59例大前庭导水管综合征<i>SLC26A4</i>基因突变频率及新发突变位点分析

苏栋; 娄凡; 黄锐; 李霞; 林垦; 李果; 马静

doi:10.13201/j.issn.2096-7993.2023.11.010

59例大前庭导水管综合征SLC26A4基因突变频率及新发突变位点分析

- 昆明市儿童医院耳鼻咽喉头颈外科昆明市儿童先天出生缺陷防控研究重点实验室云南省儿童重大疾病研究重点实验室(昆明，650228)
基金项目:
云南省王海波专家工作站(No：202105AF150056)；云南省中青年学术和技术带头人后备人才培养项目(No：2019HB102)；云南省科技厅科技计划项目(No：202001AY070001-170)

详细信息

通讯作者: 马静，E-mail: majing@etyy.cn

中图分类号: R764.3

收稿日期: 2023-06-19

修回日期: 2023-09-13

刊出日期: 2023-11-03

Analysis of 59 cases of large vestibular aqueduct syndrome SLC26A4 gene mutation frequency and new mutation sites

- Department of Otorhinolaryngology Head and Neck Surgery, Kunming Children's Hospital, Kunming Key Laboratory for Prevention and Control of Congenital Birth Defects of Children, Yunnan Key Laboratory of Children's Major Disease Research, Kunming, 650228, China

More Information

Corresponding author: MA Jing, E-mail: majing@etyy.cn

Received Date: 19 June 2023

Revised Date: 13 September 2023

Publish Date: 03 November 2023

摘要

摘要: 目的研究云南地区前庭导水管扩大耳聋患儿SLC26A4基因突变位点频率，报道SLC26A4基因新发突变位点，进一步明确SLC26A4基因突变谱，探讨SLC26A4基因的双等位、单等位基因突变与内耳CT表型的关联，为耳聋的临床和基因诊断提供依据。方法回顾2016年8月至2021年9月昆明市儿童医院耳鼻喉科390例人工耳蜗术后患儿颞骨CT检查结果，对59例发现前庭导水管扩大的患儿进行对SLC26A4基因Sanger测序，并将行颞骨CT检查的患儿按照基因检测结果分为SLC26A4双等位基因突变组(纯合突变和复合杂合突变)、单等位基因突变组，分析与内耳CT表型的关联，并对新发位点进行总结分析。结果在390例人工耳蜗植入患儿中，发现前庭导水管扩大患儿59例，其中48例(81.4%)检测出SLC26A4基因突变，其中SLC26A4双等位基因突变46例，包括纯合突变16例，复合杂合突变30例，SLC26A4单等位基因突变2例；11例(18.6%)前庭导水管扩大患儿未检测到SLC26A4基因突变。在48例可检测到SLC26A4基因突变的前庭导水管扩大耳聋患儿中，共检出36种SLC26A4基因突变，突变位点以c.919-2A>G最为常见，其次常见位点为c.1174A>T、c.2168A>G。检测发现3个突变位点为国际上尚未报道的突变，分别为c.312_322delATATGCCCTAC 2例，c.304+3A>C 1例，c.100C>T 1例，突变类型分别为移码突变、剪切突变、无义突变，突变位点均经Sanger测序证实。48例前庭导水管扩大患儿颞骨CT检查，大前庭水管综合征(enlarged vestibular aqueduct syndrome，EVAS)合并Mondini畸形33例，单独EVAS 15例，未见单独Mondini畸形。结论伴SLC26A4基因突变的前庭导水管扩大耳聋患儿以c.919-2A>G突变最常见，发现3种SLC26A4基因的新变异；CT检查联合基因检测发现，一部分前庭导水管扩大患儿与SLC26A4单等位基因突变或未检测到SLC26A4基因突变相关，提示有待研究EVAS的其他致病机制。
- SLC26A4基因 /
- 大前庭水管综合征 /
- Mondini畸形 /
- 基因突变
Abstract: Objective To study the frequency of SLC26A4 gene mutation sites in children with enlarged vestibular aqueduct deafness in Yunnan, report the new mutation sites of SLC26A4 gene, further clarify the mutation spectrum of SLC26A4 gene, and explore the association between biallelic and monoallelic mutations of SLC26A4 gene and CT phenotype of inner ear, so as to provide basis for clinical and genetic diagnosis of deafness.Methods Review the results of temporal bone CT examination of 390 children after cochlear implantation in the Department of Otolaryngology, Kunming Children's Hospital from August 2016 to September 2021. Sanger sequencing of SLC26A4 gene was performed in 59 children with enlarged vestibular aqueduct. According to the genetic test results, the children who underwent temporal bone CT examination were divided into two groups: SLC26A4 biallelic mutation group(homozygous mutation and compound heterozygous mutation), monoallelic mutation group, and the association with inner ear CT phenotype was analyzed, and the new sites were summarized and analyzed.Results The c.919-2a>g mutation was the most common mutation in children with enlarged vestibular aqueduct with SLC26A4 gene mutation. Three new variants of SLC26A4 gene were found; CT examination combined with genetic testing found that a part of children with enlarged vestibular aqueduct was associated with SLC26A4 monoallelic mutation or no SLC26A4 gene mutation was detected. Further research is needed to investigate the involvement of other pathogenic factors in the pathogenesis of EVA.
- SLC26A4 gene /
- large vestibular aqueduct syndrome /
- mondini dysplasia /
- gene mutation

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图 1 EVAS+Mondini畸形的轴位CT

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图 2 Mondini畸形的冠状位CT

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图 3 先证者1、2、3、4的家系图

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图 4 家系1 Sange测序图

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图 5 家系2 Sange测序图

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图 6 家系3 Sange测序图

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图 7 家系4 Sange测序图

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表 1 48例SLC26A4基因突变患儿的基因型

内耳畸形	基因型	分类	受试数量
EVAS	c.919-2A>G/c.919-2A>G	Splicing	3
	c.754T>C/c.754T>C	Missense	1
	c.919-2A>G /c.281C>T	Splicing/（p.T94I）	1
	c.919-2A>G/c.1707+5G>A	Splicing	1
	c.919-2A>G /c.311_321del	Splicing/ p.Y105Sfs*73	1
	c.919-2A>G/c.589G>A	Splicing/ p.G197R	1
	c.919-2A>G/ c.1226G>A	Splicing/ p.R409H	1
	c.919-2A>G/c.2027T>A	Splicing/ p.L676Q	1
	c.919-2A>G/c.1433A>T	Splicing/ p.D478V	1
	c.1520delT/c.1262A>C	p.L507X/p.Q421P	1
	c.1173C>A/c.1547dupC	p.S391R /p.S517Ffs*10	1
	c.100C>T；c.2168A>G	p.Gln34*；Splicing	1
	c.2029C>T/ del	p.R677W/-	1
	c.919-2A>G/c.919-2A>G	Splicing	9
EVAS+MD	c.86A>G/ c.86A>G	p.E29G	1
	c.1264-12T>A/ c.1264-12T>A	-	1
	c.1174A>T/ c.1174A>T	p.（Asn392Tyr）	1
	c.919-2A>G/c.2168A>G	Splicing/p.H723R	2
	c.919-2A>G /c.916dupG	Splicing/ p.V306Gfs*24	1
	c.919-2A>G/c.716T>A	Splicing/ p.V239D	1
	c.919-2A>G/c.1433A>T	Splicing/ p.D478V	1
	c.919-2A>G/c.281C>T	Splicing/ p.T94I	1
	c.919-2A>G/c.1519delT	Splicing/ p.L507X	1
	c.919-2A>G/c.1226G>A	Splicing/ p.R409H	1
	c.919-2A>G/c.2027T>A	Splicing/ p.L676Q	1
	c.919-2A>G/c.1520delT	Splicing/ p.L507X	1
	c.919-2A>G/c.2086C>T	Splicing/ p.Q696X	1
	c.1229C>T；c.304+3A>C	p.T410M；splicing	1
	c.2086C>T/c.312_322delATATGCCCTAC	p.Q696X/p.Y105Sfs*73	1
	c.1173C>A/c.2086C>T	p.S391R/ p.Q696X	1
	c.2000T>C/c.2168A>G	p.F667S/p.H723R	1
	c.1520delT/c.1262A>C	p.L507X/p.Q421P	1
	c.1343_1355dupCGGTCTTGGCAGC/c.1174A>T	p.V453Gfs*19/p.N392Y	1
	c.1546dupC/c.2086C>T	p.S517Ffs*10/p.Q696X	1
	c.1173C>A/c.1547dupC	p.S391R/p.S517Ffs*10	1
	c.589G>A/	p.G197R/	1
	c.2162C>T/	p.T721M/	1

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表 2 新发突变位点信息汇总

序号	外显子/内含子	cDNA	Protein	变异分类	致病性	CT表型
先证者1	Exon4；Exon18	c.312_322delATATGCCCTACc.2086C>T	p.Y105Sfs*73；p.Q696X	Frameshift；Nonsense	P；P	E+M
先证者2	Exon4；Intron8	c.312_322delATATGCCCTAC；c.919-2A>G	p.Y105Sfs*73；Splicing	Frameshift；Splicing	P；P	E
先证者3	Exon10；Intron3	c.1229C>T；c.304+3A>C	p.T410M；Splicing	Missense；Splicing	P；U	E+M
先证者4	Exon2；Intron3	c.100C>T；c.2168A>G	p.Gln34*；Splicing	Nonsense；Missense	LP；P	E
P: 致病性; LP: 疑似致病性; U: 临床意义未明。

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参考文献(22)

[1]	陈睿尧, 卢宇, 程静, 等. 湖北481例耳聋患者GJB2、SLC26A4及12S rRNA基因突变分析[J]. 中华耳科学杂志, 2022, 20(5): 761-765. https://www.cnki.com.cn/Article/CJFDTOTAL-ZHER202205010.htm
[2]	李秀萍, 王俊玲, 马蓉, 等. 曲靖地区7246例新生儿耳聋基因筛查结果分析[J]. 中国妇幼保健, 2023, 38(13): 2463-2466. https://www.cnki.com.cn/Article/CJFDTOTAL-ZFYB202313034.htm
[3]	Rose J, Muskett JA, King KA, et al. Hearing loss associated with enlarged vestibular aqueduct and zero or one mutant allele of SLC26A4[J]. Laryngoscope, 2017, 127(7): E238-E243.
[4]	El-Badry MM, Osman NM, Mohamed HM, et al. Evaluation of the radiological criteria to diagnose large vestibular aqueduct syndrome[J]. Int J Pediatr Otorhinolaryngol, 2016, 81: 84-91. doi: 10.1016/j.ijporl.2015.12.012
[5]	Madden C, Halsted M, Meinzen-Derr J, et al. The influence of mutations in the SLC26A4 gene on the temporal bone in a population with enlarged vestibular aqueduct[J]. Arch Otolaryngol Head Neck Surg, 2007, 133(2): 162-168. doi: 10.1001/archotol.133.2.162
[6]	Richards S, Aziz N, Bale S, et al. Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendatio n of the American College of Medical Genetics and Genomics and the Association for Molecular Patholo gy[J]. Genet Med, 2015, 17(5): 405-424. doi: 10.1038/gim.2015.30
[7]	Valvassori GE, Clemis JD. The large vestibular aqueduct syndrome[J]. Laryngoscope, 1978, 88(5): 723-728. doi: 10.1002/lary.1978.88.5.723
[8]	Yuan Y, Guo W, Tang J, et al. Molecular epidemiology and functional assessment of novel allelic variants of SLC26A4 in non-syndromi c hearing loss patients with enlarged vestibular aqueduct in China[J]. PLoS One, 2012, 7(11): e49984. doi: 10.1371/journal.pone.0049984
[9]	赵质彬, 祝园平, 符一飞, 等. 前庭水管扩大患者颞骨高分辨率CT和SLC26A4基因及听力损失的相关性研究[J]. 临床耳鼻咽喉头颈外科杂志, 2022, 36(10): 736-740. https://lceh.whuhzzs.com/article/doi/10.13201/j.issn.2096-7993.2022.10.002
[10]	Anwar S, Riazuddin S, Ahmed ZM, et al. SLC26A4 mutation spectrum associated with DFNB4 deafness and Pendred's syndrome in Pakistanis[J]. J Hum Genet, 2009, 54(5): 266-270. doi: 10.1038/jhg.2009.21
[11]	Chen N, Tranebjærg L, Rendtorff ND, et al. Mutation analysis of SLC26A4 for Pendred syndrome and nonsyndromic hearing loss by high-resolution melting[J]. J Mol Diagn, 2011, 13(4): 416-426. doi: 10.1016/j.jmoldx.2011.03.003
[12]	Wu L, Liu Y, Wu J, et al. Study on the relationship between the pathogenic mutations of SLC26A4 and CT phenotypes of inner ear in patient with sensorineural hearing loss[J]. Biosci Rep, 2019, 39(3): BSR20182241. doi: 10.1042/BSR20182241
[13]	孙宝春, 周成勇, 代志瑶. SLC26A4基因检测在儿童先天性内耳畸形早期诊断中的作用[J]. 临床耳鼻咽喉头颈外科杂志, 2014, 28(22): 1741-1744. https://lceh.whuhzzs.com/article/doi/10.13201/j.issn.1001-1781.2014.22.004
[14]	赵晓云, 胡健, 仵倩, 等. 人工耳蜗植入患者常见聋病相关基因突变与内耳畸形的相关性分析[J]. 中华耳科学杂志, 2019, 17(5): 643-649. https://www.cnki.com.cn/Article/CJFDTOTAL-ZHER201905007.htm
[15]	薛文悦, 陈正侬. SLC26A4基因突变致前庭水管扩大听力损失机制的研究进展[J]. 中华耳科学杂志, 2019, 17(5): 768-772. https://www.cnki.com.cn/Article/CJFDTOTAL-ZHER201905029.htm
[16]	Ito T, Muskett J, Chattaraj P, et al. SLC26A4 mutation testing for hearing loss associated with enlargement of the vestibular aqueduct[J]. World J Otorhinolaryngol, 2013, 3(2): 26-34.
[17]	于晓宇, 林妘, 许军, 等. 135例大前庭导水管耳聋患者SLC26A4基因突变分析[J]. 中华耳科学杂志, 2018, 16(2): 160-164. https://www.cnki.com.cn/Article/CJFDTOTAL-ZHER201802008.htm
[18]	刘亚青, 黄正华, 孙晨, 等. SLC26A4基因在大前庭水管综合征和/或Mondini畸形患儿中突变频率的观察[J]. 临床耳鼻咽喉头颈外科杂志, 2021, 35(10): 891-895. https://lceh.whuhzzs.com/article/doi/10.13201/j.issn.2096-7993.2021.10.006
[19]	Smits JJ, de Bruijn SE, Lanting CP, et al. Exploring the missing heritability in subjects with hearing loss, enlarged vestibular aqueducts, and a single or no pathogenic SLC26A4 variant[J]. Hum Genet, 2022, 141(3-4): 465-484.
[20]	Mey K, Muhamad AA, Tranebjaerg L, et al. Association of SLC26A4 mutations, morphology, and hearing in pendred syndrome and NSEVA[J]. Laryngoscope, 2019, 129(11): 2574-2579.
[21]	Yang T, Gurrola JG, Wu H, et al. Mutations of KCNJ10 together with mutations of SLC26A4 cause digenic nonsyndromic hearing loss associ ated with enlarged vestibular aqueduct syndrome[J]. Am J Hum Genet, 2009, 84(5): 651-657.
[22]	Zhao J, Yuan Y, Huang S, et al. KCNJ10 may not be a contributor to nonsyndromic enlargement of vestibular aqueduct(NSEVA)in Chinese subjects[J]. PLoS One, 2014, 9(11): e108134.