Serpin neuropathology in the P497S UBQLN2 mouse model of ALS/FTD.
Serpin neuropathology in the P497S UBQLN2 mouse model of ALS/FTD.
复制标题
肌萎缩侧索硬化症/FTD P497S UBQLN2小鼠模型的Serpin神经病理学。
DOI：
10.1111/bpa.12948
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发表时间：
2021-09
期刊：
影响因子：
--
通讯作者：
Monteiro MJ
中科院分区：
文献类型：
--
作者：
Higgins NR;Greenslade JE;Wu JJ;Miranda E;Galliciotti G;Monteiro MJ
越来越多的证据表明，UBQLN2的X连锁显性突变通过功能丧失和功能获得机制导致肌萎缩侧索硬化症（ALS）伴额颞叶痴呆（FTD）。然而，突变导致疾病的机制仍不清楚。该研究的目的是揭示UBQLN2突变导致ALS/FTD的可能病理机制。通过分析神经元组织中的蛋白质组学变化，鉴定P497S UBQLN2转基因ALS/FTD小鼠模型中积累改变的蛋白质。然后，我们使用免疫细胞化学和生化技术来确认突变P497S小鼠的蛋白质变化。此外，我们使用UBQLN2表达失活的细胞系来确定其缺失是否导致P497S小鼠中所见的蛋白改变。蛋白质组筛选鉴定出突变体P497S动物中丝氨酸蛋白酶抑制剂（serpin）蛋白的显著改变。突变小鼠和对照小鼠的脑和脊髓组织的双重免疫荧光染色显示，突变P497S小鼠的斑点中Serpin A1、C1和I1的积累存在年龄依赖性变化，其染色与UBQLN2斑点共定位。通过生化提取和过滤延迟实验证实了P497S动物中Serpin A1的聚集。在UBQLN2表达失活的HeLa和NSC34运动神经元细胞中也发现了类似的serpin蛋白聚集现象。我们在ALS/FTD的P497S UBQLN2小鼠模型中发现了serpin蛋白的异常聚集，特别是serpin A1。在UBQLN2敲除细胞中发现了类似的蛇形蛋白聚集，这表明突变P497S动物的蛇形蛋白聚集可能源于UBQLN2功能的丧失。由于已知serpin聚集通过功能丧失和功能获得机制引起疾病，我们推测它们在P497S ALS/FTD小鼠模型中的积累可能通过类似机制促进疾病发病。我们发现，在ALS/FTD的P497S UBQLN2小鼠模型中，serpin蛋白，特别是serpin A1，与UBQLN2包裹体在脑和脊髓中异常积累和共聚集。在UBQLN2敲除细胞中发现了类似的蛇形蛋白聚集，这表明蛇形蛋白聚集可能源于UBQLN2功能的丧失。已知Serpin聚集通过功能获得和功能丧失机制引起疾病，我们推测它们在P497S小鼠中的积累可能通过类似机制促进疾病发病。
Accumulating evidence suggests X‐linked dominant mutations in UBQLN2 cause amyotrophic lateral sclerosis (ALS) with frontotemporal dementia (FTD) through both loss‐ and gain‐of‐function mechanisms. However, the mechanisms by which the mutations cause disease are still unclear. The goal of the study was to uncover the possible pathomechanism(s) by which UBQLN2 mutations cause ALS/FTD. An analysis of proteomic changes in neuronal tissue was used to identify proteins with altered accumulation in the P497S UBQLN2 transgenic mouse model of ALS/FTD. We then used immunocytochemistry and biochemical techniques to confirm protein changes in the mutant P497S mice. Additionally, we used cell lines inactivated of UBQLN2 expression to determine whether its loss underlies the alteration in the proteins seen in P497S mice. The proteome screen identified a dramatic alteration of serine protease inhibitor (serpin) proteins in the mutant P497S animals. Double immunofluorescent staining of brain and spinal cord tissues of the mutant and control mice revealed an age‐dependent change in accumulation of Serpin A1, C1, and I1 in puncta whose staining colocalized with UBQLN2 puncta in the mutant P497S mice. Serpin A1 aggregation in P497S animals was confirmed by biochemical extraction and filter retardation assays. A similar phenomenon of serpin protein aggregation was found in HeLa and NSC34 motor neuron cells with inactivated UBQLN2 expression. We found aberrant aggregation of serpin proteins, particularly Serpin A1, in the brain and spinal cord of the P497S UBQLN2 mouse model of ALS/FTD. Similar aggregation of serpin proteins was found in UBQLN2 knockout cells suggesting that serpin aggregation in the mutant P497S animals may stem from loss of UBQLN2 function. Because serpin aggregation is known to cause disease through both loss‐ and gain‐of‐function mechanisms, we speculate that their accumulation in the P497S mouse model of ALS/FTD may contribute to disease pathogenesis through similar mechanism(s). We found aberrant accumulation and co‐aggregation of serpin proteins, particularly Serpin A1, with UBQLN2 inclusions in the brain and spinal cord of the P497S UBQLN2 mouse model of ALS/FTD. Similar aggregation of serpin proteins was found in UBQLN2 knockout cells suggesting serpin aggregation may stem from a loss of UBQLN2 function. Serpin aggregation is known to cause disease through both gain‐ and loss‐of‐function mechanisms, and we speculate that their accumulation in P497S mice may contribute to disease pathogenesis through similar mechanisms.
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影响因子：
4.8
作者：
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DOI：
10.1073/pnas.1603386113
发表时间：
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