Novel AAV Capsids and Gene Regulatory Elements for GeneExpression in Microglia

用于小胶质细胞基因表达的新型 AAV 衣壳和基因调控元件

基本信息

批准号：
10195876
负责人：
Benjamin E Deverman
金额：
$ 24万
依托单位：
BROAD INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10195876
关键词：
ATAC-seq Affect Alzheimer&apos s Disease Alzheimer&apos s disease risk Alzheimer&apos s disease therapy Automobile Driving Binding Blood - brain barrier anatomy Brain Brain Diseases Capsid Cell Nucleus Cell surface Cells Chromatin Code Communities DNA Data Degradation Pathway Dependovirus Development Disease Elements Engineering Enzymes Exhibits Gene Expression Genes Genetic study Genome Goals Immunohistochemistry In Situ Hybridization In Vitro Individual Injections Label Late Onset Alzheimer Disease Lentivirus Libraries Link Mediating Microglia Mus Neurosciences Neurosciences Research Outcome Play Publishing RNA Rare Diseases Regulator Genes Regulatory Element Resistance Role Safety Schizophrenia Slice Synapses Testing Time Transgenic Animals Transgenic Mice Variant Viral Viral Genome Viral Vector Virus Work adeno-associated viral vector blood-brain barrier crossing brain cell cell type complement system gene therapy human embryonic stem cell improved in vivo in vivo evaluation inhibitor/antagonist macrophage novel particle prevent protein degradation risk variant tool trafficking transgene expression vector genome

项目摘要

Project Summary: Microglia, the resident macrophages of the brain, assume a multitude of functions during brain development and disease. Long known to react to changes in brains affected by Alzheimer’s disease (AD), genetic studies of late- onset AD indicate that AD risk variants are commonly found in or near genes that are specifically expressed in microglia, suggesting that microglia play an active role in driving AD. In addition, microglia exhibit various functions during brain development, including synapse pruning. Genes coding for components of the complement system, which mediates synapse pruning by microglia, have been linked to schizophrenia. However, a major bottleneck in studying the roles of microglia in normal and diseased brains is the lack of viral tools for rapidly manipulating their gene expression. Viral vectors would also benefit studies where transgenic animals are not available. Finally, viral vectors would enable studies on the potential of gene therapy for AD and rare diseases that affect microglia. However, while microglia show modest and localized transduction by lentiviruses in vivo, they are resistant to transduction by adeno-associated viruses (AAVs), the preferred viral vectors used in neuroscience research and gene therapy due to their superior spread and excellent safety profile. Here we propose to elucidate the barriers to microglia transduction by AAVs, engineer AAV capsid variants that are able to overcome these barriers and transduce microglia in vivo, and develop novel gene regulatory elements that will enable microglia-specific transgene expression from viral vectors. This will be a collaborative project between the Stevens and Deverman labs, combining their expertise in studying microglia (Stevens) and in developing novel AAV vectors (Deverman).

项目概要：小胶质细胞是大脑中常驻的巨噬细胞，在大脑发育过程中承担多种功能长期以来，人们都知道这种疾病会对受阿尔茨海默病 (AD) 影响的大脑变化做出反应，最新的基因研究表明，发病 AD 表明 AD 风险变异通常存在于特定表达的基因中或附近小胶质细胞，表明小胶质细胞在驱动 AD 中发挥着积极作用。此外，小胶质细胞表现出各种不同的特征。大脑发育过程中的功能，包括编码突触成分的基因。补体系统通过小胶质细胞介导突触修剪，与精神分裂症有关。然而，研究小胶质细胞在正常和患病大脑中的作用的一个主要瓶颈是缺乏病毒快速操纵病毒载体的工具也将有利于转基因研究。最后，病毒载体将使研究AD和AD基因治疗的潜力成为可能。然而，虽然小胶质细胞表现出适度和局部的转导，但影响小胶质细胞的罕见疾病。慢病毒在体内，它们对腺相关病毒（AAV）的转导有抵抗力，腺相关病毒是首选的病毒载体因其优越的传播性和出色的安全性而被用于神经科学研究和基因治疗。在这里，我们建议阐明 AAV 转导小胶质细胞的障碍，设计 AAV 衣壳变体能够克服这些障碍并在体内转导小胶质细胞，并开发新的基因调控元件这将使病毒载体表达小胶质细胞特异性转基因。这将是一个合作项目。史蒂文斯和德弗曼实验室之间的合作，结合了他们在研究小胶质细胞（史蒂文斯）和开发新型 AAV 载体 (Deverman)。