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: 10170260
• 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/10170260
• 关键词: AIDS clinical trial group; Advanced Malignant Neoplasm; Affect; Antigen Targeting; Antigens; Antitumor Response; Antiviral Agents; 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; Structure of molecular layer of cerebellar cortex; 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B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 尽管目前的治疗水平有所下降，但全球慢性HBV感染的负担仍保持不变。短期治疗 因此，能够产生至少慢性HBV功能性治愈是高度优先的。耗尽 免疫系统，在HBV特异性CD 4和CD 8 T细胞中最明显，是HBV持续存在的关键因素， 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。对这条通路的封锁产生了戏剧性的 在治疗晚期癌症方面的效果，并引发了一场免疫革命。然而， 已知PD-1阻断对人类慢性感染的影响、反应机制 以及重新激活的HBV免疫应答如何影响肝内HBV复制 景观我们建议全面研究PD-1在人体中的阻断如何影响细胞层， 抗病毒免疫反应的分子调节，以及这如何影响原位病毒复制。整体 该项目的假设是，阻断人类中的PD-1将改变PD-1的大小、质量、调节和 预先存在的抗病毒CD 4和CD 8 T细胞应答的组成，导致更有效的病毒控制， 调节细胞先天免疫的其他方面，特别是巨噬细胞，并将导致减少病毒 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治疗是否以及如何消除耗尽的病毒特异性CD 4和CD 8 T细胞应答 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CD 4和CD 8 T细胞。在目标2中，我们将测试PD-1治疗是否减少活动性HBV 肝细胞中的转录。使用单细胞激光捕获显微切割和 在冷冻肝组织上进行液滴数字PCR，我们将在数百个样本中定量cccDNA和前基因组HBV RNA。 在PD-1阻断之前和完成时来自参与者的个体肝细胞。最后，在目标3中， 将定义慢性HBV感染者中通过PD-1阻断巨噬细胞的恢复。 在这里，我们将使用慢性HBV患者的FNA和PBMC，评估αPD-1治疗是否改变了HBV的免疫功能。 组成的巨噬细胞群体的抗病毒表型，并测试是否功能 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，我们希望 这些数据大大增强了我们对αPD-1介导的免疫恢复机制的理解 这不仅可以用于病原体特异性免疫疗法的设计，而且还可以进一步 总体上提高免疫疗法的功效。