A comparison of the impact on neuronal transcriptome and cognition of rAAV5 transduction with three different doses in the mouse hippocampus.
A comparison of the impact on neuronal transcriptome and cognition of rAAV5 transduction with three different doses in the mouse hippocampus.
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DOI：
10.3389/fnmol.2023.1195327
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发表时间：
2023
影响因子：
4.8
通讯作者：
Chen, Yi-Hua
中科院分区：
文献类型：
--
作者：
Liu, Yi-Si;Wang, Meng-Ling;Hu, Neng-Yuan;Li, Zi-Ming;Wu, Jia-Li;Li, Hao;Li, Jing-Ting;Li, Xiao-Wen;Yang, Jian-Ming;Gao, Tian-Ming;Chen, Yi-Hua
重组腺相关病毒（rAAV）广泛用于基因治疗。AAV 5已经显示出上级转导效率，靶向灵长类动物脑中的神经元和神经胶质细胞。尽管如此，AAV 5转导对分子和行为改变的综合影响仍然未被探索。本研究的重点是评估AAV 5转导在海马中的作用，海马是记忆形成和情绪过程的关键区域。在本实验中，在输注三种不同剂量的rAAV 5-mCherry 3周后，利用荧光激活细胞分选术（FACS）分离CaMkIIα-cre小鼠海马中mCherry标记的锥体神经元，然后对其进行RNA测序（RNA-seq）以评估基因表达谱。ELASA、数字微滴PCR和免疫组化分别检测细胞因子浓度、mRNA表达和胶质细胞反应。采用旷场实验和高架十字迷宫实验检测大鼠的运动和焦虑行为，采用Y-迷宫测试大鼠的空间工作记忆。再认记忆和恐惧反应分别采用新物体再认测验和恐惧条件反射测验进行检测。我们发现，与2.88 × 109 v.g和2.88 × 108 v. g相比，2.88 × 1010 v.g rAAV 5转导显著上调海马锥体神经元免疫反应和凋亡相关基因，下调线粒体功能和突触可塑性相关基因，而不诱导神经元丢失和胶质细胞增生。此外，同样的剂量损害了工作记忆和背景恐惧记忆，对运动和焦虑相关行为没有影响。我们的研究结果强调了与中剂量或低剂量相比，高剂量给药的有害影响，导致神经脆弱性增加和记忆受损。因此，当考虑外源基因的表达有效性时，在临床环境中也考虑潜在的副作用是至关重要的。然而，高剂量rAAV 5-mCherry的这些缺点背后的精确分子机制仍需要在未来的研究中进一步研究。
Recombinant adeno-associated viruses (rAAVs) are widely used in genetic therapeutics. AAV5 has shown superior transduction efficiency, targeting neurons and glial cells in primate brains. Nonetheless, the comprehensive impact of AAV5 transduction on molecular and behavioral alterations remains unexplored. This study focuses on evaluating the effects of AAV5 transduction in the hippocampus, a critical region for memory formation and emotional processes. In this experiment, fluorescence-activated cell sorting (FACS) was utilized to isolate the mCherry-labeled pyramidal neurons in the hippocampus of CaMkIIα-cre mice following three different doses rAAV5-mCherry infusion after 3 weeks, which were then subjected to RNA sequencing (RNA-seq) to assess gene expression profiles. The cytokines concentration, mRNA expression, and glial response in hippocampi were confirmed by ELASA, digital droplet PCR and immunohistochemistry respectively. Locomotion and anxiety-like behaviors were elevated by Open Field Test and Elevated Plus Maze Test, while the Y-Maze were used to assessed spatial working memory. Recognition memory and fear responses were examined by the Novel Object Recognition Test and Fear Conditioning Test, respectively. We found that 2.88 × 1010 v.g rAAV5 transduction significantly upregulated genes related to the immune response and apoptosis, and downregulated genes associated with mitochondrial function and synaptic plasticity in hippocampal pyramidal neurons, while did not induce neuronal loss and gliosis compared with 2.88 × 109 v.g and 2.88 × 108 v.g. Furthermore, the same doses impaired working memory and contextual fear memory, without effects on locomotion and anxiety-related behaviors. Our findings highlight the detrimental impact of high-dose administration compared to median-dose or low-dose, resulting in increased neural vulnerability and impaired memory. Therefore, when considering the expression effectiveness of exogenous genes, it is crucial to also take potential side effects into account in clinical settings. However, the precise molecular mechanisms underlying these drawbacks of high-dose rAAV5-mCherry still require further investigation in future studies.
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影响因子：
5.8
作者：
Gu, Zuguang;Eils, Roland;Schlesner, Matthias
通讯作者：
Schlesner, Matthias
影响因子：
16.2
作者：
Fanselow, Michael S.;Dong, Hong-Wei
通讯作者：
Dong, Hong-Wei
DOI：
10.1097/01.jnen.0000183345.19447.8e
发表时间：
2005-10-01
影响因子：
3.2
作者：
Huang, H;Hara, A;Ohgaki, H
通讯作者：
Ohgaki, H
影响因子：
14.8
作者：
Brewer, Gregory J.;Torricelli, John R.
通讯作者：
Torricelli, John R.
影响因子：
4.4
作者：
Kashio, Y;Nakamura, K;Hirashima, M
通讯作者：
Hirashima, M