Defining and exploiting EBV-infected cell heterogeneity in non-Hodgkin lymphomas

定义和利用非霍奇金淋巴瘤中 EBV 感染细胞的异质性

基本信息

批准号：
10541348
负责人：
Micah A. Luftig
金额：
$ 55.06万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-19 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10541348
关键词：
Adult African Burkitt&apos s lymphoma Age Animal Model B lymphocyte immortalization B-Cell Lymphomas B-Cell NonHodgkins Lymphoma B-Lymphocytes Behavior Biological Markers Cell Separation Cell model Cells Clinical Trials Coupled Cytotoxic T-Lymphocytes DNA Damage Data Development Epstein-Barr Virus Infections Epstein-Barr Virus latency Exhibits Failure Frequencies Ganciclovir Gene Expression Genetic Goals Growth HIV Infections Herpesviridae Heterogeneity Histone Deacetylase Histone Deacetylase Inhibitor Human Human Herpesvirus 4 Immune Immunosuppression In Vitro Individual Infection Latent virus infection phase Lymphoma Lymphoma cell Lymphomagenesis Lytic Lytic Virus Mediating Minor Modeling Molecular Monitor Non-Hodgkin&apos s Lymphoma Oncogenic Oncogenic Viruses Organ Transplantation Outcome Pathogenesis Phenotype Phosphotransferases Plasmablast Population Predisposition Proliferating Protein Kinase Regulation Repression Research Rest Role T cell response Testing Therapeutic Therapeutic Intervention Tumor Tissue Valganciclovir Viral Viral Proteins Virus Work Xenograft Model base cell killing epstein barr virus mediated immortalization experimental study gene product in vivo infected B cell innate immune sensing large cell Diffuse non-Hodgkin&apos s lymphoma latent infection lymphoblastoid cell line mouse model neoplastic cell novel therapeutics programs response sensor single-cell RNA sequencing success transforming virus tumor tumor heterogeneity tumor progression tumorigenesis virtual

项目摘要

Epstein-Barr virus (EBV) was the first human tumor virus discovered over 50 years ago in the context of endemic African Burkitt lymphoma. However, we now know it is also a common herpesvirus that persists as a lifelong latent infection in virtually all adults worldwide. Early work in the field led to a model for EBV infection promoting B-cell lymphomas as evidenced by the growth transformation, or immortalization, of primary resting human B cells into lymphoblastoid cell lines (LCLs). In vivo, EBV latent infection is met with a robust cytotoxic T-cell response keeping most infected individuals protected from the oncogenic potential of the virus. As such, EBV-associated B-cell lymphomas occur at significantly higher rates in the setting of immune suppression. Studies of viral and cellular gene expression in EBV-infected cells in vitro and in vivo have led to a model of lymphomagenesis characterized by the full expression of EBV latency gene products. However, the phenotypes in bulk culture and tumor tissue lack the nuanced detail of cellular heterogeneity and the consequences of minor frequency phenotypes on cancer progression. Our recent single cell RNAseq experiments have characterized gene expression within individual EBV-infected B cells leading to an appreciation of cell fate trajectories and dynamic gene expression behavior of individual cells that we will integrate with human tumor analysis and mouse models of lymphomagenesis. It is our ultimate goal to define the importance of specific EBV-infected cell populations on the progression of B-cell non-Hodgkin lymphomas of the immune suppressed. In this proposal, we aim to define how EBV-infected cell heterogeneity, including innate antiviral restriction and plasmablast differentiation, impacts lymphomagenesis and can be exploited for therapy. Our central hypothesis is that EBV- infected B cells toggle between different states that can restrict or promote lymphomagenesis as well as render cells susceptible to virus-specific therapeutic intervention. We formulated our central hypothesis based on preliminary data including single-cell RNA sequencing of EBV-infected primary B cells early after infection and in LCLs as well as characterization of cell fate dynamics regulating plasmablastic differentiation and lytic reactivation. We also provide evidence supporting a recent clinical trial using the “kick and kill” strategy of promoting EBV lytic reactivation with histone deacetylase inhibition coupled with ganciclovir to kill lymphoma cells that activate viral kinases. Thus, the rationale for the proposed research is that understanding EBV regulation of infected B-cell fates will dissect mechanisms of pathogenesis and reveal new therapeutic avenues to target EBV-positive B-cell lymphomas. We plan to test our central hypothesis and complete the objectives in this proposal through the following three specific aims: i) to define the role of innate immune sensors and effectors in EBV-mediated immortalization and lymphomagenesis, ii) to determine the role of plasmablast differentiation in suppressing EBV-mediated lymphomagenesis, and iii) to define the mechanism by which HDAC inhibition promotes susceptibility of EBV+ DLBCL to killing by ganciclovir.

爱泼斯坦 - 巴尔病毒（EBV）是50年前发现的第一种人类肿瘤病毒内在非洲伯基特淋巴瘤。但是，我们现在知道这也是一种常见的疱疹病毒几乎全世界所有成年人的终生潜在感染。该领域的早期工作导致了EBV感染的模型促进B细胞淋巴瘤，这是由原发性静止的生长转化或永生化所证明的人B细胞进入淋巴母细胞系（LCLS）。在体内，EBV潜在感染具有强大的细胞毒性 T细胞反应使大多数受感染的个体免受病毒的致癌潜力。像这样，在免疫抑制的情况下，与EBV相关的B细胞淋巴瘤以明显更高的速率发生。在体外和体内对EBV感染细胞中病毒和细胞基因表达的研究导致了一种模型淋巴作用的特征是EBV潜伏基因产物的全部表达。但是，表型在散装培养和肿瘤组织中，缺乏细胞异质性的细微细节和次要的后果癌症进展的频率表型。我们最近的单细胞RNASEQ实验表征了单个EBV感染的B细胞中的基因表达，导致对细胞命运轨迹的欣赏和单个细胞的动态基因表达行为，我们将与人类肿瘤分析和小鼠整合淋巴细胞化的模型。这是我们定义特定EBV感染细胞的重要性的最终目标受免疫抑制的B细胞非霍奇金淋巴瘤进展的种群。在此提案中，我们旨在定义感染EBV的细胞异质性，包括先天抗病毒限制和plasmablast 分化，影响淋巴作用，可以进行治疗。我们的中心假设是EBV- 感染的B细胞在不同状态之间切换，这些状态可以限制或促进淋巴作用以及渲染细胞易受病毒特异性治疗干预措施。我们根据初步数据，包括感染后EBV感染的原代B细胞的单细胞RNA测序，并在LCL和细胞命运动力学的表征中重新激活。我们还提供了使用“踢和杀戮”策略的近期临床试验的证据用组蛋白脱乙酰基酶抑制与ganciclovir促进EBV裂解重新激活以杀死淋巴瘤激活病毒激酶的细胞。那就是拟议研究的理由是了解EBV 受感染的B细胞命运的调节将剖析发病机理的机制，并揭示了新的治疗途径靶向EBV阳性B细胞淋巴瘤。我们计划检验我们的中心假设并完成目标通过以下三个具体目的，该提案：i）定义先天免疫传感器的作用和 EBV介导的永生化和淋巴作用中的生效者，ii）确定浆膜的作用抑制EBV介导的淋巴作用的分化和III）定义了HDAC的机制抑制作用促进了EBV+ DLBCL对Ganciclovir杀死的敏感性。