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

基本信息

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
项目类别：
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

项目摘要

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) 尽管有乙肝疫苗和抑制性抗病毒药物，但每年仍有数百万人死亡。治疗方面，全球慢性乙型肝炎病毒感染的负担保持不变。因此，能够至少对慢性乙型肝炎产生功能性治愈是当务之急。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。 PD-1:PD-L1/2 抑制性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 细胞耗竭。对该通路的阻断产生了巨大的影响。治疗晚期癌症的效果并引发了一场免疫治疗革命，但效果甚微。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。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，并会导致病毒减少具体来说，我们将在目标 1 中测试这一假设。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 细胞反应我们将利用肝脏细针抽吸物和 PBMC 样本进行全面的分析。深入分析HBV-表型、功能、克隆组成和转录状态的变化特定的 CD4 和 CD8 T 细胞在目标 2 中，我们将测试 PD-1 疗法是否会减少活性 HBV。使用单细胞激光捕获显微切割和肝细胞转录的集成平台。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 最后，在目标 3 中，我们研究了 PD-1 阻断之前和完成时参与者的个体肝细胞。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总的来说，我们预计单核细胞/巨噬细胞的反应性与 αPD-1 治疗结果相关。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。