成人海马神经发生的减少是阿尔茨海默病 (AD) 的早期关键事件，导致进行性记忆丧失和认知能力下降。核孔蛋白 153 (Nup153) 是 NSC 干性的关键表观遗传调节因子，其水平降低表征了从 AD (3×Tg) (AD-NSC) 小鼠模型中分离出的神经干细胞，并确定了它们改变的可塑性和基因表达。Nup153 -研究了有助于NSC功能的调节机制：（1）通过蛋白质组学从AD和野生型（WT）小鼠中分离培养的NSC； (2) 通过慢病毒介导将 Nup153 或 GFP 递送到 AD 小鼠和对照小鼠的海马体内，分析神经发生和认知功能； (3) 从 AD 患者和对照受试者获得的人类 iPSC 衍生的脑类器官作为神经发育模型。蛋白质组学方法鉴定了 WT-和 AD-NSC 中的 Nup153 相互作用因子，可能与神经发生调节有关。基因本体（GO）分析表明，WT-NSC 中的 Nup153 结合蛋白参与 RNA 代谢、核输入和表观遗传机制。 AD-NSC 中的 Nup153 结合蛋白参与神经变性、线粒体功能障碍、蛋白酶体加工和 RNA 降解的途径。此外，AD-NSC 中 Nup153 水平的恢复降低了氧化应激标记物的水平并恢复了蛋白酶体活性。慢病毒介导的 AD 小鼠海马微环境中 Nup153 的递送增加了早期祖细胞的增殖，以 BrdU/DCX 和 BrdU/PSANCAM 阳性为标志，以及随后细胞颗粒层中分化神经元（BrdU/NeuN 细胞）的整合与注射 GFP 的 AD 小鼠相比。一致的是，通过莫里斯水迷宫评估，在病毒递送后 1 个月，注射 Nup153 的 AD 小鼠与 AD-GFP 小鼠相比，表现出认知能力的改善。为了验证 Nup153 在神经发生中的作用，我们利用了源自 AD-iPSC 的脑类器官，其特征是神经上皮祖细胞环较少，分化区域减少。 AD 类器官中 Nup153 的上调恢复了神经样管的形成和分化。我们的数据表明，Nup153 对神经发生的积极作用是基于由 Nup153 精心策划的复杂调节网络，并且该蛋白是一个有价值的疾病靶标。© 2024。作者。

Reduction of adult hippocampal neurogenesis is an early critical event in Alzheimer's disease (AD), contributing to progressive memory loss and cognitive decline. Reduced levels of the nucleoporin 153 (Nup153), a key epigenetic regulator of NSC stemness, characterize the neural stem cells isolated from a mouse model of AD (3×Tg) (AD-NSCs) and determine their altered plasticity and gene expression.Nup153-regulated mechanisms contributing to NSC function were investigated: (1) in cultured NSCs isolated from AD and wild type (WT) mice by proteomics; (2) in vivo by lentiviral-mediated delivery of Nup153 or GFP in the hippocampus of AD and control mice analyzing neurogenesis and cognitive function; (3) in human iPSC-derived brain organoids obtained from AD patients and control subjects as a model of neurodevelopment.Proteomic approach identified Nup153 interactors in WT- and AD-NSCs potentially implicated in neurogenesis regulation. Gene ontology (GO) analysis showed that Nup153-bound proteins in WT-NSCs were involved in RNA metabolism, nuclear import and epigenetic mechanisms. Nup153-bound proteins in AD-NSCs were involved in pathways of neurodegeneration, mitochondrial dysfunction, proteasomal processing and RNA degradation. Furthermore, recovery of Nup153 levels in AD-NSCs reduced the levels of oxidative stress markers and recovered proteasomal activity. Lentiviral-mediated delivery of Nup153 in the hippocampal niche of AD mice increased the proliferation of early progenitors, marked by BrdU/DCX and BrdU/PSANCAM positivity and, later, the integration of differentiating neurons in the cell granule layer (BrdU/NeuN+ cells) compared with GFP-injected AD mice. Consistently, Nup153-injected AD mice showed an improvement of cognitive performance in comparison to AD-GFP mice at 1 month after virus delivery assessed by Morris Water Maze. To validate the role of Nup153 in neurogenesis we took advantage of brain organoids derived from AD-iPSCs characterized by fewer neuroepithelial progenitor loops and reduced differentiation areas. The upregulation of Nup153 in AD organoids recovered the formation of neural-like tubes and differentiation.Our data suggest that the positive effect of Nup153 on neurogenesis is based on a complex regulatory network orchestrated by Nup153 and that this protein is a valuable disease target.© 2024. The Author(s).