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

• 批准号: 10706553
• 负责人: Micah A. Luftig
• 金额: $ 55.59万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 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/10706553
• 关键词: Adult; African Burkitt' 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 B cell lymphoma; 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' 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; cell killing; epstein barr virus mediated immortalization; experimental study; gene product; in vivo; infected B cell; innate immune sensing; large cell Diffuse non-Hodgkin' 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

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.

Epstein-Barr 病毒 (EBV) 是 50 多年前发现的第一种人类肿瘤病毒 非洲地方性伯基特淋巴瘤。然而，我们现在知道它也是一种常见的疱疹病毒，作为一种持续存在的病毒。 全世界几乎所有成年人都存在终生潜伏感染。该领域的早期工作建立了 EBV 感染模型 促进 B 细胞淋巴瘤，如原发性静息的生长转化或永生化所证明的 人类 B 细胞转化为类淋巴母细胞系 (LCL)。在体内，EBV 潜伏感染具有强大的细胞毒性 T 细胞反应使大多数感染者免受病毒潜在的致癌性影响。像这样， 在免疫抑制的情况下，EBV 相关 B 细胞淋巴瘤的发生率明显更高。 对 EBV 感染细胞中病毒和细胞基因表达的体外和体内研究已经建立了一个模型 淋巴瘤发生的特征是 EBV 潜伏基因产物的完全表达。然而，表型 在大量培养物和肿瘤组织中缺乏细胞异质性的细微差别以及轻微的后果 癌症进展的频率表型。我们最近的单细胞 RNAseq 实验已表征 单个 EBV 感染的 B 细胞内的基因表达导致细胞命运轨迹的评估 我们将把单个细胞的动态基因表达行为与人类肿瘤分析和小鼠整合 淋巴瘤发生模型。我们的最终目标是确定特定 EBV 感染细胞的重要性 人群对B细胞非霍奇金淋巴瘤进展的免疫抑制。在这个提案中， 我们的目标是定义 EBV 感染的细胞异质性，包括先天抗病毒限制和浆母细胞 分化，影响淋巴瘤发生并可用于治疗。我们的中心假设是 EBV- 受感染的 B 细胞在不同状态之间切换，可以限制或促进淋巴瘤发生并渲染 细胞对病毒特异性治疗干预敏感。我们基于以下基础制定了我们的中心假设： 初步数据，包括感染后早期 EBV 感染的原代 B 细胞的单细胞 RNA 测序，以及 LCL 中的作用以及调节浆母细胞分化和裂解的细胞命运动力学特征 重新激活。我们还提供证据支持最近使用“踢杀”策略的临床试验 通过组蛋白脱乙酰酶抑制联合更昔洛韦促进 EBV 溶解再激活以杀死淋巴瘤 激活病毒激酶的细胞。因此，本研究的基本原理是了解 EBV 受感染 B 细胞命运的调节将剖析发病机制并揭示新的治疗途径 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 该提案通过以下三个具体目标：i）定义先天免疫传感器的作用和 EBV 介导的永生化和淋巴瘤发生中的效应子，ii) 确定浆母细胞的作用 抑制 EBV 介导的淋巴瘤发生的分化，以及 iii) 定义 HDAC 的机制 抑制作用促进 EBV+ DLBCL 对更昔洛韦杀伤的敏感性。