MicroRNA-mediated dysregulation of neural developmental genes in HPRT deficiency: clues for Lesch-Nyhan disease?
MicroRNA-mediated dysregulation of neural developmental genes in HPRT deficiency: clues for Lesch-Nyhan disease?
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DOI：
10.1093/hmg/ddr495
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发表时间：
2012-02-01
影响因子：
3.5
通讯作者：
Friedmann, Theodore
中科院分区：
文献类型：
--
作者：
Guibinga, Ghiabe-Henri;Hrustanovic, Gorjan;Friedmann, Theodore
编码嘌呤生物合成酶次黄嘌呤-鸟嘌呤磷酸核糖转移酶（HPRT）的基因突变导致与脑多巴胺通路异常发育相关的难治性神经发育性莱施-尼汉病（LND）。在目前的研究中，我们已经确定了在HPRT缺陷的人多巴胺能SH-SY 5 Y神经母细胞瘤细胞中microRNA miR 181 a的表达增加。在miR 181 a可能调控的基因中，有几个已知是神经发育所需的基因，包括Engrailed 1（En 1）、Engrailed 2（En 2）、Lmx 1a和Brn 2。我们证明这些基因在HPRT缺陷的SH-SY 5 Y细胞中下调，并且miR 181 a的过表达显著降低了这些基因的内源性表达，并抑制了携带En 1/2或Lmx 1a 3' UTR miRNA结合元件的荧光素酶质粒的翻译。相反，miR 181 a的抑制增加了这些基因的表达，并增强了带有En 1/2和Lmx 1a 3' UTR miRNA结合序列的荧光素酶构建体的翻译。我们还证明了关键的神经发育基因（e。G. Nurr 1、Pitx 3、Wnt 1和Mash 1）在过表达miR 181 a的SH-SY 5 Y细胞和HPRT缺陷细胞中也显著下调。我们在SH-SY 5 Y细胞中的发现表明，HPRT缺陷伴随着调节多巴胺能神经元发育的一些重要途径和多巴胺途径的失调，并且这种缺陷与miR 181 a的异常表达相关，并且可能至少部分是由于miR 181 a的异常表达。由于miR 181 a的异常表达在HPRT缺陷的LND成纤维细胞中并不明显，因此SH-SY 5 Y神经母细胞瘤细胞与人类疾病的相关性仍有待证实。然而，我们认为miR 181 a的这些多效性神经发育效应可能在LND的发病机制中发挥作用。
Mutations in the gene encoding the purine biosynthetic enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) cause the intractable neurodevelopmental Lesch-Nyhan disease (LND) associated with aberrant development of brain dopamine pathways. In the current study, we have identified an increased expression of the microRNA miR181a in HPRT-deficient human dopaminergic SH-SY5Y neuroblastoma cells. Among the genes potentially regulated by miR181a are several known to be required for neural development, including Engrailed1 (En1), Engrailed2 (En2), Lmx1a and Brn2. We demonstrate that these genes are down-regulated in HPRT-deficient SH-SY5Y cells and that over-expression of miR181a significantly reduces endogenous expression of these genes and inhibits translation of luciferase plasmids bearing the En1/2 or Lmx1a 3' UTR miRNA-binding elements. Conversely, inhibition of miR181a increases the expression of these genes and enhances translation of luciferase constructs bearing the En1/2 and Lmx1a 3' UTR miRNA-binding sequences. We also demonstrate that key neurodevelopmental genes (e. g. Nurr1, Pitx3, Wnt1 and Mash1) known to be functional partners of Lmx1a and Brn2 are also markedly down-regulated in SH-SY5Y cells over-expressing miR181a and in HPRT-deficient cells. Our findings in SH-SY5Y cells demonstrate that HPRT deficiency is accompanied by dysregulation of some of the important pathways that regulate the development of dopaminergic neurons and dopamine pathways and that this defect is associated with and possibly due at least partly to aberrant expression of miR181a. Because aberrant expression of miR181a is not as apparent in HPRT-deficient LND fibroblasts, the relevance of the SH-SY5Y neuroblastoma cells to human disease remains to be proven. Nevertheless, we propose that these pleiotropic neurodevelopment effects of miR181a may play a role in the pathogenesis of LND.