Department of Physiology, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582, Japan; Laboratory for Proteolytic Neuroscience, RIKEN Center for Brain Science, Wako City, Saitama 351-0198, Japan; Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako City, Saitama 351-0198, Japan.
Department of Physiology, School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582, Japan.
Neurosci Res. 2022 Dec;185:49-61. doi: 10.1016/j.neures.2022.08.008. Epub 2022 Sep 6.
Alzheimer's disease (AD) is the leading cause of dementia which afflicts tens of millions of people worldwide. Despite many scientific progresses to dissect the AD's molecular basis from studies on various mouse models, it has been suffered from evolutionary species differences. Here, we report generation of a non-human primate (NHP), common marmoset model ubiquitously expressing Amyloid-beta precursor protein (APP) transgenes with the Swedish (KM670/671NL) and Indiana (V717F) mutations. The transgene integration of generated two transgenic marmosets (TG1&TG2) was thoroughly investigated by genomic PCR, whole-genome sequencing, and fluorescence in situ hybridization. By reprogramming, we confirmed the validity of transgene expression in induced neurons in vitro. Moreover, we discovered structural changes in specific brain regions of transgenic marmosets by magnetic resonance imaging analysis, including in the entorhinal cortex and hippocampus. In immunohistochemistry, we detected increased Aβ plaque-like structures in TG1 brain at 7 years old, although evident neuronal loss or glial inflammation was not observed. Thus, this study summarizes our attempt to establish an NHP AD model. Although the transgenesis approach alone seemed not sufficient to fully recapitulate AD in NHPs, it may be beneficial for drug development and further disease modeling by combination with other genetically engineered models and disease-inducing approaches.
阿尔茨海默病(AD)是导致痴呆的主要原因,全球有数千万人受到影响。尽管通过对各种小鼠模型的研究在剖析 AD 的分子基础方面取得了许多科学进展,但它一直受到进化物种差异的困扰。在这里,我们报告了一种非人类灵长类动物(NHP),普通狨猴模型的产生,该模型普遍表达具有瑞典(KM670/671NL)和印第安纳(V717F)突变的淀粉样前体蛋白(APP)转基因。通过基因组 PCR、全基因组测序和荧光原位杂交,对生成的两种转基因狨猴(TG1&TG2)的转基因整合进行了彻底研究。通过重编程,我们在体外证实了转基因在诱导神经元中的表达的有效性。此外,我们通过磁共振成像分析发现了转基因狨猴特定脑区的结构变化,包括内嗅皮层和海马体。在免疫组织化学中,我们在 7 岁的 TG1 大脑中检测到了增加的 Aβ斑块样结构,尽管没有观察到明显的神经元丢失或神经胶质炎症。因此,本研究总结了我们建立非人类灵长类 AD 模型的尝试。尽管转基因方法本身似乎不足以完全再现 NHPs 中的 AD,但它可能通过与其他基因工程模型和疾病诱导方法结合,有益于药物开发和进一步的疾病建模。