Combined effect of Methamphetamine, HIV and HAART on neurons and macrophages

描述(由申请人提供)：尽管出现了高效抗逆转录病毒疗法(HAART)，但感染HIV-1往往会导致神经问题的发展。此外，艾滋病毒感染常常与使用甲基苯丙胺(冰毒)等成瘾药物有关。虽然HIV-1和冰毒都可以引起行为、神经认知和组织病理学改变，但病毒、HAART和冰毒之间的潜在相互作用尚不清楚，特别是在细胞和分子水平上，这里将对其进行研究。冰毒和艾滋病毒损害了几个神经递质系统的功能。HIV-1包膜蛋白gp120通过CD4和趋化因子共受体与巨噬细胞和小胶质细胞结合。HIV/gp120在体外和体内都会造成神经元的损伤和死亡，在他们的大脑中表达gp120的转基因小鼠表现出与艾滋病患者相似的神经病理特征。因此，中心假说是，冰毒的使用加剧了HIV-1感染的神经毒性，从而损害了HAART对HIV感染和HIV相关神经认知障碍发展的有益效果。长期目标是改善对神经艾滋病的治疗干预。其具体目的是：表征甲基苯丙胺、HIV-1和HAART对神经元生物学功能和存活的联合影响。为此，我们将使用METH、HAART、HIVgp120和赋形剂对照的组合治疗大脑皮层神经元，并评估神经元损伤、丢失和存活、电生理功能、兴奋性神经递质反应的细胞内钙离子以及神经元对兴奋性毒性损伤的易感性。神经元的死亡和丢失将使用TUNEL染色的核DNA显微镜结合神经元标记物的免疫染色进行监测。蛋白质、脂肪和DNA的氧化，以及活性的Caspase3将被监测为细胞应激和死亡信号的标志。METH、HAART和gp120在不同的组合中对应激激酶p38MAPK和促存活激酶Akt信号转导的潜在影响可能是增强HIV/gp120诱导的神经毒性的关键，将使用这两种激酶的显性负突变来研究。 公共卫生相关性：感染HIV-1通常会导致神经系统并发症，尽管进行了高效的抗逆转录病毒治疗(HAART)。此外，艾滋病毒感染经常与接触甲基苯丙胺(冰毒)等成瘾药物有关，两者都是主要的公共卫生问题。我们的研究将提高对冰毒和HIV感染在HAART存在的情况下造成脑损伤的理解。

DESCRIPTION (provided by applicant): Infection with HIV-1 often leads to the development of neurological problems despite the advent of highly active antiretroviral therapy (HAART). In addition, HIV-infection is frequently associated with the use of addictive drugs, such as Methamphetamine (METH). While both HIV-1 and METH can cause behavioral, neurocognitive and histopathological changes, the potential interaction of virus, HAART and METH is poorly understood, in particular at the cellular and molecular level, and will be studied here. METH and HIV compromise the function of several neurotransmitter systems. HIV-1 envelope protein gp120 binds to macrophages and microglia via CD4 and chemokine co-receptors. HIV/gp120 produces in vitro and in vivo neuronal injury and death, and transgenic mice expressing gp120 in their brain develop neuropathological features similar to AIDS patients. Thus, the central hypothesis is that use of METH aggravates the neurotoxicity of HIV-1 infection and thus compromises the beneficial effect of HAART against HIV infection and the development of HIV-associated neurocognitive disorders. The long-term objective is to improve therapeutic intervention for neuroAIDS. The specific aim is: To characterize the combined impact of Methamphetamine, HIV-1 and HAART on the biological function and survival of neurons. For that purpose we will treat cerebrocortical neurons with combinations of METH, HAART, HIVgp120 and vehicle controls and assess neuronal injury, loss and survival, electrophysiological function, intracellular Ca2+ in response to excitatory neurotransmitter and neuronal vulnerability to excitotoxic insult. Neuronal death and loss will be monitored using microscopy of TUNEL stained nuclear DNA in combination with immunostaining for neuronal markers. Oxidation of protein, lipid and DNA, and active Caspase 3 will be monitored as markers of cellular stress and death signaling. The potential effect of METH, HAART and gp120 in various combinations on signal transduction via stress kinase p38 MAPK and pro-survival kinase Akt may be crucial to an enhancement of HIV/gp120-induced neurotoxicity and will be studied using dominant negative mutants of both kinases. PUBLIC HEALTH RELEVANCE: Infection with HIV-1 is often leads to neurological complications despite highly active antiretroviral therapy (HAART). Moreover, HIV infection is frequently associated with exposure to addictive drugs, such as Methamphetamine (METH), and both are major public health concerns. Our studies will improve the understanding of brain injury caused by the combination of METH and HIV infection in the presence of HAART.