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.

EB病毒（EBV）是50多年前在人类肿瘤背景下发现的第一种人类肿瘤病毒。 地方性非洲伯基特淋巴瘤然而，我们现在知道它也是一种常见的疱疹病毒， 全世界几乎所有成年人的终身潜伏感染。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，如通过原发性静息细胞的生长转化或永生化所证明的 人B细胞转化为淋巴母细胞样细胞系（LCL）。在体内，EBV潜伏感染与强的细胞毒性反应有关。 T细胞反应使大多数感染者免受病毒致癌潜力的影响。因此，在本发明的一个方面， EBV相关B细胞淋巴瘤在免疫抑制的情况下发生率显著较高。 在体外和体内对EBV感染细胞中的病毒和细胞基因表达的研究已经导致了一种模型， 以EBV潜伏基因产物的完全表达为特征的淋巴瘤发生。然而，表型 在大量培养和肿瘤组织中缺乏细胞异质性的细微细节和微小的细胞异质性的后果。 频率表型对癌症进展的影响我们最近的单细胞RNAseq实验表征了 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， 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肿瘤分析相结合。 淋巴瘤模型。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的进展的免疫抑制。在这一提议中， 我们的目标是确定EB病毒感染的细胞异质性，包括先天性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。我们的核心假设是EB病毒- 受感染的B细胞在不同的状态之间切换，这些状态可以限制或促进淋巴瘤的发生， 对病毒特异性治疗干预敏感的细胞。我们的核心假设是基于 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， 在LCL中以及表征调节浆细胞分化和溶解的细胞命运动力学 重新激活我们还提供了证据支持最近的一项临床试验，该试验使用了"踢死"策略， 组蛋白去乙酰化酶抑制剂联合更昔洛韦促进EBV裂解性再活化杀伤淋巴瘤 激活病毒激酶的细胞。因此，拟议研究的基本原理是， 受感染的B细胞命运的调节将剖析发病机制，并揭示新的治疗途径 针对EB病毒阳性B细胞淋巴瘤我们计划测试我们的中心假设，并完成目标， 该建议通过以下三个具体目标：i）定义先天免疫传感器的作用， EBV介导的永生化和淋巴瘤发生中的效应物，ii）确定浆母细胞的作用 在抑制EBV介导的淋巴瘤发生中的分化，和iii）定义HDAC 抑制增强了EBV + DLBCL对更昔洛韦杀伤的敏感性。