Harnessing the CARD8 Inflammasome for HIV Reservoir Elimination

利用 CARD8 炎症小体消除 HIV 病毒库

• 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
• 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

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.

项目摘要 HIV-1储存库是一个稳定的潜伏感染的CD 4 + T细胞库，即使在感染后， 抗逆转录病毒疗法（ART）。ART本身并不能治愈，需要终身治疗的人生活 HIV-1（PLWH）。迄今为止，试图消除HIV-1感染细胞的主要策略面临多重挑战。 挑战，包括逃逸变体的选择，抗凋亡，T细胞耗竭，下调 MHC-I的HIV-1，和本地化的感染细胞在免疫避难所。这项研究计划有很长的- 长期目标是开发新的治疗方法，以消除或控制艾滋病毒水库，导致药物- 免费减免。我们最近发现，炎性小体蛋白CARD 8可以感受到 HIV-1蛋白酶我们证明了非核苷类逆转录酶抑制剂（NNRTI），如 依法韦仑（EFV）促进Gag-Pol的二聚化并引起蛋白酶的过早胞内活化。艾滋病毒- 1蛋白酶切割CARD 8，驱动半胱天冬酶-1依赖性炎性小体和焦亡的形成。所有 尽管存在病毒多样性，但临床HIV-1分离株可被CARD 8检测到，因为它识别必需的蛋白酶 功能协调发展的本申请的总体目标是利用CARD 8炎性小体来开发一种新的免疫抑制剂。 一种增强HIV-1储库消除的方法，不依赖于CTL或抗体，并触发细胞杀伤 通过非凋亡性细胞死亡。潜在的中心假设是EFV诱导的CARD 8激活， 与CARD 8增强药物组合，可以清除具有转录活性前病毒的细胞，减少病毒感染， 储存库扩大，并逐渐去除整合到常染色质区域的前病毒。的理由 项目是，增强负选择力将重塑前病毒景观，加速其衰变， 并减少病毒再激活。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CARD 8活化和在同源物刺激后对扩增的HIV储库细胞的杀伤 抗原离体; 2）确定EFV对人源化小鼠中HIV-1储库的影响;和3）研究EFV对人源化小鼠中HIV-1储库的影响。 通过研究临床试验的库存样本，研究EFV对PLWH体内HIV-1储库的影响。研究 在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 独立于HIV-1多样性和细胞对凋亡的易感性。所提出的研究是有意义的 因为它被期望为可扩展的新治疗策略的开发提供基础， 广泛适用于临床研究的不同背景，并且可以很容易地与其他干预措施相结合 旨在达到HIV-1的缓解。完成后，拟议的工作将为未来的临床前和临床 研究开发新的抗逆转录病毒化合物，可以有效地消除HIV-1感染的细胞 通过激活HIV-1蛋白酶活性的CARD 8感应。