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: 10673154
• 负责人: Hajime Hirase
• 金额: $ 13.59万
• 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/10673154
• 关键词: Acceleration; 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 Science Core; Dependovirus; Dermatology; Documentation; Engineering; Ependymal Cell; Genes; Genetic; 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; Transfection; Transgenic Mice; Variant; Viral; Viral Vector; Visualization; Visualization software; adeno-associated viral vector; 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; neural circuit; neurotransmission; neurovascular unit; new technology; noradrenergic; novel; optical imaging; programs; promoter; receptor; recombinant viral vector; 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.

病毒式工具开发核心-摘要 基因编码生物传感器的表达和特定细胞类型的光/药物遗传操作 提供了有效的方法来揭示睡眠期间脑液动力学背后的机制。最新版本 用于标记脑循环的技术是侵入性的，并改变脑脊液(CSF)的流动动力学。这个 病毒工具开发核心公司将开发新的微创方法，以改变对脑脊液流动的研究。 核心将通过提供旨在标记关键液体隔室的病毒载体来支持项目2和3， 特别是间质液体、脑脊液和血液。这将通过衣壳、推动者、信号的组合来实现 多肽，以及在腺相关病毒载体(AAV)的情况下的基因陷阱。其他重组病毒载体将 根据需要进行设计。在目标1中，我们将迅速建立病毒工具开发管道，以提供体内 验证了将进行优化和验证的病毒载体。在目标2中，我们将设计一套病毒工具， 通过转染肝表达荧光白蛋白，允许在单次 静脉注射。在目标3中，我们将通过安装生物传感器或照片来扩展血液探头的用途- 可激活的荧光蛋白，用于报告血液状况或可视化脑脊液。在目标4中，我们将设计一种敲击- 在AAV载体中掺入成功的探针进行永久表达。 创新探针的病毒介导性表达将允许在现有的 转基因小鼠不需要进一步繁殖，因此为这一问题提供了经济和及时的解决方案 研究计划。虽然核心将与项目2和3密切互动，但新颖的体内测试结果 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 成像可以特别从理论(方案1)和人脑(方案4)的角度提供。