HIV reactivation from latency - role of CD4 T cell maturation phenotype

HIV 从潜伏期重新激活 - CD4 T 细胞成熟表型的作用

• 批准号: 9323817
• 负责人: Nadejda S Beliakova-Bethell
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9323817
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Alpha Cell; Anti-Retroviral Agents; Antiviral Agents; Applications Grants; Area; Award; Bioinformatics; Biological Models; CD4 Positive T Lymphocytes; California; Cell Maturation; Cell Separation; Cells; Centers for Disease Control and Prevention (U.S.); Chronic; Clinical Trials; Data; Diagnosis; Dimethyl Sulfoxide; Enrollment; Ensure; Environment; Exposure to; Flow Cytometry; Funding; Future; Gene Expression; Gene Proteins; Genes; Genetic Transcription; Goals; HIV; HIV Infections; Healthcare Systems; Histone Deacetylase Inhibitor; Human; Immune system; Immunology; In Vitro; Individual; Infection; Intervention; Label; Mass Spectrum Analysis; Measures; Memory; Mentors; Methods; Modeling; Patients; Phenotype; Proteins; Proteome; Proteomics; RNA; RNA Splicing; Research; Research Personnel; Resolution; Role; Scientist; Shock; Solid; Solvents; System; Techniques; Testing; Time; Toxic effect; Training; Translational Research; United States; Universities; Veterans; Viral reservoir; Virus Activation; Vorinostat; Work; antiretroviral therapy; base; career; career development; career networking; cell type; clinically relevant; cost; digital; disorder prevention; experimental study; gene function; gene product; improved; in vitro testing; killings; knock-down; liquid chromatography mass spectrometry; memory CD4 T lymphocyte; overexpression; protein expression; public health relevance; purge; reactivation from latency; response; skills; small molecule; synergism; therapeutic candidate; therapeutic target; transcriptome sequencing; treatment strategy; validation studies; viral rebound

 DESCRIPTION (provided by applicant): The "shock and kill" treatment strategy has been envisioned as a controlled induction of virus reactivation in the presence of combination antiretroviral therapy (cART) to reveal latently infected cells for immune system recognition and destruction. Histone deacetylase inhibitors (HDACis) have potential for HIV reactivation in patients; however, further optimization of these strategies is needed to effectively reduce the size of latent reservoir. Emerging evidence from gene expression studies points to existence of secondary mechanisms of action of HDACis that may interfere with induction of HIV expression. Based on differences in the degree of HIV reactivation in response to HDACis in model systems that use different cell types, we propose that these inhibitory effects may vary among CD4+ T cells of different maturation phenotypes. The optimal reactivation strategy would be able to induce HIV expression from all CD4+ T cells with a minimal number of additional interventions; however, it is possible that maturation phenotype-specific interventions would be required. The objective of this proposal is to identify host genes and proteins modulated by HDACis in CD4+ T cells of various maturation phenotypes that can be therapeutically targeted to enhance the activation effect of HDACis. The central hypothesis is that HIV is reactivated by HDACis with different potencies and via different mechanisms depending on CD4+ T cell maturation state, and that reactivation may be augmented by counteracting the secondary effects of HDACis that interfere with HIV expression. The rationale for this study is to improve on existing strategies for HIV reactivation by identifying possible targets to counteract inhibitory effects of HDACis across cells of all majo maturation phenotypes to ensure HIV eradication from the entire CD4+ T cell reservoir. To achieve our goals, we will measure induction of HIV expression in naïve, central memory and effector memory cells using the in vitro latency model developed by Dr. Spina. We will then characterize global gene and protein expression changes induced by HDACis by methods of RNA-Seq and liquid chromatography mass spectrometry (LC-MS), and identify effects that may specifically interfere with HIV expression in CD4+ T cell subsets. Based on RNA-Seq and LC-MS data, a set of targets will be selected for validation studies, including validating changes in expression in a time course and in another latency model (developed by Dr. Karn), and knockdown / overexpression studies in the presence of HDACi to test for synergy. The expected positive impact of this work is identification of therapeutic candidates with strong potential to improve HIV reactivation currently achieved. Dr. Beliakova-Bethell will work with one primary mentor and three co-mentors, who are leaders in the field of HIV research. Exposure to a collaborative and diverse scientific environment, as part of the VA Healthcare System HIV research group, as well as the Center for AIDS Research at the University of California San Diego, will advance her career development. Her long-term career goal is to become a leader in the field of HIV-host interactions, HIV latency and immunology, and a successful VA scientist. During the period of the award, she will focus on obtaining training in: (1) HIV latency and human immunology; (2) proteomics; (3) translational research; and on developing skills necessary to build a solid, independently funded clinically relevant research career. To accomplish these goals, her career plan includes regular interactions with mentors, participation in specific didactic courses in career development, and acquiring expertise in new experimental techniques. Her mentors will help her establish a professional network through introductions to collaborators and will support transitioning to an independent career by serving as co-investigators on future grant proposals. Dr. Beliakova-Bethell will be well suited for independent research career within the VA System, with research focused in areas of importance to the VA.

 描述（由申请人提供）： “休克和杀死”治疗策略被设想为在联合抗逆转录病毒疗法（cART）的存在下受控诱导病毒再活化，以暴露潜伏感染的细胞用于免疫系统识别和破坏。组蛋白去乙酰化酶抑制剂（HDACis）有可能在患者中重新激活HIV;然而，需要进一步优化这些策略，以有效地减少潜伏库的大小。来自基因表达研究的新证据指出HDACis存在可能干扰HIV表达诱导的次要作用机制。基于在使用不同细胞类型的模型系统中响应HDACis的HIV再活化程度的差异，我们提出这些抑制作用在不同成熟表型的CD 4 + T细胞之间可能会有所不同。最佳的再活化策略将能够诱导所有CD 4 + T细胞的HIV表达，并且需要最少数量的额外干预;然而，可能需要成熟表型特异性干预。本提案的目的是鉴定在各种成熟表型的CD 4 + T细胞中由HDACis调节的宿主基因和蛋白质，其可以在治疗上靶向以增强HDACis的活化作用。中心假设是，HIV通过具有不同效力的HDACis并通过取决于CD 4 + T细胞成熟状态的不同机制被再激活，并且再激活可以通过抵消HDACis干扰HIV表达的次级效应来增强。本研究的基本原理是通过鉴定可能的靶点来抵消HDACis对所有主要成熟表型细胞的抑制作用，以确保从整个CD 4 + T细胞库中根除HIV，从而改善现有的HIV再活化策略。为了实现我们的目标，我们将使用Spina博士开发的体外潜伏期模型来测量幼稚、中枢记忆和效应记忆细胞中HIV表达的诱导。然后，我们将通过RNA-Seq和液相色谱质谱（LC-MS）方法表征HDACis诱导的全球基因和蛋白质表达变化，并确定可能特异性干扰HIV在CD 4 + T细胞亚群中表达的影响。基于RNA-Seq和LC-MS数据，将选择一组靶标用于验证研究，包括验证在时间过程中和另一个潜伏期模型（由Karn博士开发）中的表达变化，以及在HDACi存在下的敲减/过表达研究以测试协同作用。这项工作的预期积极影响是确定具有很大潜力的候选治疗方法，以改善目前实现的艾滋病毒重新激活。Beliakova-Bethell博士将与一名主要导师和三名共同导师合作，他们是艾滋病毒研究领域的领导者。作为VA医疗保健系统艾滋病研究小组的一部分，以及加州圣地亚哥大学艾滋病研究中心的一部分，接触到协作和多样化的科学环境将促进她的职业发展。她的长期职业目标是成为艾滋病毒-宿主相互作用，艾滋病毒潜伏期和免疫学领域的领导者，以及成功的VA科学家。在获奖期间，她将专注于获得以下方面的培训：（1）艾滋病毒潜伏期和人类免疫学;（2）蛋白质组学;（3）转化研究;以及培养建立坚实的，独立资助的临床相关研究职业所需的技能。为了实现这些目标，她的职业规划包括定期与导师互动，参加职业发展的特定教学课程，并获得新实验技术的专业知识。她的导师将通过介绍合作者帮助她建立专业网络，并将通过担任未来赠款提案的共同调查员来支持向独立职业的过渡。Beliakova-Bethell博士将非常适合VA系统内的独立研究生涯，研究重点是VA的重要领域。