Effects of HIV PIs on CNS endothelial cells in NeuroAIDS

描述（由申请人提供）：长期职业目标是：1)开发一个研究项目，研究与神经精神疾病（如hiv相关痴呆（HAD））相关的宿主-病原体相互作用引发的细胞信号改变；2)成为学术机构的独立生物医学科学家。详细的职业发展和科学计划解决高活性抗逆转录病毒治疗（HAART）对神经艾滋病的潜在贡献提出。在血清转化后不久，HIV患者可能会被开以蛋白酶抑制剂（PI）为基础的HAART治疗方案。HAART已被证明在抑制全身病毒负担方面非常成功；然而，atp依赖的外排转运泵p -糖蛋白（P-gp）对神经aids的PI治疗施加了严重的限制。由于病毒反弹、耐药突变以及PI与HIV蛋白和脑内皮细胞（CEC）的潜在相互作用，本研究的主要目的是确定慢性PI暴露对CEC对宿主来源的生长因子（如成纤维细胞生长因子2 (FGF2)）的反应能力的影响，这些生长因子是在病毒反弹时作为防御血流中HIV蛋白的防御机制而产生的。我们假设长期暴露于沙奎那韦（SQV）、茚地那韦（INV）、奈非那韦（NFV）和/或利托那韦（RTV）等pi会改变P-gp外排依赖的表达和活性，从而改变fgf2介导的caveolin/ERK/NO系统中P-gp外排独立的信号通路。为此，AIM I将确定慢性PI治疗对CEC适应度的影响，以及FGF2通过与P-gp和小窝蛋白的相互作用介导的信号传导。CEC适应度将测试四个方面：1)活力，2)P-gp的表达和活性，3)P-gp介导的caveolin和内皮型一氧化氮合酶（eNOS）信号和一氧化氮（NO）的产生，4)fgf2介导的细胞外调节激酶（ERK）信号和血管生成能力。AIM II将在体外研究慢性暴露于PI的CEC如何破坏P-gp/小窝蛋白/ fgf2依赖性HIV蛋白gp120的保护。使用MSR-gp120和GFAPFGF2转基因小鼠，AIM III将在体内研究慢性PI治疗在与HIV蛋白相互作用过程中破坏血脑屏障（BBB）信号传导的长期影响。了解慢性PI暴露后通过P-gp/小泡介导的FGF2信号改变的机制，对于确定在病毒反弹或病毒学失败期间可能导致与神经aids相关的神经和神经行为改变进展的因素非常重要。

DESCRIPTION (provided by applicant): Long-term career goals are to: 1) develop a research program to investigate the cellular signaling alterations triggered by host-pathogen interactions relevant to neuro-psychiatric disorders such as HIV-associated Dementia (HAD) and 2) become an independent bio-medical scientist at an academic institution. Detailed career development and scientific plans addressing the potential contribution of highly active anti-retroviral therapy (HAART) to NeuroAIDS is proposed. Shortly after sero-conversion, HIV patients may be prescribed protease inhibitor (PI)-based HAART regimens. HAART has proven highly successful in suppressing systemic viral burden; however, the ATP-dependent efflux transport pump, P-glycoprotein (P-gp), imposes a serious constraint on PI treatment of NeuroAIDS. Because of viral rebound, resistance mutations and the potential interactions of PIs with HIV proteins and cerebral endothelial cells (CEC), the main objective of this proposal is to determine the effects of chronic PI exposure on the CEC's ability to respond to host-derived growth factors, such as fibroblast growth factor 2 (FGF2), that are generated as defense mechanisms against HIV proteins _resent in the blood stream at viral rebound. We hypothesize that long-term exposure to PIs such as saquinavir (SQV), indinavir (INV), nelfinavir (NFV) and/or ritonavir (RTV) will modify P-gp efflux-dependent expression and activity, thereby altering P-gp efflux-independent signaling pathways in the FGF2-mediated caveolin/ERK/NO systems. For this purpose, AIM I will determine the effects of chronic PI treatment on CEC fitness and signaling mediated by FGF2 via interactions with P-gp and caveolin. Four aspects of CEC fitness will be tested: 1) viability, 2) P-gp expression and activity, 3) P-gp-mediated caveolin and endothelial nitric oxide synthase (eNOS) signaling and nitric oxide (NO) production, 4) FGF2-mediated extracellular regulated kinase (ERK) signaling and angiogenic capacity. AIM II will address in vitro, how chronic exposure of CEC to PI disrupts P-gp/caveolin/FGF2-dependent protection from the HIV protein, gp120. Using MSR-gp120 and GFAPFGF2 transgenic mice, AIM III will investigate, in vivo, the long-term effects of chronic PI treatment in disrupting signaling at the blood-brain barrier (BBB) during interaction with HIV proteins. Understanding the mechanisms responsible for alterations in FGF2 signaling via P-gp/caveolae after chronic PI exposure is important for identifying factors that may contribute to the progression of neurological and neurobehavioral alterations associated with NeuroAIDS during viral rebound or at virologic failure.