Novel approach to suppress HIV-1 innate inflammation

项目总结/摘要 拟议研究的重点是HIV-1感染引起的先天免疫激活。先天性炎症是由HIV-1复制和/或微生物Toll样受体[TLR]配体（例如脂多糖）从肠腔泄漏到循环中（微生物易位）引起的。HIV-1相关先天性炎症的后果尚不清楚，尽管HIV-1复制增强和一系列后遗症（包括HIV相关神经认知障碍、内皮功能障碍、心血管疾病、癌症或凝血病）与免疫激活相关。长期目标是通过开发靶向免疫激活的新方法来改善HIV-1相关先天性炎症的治疗。我推测，HIV-1和TLR配体通过新型信号网络协同介导经典的巨噬细胞活化，这些信号网络可以被HIV-1蛋白酶抑制剂靶向抑制。 HIV-1通过与干扰素引发表型相似的引发效应增强脂多糖诱导的巨噬细胞的经典活化，尽管具有更大的分子复杂性。目的1用系统生物学方法研究HIV-1诱导人巨噬细胞启动的分子机制。鉴定有助于HIV-1诱导的巨噬细胞活化的分子生物特征、候选基因/蛋白或细胞过程将是研究HIV-1感染免疫发病机制的长期方法的第一步。目的1是为进一步研究HIV-1诱导的先天性免疫激活奠定基础。研究结果还将提供新的见解HIV-1/宿主相互作用，并将提供基础的研究HIV-1复制/持久性在巨噬细胞。 目的2研究两种HIV-1蛋白酶抑制剂药物奈非那韦和替拉那韦的抗炎特性，独立于抗病毒作用。初步数据表明，奈非那韦和替拉那韦，独特的其他HIV-1蛋白酶抑制剂，对巨噬细胞发挥抗炎作用。目标2将确定抑制的分子基础，长期目标是鉴定药物的特定细胞蛋白质靶标，可能是宿主细胞蛋白酶。在这里，创新的蛋白质组学方法将被应用于询问主要脂多糖受体TLR 4下游的细胞信号传导事件。在这方面，奈非那韦和替拉那韦将不仅作为潜在的治疗药物，而且作为研究工具，解剖炎症细胞信号事件与HIV-1感染进行评估。 这项研究的结果将促进对HIV-1免疫发病机制的理解，并改善HIV-1感染炎症并发症的治疗/干预。这项研究不仅是为了实现科学目标，也是为了实现我独立职业发展的目标。具体目标的实现将为两个NIH R 01申请奠定基础，并增强机构对我的研究计划的承诺。

PROJECT SUMMARY / ABSTRACT The focus of the proposed study is innate immune activation by HIV-1 infection. Innate inflammation is caused by HIV-1 replication and/or microbial toll-like receptor [TLR] ligands (e.g. lipopolysaccharide) that leak from the intestinal lumen into the circulation (microbial translocation). The consequences of HIV-1-associated innate inflammation remain unclear, although enhanced HIV-1 replication and a spectrum of sequelae including HIV-associated neurocognitive impairment, endothelial dysfunction, cardiovascular disease, cancer, or coagulopathy are associated with immune activation. The long-term objective is to improve treatment of HIV-1-associated innate inflammation by developing novel approaches to target immune activation. I hypothesize that HIV-1 and TLR ligands cooperatively mediate classical macrophage activation through novel signaling networks that can be targeted for inhibition by HIV-1 protease inhibitors. HIV-1 augments lipopolysaccharide-induced classical activation of macrophages via a priming effect phenotypically similar to interferon-γ priming, although with greater molecular complexity. Aim 1 will determine the molecular mechanism of HIV-1 induced priming of human macrophages using a systems biology approach. Identification of molecular bioprofiles, candidate genes/proteins, or cellular processes that contribute to HIV-1- induced macrophage activation will be the first step in a long-term approach to study the immunopathogenesis of HIV-1 infection. Aim 1 is designed to lay the groundwork for future studies that investigate HIV-1-induced innate immune activation in vivo. Findings will also provide novel insights into the HIV-1/host interaction and will provide the basis for studies of HIV-1 replication/persistence in macrophages. Aim 2 will investigate anti-inflammatory properties of two HIV-1 protease inhibitor drugs nelfinavir and tipranavir, independent of anti-viral effects. Preliminary data demonstrates that nelfinavir and tipranavir, unique from other HIV-1 protease inhibitors, exert anti-inflammatory effects upon macrophages. Aim 2 will determine the molecular basis of inhibition with the long term goal of identifying a specific cellular protein target(s) of the drugs, potentially a host cell protease(s). Here innovative proteomics approaches will be applied to interrogate cell signaling events downstream of the primary lipopolysaccharide receptor TLR4. In this regard, nelfinavir and tipranavir will be evaluated not only as potential therapeutic agents, but also as investigative tools for dissecting inflammatory cell signaling events related to HIV-1 infection. Outcomes of this study will advance understanding of HIV-1 immune pathogenesis and lead to improved treatments/interventions for inflammatory complications of HIV-1 infection. The study is poised to achieve not only scientific goals, but also goals for my independent career development. Achieving the objectives of the Specific Aims will lay the groundwork for two NIH R01 applications and enhance institutional commitment to my research program.