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治疗（HAART），但HIV-1感染经常导致神经系统问题的发展。此外，艾滋病毒感染通常与使用甲基苯丙胺（冰毒）等成瘾药物有关。虽然HIV-1和甲基苯丙胺都能引起行为、神经认知和组织病理学改变，但人们对病毒、HAART和甲基苯丙胺的潜在相互作用知之甚少，特别是在细胞和分子水平上，本文将对此进行研究。甲基苯丙胺和艾滋病毒会损害几种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在体外和体内产生神经元损伤和死亡，在其大脑中表达gp120的转基因小鼠出现与艾滋病患者相似的神经病理特征。因此，中心假设是冰毒的使用加重了HIV-1感染的神经毒性，从而损害了HAART治疗HIV感染的有益效果和HIV相关神经认知障碍的发展。长期目标是改善对神经艾滋病的治疗干预。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，我们将联合使用冰毒、HAART、HIVgp120和载体对照治疗脑皮质神经元，并评估神经元损伤、丢失和存活、电生理功能、细胞内Ca2+对兴奋性神经递质的反应以及神经元对兴奋性毒性损伤的易感性。使用TUNEL染色的核DNA显微镜结合神经元标记物的免疫染色来监测神经元的死亡和丢失。蛋白质、脂质和DNA的氧化以及活性Caspase 3将作为细胞应激和死亡信号的标志进行监测。冰毒、HAART和gp120在不同组合下对应激激酶p38 MAPK和促生存激酶Akt信号转导的潜在影响可能对增强HIV/gp120诱导的神经毒性至关重要，将使用两种激酶的显性阴性突变体进行研究。

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.