Inhibitors of host kinases as molecular targets for immune defense against HIV-1

宿主激酶抑制剂作为 HIV-1 免疫防御的分子靶点

• 批准号: 8702919
• 负责人: Mathias Lichterfeld
• 金额: $ 43.5万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8702919
• 关键词: ALVAC; Acute; Affect; B-Lymphocytes; Biochemical; Biological Assay; Biological Models; CD4 Positive T Lymphocytes; Cells; Cellular Assay; Clinical; Communicable Diseases; Consensus; Cyclin-Dependent Kinases; Data; Defense Mechanisms; Disease; Disease remission; Enzymes; Epigenetic Process; Evaluation; Family; Genetic Transcription; Goals; HIV; HIV vaccine; HIV-1; Highly Active Antiretroviral Therapy; Host resistance; Human; Immune; Immunologics; Immunologist; Infection; Infection prevention; Integration Host Factors; Intercept; Investigation; Life Cycle Stages; Mediating; Messenger RNA; MicroRNAs; Molecular; Molecular Profiling; Molecular Target; Molecular and Cellular Biology; Monitor; Pathogenesis; Pathway interactions; Patients; Pattern; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phosphotransferases; Physicians; Physiological; Predisposition; Prevention strategy; Process; Protein Biochemistry; Proteins; Proviruses; RNA-Directed DNA Polymerase; Regulation; Resistance; Reverse Transcription; Sampling; Scientist; Testing; Time; Transcript; Up-Regulation; Viral; Viral Load result; Virus; base; cell mediated immune response; cohort; experience; in vivo; inhibitor/antagonist; innovation; novel; oncoprotein p21; pathogen; patient population; prevent

DESCRIPTION (provided by applicant): Elite controllers are HIV-1 infected persons with undetectable viral loads in the absence of HAART, and the identification of immune defense mechanisms in these patients may reveal critical new information for increasing immune defense against HIV-1 in a broader HIV-1 patient population. Previous studies in elite controllers have mostly focused on classical T- and B-cell mediated immune responses against HIV-1. However, it has been shown that such adaptive immune activity may not be a sufficient, and sometimes not even a necessary component of effective immune protection against HIV-1 in these patients. In preliminary studies, we found that CD4 T cells from elite controllers are substantially less susceptible to HIV-1 replication. This partial resistance of CD4 T cells from elite controllers was mediated by a strong upregulation of the cyclin- dependent kinase inhibitor p21 (waf-1/cip-1), which was able to independently suppress HIV-1 reverse transcription and mRNA transcription. To our knowledge, these data represent the first description of a molecular HIV-1 restriction factor that actively inhibits HIV-1 replication in vivo in elite controllers and an therefore serve as a direct molecular target for enhancing host resistance to HIV-1. Here, we propose to extend our investigations of p21-mediated HIV-1 restriction by strategically focusing on three specific aims: To analyze molecular pathways responsible for the protective high-level expression of p21 in CD4 T cells from elite controllers, we will investigate how miRNAs regulate p21 gene transcription in CD4 T cells, using longitudinal samples collected from a unique cohort of HIV-1 elite controllers for whom PBMC samples are available from the time of acute infection (specific aim 1). In specific aim 2, we propose to mechanistically investigate how p21 affects HIV-1 reverse transcription, one of the earliest and most important HIV-1 replication steps. We hypothesize that p21 can indirectly inhibit HIV-1 reverse transcriptase by blocking host proteins from the cyclin-dependent kinase family that activate reverse transcriptase. If these studies are successful, we will identify a new aspect of HIV-1 host-pathogen interactions that can intercept an early HIV-1 replication step and may therefore be particularly attractive for clinical approaches to increase host resistance to HIV-1. Finally, we will mechanistically investigate how p21 may contribute to mechanisms of HIV-1 latency by affecting host factors involved in transcriptional elongation of HIV-1 mRNA transcripts; these investigations may provide important new information for inducing a long-term drug-free remission of HIV-1 infection by preventing viral outgrowth from latently infected cells (specific aim 3). Overall, these studies represent a highly interdisciplinary investigation integrating immunologic, virologic and biochemical approaches for the mechanistic investigation of a novel HIV-1 restriction factor that is effective at limiting the susceptibility to HIV-1 infection in vivo in elite controllers; if sucessful, these studies will reveal important new information for HIV-1 pathogenesis, immune defense and clinical strategies for prevention and treatment of HIV-1 infection.

描述（由申请人提供）：精英控制者是在没有HAART的情况下具有无法检测到的病毒载荷的HIV-1感染者，并且这些患者中免疫防御机制的鉴定可能会揭示关键的新信息，以增加更广泛的HIV HIV-HIV-1患者对HIV-1的免疫防御。先前在精英控制器的研究主要集中于经典的T和B细胞介导的针对HIV-1的免疫反应。但是，已经表明，这种适应性免疫活性可能不够，有时甚至不是在这些患者中有效保护HIV-1的必要组成部分。在初步研究中，我们发现来自精英控制器的CD4 T细胞易受HIV-1复制的易感性。 CD4 T细胞从精英控制器中的这种部分电阻是通过对细胞周期蛋白依赖性激酶抑制剂p21（WAF-1/CIP-1）的强上调介导的，该激酶抑制剂P21（WAF-1/CIP-1）能够独立地抑制HIV-1逆转录和mRNA转录。据我们所知，这些数据代表了分子HIV-1限制因子的第一个描述，该因子积极抑制Elite控制器中体内HIV-1复制，因此作为增强宿主对HIV-1抗性的直接分子靶标。在这里，我们建议通过策略性地关注三个特定目的来扩展对P21介导的HIV-1限制的研究：分析负责从精英控制器的CD4 T细胞中p21保护性高级表达的分子途径，我们将研究MiRNA在CD4 T细胞中调节p21基因在CD4 T细胞中的p21基因转录，该p21基因在CD4 T细胞中如何用于p21 p21，该途径是使用CD4 T细胞中的p21基因转录的，该p21是从CD4 T细胞中进行了p21，该途径是从longivent plosection proctions plote plotime plote ploti formection p21。从急性感染开始就可以使用样品（特定目标1）。在特定的目标2中，我们建议从机械上研究p21如何影响HIV-1逆转录，这是最早，最重要的HIV-1复制步骤之一。我们假设p21可以通过阻断激活逆转录酶的细胞周期蛋白依赖性激酶家族的宿主蛋白间接抑制HIV-1逆转录酶。如果这些研究成功，我们将确定HIV-1宿主 - 病原体相互作用的新方面，该方面可以拦截早期的HIV-1复制步骤，因此对于提高宿主对HIV-1的耐药性的临床方法可能特别有吸引力。最后，我们将通过影响涉及HIV-1 mRNA转录的转录伸长的宿主因子来机械地研究p21如何促进HIV-1潜伏期的机制；这些研究可能会提供重要的新信息，以通过防止潜在感染细胞的病毒生长来诱导HIV-1感染长期无毒的缓解（特定目标3）。总体而言，这些研究代表了一项高度跨学科的研究，该研究整合了免疫，病毒学和生化方法，用于对新型HIV-1限制因子进行机械研究，该研究有效地限制了精英控制器中体内HIV-1感染的易感性；如果成功，这些研究将揭示有关HIV-1发病机理，免疫防御和临床策略的重要新信息，以预防和治疗HIV-1感染。