DNA Methyltransferase 1 Controls Nephron Progenitor Cell Renewal and Differentiation.
DNA Methyltransferase 1 Controls Nephron Progenitor Cell Renewal and Differentiation.
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DOI：
10.1681/asn.2018070736
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发表时间：
2018-12
期刊：
影响因子：
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通讯作者：
Nicola Wanner;J. Vornweg;A. Combes;Sean B. Wilson;J. Plappert;G. Rafflenbeul;Victor G. Puelles;Raza-Ur Rahman;T. Liwinski;Saskia Lindner;F. Grahammer;O. Kretz;M. Wlodek;T. Romano;K. Moritz;M. Boerries;H. Busch;Stefan Bonn;M. Little;W. Bechtel-Walz;T. Huber
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文献类型：
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作者：
Nicola Wanner;J. Vornweg;A. Combes;Sean B. Wilson;J. Plappert;G. Rafflenbeul;Victor G. Puelles;Raza-Ur Rahman;T. Liwinski;Saskia Lindner;F. Grahammer;O. Kretz;M. Wlodek;T. Romano;K. Moritz;M. Boerries;H. Busch;Stefan Bonn;M. Little;W. Bechtel-Walz;T. Huber
背景：肾元数量是长期肾功能和心血管风险的主要决定因素。观察性研究表明，母体营养和代谢因素在妊娠期间有助于肾单位禀赋的高度变异性。然而，潜在的分子机制尚不清楚。方法我们使用小鼠模型，包括DNA甲基转移酶（Dnmt1， Dnmt3a和Dnmt3b）敲除小鼠，光学投影断层扫描，肾形成生态位的三维重建，转录组和DNA甲基化分析来表征DNA甲基化在肾脏发育中的作用。结果表明，DNA低甲基化是体外和体内营养性肾脏生长受限的关键特征，DNA甲基转移酶Dnmt1和Dnmt3a在发育中的肾脏肾原区高度富集。肾元祖细胞中Dnmt1的缺失（与Dnmt3a或Dnm3b的缺失相反）模拟了肾脏生长受限的营养模型，并导致出生时肾元数量的大幅减少和肾脏发育不全。在dnmt1缺陷小鼠中，光学投影断层扫描和三维重建发现干细胞龛和祖细胞显著减少。RNA测序分析显示，全局DNA低甲基化干扰祖细胞调控网络，导致对肾形成起始至关重要的基因Wt1及其靶基因Wnt4下调。种系基因、原钙粘蛋白、Rhox基因和内源性逆转录病毒元件的抑制导致IFN靶点和细胞周期进展抑制剂的上调。结论：这些发现表明DNA甲基化是产前肾规划的关键调控事件，可能代表了妊娠期间母亲营养因子与肾单位数量减少之间的根本联系。
BACKGROUND Nephron number is a major determinant of long-term renal function and cardiovascular risk. Observational studies suggest that maternal nutritional and metabolic factors during gestation contribute to the high variability of nephron endowment. However, the underlying molecular mechanisms have been unclear. METHODS We used mouse models, including DNA methyltransferase (Dnmt1, Dnmt3a, and Dnmt3b) knockout mice, optical projection tomography, three-dimensional reconstructions of the nephrogenic niche, and transcriptome and DNA methylation analysis to characterize the role of DNA methylation for kidney development. RESULTS We demonstrate that DNA hypomethylation is a key feature of nutritional kidney growth restriction in vitro and in vivo, and that DNA methyltransferases Dnmt1 and Dnmt3a are highly enriched in the nephrogenic zone of the developing kidneys. Deletion of Dnmt1 in nephron progenitor cells (in contrast to deletion of Dnmt3a or Dnm3b) mimics nutritional models of kidney growth restriction and results in a substantial reduction of nephron number as well as renal hypoplasia at birth. In Dnmt1-deficient mice, optical projection tomography and three-dimensional reconstructions uncovered a significant reduction of stem cell niches and progenitor cells. RNA sequencing analysis revealed that global DNA hypomethylation interferes in the progenitor cell regulatory network, leading to downregulation of genes crucial for initiation of nephrogenesis, Wt1 and its target Wnt4. Derepression of germline genes, protocadherins, Rhox genes, and endogenous retroviral elements resulted in the upregulation of IFN targets and inhibitors of cell cycle progression. CONCLUSIONS These findings establish DNA methylation as a key regulatory event of prenatal renal programming, which possibly represents a fundamental link between maternal nutritional factors during gestation and reduced nephron number.