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.

项目概要 尽管存在，但从宿主基因组中持续消除整合的、具有复制能力的 HIV-1 前病毒 抑制性抗逆转录病毒治疗（ART）是功能性治愈的主要障碍。含有这些类型的细胞 的原病毒产生边际水平的病毒产物，从而对免疫监视产生抵抗力 机制。除了其生长潜力增加之外，还缺乏免疫系统的检测，这是由于 稳态增殖和克隆扩增，延长潜伏感染细胞的寿命，产生 持久性水库。人们对针对潜在疾病的精准治疗的潜力抱有极大的热情 临床环境中的储存库。为了实现这一重大生物医学目标，我们必须首先发现主宰的宿主因素 在我们可以利用这些知识进行病毒库持久性和病毒潜伏期维持和重新激活之前 临床干预。虽然之前的研究使用了几种遗传方法来检查宿主因素 参与，他们面临着不允许区分直接影响和间接影响的内在问题。 这是一个重要的问题，因为我们必须首先定义宿主因子在 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。 在这个探索性和开发性的 R21 拨款申请中，我们通过以下方式规避了以前的问题： 实施一种新的化学遗传学（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）评估其在 HIV-1 原病毒潜伏期中的作用 维护和重新激活。我们将首先在 CD4+ T 细胞潜伏期模型中内源性地标记这些因素， 概括病毒在患者体内持续存在的生物学特性。然后，我们将选择优先候选人进行交叉 对从 UT 获得的潜伏期和无病毒血症患者样本进行原代 CD4+ T 细胞模型验证 西南/帕克兰 HIV-1 诊所。如果成功，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并阐明最合适的组蛋白赖氨酸甲基来研究生物学 用于试点转化研究的转移酶。超出本重点拨款申请范围的未来研究 将检查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 干预措施。