Genome‐wide identification of microRNAs regulating the human prion protein
Genome‐wide identification of microRNAs regulating the human prion protein
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调节人类朊病毒蛋白的 microRNA 的全基因组鉴定
DOI：
10.1111/bpa.12679
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发表时间：
2018
期刊：
影响因子：
6.4
通讯作者：
A. Aguzzi
中科院分区：
文献类型：
--
作者：
D. Pease;C. Scheckel;Elke Schaper;V. Eckhardt;M. Emmenegger;I. Xenarios;A. Aguzzi
细胞朊病毒蛋白（PrPC）以其错误折叠的致病构象体PrPSc而闻名。由于PrPC的可用性通常限制朊病毒的传播，因此了解其调节可能指向可能的治疗靶点。我们试图确定人类microRNAome通过直接或间接途径参与调节PrPC水平的程度。我们使用稳健的时间分辨荧光共振筛选测定法探测了细胞中的PrPC蛋白水平，该细胞经历了包含2019种miRNA模拟物的全基因组文库。在三种人神经外胚层细胞系中进行筛选：U-251 MG、CHP-212和SH-SY 5 Y。三次筛选产生了17个重叠的高置信度miRNA模拟物命中，其中13个被发现通过与PRNP 3 'UTR结合直接调节PrPC生物合成，从而诱导转录物降解。其余四个命中（miR-124 - 3 p、192 - 3 p、299 - 5 p和376 b-3 p）不结合PRNP的3 'UTR或CDS，因此被认为是PrPC的间接调节剂。我们的研究结果表明，多种miRNA直接和间接地调节PrPC水平。这些发现可能对朊病毒疾病的发病机制及其治疗具有深远的意义。此外，PrPC作为Aβ毒性介质的可能作用表明其通过miRNA的调节也可能影响阿尔茨海默病。
The cellular prion protein (PrPC) is best known for its misfolded disease‐causing conformer, PrPSc. Because the availability of PrPC is often limiting for prion propagation, understanding its regulation may point to possible therapeutic targets. We sought to determine to what extent the human microRNAome is involved in modulating PrPC levels through direct or indirect pathways. We probed PrPC protein levels in cells subjected to a genome‐wide library encompassing 2019 miRNA mimics using a robust time‐resolved fluorescence‐resonance screening assay. Screening was performed in three human neuroectodermal cell lines: U‐251 MG, CHP‐212 and SH‐SY5Y. The three screens yielded 17 overlapping high‐confidence miRNA mimic hits, 13 of which were found to regulate PrPC biosynthesis directly via binding to the PRNP 3’UTR, thereby inducing transcript degradation. The four remaining hits (miR‐124‐3p, 192‐3p, 299‐5p and 376b‐3p) did not bind either the 3’UTR or CDS of PRNP, and were therefore deemed indirect regulators of PrPC. Our results show that multiple miRNAs regulate PrPC levels both directly and indirectly. These findings may have profound implications for prion disease pathogenesis and potentially also for their therapy. Furthermore, the possible role of PrPC as a mediator of Aβ toxicity suggests that its regulation by miRNAs may also impinge on Alzheimer’s disease.
影响因子：
6.7
作者：
Majer A;Medina SJ;Niu Y;Abrenica B;Manguiat KJ;Frost KL;Philipson CS;Sorensen DL;Booth SA
通讯作者：
Booth SA