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，病原体如乙肝病毒(乙肝病毒) 每年造成数百万人死亡。尽管有了乙肝疫苗和抑制性抗病毒药物 治疗后，世界范围内慢性乙肝病毒感染的负担仍未改变。短期治疗 因此，能够产生至少治疗慢性乙肝的功能性治疗是一个高度优先的问题。精疲力竭的 免疫系统，最明显的是在乙肝病毒特异性的CD4和CD8T细胞，是一个关键因素，在乙肝持续和 疾病。PD-1：PD-L1/2抑制受体通路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细胞衰竭。对这条道路的封锁已经产生了戏剧性的 在治疗晚期癌症方面的效果，并引发了一场免疫治疗革命。然而，几乎没有什么是 已知PD-1阻断在人类慢性感染中的作用，反应的机制 被激活，以及重新激活的乙肝免疫反应如何影响肝脏内的乙肝复制 风景。我们建议全面研究PD-1在人类体内的阻断如何影响 抗病毒免疫反应的分子调控，以及这如何影响病毒的原位复制。整体而言 该项目的假设是，在人类中阻断PD-1将改变其大小、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，并将导致病毒减弱 在肝细胞中复制。我们将通过以下目标来检验这一假设。具体地说，在目标1中，我们将 测试PD-1疗法是否以及如何激活疲惫的病毒特异性CD4和CD8T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 深入分析乙肝病毒的表型、功能、克隆组成和转录状态的变化 特异性的CD4和CD8 T细胞。在目标2中，我们将测试PD-1治疗是否减少活动性乙肝病毒 肝细胞中的转录。利用单细胞激光捕获显微切割的集成平台 液滴数字聚合酶链式反应在冰冻肝组织上的应用，我们将对数百个 受试者PD-1阻断前和阻断完成时的单个肝细胞。最后，在目标3中，我们 将确定通过PD-1阻断慢性乙肝感染者巨噬细胞的恢复。 在这里，我们将使用慢性乙肝患者的FNA和PBMC来评估αPD-1治疗是否改变了 巨噬细胞群体的组成向抗病毒表型转变，并测试其功能 单核/巨噬细胞的反应性与αPD-1治疗结果相关。总体而言，我们期待 这些数据极大地提高了我们对αPD-1介导的免疫恢复机制的理解 这不仅可以用于病原体特异性免疫疗法的设计，还可以使进一步的 免疫疗法的疗效总体上有所改善。