先天性眼外肌纤维化家系基因定位、临床分型及静息态功能磁共振的研究

发布时间：2018-04-21 01:43

本文选题：先天性眼外肌纤维化 + 常染色体显性遗传　；参考：《南昌大学》2016年博士论文

【摘要】：目的:对中国先天性眼外肌纤维化(congenital fibrosis of the extraocular muscles,CFEOM)家系进行致病基因定位及疾病分型,研究其中枢神经系统功能性改变,进一步认识和理解其神经源性的病变机制,为尚处初级研究阶段的CFEOM患者大脑fMRI变化规律提供更多临床依据。方法:1、致病基因定位与疾病分型1.1对2个CFEOM家系9名患者采集病史、体格检查、抽取外周静脉血5-8ml并提取基因组DNA。首先通过候选基因法对己知的主要基因突变位点进行探查,采取直接DNA序列分析,再通过SSCP证实突变位点,随机选取100名健康且无血缘关系中国人作对照组,以排除基因多态性可能。若仍未发现,则检测范围扩大到目前已报道的其它基因突变位点,或再运用连锁分析及全外显子测序进行检测。1.2通过查找致病基因发现已知或未知的突变位点,结合CFEOM家系患者临床表型进行疾病分型。2、神经功能检测与分析对2个家系典型CFEOM患者和正常对照组行全脑静息态功能磁共振检查及三维脑结构成像。正常组与病人组根据年龄及性别按2:1匹配。2.1首先对所有受试者进行全脑常规横断位MRI扫描以排除颅脑器质性病变。常规颅脑MRI扫描未见异常者继续行rs-fMRI扫描。2.2 rs-fMRI扫描:采用实时功能成像程序(Real-Time Imaging Processing,RTIP)方式及梯度回波-回波平面成像(Gradient-Recalled Echo-Planar Imaging,GRE-EPI)序列。2.3观察指标与分析:首先对采集的数据运用RESET软件进行预处理及校正,使用SPM8软件进行统计分析。对病变组与对照组的低频振幅(ALFF)、局域一致性(ReHo)、功能连接(FC)以及基于体素的形态学分析法(VBM)的脑结构变化进行比较分析。结果:1、临床研究1.1家系1五代中9人患病,均具有典型cfeom外观特征。致病基因位于kif21a基因外显子21,2860ct(r954w),为杂合错义突变。1.2家系2四代中4人患病,除一人临床表型不典型外,其他三人都具有典型的cfeom外观特征。致病基因位于tbuu3基因外显子4,1249ga(d417n),为杂合错义突变。1.3两个家系所有患者神经影像检查头颅mri形态上未见明显发育不良。2、静息态功能磁共振研究病患组8人:男6人,女2人,年龄13-58岁,平均年龄35.38岁(sd=14.02),正常对照组16人:男12人,女4人,年龄12-58岁,平均年龄36.26岁(sd=15.64),两组性别(chi-squaretestp=0.925)、年龄(p=0.822)比较无统计学差异。2.1rs-fmri分析结果:2.1.1alff:与正常对照组比较,病患组alff增强的脑区:小脑扁桃体、左颞下回;减弱的脑区:右顶上小叶。2.1.2reho:与正常对照组比较,病患组reho增强的脑区:右顶叶角回、左舌回、左岛叶中央沟盖、右中央前回;减弱的脑区:左小脑后叶。2.1.3fc:与正常对照组比较,以右额中回后部为种子区(roi)时,病患组无fc增加的脑区,减弱的脑区:右中央后回、左顶上小叶;以左额中回后部为种子区时,fc增加的脑区:右小脑前叶、小脑蚓部,无减弱的脑区。2.2vbm全脑灰、白质体积分析结果:2.2.1灰质体积:与正常对照组相比,病患组灰质体积增加的脑区:左辅助运动区、右顶下小叶、右额下回眶部;体积减少区:右额下回、左额上回。2.2.2白质体积:与正常对照组相比,病患组的白质无体积增加的脑区;减少的脑区:左丘脑、右枕下回、右杏仁核、胼胝体压部、右顶叶角回、右颞上回。结论:1、运用候选基因法确定2个cfeom家系致病基因定位分别为kif21a基因2860ct(r954w)和tubb3基因1249ga(d417n),均为杂合错义突变。2、cfeom1与cfeom3临床表型存在交叉,1型具更稳定的cfeom临床特征,3型表型更多样及外显不全。家系1属于cfeom1a型,遗传方式为常染色体完全外显的显性遗传;家系2属于cfeom3a型,遗传方式为常染色体不完全外显的显性遗传。3、2个CFEOM家系患者头颅常规MRI形态上均未见明显发育不良。4、全脑静息态功能磁共振研究显示CFEOM患者存在多个皮质中枢信号序列的变化,与患者异常心理和情绪、理解和辨别、运动和控制等脑区功能改变有关。以不同半球额中回后部(侧目中枢)为种子区的功能连接异常区域显示与患者视觉感知、运动学习及环境适应等相关。5、基于体素的形态学分析研究显示CFEOM患者灰、白质体积改变范围较广,涉及多个中枢神经功能区域,主要为边缘系统皮质与皮质下结构;大脑白质存在广泛体积减少区,提示颅内神经纤维传导功能下降,符合中枢神经源性肌病特征。
[Abstract]:Objective: To study the gene localization and disease classification of congenital fibrosis of the extraocular muscles (CFEOM) family in China, study the functional changes of the armature nervous system, and further understand and understand the mechanism of the neurogenic pathological changes for the changes of the fMRI changes in the brain of CFEOM patients at the primary stage of study. The rules provided more clinical basis. Methods: 1, the pathogenic gene location and the disease classification 1.1 pairs of 2 CFEOM families 9 patients to collect medical history, physical examination, extraction of peripheral venous blood 5-8ml and extract the genomic DNA. first through the candidate gene method to explore the known major gene mutation site, direct DNA sequence analysis, and then through the SSCP syndrome 100 healthy and unrelated Chinese people were randomly selected as the control group to exclude genetic polymorphisms. If not found, the detection range was extended to other gene mutations at present, or the detection of.1.2 by linkage analysis and exon sequencing was used to detect the known or not. The known mutation site, combined with the clinical phenotype of the CFEOM family with the clinical phenotype of the disease classification.2, and the neural function test and analysis of 2 families with typical CFEOM patients and the normal control group, the whole brain resting state functional magnetic resonance imaging (fMRI) and the three-dimensional brain structure imaging were performed. The normal group and the patient group were based on the age and sex of the 2:1 matching.2.1 to all the subjects first. General cerebral conventional transversal MRI scan was performed to eliminate craniocerebral organic lesions. Rs-fMRI scanning.2.2 rs-fMRI scan was continued without abnormal brain MRI scan: the real-time functional imaging program (Real-Time Imaging Processing, RTIP) and gradient echo echo plane imaging (Gradient-Recalled Echo-Planar Imaging) sequence .2.3 observation and analysis: first, the collected data were pre processed and corrected by RESET software, and the SPM8 software was used for statistical analysis. The low frequency amplitude (ALFF), local conformance (ReHo), functional connectivity (FC), and the morpheme based morphologic analysis (VBM) were compared and analyzed. The results were as follows: 1, 9 of the 1.1 families and 1 five generations of the clinical study have a typical CFEOM appearance. The pathogenic gene is located in the exon 212860ct (r954w) of the KIF21A gene, and 4 in the.1.2 family of the heterozygous mutation.1.2 family, and the other three people have typical CFEOM appearance except one person's clinical phenotype. The pathogenic gene is located outside the tbuu3 gene. 41249ga (d417n), for all patients with heterozygous missense mutation.1.3, all patients with two families had no apparent dysplasia.2 in the head MRI morphology. Resting state function magnetic resonance (fMRI) was used to study 8 patients: 6 men, 2 women, 13-58 years old, 35.38 years old (sd=14.02), and 16 men in normal control group: 12 men, 4 women, and average age 12-58 years old. Average age was 12-58 years old. Age 36.26 (sd=15.64), two groups of sex (chi-squaretestp=0.925), age (p=0.822) no statistical difference.2.1rs-fmri analysis results: 2.1.1alff: and normal control group, Alff enhanced brain area of the patient group: cerebellar tonsil, left temporal gyrus, and weakened brain area: right upper lobule.2.1.2reho: compared with normal control group, ReHo group ReHo The enhanced brain area: right parietal gyrus, left lingual gyrus, left central sulcus cover, right central anterior gyrus; weakened brain area: the left posterior lobe.2.1.3fc: was compared with the normal control group, with the right middle back and posterior part of the seed region (ROI), the patient group had no FC increase in the brain area and the weakened brain area: right central posterior gyrus and left upper lobule; the left middle posterior part was the seed area. At the time, FC increased brain area: right cerebellar anterior lobe, cerebellar vermis, no weakened brain.2.2vbm whole brain ash, white matter volume analysis results: 2.2.1 gray matter volume: compared with normal control group, the volume of gray matter volume increased in the patient group: left auxiliary motor area, right inferior lobule, right inferior frontal gyrus; right frontal gyrus, left upper.2.2.2 white matter Volume: compared with the normal control group, the white matter had no volume increase in the brain area; the reduced brain area: left thalamus, right occipital gyrus, right amygdala, corpus callosum pressure, right parietal gyrus, right temporal gyrus. Conclusion: 1, KIF21A gene 2860ct (r954w) and tubb3 gene 1249ga (d41) were identified by candidate gene method. 7n) were heterozygous mutation.2, CFEOM1 and CFEOM3 clinical phenotype intersecting, the 1 type has more stable CFEOM clinical features, 3 phenotype diversity and exotoxicity. Family 1 belongs to cfeom1a type, hereditary mode is autosomal explicit explicit inheritance; family 2 belongs to cfeom3a type, hereditary mode is autosomal incomplete explicit dominance There was no obvious dysplasia.4 in the routine MRI morphology of the patients with.3,2 CFEOM family, and the study of resting state functional magnetic resonance (fMRI) showed that there were many changes of the central signal sequence of the cortex in the patients with CFEOM, which was related to the abnormal mental and emotional, understanding and discrimination, movement and control of brain function changes in patients with different hemispheres. The posterior (lateral center) area of the functional connectivity of the seed region shows.5 related to visual perception, exercise learning and environmental adaptation. The morphological analysis of voxel shows that CFEOM patients are grey with a wide range of white matter volume, involving several central nervous power areas, mainly the cortex of the marginal system and subcortical structure. There is a wide volume reduction area in the white matter of the brain, suggesting that the decrease of intracranial nerve fiber conduction function is consistent with the characteristics of central neurogenic myopathy.

【学位授予单位】：南昌大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R777.4

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