (7) the iris tissue of congenital aniridia patients with pupillary membrane for the pathogenesis of the atrophy of too much theory to provide support.
The second chapter of an autosomal dominant hereditary inirinosus family PAX6 gene sequence and the determination of the exon copy number
Objective to determine whether there is genetic variation in the promoter region of PAX6 gene, exon, exon and intron in Chinese family with autosomal dominant congenital irirma, and whether exon copy number is changed.
A method of extracting China autosomal dominant congenital aniridia proband ANL III: 9 genomic DNA 2 promoter region was amplified by PCR direct sequencing method for the determination of the PAX6 gene of 3 transcripts, 5 'non encoding region (5'-untranslated, region, 5'-UTR (coding) encoding region region), 3' non encoding region (3'-untranslated region, 3'-UTR) and splice sites of DNA sequences, using quantitative real-time PCR (real-time quantitative PCR, qPCR) method for the determination of 3 transcripts of all exon copy number of qPCR, adopting the analysis method of relative quantitative data of delta delta Ct.
Results the proband III: PAX6 9 gene transcription in the detection of 3 isomers of the promoter region to 4 reported single nucleotide polymorphisms (Single, nucleotide polymorphism, SNP) rs1806155, rs5790870, rs1806156, rs1806157 and rs1806180, and detected 1 reports for SNP but not with the disease phenotype without co segregation phenomenon base substitution g.-1217CT; in intron twelfth was detected in 1 SNP rs3026393; in the 3 '-UTR detected 1 SNP rs1506. transcripts of 1 and 2 transcripts of the promoter region due to the high content of GC to PCR by optimizing conditions still failed to amplify the.PAX6 gene transcripts of the 3 exon copy the number was two copies.
Conclusion the pathogenesis of congenital ironic pedigree ANL is independent of PAX6 exon and exon intron junction sequence and copy number variation, but has nothing to do with the promoter region of transcriptional isomer 3, but it cannot be excluded whether it is related to the promoter region of transcriptional isomer 1 and transcriptional isomer 2.
The third chapter of gene linkage localization in an autosomal dominant congenital inininosinic family
Objective whether the pathogenic gene of ANL in the congenital iririd family is related to the PAX6 gene.
A method of extracting Chinese autosomal dominant congenital aniridia family ANL of the 2 branches of the existing 21 human genomic DNA, and selected 6 microsatellite markers D11S904, D11S1324, D11S914, D11S1776 in D11S907 and D11S935 near the PAX6 gene, linkage analysis, the LOD value of two-point linkage analysis and haplotype.
The results of 6 microsatellite markers in the downstream of PAX6 gene microsatellite marker D11S1324 has a maximum value of 3.16 LOD (0 =0), and near the marker LOD was obtained in theta =0 greater than 1 positive. Haplotype analysis showed that 6 microsatellite markers of the region showing co segregated with the disease phenotype the phenomenon in the family.
Conclusion the pathogenicity of ANL in the congenital irinic family is located near the PAX6 gene, and the possibility of the downstream of the PAX6 gene is great.

【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R773.1

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