Microglia, HIV and drugs of abuse

我们正在研究啮齿动物小胶质细胞在慢性和急性暴露模型中对甲基苯丙胺的反应。正在进行的工作已经确定了甲基苯丙胺的剂量，这种剂量会导致纹状体小胶质细胞的独特激活，这可以用来研究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，它将修饰的HIV前病毒插入到基因组中。这种前病毒缺乏Gag和Pol1基因，但表达一种荧光素酶，可用于监测前病毒转录活性的效应因子。我们已经使用CRISPR在该细胞系中靶向HIV的长末端重复序列(LTR)启动子，并正在使用它来评估基于CRISPR的改变小胶质细胞中HIV前病毒活性的方法。 众所周知，小胶质细胞很难用转基因材料作为靶点。我们正在研究一种非病毒方法，将转基因材料(DNA、RNA、CRISPR核糖核蛋白)输送到小胶质细胞。 最后，使用上述转基因构建物的修改形式用于细胞系，我们正在开发小胶质细胞和星形胶质细胞中HIV致病的转基因大鼠模型。

We are examining how rodent microglia respond to 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 are working on a non-viral approach to delivery transgenic material (DNA, RNA, CRISPR ribonucleoproteins) to microglia. Finally, using a modified form of the transgenic construct described above for the cell line, we are developing a transgenic rat model of HIV pathogenesis in microglia and astrocytes.