Microglia, HIV and drugs of abuse

我们正在研究啮齿动物小胶质细胞如何在慢性和急性暴露模型中对精神兴奋剂（如甲基苯丙胺）作出反应。 正在进行的工作已经确定了甲基苯丙胺的剂量，导致纹状体中小胶质细胞的独特激活，可用于检查激活的小胶质细胞如何影响周围的神经元。 我们最近开发了一种新的小胶质细胞系，该细胞系在基因组中插入了一种经修饰的HIV前病毒。 该前病毒缺乏gag和pol基因，但表达可用于监测前病毒转录活性的效应子的荧光素酶。 我们已经使用CRISPR靶向该细胞系中HIV的长末端重复序列（LTR）启动子，并使用它来评估基于CRISPR的方法来改变小胶质细胞中的HIV前病毒活性。 众所周知，小胶质细胞很难用转基因材料靶向。 我们最近鉴定了一种能够在大鼠小胶质细胞中表达的AAV载体。 我们还在研究一种非病毒方法，将转基因材料（DNA，RNA，CRISPR核糖核蛋白）递送到小胶质细胞。这些工具将使我们能够更容易地检查小胶质细胞产生HIV蛋白的影响，以及暴露于成瘾物质时对神经元周围的影响。

We are examining how rodent microglia respond to psychostimulants such as methamphetamine in both a chronic and acute exposure model. Ongoing work has identified dosing of methamphetamine that causes unique activation of microglia in the striatum that can be used to examine how activated microglia can affect the surround neurons. We have recently developed a novel microglial cell line that has a modified HIV provirus inserted into the genome. This provirus lacks the gag and pol genes but expresses a luciferase which can be used to monitor effectors of proviral transcriptional activity. We have used CRISPR to target the long terminal repeat (LTR) promoter of HIV in this cell line and are using it to evaluate CRISPR-based approaches to alter HIV proviral activity in microglia. Microglia are notoriously difficult to target with transgenic material. We have recently identified an AAV vector capable of expressing in rat microglia. We are also working on a non-viral approach to delivery transgenic material (DNA, RNA, CRISPR ribonucleoproteins) to microglia. These tools will allow us to more readily examine the effect of microglia production of HIV proteins and the effects on surrounds in neurons when exposed to substances of addiciton.