Harnessing the CARD8 Inflammasome for HIV Reservoir Elimination

利用 CARD8 炎症小体消除 HIV 病毒库

基本信息

批准号：
10676618
负责人：
LIANG SHAN
金额：
$ 81.21万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-10 至 2028-02-29
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10676618
关键词：
AIDS clinical trial group Acceleration Acquired Immunodeficiency Syndrome Aftercare Anti-Retroviral Agents Antibodies Antigens Apoptosis Automobile Driving CASP1 gene CD28 gene CD4 Positive T Lymphocytes Cell Death Cells Chromatin Clear Cell Clinical Clinical Research Clinical Trials Clonal Expansion Clone Cells DNA Development Dimerization Dipeptidyl Peptidases Disease remission Down-Regulation Epidemic Euchromatin Face Future Genetic Transcription Genomic Segment Genomics Goals HIV HIV-1 HIV-1 protease Human Immune Immune system Inflammasome Intervention Knock-in Mouse Measures Mediating Mission Monitor Mus Mutation Names Participant Pattern recognition receptor Peptide Hydrolases Peripheral Blood Mononuclear Cell Persons Pharmaceutical Preparations Phase Plasma Population Population Dynamics Predisposition Production Productivity Proliferating Proteins Provirus Integration Proviruses RNA RNA Sequences Randomized Regimen Research Resistance Sampling Shapes Shock T-Lymphocyte Testing Time Tissues United States National Institutes of Health Variant Viral Viral Cytopathogenic Effect Viral Load result Viral reservoir Virion Virus Virus Replication Work antiretroviral therapy cell killing cytokine decay acceleration efavirenz exhaustion experimental study humanized mouse immune clearance in vivo inhibitor innovation integration site latent HIV reservoir memory CD4 T lymphocyte mouse model non-nucleoside reverse transcriptase inhibitors novel novel strategies novel therapeutic intervention pre-clinical research premature pressure programs viral DNA viral RNA viral rebound

项目摘要

PROJECT SUMMARY The HIV-1 reservoir is a stable pool of latently infected CD4+ T cells that rekindles viral replication even after decades of antiretroviral therapy (ART). ART alone is not curative, requiring life-long treatment for people living with HIV-1 (PLWH). The main strategies attempted so far to eliminate HIV-1-infected cells face multiple challenges, including the selection of escape variants, resistance to apoptosis, T cell exhaustion, downregulation of MHC-I by HIV-1, and localization of infected cells in immune sanctuaries. This research program has the long- term goal of developing new therapeutic approaches to eliminate or control the HIV reservoir, leading to a drug- free remission. We recently discovered that the inflammasome protein CARD8 senses the enzymatic activity of the HIV-1 protease. We demonstrated that non-nucleoside reverse transcriptase inhibitors (NNRTIs) such as efavirenz (EFV) promote dimerization of Gag-Pol and cause premature intracellular activation of protease. HIV- 1 protease cleaves CARD8 driving the formation of caspase-1-dependent inflammasome and pyroptosis. All clinical HIV-1 isolates can be sensed by CARD8 despite viral diversity because it recognizes essential protease functions. The overall objective of this application is to harness CARD8 inflammasome to develop a novel approach to enhance HIV-1 reservoir elimination independently of CTLs or antibodies and trigger cell killing through non-apoptotic cell death. The underlying central hypothesis is that EFV-induced activation of CARD8, in combination with CARD8-enhancing drugs, can clear cells with transcriptionally active proviruses, reduce viral reservoir expansion, and gradually remove proviruses integrated into euchromatin regions. The rationale for the project is that enhancing the negative selection forces will reshape the proviral landscape, accelerate its decay, and reduce viral reactivation. The central hypothesis will be tested by pursuing three specific aims: 1) Establish EFV-induced CARD8 activation and killing of expanding HIV reservoir cells upon stimulation with cognate antigens ex vivo; 2) Determine the impact of EFV on HIV-1 reservoirs in humanized mice; and 3) Investigate the impact of EFV on HIV-1 reservoirs in PLWH in vivo by studying banked samples from clinical trials. The research proposed in this application is innovative because, compared to the status quo, it focuses on a new mechanism independent from HIV-1 diversity and cell susceptibility to apoptosis. The proposed research is significant because it is expected to provide the groundwork for the development of a new curative strategy that is scalable, broadly applicable in different contexts of clinical research, and can be easily combined with other interventions aiming to achieve HIV-1 remission. At completion, the proposed work will inform future pre-clinical and clinical research on the development of new antiretroviral compounds that can potently eliminate HIV-1-infected cells by activating CARD8-sensing of HIV-1 protease activity.

项目概要 HIV-1 储存库是一个稳定的潜伏感染 CD4+ T 细胞池，即使在感染后仍能重新点燃病毒复制。数十年的抗逆转录病毒治疗（ART）。单独的抗逆转录病毒疗法并不能治愈，需要对活着的人进行终生治疗 HIV-1（艾滋病病毒感染者）。迄今为止，消除 HIV-1 感染细胞的主要策略面临多种挑战。挑战，包括逃逸变异的选择、细胞凋亡抵抗、T 细胞耗竭、下调 HIV-1 对 MHC-I 的影响，以及受感染细胞在免疫避难所中的定位。该研究计划具有长期开发新的治疗方法来消除或控制艾滋病毒储存库的长期目标，从而导致药物免费缓解。我们最近发现炎性体蛋白 CARD8 能够感知 HIV-1 蛋白酶。我们证明了非核苷逆转录酶抑制剂（NNRTI），例如依非韦伦 (EFV) 促进 Gag-Pol 的二聚化并导致细胞内蛋白酶过早激活。艾滋病病毒- 1 蛋白酶裂解 CARD8，驱动 caspase-1 依赖性炎症小体的形成和细胞焦亡。全部尽管病毒具有多样性，但 CARD8 仍可检测临床 HIV-1 分离株，因为它可识别必需的蛋白酶功能。该应用的总体目标是利用 CARD8 炎性体开发一种新型独立于 CTL 或抗体增强 HIV-1 储存库消除并触发细胞杀伤的方法通过非凋亡细胞死亡。潜在的中心假设是 EFV 诱导的 CARD8 激活，与CARD8增强药物联合使用，可以清除具有转录活性原病毒的细胞，减少病毒病毒库扩大，并逐渐清除整合到常染色质区域的原病毒。理由如下：项目是增强负选择力将重塑原病毒景观，加速其衰退，并减少病毒再激活。将通过追求三个具体目标来检验中心假设：1）建立 EFV 诱导的 CARD8 激活并在同源刺激后杀死正在扩张的 HIV 储存细胞离体抗原； 2) 确定 EFV 对人源化小鼠中 HIV-1 储存库的影响； 3) 调查通过研究临床试验的储存样本，了解 EFV 对 PLWH 体内 HIV-1 储存库的影响。研究本申请中提出的方案具有创新性，因为与现状相比，它侧重于一种新的机制独立于 HIV-1 多样性和细胞凋亡敏感性。拟议的研究意义重大因为它有望为制定可扩展的新治疗策略奠定基础，广泛适用于临床研究的不同背景，并且可以轻松与其他干预措施相结合旨在实现 HIV-1 缓解。完成后，拟议的工作将为未来的临床前和临床提供信息研究开发可有效消除 HIV-1 感染细胞的新型抗逆转录病毒化合物通过激活 CARD8 感应 HIV-1 蛋白酶活性。