Identification of hub molecules of FUS-ALS by Bayesian gene regulatory network analysis of iPSC model: iBRN
Identification of hub molecules of FUS-ALS by Bayesian gene regulatory network analysis of iPSC model: iBRN
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DOI：
10.1016/j.nbd.2021.105364
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发表时间：
2020-05
影响因子：
6.1
通讯作者：
M. Nogami;Mitsuru Ishikawa;Atsushi Doi;Osamu Sano;T. Sone;T. Akiyama;M. Aoki;Atsushi Nakanishi;K. Ogi;M. Yano;H. Okano
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文献类型：
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作者：
M. Nogami;Mitsuru Ishikawa;Atsushi Doi;Osamu Sano;T. Sone;T. Akiyama;M. Aoki;Atsushi Nakanishi;K. Ogi;M. Yano;H. Okano
融合于肉瘤/翻译于脂肪肉瘤（FUS）是肌萎缩侧索硬化症（ALS）的致病基因。突变的FUS引起DNA损伤的积累和胞质应激颗粒（SG）的形成，从而导致运动神经元（MN）死亡。然而，关键的分子病因仍不清楚。在这里，我们应用了一种新的平台技术，iBRN，基于诱导多能干细胞（iPSC）衍生的细胞模型的“无偏“贝叶斯基因调控网络分析，以使用携带FUSH 517 D的iPSC衍生的MN的转录组来阐明分子病因。iBRN揭示了影响转录组网络的“枢纽分子”，如miR-125 b-5 p-TIMELESS轴和PRKDC等。接下来，我们在FUSH 517 DMNs中证实了miR-125 b-5 p-TIMELESS轴，因此miR-125 b-5 p调节了包括TIMELESS在内的几个DNA修复相关基因。此外，我们在细胞培养模型中验证了miR-125 b-5 p的引入和TIMELESS的敲低均引起DNA损伤。此外，PRKDC与在受损的DNA-PK活性下通过DNA损伤导致的FUS错误定位到SG中密切相关。总的来说，我们的iBRN策略为阐明神经退行性疾病的分子病因提供了第一个令人信服的证据。
Fused in sarcoma/translated in liposarcoma (FUS) is a causative gene of amyotrophic lateral sclerosis (ALS). Mutated FUS causes accumulation of DNA damage and cytosolic stress granule (SG) formation, thereby motor neuron (MN) death. However, key molecular aetiology remains unclear. Here, we applied a novel platform technology, iBRN, “Non- biased”Bayesian gene regulatorynetwork analysis based oninduced pluripotent stem cell (iPSC)-derived cell model, to elucidate the molecular aetiology using transcriptome of iPSC-derived MNs harboring FUSH517D. iBRN revealed “hub molecules”, which strongly influenced transcriptome network, such as miR-125b-5p-TIMELESSaxis andPRKDCfor the molecular aetiology. Next, we confirmed miR-125b-5p-TIMELESS axis inFUSH517DMNs such that miR-125b-5p regulated several DNA repair-related genes includingTIMELESS. In addition, we validated both introduction of miR-125b-5p and knocking down ofTIMELESScaused DNA damage in the cell culture model. Furthermore, PRKDC was strongly associated with FUS mis-localization into SGs by DNA damage under impaired DNA-PK activity. Collectively, our iBRN strategy provides the first compelling evidence to elucidate molecular aetiology in neurodegenerative diseases.