DNA Damage Increases Secreted Aβ40 and Aβ42 in Neuronal Progenitor Cells: Relevance to Alzheimer's Disease.
DNA Damage Increases Secreted Aβ40 and Aβ42 in Neuronal Progenitor Cells: Relevance to Alzheimer's Disease.
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DOI：
10.3233/jad-220030
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发表时间：
2022
影响因子：
4
通讯作者：
Levine, Arthur Samuel
中科院分区：
文献类型：
--
作者：
Welty, Starr;Thathiah, Amantha;Levine, Arthur Samuel
关键词：
最近的研究表明，在阿尔茨海默病（AD）中，神经元DNA损伤、淀粉样蛋白-β (a β)水平升高和大脑退化区域之间存在密切关联。为了研究这种关联的本质，我们测试了广泛的DNA损伤导致Aβ40和Aβ42代增加的假设。我们使用永生化的人类神经祖细胞系（npc）， ReN VM GA2。用过氧化氢或依托泊苷处理npc或分化20天的神经元，并在指定时间内恢复。采用夹心ELISA法测定Aβ40和Aβ42的分泌量。Western blotting、免疫染色和中性彗星试验用于评估DNA损伤反应和指示AD病理的过程。我们发现，过氧化氢损伤导致ReN GA2 npc在治疗24小时后细胞Aβ40和Aβ42分泌增加。同样，DNA双链断裂（DSB）特异性依托opo苷损伤导致ReN GA2 npc在治疗后2小时和4小时Aβ40和Aβ42分泌增加。与此相反，依托坡苷损伤并未增加有丝分裂后ReN GA2神经元Aβ40和Aβ42的分泌。这些发现提供了证据，在我们的模型中，DNA损伤与神经元祖细胞中Aβ分泌的增加有关，这可能有助于阿尔茨海默病的早期神经元病理。
Recent studies suggest a strong association between neuronal DNA damage, elevated levels of amyloid-β (Aβ), and regions of the brain that degenerate in Alzheimer’s disease (AD). To investigate the nature of this association, we tested the hypothesis that extensive DNA damage leads to an increase in Aβ40 and Aβ42 generation. We utilized an immortalized human neuronal progenitor cell line (NPCs), ReN VM GA2. NPCs or 20 day differentiated neurons were treated with hydrogen peroxide or etoposide and allowed to recover for designated times. Sandwich ELISA was used to assess secreted Aβ40 and Aβ42. Western blotting, immunostaining, and neutral comet assay were used to evaluate the DNA damage response and processes indicative of AD pathology. We determined that global hydrogen peroxide damage results in increased cellular Aβ40 and Aβ42 secretion 24 h after treatment in ReN GA2 NPCs. Similarly, DNA double strand break (DSB)-specific etoposide damage leads to increased Aβ40 and Aβ42 secretion 2 h and 4 h after treatment in ReN GA2 NPCs. In contrast, etoposide damage does not increase Aβ40 and Aβ42 secretion in post-mitotic ReN GA2 neurons. These findings provide evidence that in our model, DNA damage is associated with an increase in Aβ secretion in neuronal progenitors, which may contribute to the early stages of neuronal pathology in AD.