New AAV tools fail to detect Neurod1-mediated neuronal conversion of Müller glia and astrocytes in vivo.
New AAV tools fail to detect Neurod1-mediated neuronal conversion of Müller glia and astrocytes in vivo.
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DOI：
10.1016/j.ebiom.2023.104531
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发表时间：
2023-04
期刊：
影响因子：
11.1
通讯作者：
Chen B
中科院分区：
文献类型：
--
作者：
Xie Y;Zhou J;Wang LL;Zhang CL;Chen B
重新编程驻留的神经胶质细胞以将其在体内转化为神经元代表了一种潜在的治疗策略，可以补充丢失的神经元，修复受损的神经回路并恢复功能。基于腺相关病毒（AAV）的表达系统是在胶质细胞-神经元重编程中进行体内基因递送的有力工具，然而，最近的研究表明，基于AAV的Neurod 1基因递送到小鼠脑中可导致严重的内源性神经元表达渗漏，从而导致对胶质细胞-神经元转化的误解。对基于AAV的递送系统进行了修改，以改善成年小鼠视网膜和脑中Neurod 1、Math 5、Ascl 1和Neurog 2的体内递送。为了检查是否发生真正的神经胶质-神经元转化，进行严格的命运作图实验以追踪神经胶质细胞的谱系。神经元渗漏在AAV-GFAP介导的Neurod 1、Math 5、Ascl 1和Neurog 2递送后普遍存在。在降低AAV剂量后，使用替代的AAV血清型或注射途径不能校正转基因依赖性渗漏。重要的是，我们报告了两种新的基于AAV的工具的开发，它们可以显着减少神经元渗漏。使用新的基于AAV的工具，我们提供了证据表明Neurod 1基因转移未能将谱系追踪的神经胶质细胞转化为神经元。独立于基于AAV的表达系统的严格的命运作图技术是在神经元重编程期间追踪神经胶质细胞命运的金标准。新开发的基于AAV的系统是体内胶质细胞到神经元重编程的宝贵工具。Chen实验室的工作得到了（NIH）赠款R 01 EY 024986和R 01 EY 028921的支持，这是一项来自、和的无限制挑战资助。Zhang实验室的工作得到（R 01 NS 127375和R 01 NS 117065）和的支持。
Reprogramming resident glial cells to convert them into neurons in vivo represents a potential therapeutic strategy that could replenish lost neurons, repair damaged neural circuits, and restore function. AAV (adeno-associated virus)-based expression systems are powerful tools for in vivo gene delivery in glia-to-neuron reprogramming, however, recent studies show that AAV-based gene delivery of Neurod1 into the mouse brain can cause severe leaky expression into endogenous neurons leading to misinterpretation of glia-to-neuron conversion. AAV-based delivery systems were modified for improved in vivo delivery of Neurod1, Math5, Ascl1, and Neurog2 in the adult mouse retina and brain. To examine whether bona fide glia-to-neuron conversion occurs, stringent fate mapping experiments were performed to trace the lineage of glial cells. The neuronal leakage is prevalent after AAV-GFAP-mediated delivery of Neurod1, Math5, Ascl1, and Neurog2. The transgene-dependent leakage cannot be corrected after lowering the AAV doses, using alterative AAV serotypes or injection routes. Importantly, we report the development of two new AAV-based tools that can significantly reduce neuronal leakage. Using the new AAV-based tools, we provide evidence that Neurod1 gene transfer fails to convert lineage traced glial cells into neurons. Stringent fate mapping techniques independently of an AAV-based expression system are the golden standard for tracing the fate of glia cells during neuronal reprogramming. The newly developed AAV-based systems are invaluable tools for glia-to-neuron reprogramming in vivo. The work in Chen lab was supported by (NIH) grants R01 EY024986 and R01 EY028921, an unrestricted challenge grant from , the , and . The work in Zhang lab was supported by (R01 NS127375 and R01 NS117065) and .
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64.8
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