HIV-CRISPR: A novel approach to the comprehensive discovery of HIV latency factors

HIV-CRISPR：全面发现HIV潜伏因子的新方法

• 批准号: 10371192
• 负责人: Michael Emerman
• 金额: $ 88万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10371192
• 关键词: AIDS/HIV problem; Adopted; Antiviral Agents; Antiviral Therapy; Basic Science; Cataloging; Cell model; Cells; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Combination Drug Therapy; Communities; Dose; Genes; Genetic; Genetic Screening; Genetic screening method; Goals; HIV; HIV Budding; HIV Genome; HIV Infections; Human; Immune system; Immunologics; Individual; Intervention; Life; Maintenance; Methods; Modeling; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Proviruses; Research; Specificity; Techniques; Technology; Time; Virus; Virus Latency; antiretroviral therapy; base; burden of illness; cell killing; cell type; drug of abuse; high throughput technology; in vivo; innovation; new technology; novel; novel strategies; screening; synergism; transmission process

1 Project Summary/Abstract The availability of simplified and better-tolerated antiretroviral therapy (ART) drug combinations has decreased disease burden in infected individuals and reduced transmission to their contacts. However, treatment for HIV infections requires life-long antiviral therapy since current drug treatments do not lead to “cures”. The single most significant obstacle to total HIV clearance is that of viral latency since the presence of a silent HIV genome in human cells is not recognized as infected by the immune system or by any available antiviral drugs. Thus, the largest challenge to the HIV/AIDS basic sciences research community today is to determine targets and strategies to fully eradicate virus from the body by eliminating this reservoir pool of latently infected cells. One strategy to eliminate latently infected cells involves activating the latent provirus with drugs called latency reversal agents (LRAs) in the presence of suppressive ART, and then using immunological methods to kill the cells newly-producing virus. However, we lack a comprehensive and complete understanding of how HIV latency is established and maintained, and how reactivation occurs across cell types. Importantly, no single genetic or chemical intervention has been able to induce activation of all latent proviruses (or even the majority of them). This proposal describes a novel and powerful approach to understanding HIV latency based on an innovative genetic screening method that we have developed called HIV-CRISPR that uses the packaging of CRISPR guides into budding HIV to serve as a readout for the effects of host genes on HIV. This technique is both high-throughput and comprehensive and our plan is to adapt to models of HIV latency. Because the screening technology is versatile and can be done in many iterations, we will be able to adopt a strategy based on combining genetic screens with low doses candidate latency reversal agents to identify synergism between different pathways of HIV latency maintenance in order to find targets that more broadly activate HIV from latency while at the same time providing a greater degree of specificity for the HIV LTR. Moreover, we will develop methods this technique to primary cell models of HIV latency to better mimic how to reverse latency in vivo. Finally, will also apply this screen to understand how drugs of abuse influence HIV latency pathways.

1 项目摘要/摘要 简化且耐受性更好的抗逆转录病毒疗法（ART）药物组合的可用性具有 疾病减少了感染的个体燃烧，并减少了向其接触的传播。然而， 艾滋病毒感染的治疗需要终身抗病毒治疗，因为目前的药物治疗不会导致 “治愈”。全艾滋病毒完全清除的最重要的障碍是病毒潜伏期，因为存在 人类细胞中的无声艾滋病毒基因组未被免疫系统或任何可用的感染。 抗病毒药物。这是当今艾滋病毒/艾滋病基础科学研究社区最大的挑战 通过消除该储层池，确定从体内完全放射性病毒的目标和策略 潜在感染的细胞。 消除潜在感染细胞的一种策略涉及用药物激活潜在的病毒 在存在抑制性艺术的情况下，称为潜伏期逆转剂（LRA），然后使用免疫学 杀死新产生病毒细胞的方法。但是，我们缺乏全面而完整的 了解如何建立和维持艾滋病毒潜伏期，以及如何在细胞之间进行重新激活 类型。重要的是，尚无单一遗传或化学干预能够诱导所有潜在的激活 病毒（甚至大多数）。 该提案描述了一种新颖而有力的方法来理解基于一个的艾滋病毒潜伏期 我们开发的创新基因筛选方法，称为HIV-Crispr，使用的包装 CRISPR引导为崭露头角的艾滋病毒，以作为宿主基因对艾滋病毒的影响的读数。这种技术是 高通量和全面，我们的计划是适应艾滋病毒潜伏期的模型。因为 筛选技术用途广泛，可以在许多迭代中完成，我们将能够采用策略 基于将遗传筛选与低剂量的候选潜伏期反向剂相结合以识别协同作用 在艾滋病毒潜伏期维持的不同途径之间，以找到更广泛激活艾滋病毒的目标 从延迟开始，同时为HIV LTR提供了更高程度的特异性。 此外，我们将将这种技术开发为艾滋病毒潜伏期的主要细胞模型，以更好地模仿如何如何模仿 体内反向潜伏期。最后，还将应用此屏幕以了解滥用药物如何影响艾滋病毒 延迟路径。