HIV-induced monocyte and macrophage perturbations driving liver fibrogenesis

HIV诱导的单核细胞和巨噬细胞扰动驱动肝纤维化

• 批准号: 10647917
• 负责人: Nadia Alatrakchi
• 金额: $ 65.1万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10647917
• 关键词: Anti-Inflammatory Agents; Antiinflammatory Effect; Automobile Driving; B-Lymphocytes; Biological Assay; Blocking Antibodies; Cell Line; Cells; Chronic; Cicatrix; Data; Environment; Excision; Exposure to; Fatty Acids; Fine needle aspiration biopsy; Grant; HIV; HIV Infections; HIV/HCV; Hepatic; Hepatic Fibrogenesis; Hepatic Stellate Cell; Hepatitis B Virus; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; Immune; Immunosuppressive Agents; In Vitro; Infection; Inflammation; Inflammatory; Kupffer Cells; Light; Liver; Liver Fibrosis; Liver diseases; Molecular Target; Myeloid Cells; Myeloid-derived suppressor cells; Natural Immunity; Pathogenesis; Pathologic; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Phenotype; Plasma; Play; Population; Proteins; Proteomics; Public Health; Recombinants; Residual state; Role; S100A9 gene; Serum; Testing; Viral; Viral Antigens; Virus; antiretroviral therapy; cell type; cellular targeting; chronic liver disease; cohort; comparative; fibrogenesis; immune activation; in vivo; liver injury; macrophage; macrophage product; monocyte; nonalcoholic steatohepatitis; peripheral blood; scavenger receptor; simple steatosis; systemic inflammatory response; transcriptomics

Abstract Liver disease in patients with HIV infection remains a major public health problem despite effective combination antiretroviral therapy (cART), and cART does not fully reverse HIV-related immune activation. Monocyte and macrophage populations play a central role in inflammation and liver pathogenesis and are triggered during HIV, even in the absence of known causes of liver injury. Chronically activated monocytes/macrophages polarize to a spectrum of pro-inflammatory and anti-inflammatory states which contribute to hepatic fibrogenesis. Our preliminary study of the immune cell landscape in liver fine needles aspirates (FNAs) and corresponding peripheral blood mononuclear cells (PBMCs) from patients with HIV infection prior to and during the first 6- months of cART, uncovered 1) the presence of monocytic myeloid-derived suppressor cells (M-MDSC) that persist on cART despite virus control; and 2) increased serum levels of sCD5L, the soluble scavenger receptor secreted by macrophages, which does not decrease on cART. Both M-MDSCs and sCD5L are described to expand in inflammatory conditions, have anti-inflammatory effects, and recently have been shown to promote the anti-inflammatory/profibrotic macrophage phenotype. We have also observed similar findings in HCV infection and NASH. Together these findings may constitute a causative mechanism driving a profibrotic milieu, particularly in the liver. We hypothesize that HIV-induced M-MDSCs and CD5L remain upregulated on suppressive ART and contribute to a persistent profibrotic liver milieu. Using patient derived human material, we will test this hypothesis through the following aims: (1) Define the role of M-MDSCs in driving fibrogenesis in the setting of HIV infection. Using M-MDSCs from healthy donor PBMC pre/post exposure to HIV, we will assess whether HIV-induced M-MDSCs drive the profibrotic profile in macrophages and hepatic stellate cells (HSCs) and will explore contributory mechanisms using blocking/silencing functional assays. Additionally, to confirm whether M-MDSCs persist in suppressed HIV in vivo, we will comprehensively characterize the transcriptomic/proteomic single cell landscape in HIV patient PBMCs, pre-ART/at several timepoints of cART. (2) Define the role of CD5L in driving fibrogenesis in the setting of HIV infection. Using recombinant (r)CD5L we will determine whether this product promotes the profibrogenic profile in HIV-triggered healthy donor monocyte derived macrophages and M-MDSCs as well as in HIV-exposed primary HSCs and/or hepatocytes. Additionally, we will determine if plasma from HIV patients (pre/during cART) with or without CD5L blocking antibodies induces a profibrotic phenotype. (3) Determine how HIV impacts M-MDSC and CD5L function in the setting of co-existing liver disease. We will comparatively explore M-MDSCs and CD5L by single cell transcriptomics/proteomics in cohorts of patients with HIV, HIV/HCV, HIV/HBV and HIV/NASH in liver FNAs, and we will validate suggested profibrogenic mechanisms in vitro using primary/cell lines co-exposed to these insults. Together these studies will shed great light on the contribution of monocytes and macrophages to HIV-associated liver disease.

摘要 尽管有效的联合治疗，HIV感染患者的肝脏疾病仍然是一个主要的公共卫生问题 抗逆转录病毒疗法（cART），并且cART不能完全逆转HIV相关的免疫激活。单核细胞和 巨噬细胞群体在炎症和肝脏发病机制中起着中心作用并且在HIV期间被触发， 即使没有已知的肝损伤原因。慢性活化单核细胞/巨噬细胞 导致肝纤维化的一系列促炎和抗炎状态。我们 肝脏细针穿刺物（FNAs）中免疫细胞景观的初步研究 外周血单个核细胞（PBMC）从HIV感染患者之前和期间的第一个6- 月的cART，发现1）单核细胞髓源性抑制细胞（M-MDSC）的存在， 尽管病毒得到控制，但仍持续接受cART; 2）可溶性清道夫受体sCD 5L的血清水平升高 由巨噬细胞分泌，在cART中不会减少。M-MDSC和sCD 5L均被描述为 在炎症条件下扩张，具有抗炎作用，最近已被证明可以促进 抗炎/促纤维化巨噬细胞表型。我们在HCV中也观察到了类似的发现 感染和NASH。这些发现一起可能构成驱动促纤维化环境的致病机制， 特别是在肝脏中。我们假设HIV诱导的M-MDSC和CD 5L在 抑制性ART并导致持续的促纤维化肝环境。使用患者来源的人体材料，我们 我们将通过以下目的来检验这一假设：（1）确定M-MDSC在驱动肝纤维化中的作用。 艾滋病毒感染的背景。使用来自暴露于HIV前/后的健康供体PBMC的M-MDSC，我们将评估 HIV诱导的M-MDSC是否驱动巨噬细胞和肝星状细胞（HSC）中的促纤维化特征 并将使用阻断/沉默功能测定来探索贡献机制。此外，为了确认 无论M-MDSC是否在体内持续抑制HIV，我们将全面表征 在ART前/cART的几个时间点，HIV患者PBMC中的转录组/蛋白质组单细胞景观。 (2)定义CD 5L在HIV感染背景下驱动纤维化中的作用。使用重组（r）CD 5L， 将确定该产品是否促进HIV触发的健康供体单核细胞中的促纤维化特征 衍生的巨噬细胞和M-MDSC以及暴露于HIV的原代HSC和/或肝细胞中。此外，本发明还 我们将确定来自HIV患者（cART前/期间）的血浆（有或没有CD 5L阻断抗体）是否诱导 促纤维化表型。(3)确定HIV如何影响M-MDSC和CD 5L功能， 肝脏疾病我们将通过单细胞转录组学/蛋白质组学比较研究M-MDSCs和CD 5L， 在肝脏FNAs中的HIV、HIV/HCV、HIV/HBV和HIV/NASH患者队列中，我们将验证建议 使用共暴露于这些损伤的原代/细胞系在体外研究促纤维化机制。这些研究 将有助于了解单核细胞和巨噬细胞对HIV相关肝病的作用。