WT1 regulates expression of DNA repair gene Neil3 during nephrogenesis.

IF 3.7 2区 医学 Q1 PHYSIOLOGY American Journal of Physiology-renal Physiology Pub Date : 2023-03-01 DOI:10.1152/ajprenal.00207.2022
Kyle Dickinson, Leah Hammond, Murielle Akpa, Lee Lee Chu, Caleb Tse Lalonde, Alexandre Goumba, Paul Goodyer
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Abstract

Mammalian nephrons arise from a population of nephron progenitor cells (NPCs) expressing the master transcription factor Wilms tumor-1 (WT1), which is crucial for NPC proliferation, migration, and differentiation. In humans, biallelic loss of WT1 precludes nephrogenesis and leads to the formation of Wilms tumor precursor lesions. We hypothesize that WT1 normally primes the NPC for nephrogenesis by inducing expression of NPC-specific DNA repair genes that protect the genome. We analyzed transcript levels for a panel of DNA repair genes in embryonic day 17.5 (E17.5) versus adult mouse kidneys and noted seven genes that were increased >20-fold. We then isolated Cited1+ NPCs from E17.5 kidneys and found that only one gene, nei-like DNA glycosylase 3 (Neil3), was enriched. RNAscope in situ hybridization of E17.5 mouse kidneys showed increased Neil3 expression in the nephrogenic zone versus mature nephron structures. To determine whether Neil3 expression is WT1 dependent, we knocked down Wt1 in Cited1+ NPCs (60% knockdown efficiency) and noted a 58% reduction in Neil3 transcript levels. We showed that WT1 interacts with the Neil3 promoter and that activity of a Neil3 promoter-reporter vector was increased twofold in WT1+ versus WT1- cells. We propose that Neil3 is a WT1-dependent DNA repair gene expressed at high levels in Cited1+ NPCs, where it repairs mutational injury to the genome during nephrogenesis. NEIL3 is likely just one of many such lineage-specific repair mechanisms that respond to genomic injury during kidney development.NEW & NOTEWORTHY We studied the molecular events leading to Wilms tumors as a model for the repair of genomic injury. Specifically, we showed that WT1 activates DNA repair gene Neil3 in nephron progenitor cells. However, our observations offer a much broader principle, demonstrating that the embryonic kidney invests in lineage-specific expression of DNA repair enzymes. Thus, it is conceivable that failure of these mechanisms could lead to a variety of "sporadic" congenital renal malformations and human disease.

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在肾形成过程中,WT1调控DNA修复基因Neil3的表达。
哺乳动物的肾元来源于一群表达主转录因子Wilms tumor-1 (WT1)的肾元祖细胞(NPC),该因子对NPC的增殖、迁移和分化至关重要。在人类中,WT1双等位基因的缺失阻止了肾形成,并导致肾母细胞瘤前体病变的形成。我们假设WT1通常通过诱导NPC特异性DNA修复基因的表达来启动NPC的肾形成。我们分析了胚胎17.5天(E17.5)与成年小鼠肾脏的一组DNA修复基因的转录水平,并注意到7个基因增加了>20倍。然后,我们从E17.5的肾脏中分离出Cited1+ npc,发现只有一个基因富集,即nei-like DNA糖基酶3 (Neil3)。E17.5小鼠肾脏的RNAscope原位杂交显示,与成熟肾元结构相比,肾原区Neil3表达增加。为了确定Neil3的表达是否依赖于WT1,我们在Cited1+ npc中敲除了WT1(敲除效率为60%),并注意到Neil3转录水平降低了58%。我们发现WT1与Neil3启动子相互作用,并且与WT1-细胞相比,WT1+细胞中Neil3启动子报告载体的活性增加了两倍。我们认为Neil3是一种依赖wt1的DNA修复基因,在Cited1+ npc中高水平表达,在肾脏形成过程中修复基因组的突变损伤。NEIL3可能只是肾脏发育过程中对基因组损伤作出反应的许多谱系特异性修复机制之一。我们研究了导致Wilms肿瘤的分子事件,作为基因组损伤修复的模型。具体来说,我们发现WT1激活了肾元祖细胞中的DNA修复基因Neil3。然而,我们的观察提供了一个更广泛的原理,证明胚胎肾脏投资于DNA修复酶的谱系特异性表达。因此,可以想象,这些机制的失败可能导致各种“散发性”先天性肾脏畸形和人类疾病。
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来源期刊
CiteScore
8.40
自引率
7.10%
发文量
154
审稿时长
2-4 weeks
期刊介绍: The American Journal of Physiology - Renal Physiology publishes original manuscripts on timely topics in both basic science and clinical research. Published articles address a broad range of subjects relating to the kidney and urinary tract, and may involve human or animal models, individual cell types, and isolated membrane systems. Also covered are the pathophysiological basis of renal disease processes, regulation of body fluids, and clinical research that provides mechanistic insights. Studies of renal function may be conducted using a wide range of approaches, such as biochemistry, immunology, genetics, mathematical modeling, molecular biology, as well as physiological and clinical methodologies.
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