HIV and Methamphetamine-Associated Alterations of Behavior and Neural Networks

HIV-1和METH都可以引起行为改变和脑损伤，但它们之间的相互作用知之甚少。在当前的资助期内，我们观察到一次METH狂欢仅在HIV/gp 120转基因（tg）小鼠而不是对照小鼠中引起海马强直后增强的减少。RNA表达研究表明，HIVgp 120和METH都触发了多巴胺能和GABA能神经传递的显著变化。体外研究表明，100 pM的METH增加了HIVgp 120和兴奋毒性NMDA的神经毒性。METH还增加了人类巨噬细胞的HIV感染，这与至少四种干扰素诱导基因的下调有关。因此，总的假设是，使用METH会引起与HIV-1感染相关的行为障碍和神经毒性。我们的具体目标是：（1）通过研究与METH在HlVgpl 20-和iTat-tg和非tg对照小鼠中的长期作用相关的基因表达来鉴定受影响的神经网络，并将这种基因表达变化与行为表现相关联;（2）确定METH是否干扰cART药物减少病毒复制和受感染的单核细胞/巨噬细胞产生神经毒性产物的功效。对于目标1，gp 120-和iTat-tg和非tg对照小鼠将用新的慢性低剂量12周METH方案治疗。在5个月的戒断期和行为测试后，将使用去卷积显微镜、微阵列和qRT-PCR分析脑组织中的神经元和神经胶质损伤和基因表达。将分别分析皮质、海马、纹状体和其他脑区的基因表达数据，并将其与行为结果和神经病理学相关联。基因表达分析的后续研究将使用生物化学和显微镜方法探测蛋白质表达、定位和功能。对于特定目标2，感染HIV的巨噬细胞将用ARV组合和不同浓度的METH处理。将在大鼠小胶质细胞耗尽的混合神经元胶质细胞-皮质细胞上测试来自这种巨噬细胞实验的上清液的神经毒性。长期目标是确定作为治疗干预潜在靶点的神经毒性机制。

Both HIV-1 and METH can cause behavioral changes and brain injury, but their interaction is poorly understood. In the current funding period, we observed that a one-time METH binge caused a reduction in hippocampal post-tetanic potentiation only in HlV/gp120-transgenic (tg) but not control mice. RNA expression studies indicated that both HIVgp120 and METH trigger significant changes of glutamatergic and GABAergic neurotransmission. In vitro studies showed that METH at 100 pM increased neurotoxicity of HIVgp120 and excitotoxic NMDA. METH also increased HIV infection of human macrophages in association with down-regulation of at least four interferon-inducible genes. Therefore, the overall hypothesis is that use of METH aggravates behavioral disturbances and neurotoxicity associated with HIV-1 infection. Our Specific Aims are to: (1) identify affected neural networks by studying gene expression associated with long-term effects of METH in HlVgp120- and iTat-tg and non-tg control mice and to relate such gene expression changes to behavioral performance; (2) determine whether METH interferes with the efficacy of cART drugs to reduce viral replication and production of neurotoxic products by infected monocytes/macrophages. For Aim 1, gp120- and iTat-tg and non-tg control mice will be treated with a novel chronic, low dose 12-week METH regimen. Following a 5-month abstinence period and behavioral testing, neuronal and glial injury and gene expression will be analyzed in brain tissue using deconvolution microscopy, microarray and qRT-PCR. Gene expression data will be analyzed separately for cortex, hippocampus, striatum and other brain regions, and correlated with behavioral outcomes and neuropathology. Follow-up studies to gene expression analyses will probe protein expression, localization and function using biochemical and microscopy approaches. For Specific Aim.2, macrophages infected with HIV will be treated with ARV combinations and varying METH concentrations. The neurotoxicity of supernatents from such macrophage experiments will be tested on rat microglia depleted mixed neuronal glial cerebrocortical cells. The long-term objective is to identify neurotoxic mechanisms that are potential targets for therapeutic intervention.