Viral Tool Development Core: Visualization and manipulation of brain fluid dynamics by recombinant viral vectors

病毒工具开发核心：重组病毒载体对脑液动力学的可视化和操纵

• 批准号: 10516500
• 负责人: Hajime Hirase
• 金额: $ 15.09万
• 依托单位: UNIVERSITY OF ROCHESTER
• 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 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10516500
• 关键词: Affect; Albumins; Astrocytes; Biomedical Research; Biosensor; Blood; Blood - brain barrier anatomy; Blood Vessels; Blood flow; Brain; Brain imaging; Breeding; CRISPR/Cas technology; Capsid; Cells; Cerebrospinal Fluid; Cerebrovascular Circulation; Chronic; Complement; Cyclic AMP; Data Analyses; Data Science Core; Dependovirus; Dermatology; Documentation; Engineering; Ependymal Cell; Genes; Goals; Hepatocyte; Human; Image; Imaging Device; Immune response; Inflammation; Injections; Intercellular Fluid; Ischemia; Knock-in; Label; Liquid substance; Liver; Malignant Neoplasms; Measurement; Mediating; Methods; Molecular; Nephrology; Neuroglia; Neurons; Optics; Peptide Signal Sequences; Pharmacogenetics; Physiologic Monitoring; Physiological; Plasma; Process; Production; Proteins; Publications; Published Comment; Recombinants; Reporting; Research; Resource Sharing; Resources; Rodent; Second Messenger Systems; Signal Transduction; Sleep; Specificity; Structure of choroid plexus; TNFSF5 gene; Techniques; Technology; Test Result; Testing; Time; Toxic effect; Tracer; Transgenic Mice; Variant; Viral; Viral Vector; Visualization; Visualization software; adeno-associated viral vector; base; brain circulation; capillary bed; cell type; cerebrospinal fluid flow; cholinergic; cost; design; experimental study; genome editing; imaging study; in vivo; in vivo evaluation; innovation; interest; intravenous injection; minimally invasive; neural circuit; neurotransmission; neurovascular unit; new technology; noradrenergic; novel; optical imaging; programs; promoter; receptor; recombinant viral vector; relating to nervous system; serial imaging; solute; tool; tool development; vector

Viral Tool Development Core - Abstract Expression of genetically encoded biosensors and cell-type-specific opto-/pharmacogenetic manipulation represent effective approaches to reveal the mechanisms behind brain fluid dynamics during sleep. Most current technologies for labeling brain circulation are invasive and alter dynamics of cerebrospinal fluid (CSF) flow. The Viral Tool Development Core will develop novel, minimally-invasive methods to transform the study of CSF flow. The Core will support Projects 2 and 3 by providing viral vectors designed to label key fluid compartments, specifically interstitial fluid, CSF, and blood. This will be achieved by a combination of capsids, promoters, signal peptides, and gene traps in the case of adeno-associated viral vector (AAV). Other recombinant viral vectors will be designed as needed. In Aim 1, we will swiftly establish a viral tool development pipeline to provide in vivo verified viral vectors that will be optimized and validated. In Aim 2, we will engineer a set of viral tools that will express fluorescent albumin by transfecting the liver, allowing longitudinal vascular imaging after a single intravenous injection. In Aim 3, we will extend the utility of the blood probe by attaching biosensors or photo- activatable fluorescent proteins to report blood conditions or visualize CSF. In Aim 4, we will engineer a knock- in AAV vector to incorporate successful probes for permanent expression. The virally mediated expression of the innovative probes will allow for longitudinal imaging in existing transgenic mice without a need for further breeding, hence provides cost- and time-effective solutions for this research program. While the core will closely interact with Projects 2 and 3, the in vivo testing results for novel viral vectors will be shared and analyzed with the Data Science Core for comparisons with conventional probes. New probe ideas will be discussed with all research teams to determine the critical measurements that optical imaging can provide particularly from theoretical (Project 1) and human brain (Project 4) viewpoints.

病毒工具开发核心-摘要 基因编码的生物传感器的表达和细胞类型特异性光/药物遗传学操作 代表了揭示睡眠期间脑流体动力学背后机制的有效方法。最新 用于标记脑循环的技术是侵入性的并且改变脑脊液（CSF）流动的动力学。的 Viral Tool Development Core将开发新的微创方法来改变CSF流动的研究。 核心将通过提供病毒载体来支持项目2和3，这些病毒载体被设计用于标记关键的流体隔室， 特别是间质液脑脊液和血液这将通过衣壳、启动子、信号转导子和蛋白质的组合来实现。 肽，和在腺相关病毒载体（AAV）的情况下的基因陷阱。其它重组病毒载体将 根据需要设计。在目标1中，我们将迅速建立病毒工具开发管道， 将被优化和验证的已验证病毒载体。在目标2中，我们将设计一套病毒工具， 通过切除肝脏表达荧光白蛋白，允许在单次 静脉注射在目标3中，我们将通过连接生物传感器或光传感器来扩展血液探针的实用性。 可激活的荧光蛋白报告血液状况或可视化CSF。在目标4中，我们将设计一个敲门声- 在AAV载体中掺入成功的探针用于永久表达。 创新探针的病毒介导的表达将允许在现有技术中进行纵向成像。 转基因小鼠而不需要进一步育种，因此为此提供了成本和时间有效的解决方案。 研究计划。虽然核心将与项目2和3密切相互作用，但新的体内测试结果表明， 病毒载体将与数据科学核心共享和分析，以与传统探针进行比较。 将与所有研究团队讨论新的探测想法，以确定光学 成像可以提供特别是从理论（项目1）和人脑（项目4）的观点。