HIV and Methamphetamine-Associated Alterations of Behavior and Neural Networks

HIV-1和冰毒都会导致行为改变和脑损伤，但它们之间的相互作用尚不清楚。在目前的资助期间，我们观察到，一次性冰毒暴饮暴食只导致HLV/gp120转基因(TG)小鼠的海马区强直后增强减少，而不是对照组小鼠。RNA表达研究表明，HIVgp120和METH都能引起谷氨酸和GABA能神经传递的显著改变。体外研究表明，100 PM的冰毒增加了HIVgp120的神经毒性和兴奋性毒性NMDA。METH还增加了人类巨噬细胞的HIV感染，这与至少四个干扰素诱导基因的下调有关。因此，总体假设是，冰毒的使用加剧了与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)通过研究与冰毒长期作用相关的基因在HlVgp120、ITAT-TG和非甘油三酯对照组小鼠中的表达，确定受影响的神经网络，并将这些基因表达的变化与行为表现联系起来；(2)确定冰毒是否干扰CART药物的疗效，以减少受感染的单核/巨噬细胞的病毒复制和神经毒性产物的产生。对于目标1，gp120-、ITAT-TG和非甘油三酯对照组小鼠将接受一种新的慢性、低剂量12周冰毒疗法。经过5个月的戒断期和行为学测试，将使用去卷积显微镜、基因芯片和qRT-PCR分析脑组织中神经元和胶质细胞的损伤和基因表达。基因表达数据将分别分析皮质、海马体、纹状体和其他大脑区域，并与行为结果和神经病理学相关。基因表达分析的后续研究将使用生化和显微镜方法探索蛋白质的表达、定位和功能。对于特定的Aim2，感染HIV的巨噬细胞将用ARV组合和不同的冰毒浓度进行治疗。这种巨噬细胞实验上清液的神经毒性将在耗尽小胶质细胞的大鼠混合神经胶质脑皮质细胞上进行测试。长期目标是确定神经毒性机制，作为治疗干预的潜在靶点。

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.