Identification and characterization of variants and a novel 4 bp deletion in the regulatory region of SIX6, a risk factor for primary open-angle glaucoma.
Identification and characterization of variants and a novel 4 bp deletion in the regulatory region of SIX6, a risk factor for primary open-angle glaucoma.
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DOI：
10.1002/mgg3.290
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发表时间：
2017-07
影响因子：
2
通讯作者：
Sundaresan P
中科院分区：
文献类型：
--
作者：
Shah MH;Tabanera N;Krishnadas SR;Pillai MR;Bovolenta P;Sundaresan P
原发性开角型青光眼（POAG）是一种多基因遗传的复杂疾病，也是最常见的青光眼亚型。SIX 6编码一种参与视网膜、视神经和垂体发育的转录因子。先前的研究表明SIX 6基因座和POAG之间存在遗传关联，确定了风险等位基因。这些等位基因是否也存在于南印度人群中尚不清楚。为了解决这个问题，SIX 6基因和一个已经表征和高度保守的SIX 6增强子（Ch 14：60974427 - 60974430）分别在两个南印度队列中进行了测序，这两个队列分别由65/65和200/200例POAG病例/年龄匹配的对照组成。接下来，我们使用基于Taqman的等位基因区分试验对两个队列中的常见变异（rs33912345：c.421A>C）和rs 1048372 SNP进行基因分型，分别由557/387和590/448例POAG病例/年龄匹配的对照组成。随后招募了另外一个包含153名POAG病例的队列，使用谱域光学相干断层扫描来评估rs33912345：c.421A>C和rs 10483727变体与两个POAG诊断参数（视网膜神经纤维层厚度和垂直杯/盘比）更显着变化的相关性。新鉴定的增强子变体的活性通过在斑马鱼中的转基因和荧光素酶测定来评估。我们确定了两个已知的罕见和两个常见的SIX 6基因座的变体和一个新的4 bp缺失的分析增强子。与先前的研究相反，我们无法在南印度种族中建立rs 10483727和rs33912345：c.421A>C变异体与PAOG之间的显著关联，但携带相应C或T风险等位基因的患者显示出视网膜神经纤维层厚度的剂量依赖性降低和垂直杯/盘比的显著增加。在斑马鱼和荧光素酶测定中的转基因表明，新鉴定的4 bp缺失显著降低了视网膜神经节和无长突层细胞中的报告基因表达，其中表达人SIX 6。总之，我们的数据进一步支持SIX 6变异作为POAG危险因素的含义，并暗示SIX 6单倍不足在POAG发病机制中的作用。
Primary open‐angle glaucoma (POAG) is a complex disease of multigenic inheritance and the most common subtype of glaucoma. SIX6 encodes a transcription factor involved in retina, optic nerve, and pituitary development. Previous studies showed a genetic association between the SIX6 locus and POAG, identifying risk alleles. Whether these alleles are present also in the south Indian population is unclear. To address this question, the SIX6 gene and an already characterized and highly conserved SIX6 enhancer (Ch14:60974427‐60974430) were sequenced in two south Indian cohorts, respectively, composed of 65/65 and 200/200 POAG cases/age‐matched controls. We next used Taqman‐based allelic discrimination assay to genotype a common variant (rs33912345: c.421A>C) and the rs1048372 SNP in two cohorts, respectively, composed of 557/387 and 590/448 POAG cases/age‐matched controls. An additional cohort of 153 POAG cases was subsequently recruited to assess the association of the rs33912345:c.421A>C and rs10483727 variants with more prominent changes in two POAG diagnostic parameters: retinal nerve fiber layer thickness and vertical cup/disc ratio, using spectral domain optical coherence tomography. The activity of the newly identified enhancer variants was assessed by transgenesis in zebrafish and luciferase assays. We identified two known rare and two common variants in the SIX6 locus and a novel 4 bp deletion in the analyzed enhancer. Contrary to previous studies, we could not establish a significant association between the rs10483727 and rs33912345:c.421A>C variants and PAOG in the south Indian ethnicity but patients carrying the corresponding C or T risk alleles exhibited a dose‐dependent reduction of the thickness of the retinal nerve fiber layer and a significant increase in the vertical cup/disc ratio. Transgenesis in zebrafish and luciferase assays demonstrated that the newly identified 4 bp deletion significantly reduced reporter expression in cells of the retinal ganglion and amacrine layers, where human SIX6 is expressed. Altogether, our data further support the implication of SIX6 variants as POAG risk factors and implicates SIX6 haploinsufficiency in POAG pathogenesis.
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影响因子：
158.5
作者：
Knowler, WC;Barrett-Connor, E;Nathan, DM
通讯作者：
Nathan, DM
影响因子：
4.2
作者：
Kuo JZ;Zangwill LM;Medeiros FA;Liebmann JM;Girkin CA;Hammel N;Rotter JI;Weinreb RN
通讯作者：
Weinreb RN
影响因子：
30.8
作者：
Gharahkhani, Puya;Burdon, Kathryn P.;Fogarty, Rhys;Sharma, Shiwani;Hewitt, Alex W.;Martin, Sarah;Law, Matthew H.;Cremin, Katie;Bailey, Jessica N. Cooke;Loomis, Stephanie J.;Pasquale, Louis R.;Haines, Jonathan L.;Hauser, Michael A.;Viswanathan, Ananth C.;McGuffin, Peter;Topouzis, Fotis;Foster, Paul J.;Graham, Stuart L.;Casson, Robert J.;Chehade, Mark;White, Andrew J.;Zhou, Tiger;Souzeau, Emmanuelle;Landers, John;Fitzgerald, Jude T.;Klebe, Sonja;Ruddle, Jonathan B.;Goldberg, Ivan;Healey, Paul R.;Mills, Richard A.;Wang, Jie Jin;Montgomery, Grant W.;Martin, Nicholas G.;Radford-Smith, Graham;Whiteman, David C.;Brown, Matthew A.;Wiggs, Janey L.;Mackey, David A.;Mitchell, Paul;MacGregor, Stuart;Craig, Jamie E.
通讯作者：
Craig, Jamie E.
影响因子：
4.6
作者：
Conte, Ivan;Marco-Ferreres, Raquel;Bovolenta, Paola
通讯作者：
Bovolenta, Paola
影响因子：
4.4
作者：
Cheng, Ching-Yu;Allingham, R. Rand;Wong, Tien Yin
通讯作者：
Wong, Tien Yin