A chemical genetics approach for studies of HIV-1 latency

研究 HIV-1 潜伏期的化学遗传学方法

• 批准号: 10711683
• 负责人: Ivan D'Orso
• 金额: $ 24.6万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711683
• 关键词: Acute; Applications Grants; Attention; Basic Science; Biology; Bypass; CD4 Positive T Lymphocytes; Cell model; Cells; Chromatin; Chronic; Clinic; Clinical; Clonal Expansion; Clustered Regularly Interspaced Short Palindromic Repeats; Deposition; Detection; Development; Epigenetic Process; Euchromatin; Event; Family; Future; Genetic Transcription; Genome; Genome engineering; Goals; Growth; HIV-1; Heterochromatin; Histones; Immune system; Immunologic Stimulation; Immunologic Surveillance; Integration Host Factors; Intervention; Knowledge; Longevity; Lysine; Maintenance; Methylation; Modeling; Molecular; Outcome; Patients; Physiological; Precision therapeutics; Proliferating; Provirus Integration; Proviruses; RNA Interference; Refractory; Regulation; Research; Role; Sampling; Shock; Source; Therapeutic; Therapeutic Intervention; Transcriptional Activation; Transferase; Validation; Viral; Virus Activation; Virus Latency; antiretroviral therapy; chemical genetics; clinically relevant; epigenetic silencing; genetic approach; histone modification; novel; novel strategies; reactivation from latency; success; targeted treatment; therapeutic target; translational study; viral rebound

PROJECT SUMMARY Elimination of integrated, replication-competent HIV-1 proviruses from host genomes persisting despite suppressive anti-retroviral therapy (ART) is the major roadblock to a functional cure. Cells harboring these types of proviruses produce marginal levels of viral products thereby becoming refractory to immune surveillance mechanisms. This lack of detection by the immune system, in addition to its increased growth potential, due to homeostatic proliferation and clonal expansion, extend the lifespan of latently infected cells generating a persistent reservoir. There is enormous enthusiasm for the potential of precision therapies targeting the latent reservoir in clinical settings. To achieve this major biomedical goal, we must first discover host factors dictating reservoir persistence and viral latency maintenance and reactivation before we can leverage this knowledge for clinical intervention. While previous studies have used several genetic approaches to examine host factor’s involvement, they hold the intrinsic problem of not allowing to distinguish between direct and indirect effects. This is a significant issue because one must first define the host factor’s primary function(s) in HIV-1 latency control to then illuminate the most appropriate approaches for therapeutic intervention. In this exploratory and developmental R21 grant application, we circumvent previous issues by implementing a novel chemical genetics (dTAG) approach to acutely eliminate the expression of a set of host chromatin regulators (Histone Lysine Methyl Transferases) to assess their roles in HIV-1 proviral latency maintenance and reactivation. We will first endogenously tag these factors in CD4+ T cell models of latency that recapitulate the biology of viral persistence in patients. We will then select prioritized candidates for cross- validation in primary CD4+ T cell models of latency and aviremic patient samples obtained from the UT Southwestern/Parkland HIV-1 Clinic. If successful, our studies will fill a void in our understanding of HIV-1 latency biology by describing new basic science and elucidating the most appropriate Histone Lysine Methyl Transferases for pilot translational studies. Future studies beyond the scope of this focused grant application will examine the clinical relevance of these host chromatin regulators and devise appropriate therapeutic interventions.

项目摘要 从宿主基因组中消除整合的、有复制能力的HIV-1前病毒， 抑制性抗逆转录病毒疗法（ART）是功能性治愈的主要障碍。这些类型的细胞 的前病毒产生边际水平的病毒产物，从而变得难以免疫监视 机制等这种缺乏免疫系统的检测，除了其增加的生长潜力，由于 稳态增殖和克隆扩增，延长潜伏感染细胞的寿命， 持久储层人们对针对潜伏性疾病的精确治疗的潜力抱有极大的热情。 临床环境中的储药器。为了实现这一主要的生物医学目标，我们必须首先发现宿主因素， 水库持久性和病毒潜伏期的维持和重新激活，然后我们才能利用这些知识， 临床干预。虽然之前的研究已经使用了几种遗传方法来检查宿主因素 由于参与，他们持有的内在问题是不允许区分直接和间接影响。 这是一个重要的问题，因为必须首先确定宿主因子在HIV-1潜伏期中的主要功能 控制，然后阐明最合适的治疗干预方法。 在这个探索性和发展性的R21赠款申请中，我们通过以下方式规避了以前的问题： 实施一种新型化学遗传学（dTAG）方法来急剧消除一组宿主的表达 染色质调节剂（组蛋白赖氨酸甲基转移酶），以评估其在HIV-1前病毒潜伏期中的作用 维护和重新激活。我们将首先在潜伏期的CD 4 + T细胞模型中内源性标记这些因子， 概括了病毒在患者体内持续存在的生物学。然后我们将选择优先的候选人进行交叉- 在从UT获得的潜伏期和病毒血症患者样本的原代CD 4 + T细胞模型中进行验证 西南/帕克兰HIV-1诊所如果成功的话，我们的研究将填补我们对HIV-1潜伏期理解的空白 通过描述新的基础科学和阐明最合适的组蛋白赖氨酸甲基生物学 用于初步翻译研究的转移酶。未来的研究超出了这个重点补助金申请的范围 将研究这些宿主染色质调节因子的临床相关性，并设计适当的治疗方法。 干预措施。