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Cell type selective viral tools to interrogate and correct non-human primate and human brain circuitry

用于询问和纠正非人类灵长类动物和人类大脑回路的细胞类型选择性病毒工具

基本信息

批准号：
10462660
负责人：
Franck K Kalume
金额：
$ 133.76万
依托单位：
ALLEN INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10462660
关键词：
Adult BRAIN initiative Biodistribution Brain Brain Diseases Capsid Cells Cerebrospinal Fluid Clinical Consumption Dependovirus Discrimination Disease Dose Engineering Enhancers Epilepsy Etiology Functional disorder Gene Delivery Gene Expression Genetic Human Individual Injections Intravenous Knowledge Lead Mediating Methods Molecular Molecular Genetics Molecular Profiling Mus Neocortex Neurologic Deficit Neurons Phenotype Physiology Positioning Attribute Primates Process Prosencephalon Reporter Resolution Role Route Safety Shapes Site Slice Sodium Channel Taxonomy Testing Therapeutic Intervention Time Tissues Transgenes Transgenic Organisms Translating Translational Research Validation Viral Viral Vector Virus Visual Visual Cortex adeno-associated viral vector base brain cell brain circuitry brain tissue cell type cellular transduction childhood epilepsy cost dravet syndrome experimental study gene replacement therapy gene therapy improved in vivo inhibitory neuron mouse genetics mouse model neocortical nonhuman primate novel recombinase restoration screening selective expression targeted treatment therapeutic gene therapeutic transgene tool transcriptomics transduction efficiency transgene delivery transgene expression vector voltage

项目摘要

Abstract: Many cell types together assemble the functional circuitry of the human brain. For over a century, neuroscientists have categorized brain cell types by their features, including shape, position, physiology, molecules, and function. Single cell transcriptomics studies are now defining molecular cell types at a resolution not previously possible, uncovering a taxonomy of hundreds to thousands of brain cell types. These studies have also revealed dramatic differences in molecular signatures of homologous cell types across species, showing decisively that the difference between mouse and human brain is not simply the total number of neurons. However, the function of each cell class or type in brain circuitry, and dysfunction in disease, is only beginning to be evaluated. To characterize the roles of human brain cell classes in normal function and disease, it is critical that tools be developed to allow genetic access to cell classes in vivo. Such tools would enable precise therapeutic gene delivery to brain cell classes, permitting targeted treatment for class-specific etiologies like some epilepsies. Few genetic tools are available to mark and manipulate cell classes and types in non-genetically tractable species like human and non-human primate (NHP). Viruses including adeno-associated viruses (AAVs), containing cell class and type selective enhancers can be leveraged to gain genetic access to, and drive gene expression in specific brain cell classes in these species. We have initiated a project through the BRAIN Initiative to generate and validate reporter AAVs to mark specific cell classes in the mouse cortex in vivo and in human neocortical tissue ex vivo. Our groups have engineered AAV vectors and optimized capsids to access neurons and express transgenes in many discrete cell classes and types in mouse and primate. New and improved AAV tools promise to fuel human brain scientific discovery and clinical progress, but one impediment has been the costly and time-consuming process of validating new vectors in primates. We present three Aims to translate these promising new AAV vectors into a high-value set of primate-optimized tools that could eventually be used for gene therapies in humans. First, we will develop a platform for screening AAV vectors in NHP ex vivo brain slices, followed by individual validation of promising vectors in NHP in vivo and human ex vivo brain slice cultures. Second, we will identify optimal AAV capsids to: a) support widespread NHP neuronal transduction in vivo when applied intravenously or to cerebrospinal fluid (CSF), two preferred routes of delivery for human CNS gene therapy, and b) support AAV transduction of human primary brain tissue ex vivo. Third, we will perform proof-of-concept experiments using cell class-selective vectors to express a therapeutic transgene in defined classes to treat a severe and intractable form of childhood epilepsy called Dravet syndrome (DS). These experiments represent a significant step towards converting cell class-selective AAVs into first-in-class viral tools optimized for in vivo NHP brain studies and human gene therapy applications.

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