Novel AAV Capsids and Gene Regulatory Elements for GeneExpression in Microglia

项目总结： 小胶质细胞是大脑的常驻巨噬细胞，在大脑发育和发育过程中承担着多种功能。 疾病。很早就知道对阿尔茨海默病(AD)影响的大脑变化做出反应，最近的基因研究- 发病阿尔茨海默病表明AD风险变异体通常在特定表达于 提示小胶质细胞在阿尔茨海默病的发病机制中起着积极作用。此外，小胶质细胞还表现出各种 大脑发育过程中的功能，包括突触修剪。编码基因成分的基因 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疾病大脑中的作用的一个主要瓶颈是缺乏病毒。 用于快速操纵其基因表达的工具。病毒载体也将有利于转基因研究 动物是不可用的。最后，病毒载体将使研究阿尔茨海默病和老年痴呆症的基因治疗的可能性成为可能。 影响小胶质细胞的罕见疾病。然而，虽然小胶质细胞表现出适度和局部的转导，但通过 慢病毒在体内，它们对腺相关病毒(AAVs)转导具有抵抗力，是首选病毒 载体因其优越的传播性和良好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对小胶质细胞转导的障碍，设计AAV衣壳变异体，使其 能够克服这些障碍，在体内转导小胶质细胞，并开发新的基因调控元件 这将使病毒载体中的小胶质细胞特异性转基因表达成为可能。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，结合他们在研究小胶质细胞方面的专业知识(史蒂文斯)和 开发新的AAV载体(Deverman)。

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).