AnteroTag, a Novel Method for Trans-Synaptic Delivery of Active Agents to Map and Modify Anterograde Populations

AnteroTag，一种跨突触传递活性剂以绘制和修改顺行群体的新方法

• 批准号: 10258693
• 负责人: Jason M Christie
• 金额: $ 230.48万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-13 至 2024-09-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10258693
• 关键词: Adopted; Alleles; Amino Acid Sequence; Animal Model; Animals; Apoptosis; Area; Axon; BRAIN initiative; Behavior Control; Benchmarking; Biological; Biological Assay; Biological Products; Biology; Brain; Brain Diseases; Brain region; Cell Count; Cell Nucleus; Cells; Cessation of life; Chronic; Complex; Cre driver; Data; Dependovirus; Development; Electrophysiology (science); Engineering; Ferrets; Functional Imaging; Future; Goals; Grant; Herpes Simplex Infections; Histology; Human; Image; Inflammation; Injections; Label; Maps; Methods; Microscopy; Modeling; Molecular; Molecular Target; Molecular Weight; Monitor; Mus; Nerve Degeneration; Neurons; Organism; Outcome; Pathway interactions; Performance; Peroxidases; Pontine structure; Population; Property; Protein Engineering; Proteins; Rabies; Reporter; Research; Research Personnel; Safety; Serotyping; Slice; Specificity; Synapses; Synaptic Vesicles; System; Target Populations; Technology; Tertiary Protein Structure; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Toxic effect; Tropism; Tupaiidae; Validation; Variant; Vesicular stomatitis Indiana virus; Viral; Viral Vector; Virus; Wheat Germ Agglutinins; Work; base; biomaterial compatibility; cell type; cohort; cytotoxic; discrete time; granule cell; improved; in vivo; insight; mossy fiber; neural circuit; neurotransmission; new technology; nonhuman primate; novel; novel therapeutics; nuclease; postsynaptic; presynaptic; protein transport; recombinase; red fluorescent protein; relating to nervous system; tool; vector; vector control; vesicular release

PROJECT SUMMARY A goal of the BRAIN initiative is to develop and validate novel tools to map and manipulate neural circuits. The definition and control of behaviorally relevant circuits requires both retrograde and anterograde trans-synaptic technologies that perform well in vivo. In this project, we pursue the long-term goal of developing a small protein tag for the anterograde delivery of cargos (an “AnteroTag”). The novel concept of AnteroTag is to leverage the endogenous biology of neurotransmission to target tagged payloads to synaptic vesicles where they undergo vesicular release, postsynaptic entry, and desired action. Our consortium of investigators proposes to utilize the pontocerebellar circuit as a model pipeline to further refine and rigorously validate AnteroTag. As the axons of basal pontine neurons (mossy fibers) make specific and quantitative synapses upon cerebellar granule cells, the pontocerebellar circuit affords an established and conserved model circuit to determine how iterations of AnteroTag alter its performance. The investigators of the consortium will leverage their respective areas of expertise to quantify the specificity and biocompatibility of AnteroTag, benchmarked against a current state-of- the-art anterograde viral vector HSV-129; these analyses in mice will include microscopy, electrophysiology, and in vivo activity imaging in behaving animals. Once the pipeline determines the most specific and safe version of AnteroTag in the pontocerebellar circuit, we will assay AnteroTag performance when delivered to diverse starter populations of neurons. The broad utility of AnteroTag will then be determined by testing its efficacy to deliver a variety of genetically encoded markers and modifiers, and by testing AnteroTag derivatives for their utility in accessing second-order anterograde populations. With safety, specificity, and utility thus rigorously demonstrated in mice, we will assay the performance of AnteroTag in the pontocerebellar circuit of higher organisms including tree shrews, ferrets, and NHPs. The outcome of this proposal will be the development, validation, and implementation of a novel technology for the trans-synaptic anterograde delivery of biological agents across multiple brain regions and multiple species. Thus, if successful, our project will ultimately aid in the development of targeted cell-type and circuit-specific therapeutics to treat brain disorders.

项目摘要 大脑计划的一个目标是开发和验证新颖的工具来绘制和操纵神经回路。这 对行为相关的电路的定义和控制需要逆行和顺行反射突触 体内表现良好的技术。在这个项目中，我们实现了开发小蛋白质的长期目标 标签赤道（“ Anterotag”）的顺行交付。 Anterotag的新颖概念是利用 神经传递的内源性生物学，将标记有效载荷靶向突触蔬菜 囊泡释放，突触后进入和所需的动作。我们的调查人员联盟提议利用 pontocerebellar电路作为模型管道，以进一步完善和严格验证匿名词。作为轴突 基本蓬丁神经元（苔藓纤维）在小脑颗粒细胞上制作特定的定量突触， Pontocerebellar Circuit提供了一个已建立且构成的模型电路，以确定如何迭代 Anterotag改变其性能。财团的调查人员将利用其各自的领域 量化Anterotag的特异性和生物相容性的专业知识，以当前的状态为基准 Art Art Arteroval病毒载体HSV-129;这些小鼠中的这些分析将包括显微镜，电生理学和 在行为动物中成像体内活动成像。一旦管道确定了最具体，最安全的版本 pontocerebellar电路中的Anterotag，我们将在交付到不同的入门器时主张Anterotag性能 神经元的种群。然后将通过测试其提供有效性来确定Anterotag的广泛效用 多种一般编码的标记和修饰符，并通过测试其在 访问二阶顺行人口。因此，严格的安全性，特异性和实用性 在小鼠中证明，我们将在较高的pontocerebellar电路中断言Anterotag的性能 包括树生，雪貂和NHP在内的生物。该提案的结果将是发展， 验证和实施一种新型技术，用于生物学的反射突触的递送 多个大脑区域和多种物种的代理。如果成功的话，我们的项目最终将有助于 靶向细胞类型和电路特异性疗法的发展，以治疗脑疾病。