Effects of HIV PIs on CNS endothelial cells in NeuroAIDS

HIV PI 对 NeuroAIDS 中中枢神经系统内皮细胞的影响

• 批准号: 7046764
• 负责人: Dianne Teresa LANGFORD
• 金额: $ 13.01万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7046764
• 关键词: AIDS /HIV neuropathy; AIDS therapy; HIV envelope protein gp120; HIV infections; P glycoprotein; SDS polyacrylamide gel electrophoresis; apoptosis; biological signal transduction; blood brain barrier; caveolins; central nervous system; clinical research; fibroblast growth factor; genetically modified animals; host organism interaction; immunocytochemistry; indinavir; laboratory mouse; mitogen activated protein kinase; nelfinavir; nitric oxide synthase; polymerase chain reaction; protease inhibitor; ritonavir; saquinavir; virus infection mechanism; western blottings

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.

描述（由申请人提供）：长期职业目标是：1）制定研究计划，以调查与与神经心理疾病有关的宿主 - 病原体相互作用触发的细胞信号变化，例如与HIV相关的痴呆（HAT）和2）成为一名学术机构的独立生物医学科学家。提出了详细的职业发展和科学计划，旨在提出高度活跃的抗逆转录病毒疗法（HAART）对神经辅助的潜在贡献。血清转化后不久，HIV患者可能是基于蛋白酶抑制剂（PI）的HAART方案。 Haart在抑制系统性病毒负担方面已被证明非常成功。然而，依赖ATP的外排运输泵P-糖蛋白（P-GP）对神经辅助的PI治疗施加了严重的限制。 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反弹。 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.为此，目的我将通过与P-gp和小窝蛋白的相互作用来确定慢性PI处理对FGF2介导的CEC适应性和信号的影响。将测试CEC适应性的四个方面：1）生存力，2）P-gp表达和活性，3）P-gp介导的小窝蛋白和内皮一氧化氮合酶（ENOS）信号传导和一氧化氮（NO）的产生（4）FGF2介导的FGF2介导的FGF2介导的FGF2介导的细胞外调节激酶（ERK）信号和远征能力。 AIM II将在体外解决，CEC慢性暴露在PI中如何破坏P-gp/Caveolin/fgf2依赖性保护与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处理在与HIV蛋白相互作用期间破坏血脑屏障（BBB）的信号传导的长期影响。了解慢性PI暴露后通过P-gp/caveolae改变FGF2信号的机制对于识别可能有助于神经和神经行为变化的因素，在病毒反弹或病毒学衰竭期间与神经辅助的发展。