Identification of two novel CAKUT-causing genes by massively parallel exon resequencing of candidate genes in patients with unilateral renal agenesis.
Identification of two novel CAKUT-causing genes by massively parallel exon resequencing of candidate genes in patients with unilateral renal agenesis.
复制标题
通过大量平行外显子在单侧肾脏发育不全的患者中重新平行候选基因来鉴定两种新型Cakut引起的基因。
DOI：
10.1038/ki.2011.315
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发表时间：
2012-01
影响因子：
19.6
通讯作者：
中科院分区：
文献类型：
--
作者：
先天性肾脏和尿路异常(CAKUT)是儿童慢性肾脏疾病最常见的病因，约占所有病例的50%。虽然许多形式的CAKUT可能是由单基因缺陷引起的，但只有很少的致病基因被发现。由于CAKUT广泛的遗传位点异质性，需要对许多候选基因进行分析以寻找新的致病基因。因此，我们应用我们新开发的DNA池与连续大规模平行外显子重测序的方法来克服这个问题。我们汇集了20个个体的DNA，通过聚合酶链式反应扩增了30个CAKUT候选基因的全部313个外显子。然后对扩增产物进行大规模平行外显子重测序。用桑格测序鉴定突变携带者。我们重复了这项实验，总共覆盖了40名患者(29名单侧肾发育不全和11名其他CAKUT表型)。我们在2个以前未涉及人类非综合征CAKUT的候选基因中检测到5个杂合性错义突变，4个FRAS1基因突变和1个FREM2突变。在96名健康对照组中，所有的突变都不存在，其PolyPhen评分为>；1.4(“可能具有破坏性”)。FRAS1和FREM2的隐性截断突变已知会导致人类和小鼠的弗雷泽综合征，而杂合子携带者的表型尚未被描述。我们在此发现FRAS1和FREM2杂合错义突变是人类非综合征CAKUT的新原因。
Congenital abnormalities of the kidney and urinary tract (CAKUT) constitute the most frequent cause of chronic kidney disease in children, accounting for ~50% of all cases. Although many forms of CAKUT are likely caused by single-gene defects, only few causative genes have been identified. To identify new causative genes many candidate genes need to be analyzed due to the broad genetic locus heterogeneity of CAKUT. We therefore applied our newly developed approach of DNA pooling with consecutive massively parallel exon resequencing to overcome this problem. We pooled DNA of 20 individuals and amplified by PCR all 313 exons of 30 CAKUT candidate genes. PCR products were then subjected to massively parallel exon resequencing. Mutation carriers were identified using Sanger sequencing. We repeated the experiment to cover 40 patients in total (29 with unilateral renal agenesis and 11 with other CAKUT phenotypes). We detected 5 heterozygous missense mutations in 2 candidate genes that were not previously implicated in non-syndromic CAKUT in humans, 4 mutations in the FRAS1 gene and 1 in FREM2. All mutations were absent from 96 healthy control individuals and had a PolyPhen score of >1.4 (“possibly damaging”). Recessive truncating mutations in FRAS1 and FREM2 were known to cause Fraser syndrome in humans and mice, whereas a phenotype in heterozygous carriers has not been described. We hereby identify heterozygous missense mutations in FRAS1 and FREM2 as a new cause of non-syndromic CAKUT in human.
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影响因子：
14.9
作者：
Ramensky, V;Bork, P;Sunyaev, S
通讯作者：
Sunyaev, S
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Woolf AS
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Vrontou, S;Petrou, P;Chalepakis, G
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Chalepakis, G
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Otto EA;Ramaswami G;Janssen S;Chaki M;Allen SJ;Zhou W;Airik R;Hurd TW;Ghosh AK;Wolf MT;Hoppe B;Neuhaus TJ;Bockenhauer D;Milford DV;Soliman NA;Antignac C;Saunier S;Johnson CA;Hildebrandt F;GPN Study Group
通讯作者：
GPN Study Group
影响因子：
30.8
作者：
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通讯作者：
Scambler, PJ