Understand the role of CARD8 inflammasome in HIV-1 infection

了解 CARD8 炎性体在 HIV-1 感染中的作用

基本信息

批准号：
10327114
负责人：
LIANG SHAN
金额：
$ 70.2万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-21 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10327114
关键词：
Adaptive Immune System Amino Acid Sequence Antibodies Apoptotic Binding Biological Assay Blood C-terminal CASP1 gene CD28 gene CD3 Antigens CD4 Positive T Lymphocytes Cell Death Cell Line Cells Chemicals Cleaved cell Combined Modality Therapy Complex DNA Dimerization Early treatment Epitopes Evaluation Frequencies Gene Expression Genome Grant HIV HIV Protease HIV-1 HIV-1 protease Human Immune Immune system In Vitro Individual Infection Inflammasome Inflammation Interleukin-1 beta Interruption Investigation Knock-in Mouse Libraries Ligands Measures Mediating Molecular Molecular Conformation Mus Mutation N-terminal NNRTI-resistance Names Patients Pattern recognition receptor Peptide Hydrolases Pharmaceutical Preparations Physiological Plasma Protein Precursors Proteins RNA Regimen Reporter Research Resistance Rest Role Shock Signal Transduction T-Lymphocyte Testing Time Tissues Variant Viral Viral Cytopathogenic Effect Viral Genes Viral Proteins Viral reservoir Virus Virus Activation Virus Latency antiretroviral therapy base clinical subtypes cytokine efavirenz high throughput screening humanized mouse immune clearance in vivo latent HIV reservoir macrophage memory CD4 T lymphocyte microbial mouse model non-nucleoside reverse transcriptase inhibitors novel strategies pol Gene Products premature prevent protein function resistance mutation response screening viral RNA viral rebound

项目摘要

Abstract Despite antiretroviral therapy (ART), HIV-1 infection is not curable due to the presence of viral latent reservoirs primarily in long-lived resting memory CD4+ T cells. The viral latent reservoirs are the major barrier to HIV-1 eradication. The “shock and kill” strategy for HIV-1 eradication involves the use of latency-reversing agents (LRAs) to induce viral gene expression, which renders the infected cells susceptible to viral cytopathic effects or immune clearance. However, in ART-treated patients, the latent HIV-1 resides in cells is resistant to viral- or immune-mediated apoptotic cell death and often carries mutations to escape recognition by T cells or antibodies. Therefore, novel approaches to target immutable components of the virus such as essential viral protein functions are needed. Inflammasome is a critical molecular complex that mediates inflammation and pyroptotic cell death in response to microbial or danger signals. In humans, the physiologic ligand(s) for the CARD8 inflammasome remains unknown. Our studies demonstrate that HIV-1 protease degrades the CARD8 N-terminal domain and releases the C-terminus for inflammasome activation. In HIV-1-infected cells, the viral protease remains inactive as a subunit of viral Gag-Pol polyprotein. After virus budding, it is activated through Gag-Pol dimerization. We show that premature activation of intracellular HIV-1 protease by non-nucleoside reverse transcriptase inhibitors (NNRTI) triggers CARD8 sensing and killing of HIV-infected macrophages and CD4+ T cells. All subtypes of HIV-1 can be sensed by CARD8 despite substantial viral diversity. Our finding suggests that targeted activation of CARD8 inflammasome is a promising strategy to eliminate latent HIV-1 reservoirs. In this proposed study, we will: 1) perform ex vivo assessment and optimization of the CARD8-based “shock and kill strategy” for clearing latent HIV-1 in patient CD4+ T cells; 2) understand the molecular mechanisms of HIV-1 protease- mediated CARD8 inflammasome activation; 3) test CARD8-based “shock and kill” strategy in humanized mouse model of HIV-1 latency. Our proposed study will advance the understanding of the physiological mechanisms of CARD8 inflammasome activation and its role in HIV-1 infection, which will provide critical implications to HIV cure research.

抽象的尽管进行了抗逆转录病毒治疗 (ART)，但由于病毒的存在，HIV-1 感染仍无法治愈。潜伏病毒库主要存在于长寿命静息记忆 CD4+ T 细胞中。是根除 HIV-1 的主要障碍根除 HIV-1 的“休克和杀戮”策略。涉及使用潜伏期逆转剂（LRAs）来诱导病毒基因表达，这使受感染的细胞容易受到病毒细胞病变效应或免疫清除的影响。在接受 ART 治疗的患者中，潜伏在细胞中的 HIV-1 对病毒或免疫介导具有抵抗力细胞凋亡，并且常常携带突变以逃避 T 细胞或抗体的识别。因此，针对病毒的不可变成分（例如基本病毒成分）的新方法炎症体是介导蛋白质功能的关键分子复合物。对微生物或危险信号做出反应的炎症和焦亡细胞死亡。我们的研究表明，CARD8 炎性体的生理配体仍然未知。 HIV-1 蛋白酶降解 CARD8 N 末端结构域并释放 C 末端在 HIV-1 感染的细胞中，炎症小体激活，病毒蛋白酶作为亚基保持失活。病毒Gag-Pol多蛋白在病毒出芽后，通过Gag-Pol二聚化被激活。表明非核苷逆转细胞内 HIV-1 蛋白酶的过早激活转录酶抑制剂 (NNRTI) 触发 CARD8 感知并杀死 HIV 感染的巨噬细胞尽管存在大量病毒，CARD8 仍可检测到 HIV-1 的所有亚型和 CD4+ T 细胞。我们的研究结果表明，CARD8 炎性体的靶向激活是一种有前途的方法。消除潜在 HIV-1 病毒库的策略在这项拟议的研究中，我们将： 1) 进行离体治疗。基于CARD8的“电击杀伤策略”清除潜伏的评估和优化患者 CD4+ T 细胞中的 HIV-1；2) 了解 HIV-1 蛋白酶的分子机制- 介导的 CARD8 炎性体激活；3）测试基于 CARD8 的“休克和杀死”策略我们提出的研究将促进对 HIV-1 潜伏期的人源化小鼠模型的理解。 CARD8炎症小体激活的生理机制及其在HIV-1中的作用感染，这将为治愈艾滋病毒研究提供重要影响。