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.

项目概要 BRAIN 计划的目标是开发和验证绘制和操纵神经回路的新工具。这 行为相关回路的定义和控制需要逆行和顺行跨突触 在体内表现良好的技术。在这个项目中，我们追求开发一种小蛋白质的长期目标 用于顺行运送货物的标签（“AnteroTag”）。 AnteroTag 的新颖概念是利用 神经传递的内源生物学将标记的有效负载靶向它们经历的突触小泡 囊泡释放、突触后进入和所需的作用。我们的研究人员联盟建议利用 脑桥小脑回路作为模型管道，进一步完善和严格验证 AnteroTag。作为轴突 基底脑桥神经元（苔藓纤维）在小脑颗粒细胞上形成特异性和定量的突触， 脑桥小脑电路提供了一个已建立且保守的模型电路，以确定如何迭代 AnteroTag 改变其性能。该联盟的研究人员将利用各自领域的研究成果 量化 AnteroTag 的特异性和生物相容性的专业知识，以当前状态为基准 最先进的顺行病毒载体HSV-129；这些对小鼠的分析将包括显微镜、电生理学和 行为动物的体内活动成像。一旦管道确定了最具体和最安全的版本 AnteroTag 在脑桥小脑回路中，我们将测试 AnteroTag 在传送到不同启动器时的性能 神经元群体。然后，将通过测试 AnteroTag 的功效来确定其广泛的实用性，以提供 各种基因编码标记和修饰符，并通过测试 AnteroTag 衍生物在 访问二阶顺行人群。具有安全性、特异性和实用性，因此严格 在小鼠中得到证实，我们将检测 AnteroTag 在高等脑桥小脑回路中的性能 生物体，包括树鼩、雪貂和 NHP。该提案的结果将是发展， 验证和实施一种用于跨突触顺行传递生物制剂的新技术 跨多个大脑区域和多个物种的代理。因此，如果成功，我们的项目最终将有助于 开发用于治疗脑部疾病的靶向细胞类型和电路特异性疗法。