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: 9137251
• 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/9137251
• 关键词: Acquired Immunodeficiency Syndrome; Affect; 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; Goals; HIV; HIV Infections; Healthcare Systems; Histone Deacetylase Inhibitor; Human; Immune system; Immunology; In Vitro; Individual; Infection; Intervention; Label; Life; 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; T-Lymphocyte Subsets; Techniques; Testing; Therapeutic; Time; Toxic effect; Training; Translational Research; United States; Universities; Veterans; Viral reservoir; Virus; Virus Activation; Vorinostat; Work; antiretroviral therapy; base; career; career development; career networking; cell type; clinically relevant; cost; digital; disorder prevention; 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; research study; response; skills; small molecule; transcriptome sequencing; treatment strategy; validation studies

 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)： “休克并杀死”治疗策略被设想为在联合抗逆转录病毒治疗(CART)的情况下控制地诱导病毒重新激活，以揭示潜伏感染的细胞，以识别和破坏免疫系统。组蛋白去乙酰酶抑制剂(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，以有效减少潜伏库的大小。来自基因表达研究的新证据表明，HDACis存在可能干扰HIV表达诱导的二级作用机制。根据使用不同细胞类型的模型系统中对HDACis反应的HIV重新激活程度的差异，我们认为这些抑制作用可能在不同成熟表型的CD4+T细胞之间有所不同。最优的再激活策略将能够在最少的额外干预次数下诱导所有CD4+T细胞表达HIV；然而，可能需要成熟表型特定的干预。本研究的目的是在不同成熟表型的CD4+T细胞中识别受HDACis调控的宿主基因和蛋白，这些基因和蛋白可以作为治疗靶点来增强HDACis的激活效果。中心假说是，HIV被HDACi以不同的效力和不同的机制重新激活，这取决于CD4+T细胞的成熟状态，并且可以通过抵消HDACis干扰HIV表达的次要效应来增强重新激活。这项研究的基本原理是通过确定可能的靶点来对抗HDACis对所有主要成熟表型细胞的抑制作用，以确保从整个CD4+T细胞库中根除HIV，从而改进现有的HIV再激活策略。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天真、中央记忆和效应器记忆细胞中HIV表达的诱导。然后，我们将通过RNA-Seq和LC-MS方法表征HDACis诱导的全球基因和蛋白质表达变化，并识别可能特异性干扰CD4+T细胞亚群中HIV表达的影响。基于RNA-Seq和LC-MS数据，将选择一组靶点进行验证研究，包括验证某个时间段和另一个潜伏期模型(由Karn博士开发)中表达的变化，以及在HDACi存在的情况下进行敲除/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目前实现的具有改善艾滋病毒重新激活的强大潜力的治疗候选者。贝利亚科娃-贝塞尔博士将与一名主要导师和三名共同导师合作，他们都是艾滋病毒研究领域的领导者。作为退伍军人管理局医疗系统艾滋病毒研究小组的一部分，以及加州大学圣地亚哥分校的艾滋病研究中心，在协作和多样化的科学环境中的接触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，以及一名成功的退伍军人事务部科学家。在获奖期间，她将重点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)翻译研究；以及发展必要的技能，以建立坚实的、独立资助的临床相关研究事业。为了实现这些目标，她的职业规划包括定期与导师互动，参加职业发展方面的特定教学课程，以及获得新的实验技术方面的专业知识。她的导师将通过向合作者介绍来帮助她建立一个专业网络，并将通过担任未来拨款提案的联合调查员来支持过渡到独立的职业生涯。Beliakova-Bethell博士将非常适合在退伍军人管理局系统内从事独立研究，专注于对退伍军人管理局重要的领域的研究。