WNT3 involvement in human bladder exstrophy and cloaca development in zebrafish
WNT3 involvement in human bladder exstrophy and cloaca development in zebrafish
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DOI：
10.1093/hmg/ddv225
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发表时间：
2015-09-15
影响因子：
3.5
通讯作者：
Nordenskjold, Agneta
中科院分区：
文献类型：
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作者：
Korberg, Izabella Baranowska;Hofmeister, Wolfgang;Nordenskjold, Agneta
膀胱外翻是一种严重的先天性泌尿系统畸形，当一个孩子出生时有一个开放的膀胱，是最常见的形式的膀胱外翻-尿道上裂综合征（BEEC）的发病率为1：30,000的白人后裔的儿童。最近的研究表明，WNT基因可能有助于膀胱外翻的病因。在这里，我们评估了WNT通路基因在20例膀胱外翻患者使用大规模平行测序。在WNT 3、WNT 6、WNT 7A、WNT 8B、WNT 10A、WNT 11、WNT 16、FZD 5、LRP 1和LRP 10基因中总共鉴定出13种变异，并预测为潜在的致病基因，其中7种变异是新的。在斑马鱼中进一步评估了在WNT 3（p.Cys91Arg）中具有从头非同义取代的患者中鉴定的一种变体。在斑马鱼中敲除wnt 3后显示出泄殖腔畸形，包括泄殖腔上皮的解体和泄殖腔腔的扩张。我们的研究表明，WNT 3 p.Cys91Arg变体的功能被改变，因为突变WNT 3 RNA的RNA过表达不会导致胚胎致死，如野生型WNT 3 mRNA所见。最后，我们还进一步在410例BEEC病例的DNA样本中筛查了WNT 3基因的突变，并确定了一个额外的突变c.638G> A（p.Gly213Asp），该突变是父系遗传的。总之，我们的数据支持WNT通路基因参与BEEC，并表明WNT 3本身是BEEC的罕见原因。
Bladder exstrophy, a severe congenital urological malformation when a child is born with an open urinary bladder, is the most common form of bladder exstrophy-epispadias complex (BEEC) with an incidence of 1: 30,000 children of Caucasian descent. Recent studies suggest that WNT genes may contribute to the etiology of bladder exstrophy. Here, we evaluated WNT-pathway genes in 20 bladder exstrophy patients using massively parallel sequencing. In total 13 variants were identified in WNT3, WNT6, WNT7A, WNT8B, WNT10A, WNT11, WNT16, FZD5, LRP1 and LRP10 genes and predicted as potentially disease causing, of which seven variants were novel. One variant, identified in a patient with a de novo nonsynonymous substitution in WNT3 (p.Cys91Arg), was further evaluated in zebrafish. Knock down of wnt3 in zebrafish showed cloaca malformations, including disorganization of the cloaca epithelium and expansion of the cloaca lumen. Our study suggests that the function of the WNT3 p.Cys91Arg variant was altered, since RNA overexpression of mutant Wnt3 RNA does not result in embryonic lethality as seen with wild-type WNT3 mRNA. Finally, we also mutation screened the WNT3 gene further in 410 DNA samples from BEEC cases and identified one additional mutation c.638G> A (p.Gly213Asp), which was paternally inherited. In aggregate our data support the involvement of WNT-pathway genes in BEEC and suggest that WNT3 in itself is a rare cause of BEEC.