Impaired APP activity and altered Tau splicing in embryonic stem cell-derived astrocytes obtained from an APPsw transgenic minipig.
Impaired APP activity and altered Tau splicing in embryonic stem cell-derived astrocytes obtained from an APPsw transgenic minipig.
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DOI：
10.1242/dmm.019489
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发表时间：
2015-10-01
影响因子：
4.3
通讯作者：
Hyttel P
中科院分区：
文献类型：
--
作者：
Hall VJ;Lindblad MM;Jakobsen JE;Gunnarsson A;Schmidt M;Rasmussen MA;Volke D;Zuchner T;Hyttel P
家族性青少年阿尔茨海默病(AD)的动物模型往往不能通过修改导致该疾病的单基因突变来产生该疾病的不同病理特征。因此，它们可能是测试和开发新药的糟糕模型。在这里，我们分析了由单个突变人类基因(APPsw)的过度表达所产生的疾病的大型哺乳动物模型的体外产生的干细胞及其衍生细胞。我们从交配的半合子小型猪的胚胎中分离出胚胎干细胞(ESCs)进行培养和分化，获得了半合子和纯合子的放射状神经胶质样细胞。这些细胞被证实共表达不同的神经标记物，包括NES，GFAP和BLBP，典型的来自颗粒下带的1型放射状胶质细胞(RGS)。这些细胞改变了CCND1和NOTCH1的表达，并降低了几个核糖体RNA基因的表达。我们发现这些细胞在定向分化后能够分化为星形胶质细胞。产生的星形胶质细胞α-和β-分泌酶活性降低，γ-分泌酶活性升高，tau剪接改变。这表明了与家族性AD星形胶质细胞的细胞更新和功能相关的早期发病机制的新方面。这些结果还强调，放射状胶质细胞可能是药物发现的潜在有用细胞群，并且可以在疾病的体外模型中检测到改变的APP表达和改变的tau磷酸化。最后，通过只插入一个突变，使用大型哺乳动物模型来模拟家族性阿尔茨海默病是可能的。摘要：在APPsw猪的体外培养系统中对星形胶质细胞和放射状胶质细胞病理的洞察。
Animal models of familial juvenile onset of Alzheimer's disease (AD) often fail to produce diverse pathological features of the disease by modification of single gene mutations that are responsible for the disease. They can hence be poor models for testing and development of novel drugs. Here, we analyze in vitro-produced stem cells and their derivatives from a large mammalian model of the disease created by overexpression of a single mutant human gene (APPsw). We produced hemizygous and homozygous radial glial-like cells following culture and differentiation of embryonic stem cells (ESCs) isolated from embryos obtained from mated hemizygous minipigs. These cells were confirmed to co-express varying neural markers, including NES, GFAP and BLBP, typical of type one radial glial cells (RGs) from the subgranular zone. These cells had altered expression of CCND1 and NOTCH1 and decreased expression of several ribosomal RNA genes. We found that these cells were able to differentiate into astrocytes upon directed differentiation. The astrocytes produced had decreased α- and β-secretase activity, increased γ-secretase activity and altered splicing of tau. This indicates novel aspects of early onset mechanisms related to cell renewal and function in familial AD astrocytes. These outcomes also highlight that radial glia could be a potentially useful population of cells for drug discovery, and that altered APP expression and altered tau phosphorylation can be detected in an in vitro model of the disease. Finally, it might be possible to use large mammal models to model familial AD by insertion of only a single mutation. Summary: Insight into astrocyte and radial glia pathology in an in vitro culture system derived from the APPsw pig.