Immunologic correlates of functional cure of HBV with immune checkpoint blockade

乙型肝炎功能性治愈与免疫检查点阻断的免疫学相关性

• 批准号: 10388224
• 负责人: RAYMOND T CHUNG
• 金额: $ 83.45万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-22 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10388224
• 关键词: AIDS clinical trial group; Advanced Malignant Neoplasm; Affect; Antigen Targeting; Antigens; Antitumor Response; Blood; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Nucleus; Cells; Cellular Immunity; Cessation of life; Chronic; Chronic Hepatitis B; Clinical Trials; Data; Disease; Epigenetic Process; Fine needle aspiration biopsy; Freezing; Funding; Genetic Transcription; Genomics; HIV; Hepatitis B Vaccines; Hepatitis B Virus; Hepatocyte; Human; Immune; Immune response; Immune system; Immunity; Immunologics; Immunotherapeutic agent; Immunotherapy; In Situ; Individual; Inflammatory; Innate Immune Response; Liver; Mediating; Mediator of activation protein; Minority; Molecular; National Institute of Allergy and Infectious Disease; Natural Immunity; PD-1 blockade; PD-1 pathway; Participant; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Persons; Phagocytes; Phenotype; Population; Population Heterogeneity; Recovery; Regulation; Sampling; T cell response; T-Lymphocyte; Testing; Therapeutic; Tissues; Transcriptional Regulation; Treatment Failure; Treatment outcome; Viral; Viral Antigens; Viral Cancer; Virus; Virus Diseases; Virus Replication; anti-PD-1; anti-PD1 therapy; antigen-specific T cells; cancer therapy; checkpoint therapy; chronic infection; design; digital; exhaust; exhaustion; global health; immune checkpoint blockade; insight; intrahepatic; laser capture microdissection; macrophage; monocyte; novel; novel therapeutics; pathogen; pgRNA; programmed cell death ligand 1; programmed cell death protein 1; receptor; response; viral RNA

Chronic viral infections remain major threats to global health, with pathogens such as hepatitis B virus (HBV) responsible for millions of deaths annually. Despite availability of an HBV vaccine and suppressive antiviral treatment, the worldwide burden of chronic HBV infection has remained unchanged. Short-term therapies capable of producing at least functional cure for chronic HBV are therefore a high priority. Exhaustion of the immune system, most obvious in HBV-specific CD4 and CD8 T cells, is a key factor in HBV persistence and disease. The PD-1:PD-L1/2 inhibitory receptor pathway regulates many key aspects of cellular immunity, including T cell exhaustion in chronic viral infection and cancer. Blockade of this pathway has produced dramatic effects in the treatment of advanced cancer and unleashed an immunotherapeutic revolution. However, little is known about the effect of PD-1 blockade in chronic infections in humans, the mechanisms by which responses are invigorated, and how a reinvigorated HBV immune response impacts the intrahepatic HBV replication landscape. We propose to comprehensively investigate how blockade of PD-1 in humans affects the layers of molecular regulation of the antiviral immune response, and how this impacts viral replication in situ. The overall hypothesis of this project is that blocking PD-1 in humans will alter the magnitude, quality, regulation and composition of pre-existing antiviral CD4 and CD8 T cell responses, leading to more effective viral control, will modulate other aspects of cellular innate immunity, notably macrophages, and will lead to diminished viral replication in hepatocytes. We will test this hypothesis through the following aims. Specifically, in Aim 1 we will test whether and how PD-1 therapy invigorates exhausted virus-specific CD4 and CD8 T cell responses in the blood and liver. We will utilize liver fine needle aspirates and PBMC samples for a comprehensive and in-depth analysis of changes in the phenotype, function, clonal composition, and transcriptional state of HBV- specific CD4 and CD8 T cells. In Aim 2 we will test whether PD-1 therapy diminishes active HBV transcription in hepatocytes. Using an integrated platform of single-cell laser capture microdissection and droplet digital PCR on frozen liver tissues, we will quantify cccDNA and pre-genomic HBV RNA in hundreds of individual hepatocytes from participants before and at the completion of PD-1 blockade. Finally, in Aim 3 we will define the recovery of macrophages through PD-1 blockade in persons with chronic HBV infection. Here using FNA and PBMCs from patients with chronic HBV, we will assess whether αPD-1 treatment shifts the composition of macrophage populations toward an antiviral phenotype and test whether the functional responsiveness of monocyte/macrophages correlates with αPD-1 treatment outcome. Collectively, we expect these data to dramatically enhance our understanding of the mechanism of αPD-1 mediated immune recovery that can be utilized not only for the design of pathogen-specific immunotherapy, but also to enable further improvements in the efficacy of immunotherapy in general.

慢性病毒感染仍然是全球健康的主要威胁，其病原体包括乙型肝炎病毒 (HBV) 每年造成数百万人死亡。尽管有乙肝疫苗和抑制性抗病毒药物 治疗后，全球范围内慢性乙型肝炎病毒感染的负担保持不变。短期治疗 因此，能够至少对慢性乙型肝炎进行功能性治愈是当务之急。精疲力尽的 免疫系统，在 HBV 特异性 CD4 和 CD8 T 细胞中最为明显，是 HBV 持续存在和传播的关键因素。 疾病。 PD-1:PD-L1/2 抑制性受体途径调节细胞免疫的许多关键方面， 包括慢性病毒感染和癌症中的 T 细胞耗竭。这条通路的封锁产生了戏剧性的后果 治疗晚期癌症的效果，并引发了一场免疫治疗革命。然而，很少的是 已知 PD-1 阻断对人类慢性感染的影响，其反应机制 被激活，以及重新激活的 HBV 免疫反应如何影响肝内 HBV 复制 景观。我们建议全面研究人类 PD-1 的阻断如何影响细胞层 抗病毒免疫反应的分子调节，以及这如何影响病毒原位复制。整体 该项目的假设是，阻断人类的 PD-1 将改变其程度、质量、调节和 预先存在的抗病毒 CD4 和 CD8 T 细胞反应的组成，导致更有效的病毒控制，将 调节细胞先天免疫的其他方面，特别是巨噬细胞，并将导致病毒减少 在肝细胞中复制。我们将通过以下目标来检验这一假设。具体来说，在目标 1 中，我们将 测试 PD-1 疗法是否以及如何激活耗尽的病毒特异性 CD4 和 CD8 T 细胞反应 在血液和肝脏中。我们将利用肝脏细针抽吸物和 PBMC 样本进行全面和 深入分析HBV-的表型、功能、克隆组成和转录状态的变化 特异性 CD4 和 CD8 T 细胞。在目标 2 中，我们将测试 PD-1 疗法是否可以减少活性 HBV 肝细胞中的转录。使用单细胞激光捕获显微切割集成平台和 对冷冻肝组织进行液滴数字 PCR，我们将定量数百个样本中的 cccDNA 和前基因组 HBV RNA PD-1 阻断之前和完成时参与者的个体肝细胞。最后，在目标 3 中，我们 将确定慢性 HBV 感染者通过 PD-1 阻断后巨噬细胞的恢复情况。 在这里，我们将使用来自慢性 HBV 患者的 FNA 和 PBMC 来评估 αPD-1 治疗是否会改变 巨噬细胞群的组成朝向抗病毒表型并测试功能是否 单核细胞/巨噬细胞的反应性与 αPD-1 治疗结果相关。总的来说，我们期望 这些数据极大地增强了我们对 αPD-1 介导的免疫恢复机制的理解 这不仅可以用于设计病原体特异性免疫疗法，还可以进一步实现 免疫疗法总体疗效的改善。