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：
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
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/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

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 - 摘要基因编码生物传感器的表达和细胞类型特异性的光/药物遗传学操作代表了揭示睡眠期间脑液动力学背后机制的有效方法。最新标记脑循环的技术是侵入性的，会改变脑脊液（CSF）流动的动力学。这病毒工具开发核心将开发新颖的微创方法来改变脑脊液流的研究。核心将通过提供设计用于标记关键流体隔室的病毒载体来支持项目 2 和 3，特别是间质液、脑脊液和血液。这将通过衣壳、启动子、信号的组合来实现肽，以及腺相关病毒载体（AAV）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，以提供体内经过验证的病毒载体将被优化和验证。在目标 2 中，我们将设计一套病毒式工具，通过转染肝脏表达荧光白蛋白，在单次注射后允许纵向血管成像静脉注射。在目标 3 中，我们将通过附加生物传感器或光电传感器来扩展血液探针的效用。可激活的荧光蛋白可报告血液状况或可视化脑脊液。在目标 4 中，我们将设计一个敲击在 AAV 载体中掺入成功的探针以实现永久表达。创新探针的病毒介导表达将允许在现有技术中进行纵向成像转基因小鼠无需进一步育种，因此为此提供了具有成本效益和时间效益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，以便与传统探针进行比较。新的探针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从理论（项目 1）和人脑（项目 4）的角度提供。