Epigenetic Small Molecules Rescue Nucleocytoplasmic Transport and DNA Damage Phenotypes in C9ORF72 ALS/FTD.
Epigenetic Small Molecules Rescue Nucleocytoplasmic Transport and DNA Damage Phenotypes in C9ORF72 ALS/FTD.
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DOI：
10.3390/brainsci11111543
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发表时间：
2021-11-20
期刊：
影响因子：
3.3
通讯作者：
Zeier Z
中科院分区：
文献类型：
--
作者：
Ramic M;Andrade NS;Rybin MJ;Esanov R;Wahlestedt C;Benatar M;Zeier Z
肌萎缩侧索硬化症 (ALS) 是一种进行性、致命性的神经退行性疾病，现有的治疗方法只能略微减缓病情进展或提高生存率。 C9ORF72 基因中的六核苷酸重复扩展突变是散发性和家族性 ALS 和额颞叶痴呆 (FTD) 病例最常见的遗传原因。 C9ORF72 扩展突变产生五个二肽重复蛋白 (DPR)，虽然 DPR 介导的神经毒性的机制决定因素仍未完全了解，但有证据表明核细胞质运输的破坏和 DNA 损伤的增加会导致病理。因此，需要表征这些干扰并确定不同 DPR 的相对贡献，以促进 C9ALS/FTD 新型疗法的开发。为此，我们生成了一系列核细胞质运输“生物传感器”，由绿色荧光蛋白（GFP）组成，融合到不同类别的核定位信号（NLS）和核输出信号（NES）。将这些生物传感器与自动显微镜结合使用，我们研究了三种最具神经毒性的 DPR（PR、GR 和 GA）对 7 个核输入和 2 个核输出途径的作用。除了其他 DPR 之外，我们发现 PR 对经典的核输出途径和几种核输入途径具有明显的抑制作用。为了鉴定能够抵消 PR 对核细胞质转运影响的化合物，我们开发了核细胞质转运测定法并筛选了几个市售化合物库，总共 2714 种化合物。除了恢复核细胞质运输效率外，屏幕上的点击还可以抵消 PR 的细胞毒性作用。随后测试了选定的命中物拯救另一种 C9ALS/FTD 表型——持续 DNA 双链断裂的能力。总体而言，我们发现 DPR 会破坏多种核细胞质转运途径，并且我们确定了抵消这些影响的小分子，从而提高了表达 PR 的细胞的活力，并减少了患者来源的运动神经元中的 DNA 损伤标记。几种 HDAC 抑制剂被验证为有效药物，支持了之前的研究，即 HDAC 抑制剂在神经退行性模型中具有治疗作用。
Amyotrophic lateral sclerosis (ALS) is a progressive and fatal neurodegenerative disease with available treatments only marginally slowing progression or improving survival. A hexanucleotide repeat expansion mutation in the C9ORF72 gene is the most commonly known genetic cause of both sporadic and familial cases of ALS and frontotemporal dementia (FTD). The C9ORF72 expansion mutation produces five dipeptide repeat proteins (DPRs), and while the mechanistic determinants of DPR-mediated neurotoxicity remain incompletely understood, evidence suggests that disruption of nucleocytoplasmic transport and increased DNA damage contributes to pathology. Therefore, characterizing these disturbances and determining the relative contribution of different DPRs is needed to facilitate the development of novel therapeutics for C9ALS/FTD. To this end, we generated a series of nucleocytoplasmic transport “biosensors”, composed of the green fluorescent protein (GFP), fused to different classes of nuclear localization signals (NLSs) and nuclear export signals (NESs). Using these biosensors in conjunction with automated microscopy, we investigated the role of the three most neurotoxic DPRs (PR, GR, and GA) on seven nuclear import and two export pathways. In addition to other DPRs, we found that PR had pronounced inhibitory effects on the classical nuclear export pathway and several nuclear import pathways. To identify compounds capable of counteracting the effects of PR on nucleocytoplasmic transport, we developed a nucleocytoplasmic transport assay and screened several commercially available compound libraries, totaling 2714 compounds. In addition to restoring nucleocytoplasmic transport efficiencies, hits from the screen also counteract the cytotoxic effects of PR. Selected hits were subsequently tested for their ability to rescue another C9ALS/FTD phenotype—persistent DNA double strand breakage. Overall, we found that DPRs disrupt multiple nucleocytoplasmic transport pathways and we identified small molecules that counteract these effects—resulting in increased viability of PR-expressing cells and decreased DNA damage markers in patient-derived motor neurons. Several HDAC inhibitors were validated as hits, supporting previous studies that show that HDAC inhibitors confer therapeutic effects in neurodegenerative models.
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影响因子：
16.2
作者：
Ash PE;Bieniek KF;Gendron TF;Caulfield T;Lin WL;Dejesus-Hernandez M;van Blitterswijk MM;Jansen-West K;Paul JW 3rd;Rademakers R;Boylan KB;Dickson DW;Petrucelli L
通讯作者：
Petrucelli L
影响因子：
25
作者：
Chou CC;Zhang Y;Umoh ME;Vaughan SW;Lorenzini I;Liu F;Sayegh M;Donlin-Asp PG;Chen YH;Duong DM;Seyfried NT;Powers MA;Kukar T;Hales CM;Gearing M;Cairns NJ;Boylan KB;Dickson DW;Rademakers R;Zhang YJ;Petrucelli L;Sattler R;Zarnescu DC;Glass JD;Rossoll W
通讯作者：
Rossoll W
影响因子：
14.5
作者：
Bruneteau, Gaelle;Simonet, Thomas;Hantai, Daniel
通讯作者：
Hantai, Daniel
DOI：
10.1093/brain/awu120
发表时间：
2014-07
期刊：
Brain : a journal of neurology
影响因子：
--
作者：
Cooper-Knock J;Walsh MJ;Higginbottom A;Robin Highley J;Dickman MJ;Edbauer D;Ince PG;Wharton SB;Wilson SA;Kirby J;Hautbergue GM;Shaw PJ
通讯作者：
Shaw PJ
影响因子：
4
作者：
Andres-Benito, Pol;Gelpi, Ellen;Ferrer, Isidro
通讯作者：
Ferrer, Isidro