Mutations in GREB1L Cause Bilateral Kidney Agenesis in Humans and Mice
Mutations in GREB1L Cause Bilateral Kidney Agenesis in Humans and Mice
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DOI：
10.1016/j.ajhg.2017.09.026
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发表时间：
2017-11-02
影响因子：
9.8
通讯作者：
Jeanpierre, Cecile
中科院分区：
文献类型：
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作者：
De Tomasi, Lara;David, Pierre;Jeanpierre, Cecile
先天性肾脏和泌尿道异常（CAKUT）是儿童慢性肾脏疾病的主要原因，占产前检测异常的20%。CAKUT包括一系列发育性肾缺陷，包括肾发育不全、发育不全以及囊性和非囊性发育不良。据报道，在受CAKUT影响的病例受试者中有50多个基因发生突变。然而，导致双侧肾发育不全（BKA）的病理生理机制仍然很难理解。对183例不相关的家族性和/或严重CAKUT影响的病例受试者（包括54例BKA胎儿）进行全外显子组或靶向外显子组测序，鉴定出GREB 1 L（乳腺癌1样雌激素生长调节）中的16种杂合变体，GREB 1 L是一种报告为视黄酸信号传导靶点的基因。鉴定出4个功能丧失和12个破坏性错义变体，其中14个缺失于GnomAD。其中12例存在于家族性或单纯BKA感染的病例受试者中。受BKA影响的雌性胎仔也显示子宫发育不全。我们证明了GREB 1 L变异体和BKA之间的显着关联。通过原位杂交，我们发现Greb 1 l在发育中的小鼠肾脏的肾发生区的表达。我们通过CRISPR-Cas9产生了Greb 1 l敲除小鼠模型。在E13.5的分析显示，雄性和雌性Greb 1 l（-/-）胚胎中缺乏肾脏和生殖道异常，Greb 1 l（+/-）胚胎中输尿管芽分支略有减少。我们发现，在mIMCD 3细胞中Greb 1 l失效影响三维胶原培养中的小管形态发生，这是一种通过表达野生型人类蛋白质而挽救的表型。这表明GREB 1 L在小鼠和人类的早期后肾和生殖器发育中起着重要作用。
Congenital anomalies of the kidney and urinary tract (CAKUT) constitute a major cause of chronic kidney disease in children and 20% of prenatally detected anomalies. CAKUT encompass a spectrum of developmental kidney defects, including renal agenesis, hypoplasia, and cystic and non-cystic dysplasia. More than 50 genes have been reported as mutated in CAKUT-affected case subjects. However, the pathophysiological mechanisms leading to bilateral kidney agenesis (BKA) remain largely elusive. Whole-exome or targeted exome sequencing of 183 unrelated familial and/or severe CAKUT-affected case subjects, including 54 fetuses with BKA, led to the identification of 16 heterozygous variants in GREB1L (growth regulation by estrogen in breast cancer 1-like), a gene reported as a target of retinoic acid signaling. Four loss-of-function and 12 damaging missense variants, 14 being absent from GnomAD, were identified. Twelve of them were present in familial or simplex BKA-affected case subjects. Female BKA-affected fetuses also displayed uterus agenesis. We demonstrated a significant association between GREB1L variants and BKA. By in situ hybridization, we showed expression of Greb1l in the nephrogenic zone in developing mouse kidney. We generated a Greb1l knock-out mouse model by CRISPR-Cas9. Analysis at E13.5 revealed lack of kidneys and genital tract anomalies in male and female Greb1l(-/-) embryos and a slight decrease in ureteric bud branching in Greb1l(+/-) embryos. We showed that Greb1l invalidation in mIMCD3 cells affected tubulomorphogenesis in 3D-collagen culture, a phenotype rescued by expression of the wild-type human protein. This demonstrates that GREB1L plays a major role in early metanephros and genital development in mice and humans.