Harnessing the CARD8 Inflammasome for HIV Reservoir Elimination

项目总结 HIV-1的储存库是一个稳定的潜伏感染的CD4T细胞池，即使在 数十年的抗逆转录病毒治疗(ART)。单靠抗逆转录病毒疗法是不能治愈的，需要对活着的人进行终身治疗。 人类免疫缺陷病毒1型(PLWH)。到目前为止，试图消灭HIV-1感染细胞的主要战略面临着多个 挑战，包括逃逸变异体的选择、对凋亡的抵抗、T细胞耗竭、下调调节 HIV-1对MHC-I的感染，以及受感染细胞在免疫保护区的定位。这个研究项目有很长的- 开发新的治疗方法以消除或控制艾滋病毒宿主的长期目标，导致一种药物- 免费减免。我们最近发现，炎症体蛋白CARD8感觉到 HIV-1蛋白水解酶。我们证明了非核苷逆转录酶抑制剂(NNRTI)，如 EFV可促进GAG-Pol的二聚化，导致细胞内蛋白水解酶过早激活。艾滋病毒-- 1酶能裂解CARD8，导致caspase-1依赖的炎性小体和下垂形成。全 尽管存在病毒多样性，但CARD8仍能感觉到HIV-1临床分离株，因为它识别必要的蛋白酶 功能。本应用程序的总体目标是利用CARD8炎症小体开发一种新的 一种增强不依赖CTL或抗体并触发细胞杀伤的HIV-1宿主清除的方法 通过非凋亡性细胞死亡。潜在的中心假设是EFV诱导CARD8，in 与增强CARD8的药物相结合，可以清除转录活性前病毒的细胞，减少病毒 储集层扩大，并逐渐清除整合成常染色质区域的前病毒。该计划的基本原理 项目是，加强负面选择力量将重塑前驱动力格局，加速其腐朽， 并减少病毒的重新激活。中心假设将通过追求三个具体目标来检验：1)建立 EFV诱导的CARD8在同源刺激下对正在扩张的HIV储存库细胞的激活和杀伤 体外抗原；2)确定EFV对人源化小鼠HIV-1储存库的影响；以及3)调查 通过研究临床试验中储存的样本，研究流行性出血热病毒对PLWH中HIV-1储存库的影响。这项研究 本申请中提出的是创新的，因为与现状相比，它侧重于一种新的机制 不受艾滋病毒-1多样性和细胞对凋亡的敏感性的影响。这项拟议的研究具有重要意义 因为它有望为开发一种可扩展的新治疗策略提供基础， 广泛适用于临床研究的不同情况，并可轻松与其他干预措施相结合 旨在实现HIV-1的缓解。完成后，拟议的工作将通知未来的临床前和临床 有效清除HIV-1感染细胞的新型抗逆转录病毒化合物的开发研究 通过激活对HIV-1蛋白酶活性的CARD8传感。

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.