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

基本信息

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
项目类别：
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

项目摘要

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 - 摘要遗传编码的生物传感器和细胞型特异性光学/药物遗传操作的表达表示有效的方法来揭示睡眠过程中脑流体动力学背后的机制。最新标记脑循环的技术是侵入性的，改变了脑脊液（CSF）流动的动力学。这病毒工具开发核心将开发新颖的，微创的方法来改变CSF流量的研究。核心将通过提供旨在标记关键流体室的病毒向量来支持项目2和3 特别是间质性液体，CSF和血液。这将通过包扎，启动子，信号的组合来实现在腺相关病毒载体（AAV）的情况下，肽和基因陷阱。其他重组病毒载体将根据需要设计。在AIM 1中，我们将迅速建立病毒工具开发管道以提供体内经过验证的病毒向量将得到优化和验证。在AIM 2中，我们将设计一组病毒工具通过转染肝脏来表达荧光白蛋白，允许一次纵向血管成像静脉注射。在AIM 3中，我们将通过连接生物传感器或照片来扩展血液探针的效用可激活的荧光蛋白报告血液状况或可视化CSF。在AIM 4中，我们将设计一个敲门声 - 在AAV载体中，以结合永久表达的成功探针。创新探针的病毒介导的表达将允许现有的纵向成像转基因小鼠无需进一步繁殖，因此为此提供了成本和时间效率的解决方案研究计划。尽管核心将与项目2和3密切相互作用，但新型的体内测试结果病毒矢量将与数据科学核心共享和分析，以与常规探针进行比较。新的探测想法将与所有研究团队讨论，以确定光学的关键测量成像可以特别来自理论（项目1）和人脑（项目4）观点。