Cell Type-specific Anterograde Circuit Mapping and Functional Control by Optimizing YFV-17D Transneuronal Systems

通过优化 YFV-17D 跨神经元系统进行细胞类型特异性顺行电路映射和功能控制

• 批准号: 10505702
• 负责人: Wei Xu
• 金额: $ 156.63万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10505702
• 关键词: Animals; Attenuated Vaccines; Brain; Brain region; Communities; Complement; Complex; Corpus striatum structure; Dissection; Distal; Engineering; Feedback; Gene Delivery; Gene Expression; Genes; Goals; Injections; Interneurons; Maps; Mediating; Midbrain structure; Mus; Neurons; Neurosciences; Neurosciences Research; Pattern; RNA Editing; Research; Route; Series; Signal Transduction; Synapses; System; Technology; Testing; Time; Travel; Vaccines; Viral; Viral Vector; Virus; Virus Replication; Work; Yellow Fever Vaccine; Yellow fever virus; adeno-associated viral vector; anatomical tracing; base; brain circuitry; cell type; clinical application; complement system; design; experimental study; fascinate; improved; interest; mind control; neuronal circuitry; neurotoxicity; postsynaptic neurons; presynaptic neurons; reconstitution; retrograde transport; sensor; success; tool; vector; Animals; Attenuated Vaccines; Brain; Brain region; Communities; Complement; Complex; Corpus striatum structure; Dissection; Distal; Engineering; Feedback; Gene Delivery; Gene Expression; Genes; Goals; Injections; Interneurons; Maps; Mediating; Midbrain structure; Mus; Neurons; Neurosciences; Neurosciences Research; Pattern; RNA Editing; Research; Route; Series; Signal Transduction; Synapses; System; Technology; Testing; Time; Travel; Vaccines; Viral; Viral Vector; Virus; Virus Replication; Work; Yellow Fever Vaccine; Yellow fever virus; adeno-associated viral vector; anatomical tracing; base; brain circuitry; cell type; clinical application; complement system; design; experimental study; fascinate; improved; interest; mind control; neuronal circuitry; neurotoxicity; postsynaptic neurons; presynaptic neurons; reconstitution; retrograde transport; sensor; success; tool; vector

Cell Type-specific Anterograde Circuit Mapping and Functional Control by Optimizing YFV-17D Transneuronal Systems Summary To elucidate the functional organization of brain circuitry we need to delineate neuronal connectivity and control the activity of neurons with specific connectivity. Both tasks have increasingly relied on transneuronal viral vectors. Anterograde transneuronal viral vectors, which spread from presynaptic neurons to the postsynaptic neurons, can reveal the neuronal projections and selectively target postsynaptic neurons. Due to the lack of ideal anterograde transneuronal viral vectors, we have spent the last few years developing a new anterograde system based on yellow fever vaccine—YFV-17D. We have successfully constructed two major anterograde tools: the packaging defective YFV∆CME for mapping neuronal projectomes, and the replication defective YFV∆NS1 (or YFV∆CMENS1) for transneuronal control of gene expression in postsynaptic neurons, which can be used for functional observation or manipulation. Neuronal toxicity can be avoided in the case of YFV∆NS1 and YFV∆CMENS1, but not in YFV∆CME. These new systems provide benefits of transneuronal efficacy, diverse applications, and ease of engineering, but they still have limitations for many experimental scenarios. Here we propose to further improve these viral systems to make them broadly applicable, powerful and safe tools for functional dissection of the brain circuits. Firstly, we will construct self- constrained YFV-17D to further minimize neuronal toxicity incurred by viral replication in neurons, which will make these vectors more useful for both research and potential clinical applications. Secondly, we will test multiple strategies to target this system to specific neurons for cell-type specific tracing or functional control in both local circuits and long-range projections. Some of the cell type specific tools will not rely on the availability of genetically modified mouse lines and can be applied to broad species. Thirdly, we will incorporate commonly used tags, sensors or effectors in the optimized transneuronal viral vectors for versatile applications. We will also apply the improved versions to delineate a selected neuronal circuit that is of great interest to neuroscientists. Therefore, this project will yield a set of highly powerful tools widely applicable to neuroscience research, and will reveal the projectomes of multiple classical neuronal types.

通过细胞类型特异性的顺行电路映射和功能控制 优化YFV-17D跨神经元系统 概括 为了阐明大脑电路的功能组织，我们需要描述神经元 连通性并控制具有特定连通性的神经元活动。这两个任务都有 越来越多地依赖跨神经元病毒载体。顺行跨神经元病毒载体，该病毒载体 从突触前神经元传播到突触后神经元，可以揭示神经元投射 并有选择地靶向突触后神经元。由于缺乏理想的顺行透性 病毒向量，我们在过去的几年中一直在开发基于 黄热病疫苗-YFV-17D。我们已经成功地构建了两个主要的正常工具： 包装有缺陷的YFVΔCME，用于映射神经元射击和复制有缺陷 YFVΔNS1（或YFVΔCMENS1）用于突触后神经元基因表达的跨神经元对照， 可用于功能观察或操纵。可以避免神经元毒性 在YFVΔNS1和YFVΔCMENS1的情况下，但不在YFVΔCME中。这些新系统提供好处 跨神经元效率，潜水应用和易于工程的效率，但它们仍然有 许多实验场景的局限性。 在这里，我们建议进一步改善这些病毒系统，以使其广泛适用， 强大且安全的工具，用于脑电路的功能解剖。首先，我们将建立自我 受限的YFV-17D进一步最小化神经元病毒复制引起的神经元毒性， 这将使这些向量对研究和潜在的临床应用更有用。 其次，我们将测试多种策略将该系统靶向细胞类型的特定神经元 在本地电路和远程预测中的特定跟踪或功能控制。一些 特定于小区类型的工具将不依赖于一般修改的鼠标线的可用性，并且可以 应用于广泛的物种。第三，我们将合并常用的标签，传感器或效果 在针对多功能应用的优化跨神经元病毒矢量中。我们还将应用 改进版本来描述选定的神经元电路，这引起了人们的极大兴趣 神经科学家。因此，该项目将产生一组非常强大的工具，可广泛适用于 神经科学研究，并将揭示多种经典神经元类型的项目组。