先天性小耳畸形的发病特征及候选基因检测研究

发布时间：2018-03-02 03:17

本文关键词： 小耳畸形伴发畸形遗传特征候选基因　出处：《北京协和医学院》2017年博士论文　论文类型：学位论文

【摘要】：目的先天性小耳畸形是体表重大畸形,严重影响患者的外貌和心理健康,进行小耳畸形发病特征和相关遗传学研究意义重大。本研究探讨中国小耳畸形患者伴发畸形的发生情况和特点,分析之间的关联;经过对家族史的详细调查,了解小耳畸形患者的遗传特征;在小耳畸形患者中进行候选基因序列变异和拷贝数变异检测,对变异进行生物信息学分析。通过以上研究逐步深化对小耳畸形的认知,可作为今后小耳畸形分子机制的研究和防治的基础。方法1.在基于医院的研究中选取2014年12月-2016年2月间小耳畸形患者,对所有非综合征型小耳畸形患者672例进行详细地体格检查,并记录相关伴发畸形。将伴发畸形根据受影响的主要器官系统进行分类,采用SPSS 19.0统计学软件分析小耳畸形与伴发畸形间的关系。并与其他国家地区的小耳伴发畸形资料对比分析中国小耳群体患病特点。2.对上述672个患者通过查体、直接询问及电话随访的方法获得患者的家系资料及耳部畸形发病情况并登记。主要调查对象为先证者、一级亲属、二级亲属、三级亲属、四级亲属,按照小耳畸形患者是否具有耳部畸形家族史背景分为家族史组和散发组,对数据进行统计学分析,探寻有价值的规律。3.随机选取200例散发小耳畸形患者对六个候选基因(HOXA1、HOXA2、EYA1、SIX1、SALL1、FGF3)进行序列变异和拷贝数变异检测,并利用生物信息学分析变异的危害。结果1.本研究显示了男性(72%)、单侧(92.99%)和右侧(63%)多发,与文献中的结果一致。我们在293(43.6%)位患者中发现了一项及以上的伴发畸形,尤其是耳面颈系统、肌肉骨骼系统、心血管系统伴发率最高。耳廓发育程度越差伴发畸形率越高。小耳不同伴发畸形比例有显著异质性,但大体在器官系统畸形方面的权重是相符的,即伴发畸形最常累及的是耳面颈系统,接下来是肌肉骨骼系统,然后是心血管系统,后面是泌尿生殖系统,呼吸系统、神经系统、消化系统等较少累及。2.本研究中耳部畸形家族史阳性率为34.1%,其中一级亲属所占比例最高,二、三、四级亲属比例依次减少,符合遗传学规律,且父母两系背景在各级小耳畸形患者人数构成比无统计学差异。有33.8%小耳畸形患者伴发附耳/瘘管,且这部分患者比单纯小耳患者有更高的家族耳部畸形发生率。3.在200例患者中,鉴定出1例患者HOXA2基因的错义突变(c.260CT,p.A87V),此突变不存在于患者父母,为新发突变,在1000 human genome、HGMD、ESP6500、dbSNP和ExAC等数据库中未查到相关记录,经生物信息学分析该突变高度保守,推测其具有致病性。检测到两例患者存在EYA1基因的拷贝数缺失,推测为小耳畸形的致病原因。结论1.本研究是中国小耳患者中第一份有关伴发畸形的详细专题性研究,小耳患者的伴发畸形率较高,且临床异质性较大,为了探索小耳畸形病因及为患者提供更好的治疗,应加大未来对相关伴发畸形的研究力度。2.本研究结合其他耳部畸形多发性,将家属耳部畸形纳入研究并证实耳部畸形家族史发生率为34.1%,且存在垂直传递、隔代遗传和家族聚集的遗传特征。在研究小耳畸形遗传特征时不应忽略亲属中的其他耳畸形。虽然小耳畸形目前发病机制不明,但遗传因素不容忽视。3.本研究分别对200例散发小耳患者六个候选基因进行了序列变异和拷贝数变异检测,成功地鉴定了一例患者存在HOXA2基因编码区的c.260CT(p.A87V)突变,此突变位于exon1,为国际首次报道,扩增了小耳畸形HOXA2基因突变谱;并且在两例患者中检测到EYA1基因拷贝数缺失。这些发现可能是小耳畸形的致病因素,其发病机制,有待下一步研究验证。
[Abstract]:The purpose of congenital microtia is a major body deformity, seriously affect the patient's appearance and mental health, study significance of microtia disease characteristics and related genetics. Major of this study was to investigate the occurrence and characteristics of deformity with Chinese microtia patients, correlation analysis between; after a detailed investigation of family history, understand the genetic characteristics of patients with microtia; and the copy number variation of candidate gene sequence variation detection in patients with microtia, bioinformatics analysis of variation. Through the above study, the gradual deepening of the microtia cognition, can be used as a basis for future research and the prevention of microtia molecular mechanism. Methods 1. in December 2014 select the -2016 in February of microtia patients in hospital in the study based on the physical examination of 672 cases with all non syndromic patients with microtia, and record The associated malformation accompanied malformations. According to the main organ system affected the classification, using SPSS 19 statistical software to analyze the relationship between microtia and associated malformations. With comparative analysis of the data Chinese small ear malformation group prevalence characteristics of.2. of the 672 patients through the examination with other countries and regions of the small ear method, direct examination and telephone follow-up for patients with family data and ear deformity incidence and registration. The main research object for the proband, first-degree relatives, two relatives, three relatives, four relatives, according to the patients with microtia is not with ear deformity family history background into family history group and the sporadic group, the statistical analysis of the data, to explore the value of.3. in 200 randomly selected patients with sporadic microtia in six candidate genes (HOXA1, HOXA2, EYA1, SIX1, SALL1, FGF3) sequence variation and copy number variation Abnormal detection, and the use of bioinformatics analysis of variation of harm. 1. results of this study shows that male (72%), unilateral (92.99%) and right (63%) multiple, consistent with the results in the literature. We in 293 (43.6%) patients were found in one or more of the associated anomalies, especially the ear is the face and neck system, musculoskeletal system, cardiovascular system and the incidence rate of the highest degree of development. The worse ear malformations was higher. Small ear malformations have significantly different proportion of heterogeneity, but generally in the organ system abnormalities weight is consistent, which is associated with the most frequently involved is deformity next is the ear of face and neck system, musculoskeletal system, and then is behind the cardiovascular system, genitourinary system, respiratory system, nervous system, digestive system and the positive rate of less involved.2. this study of middle ear malformation family history was 34.1%, of which the first-degree relatives of the highest proportion, two, three, four In order to reduce the proportion of relatives, comply with the laws of genetics, and the parental background at all levels of two-line microtia patients who showed no significant difference. There are 33.8% patients with fistula / ear hair with microtia, and these patients than microtia patients had a higher incidence of.3. family ear deformity in 200 patients, 1 patients identified missense mutations in the HOXA2 gene (c.260CT, p.A87V), this mutation does not exist in patients with parents, is a new mutation in 1000 human, genome, HGMD, ESP6500, dbSNP and ExAC in the database did not find relevant records, by bioinformatics analysis of the mutations are highly conserved, presumably with pathogenicity. Two patients have detected EYA1 gene copy number is missing, presumed cause of microtia. Conclusion: 1. this study is Chinese with the first small ear malformations with the special study, patients with small ears The malformation rate is higher, and the heterogeneity is larger, in order to explore the microtia etiology and provide better treatment for patients, should increase the future of related research efforts with.2. deformity combined with the other ear deformity of multiple family members, will be included in the study and confirmed that the ear ear deformity deformity of the family history of incidence was 34.1%. And there are vertical transmission, genetic characteristics of atavism and family aggregation. In the study of genetic characteristics of microtia should not ignore other relatives in ear microtia malformation. Although the pathogenesis is unknown, but genetic factors can not be ignored in this study were.3. in 200 cases of sporadic microtia and six candidate genes were and copy number variation detection of sequence variation, successfully identified HOXA2 gene encoding region in a patient with c.260CT (p.A87V) mutation, the mutation in exon1, reported for the first time international, the amplification of microtia H OXA2 gene mutation spectrum and EYA1 gene copy number deletion were detected in two patients. These findings may be the pathogenic factors of microtia, and its pathogenesis needs further research and verification.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R764.7

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