Role of microRNAs in B-cell dysfunction in HIV/SIV infection

microRNA 在 HIV/SIV 感染 B 细胞功能障碍中的作用

• 批准号: 10060222
• 负责人: Mahesh Mohan
• 金额: $ 94.28万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10060222
• 关键词: Acetylation; Acute; Address; Anti-Inflammatory Agents; Antibodies; Antibody Response; Antiinflammatory Effect; Apoptosis; Attenuated; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; B cell differentiation; B-Cell Activation; B-Lymphocytes; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cannabinoids; Cell Survival; Cell physiology; Cells; Chronic; Data; Defect; Disease; Disease Progression; Down-Regulation; Functional disorder; Gastrointestinal tract structure; Gene Expression Regulation; Generations; Genetic Transcription; Goals; HIV; HIV Infections; Homeostasis; Hyperactive behavior; Immune; Immune Targeting; Immunologics; Immunosuppressive Agents; Impairment; In Vitro; Infection; Inflammation; Intestines; Lead; Link; Luciferases; Lymphocyte; Lymphomagenesis; MS4A1 gene; Macaca; Macaca mulatta; Marijuana; Mediating; Memory; Memory B-Lymphocyte; MicroRNAs; Molecular; Morbidity - disease rate; Multiple Sclerosis; Pathogenesis; Patients; Plasma Cells; Play; Population; Post-Transcriptional Regulation; Property; Quantitative Reverse Transcriptase PCR; RNA Interference; Regulation; Reporter; Research; Rheumatoid Arthritis; Role; SIRT1 gene; SIV; System; Systemic Lupus Erythematosus; T-Cell Depletion; Testing; Tetrahydrocannabinol; Therapeutic; Time; Up-Regulation; Vaccines; Virus Replication; antiretroviral therapy; base; chronic inflammatory disease; cytokine; high risk; immune activation; immunoregulation; improved; inhibitor/antagonist; knock-down; lymph nodes; marijuana use; mortality; novel; p65; plasma cell differentiation; prevent; public health relevance; restoration; therapeutic target; transcription factor; transcriptome

 DESCRIPTION (provided by applicant): B-cell dysfunction is a hallmark of HIV/SIV infection that begins very early in infection even before CD4 T cell depletion. Increasing evidence suggests that heightened B-cell activation, lack of memory B-cell restoration and impaired vaccine induced antibody responses persists even in patients receiving suppressive anti- retroviral therapy. Even though our understanding of B-cell dynamics in HIV/SIV infection has significantly improved, the molecular mechanisms underlying B-cell dysfunction remain poorly understood. In addition to cytokines and transcription factors, recent studies show miRNA-mediated gene regulation to be critical for B- cell differentiation, maturation, activation and autoimmune responses. Extensive studies in autoimmune diseases associated with B-cell defects have revealed a causative role for microRNAs in disease pathogenesis resulting in their identification as promising therapeutic targets. However, the role of miRNAs in HIV induced B-cell dysfunction is lacking. Our preliminary studies for this application identified significant dysregulation of miRNAs linked to B-cell activation and lymphomagenesis in purified B-cells from chronically SIV-infected macaques. Interestingly and more importantly, chronic THC treatment to SIV-infected macaques markedly inhibited their expression suggesting their immense therapeutic potential for attenuating immune (B-cell) activation and slowing disease progression. Based on our strong preliminary data demonstrating that miRNA expression is dysregulated in B-cells in HIV/SIV infection, we hypothesize that this leads to B-cell dysfunction which contributes to immune activation and HIV disease progression. Further, we hypothesize that chronic 9- THC treatment may reduce inflammation, restore immune (B-cell) function, and slow HIV/SIV disease progression by modulating miRNA expression. A major goal of this application is to identify miRNA mechanisms associated with B-cell dysfunction and cannabinoid mediated suppression of B-cell activation. The project has three specific aims: 1) Test the hypothesis that miR-34a and miR-155 targeting of SIRT1 leads to enhanced acetylated-p65 and acetylated-FOXO3a expression, causing B-cell hyperactivity and apoptosis, respectively. 2) Identify miRNA mechanisms of cannabinoid induced suppression of B-cell activation in blood, lymph nodes, and intestine at three different time points following SIV infection. 3) Determine the effect of combination anti-retroviral treatment (cART) in conjunction with chronic 9-THC administration on inflammation, chronic immune activation, viral replication, miRNA expression, SIRT1 levels, B-cell activation and apoptosis. The proposed research is novel and applies state of the art immunological and molecular approaches to address a significant gap in our understanding of the post-transcriptional mechanisms underlying B-cell dysfunction. Additionally and more importantly, the results will deepen our understanding of the mechanisms underlying the anti-inflammatory effects of cannabinoids and have important therapeutic implications for targeted immune modulation in HIV and other autoimmune diseases with B-cell defects.

 描述（由申请人提供）：B细胞功能障碍是HIV/SIV感染的标志，甚至在CD 4 T细胞耗竭之前就在感染早期开始。 越来越多的证据表明，即使在接受抑制性抗逆转录病毒治疗的患者中，B细胞活化增强、记忆B细胞恢复缺乏和疫苗诱导的抗体应答受损也持续存在。 尽管我们对HIV/SIV感染中B细胞动力学的理解已经有了显著的提高，但B细胞功能障碍的分子机制仍然知之甚少。除了细胞因子和转录因子外，最近的研究表明，miRNA介导的基因调控对B细胞分化、成熟、活化和自身免疫应答至关重要。在与B细胞缺陷相关的自身免疫性疾病中的广泛研究已经揭示了microRNA在疾病发病机制中的致病作用，导致其被鉴定为有希望的治疗靶点。然而，miRNA在HIV诱导的B细胞功能障碍中的作用缺乏。本申请的初步研究鉴定了与来自慢性SIV感染的猕猴的纯化B细胞中的B细胞活化和淋巴瘤发生相关的miRNA的显著失调。有趣的是，更重要的是，对SIV感染的猕猴进行慢性THC治疗显著抑制了它们的表达，这表明它们在减弱免疫（B细胞）活化和减缓疾病进展方面具有巨大的治疗潜力。基于我们强有力的初步数据表明，在HIV/SIV感染中，B细胞中的miRNA表达失调，我们假设这导致B细胞功能障碍，这有助于免疫激活和HIV疾病进展。此外，我们假设，慢性THC治疗可以减少炎症，恢复免疫（B细胞）功能，并通过调节miRNA表达来减缓HIV/SIV疾病的进展。本申请的主要目标是鉴定与B细胞功能障碍和大麻素介导的B细胞活化抑制相关的miRNA机制。该项目有三个具体目标：1）测试miR-34 a和miR-155靶向SIRT 1导致乙酰化p65和乙酰化FOXO 3a表达增强，分别引起B细胞过度活跃和凋亡的假设。2)在SIV感染后的三个不同时间点，确定大麻素诱导抑制血液、淋巴结和肠道中B细胞活化的miRNA机制。3)确定联合抗逆转录病毒治疗（cART）与慢性THC给药对炎症、慢性免疫活化、病毒复制、miRNA表达、SIRT 1水平、B细胞活化和凋亡的影响。拟议的研究是新颖的，应用最先进的免疫学和分子方法来解决我们对B细胞功能障碍的转录后机制的理解中的一个重大差距。此外，更重要的是，这些结果将加深我们对大麻素抗炎作用机制的理解，并对HIV和其他B细胞缺陷自身免疫性疾病的靶向免疫调节具有重要的治疗意义。