EBV LMP1/LMP2A Proteins Promote Hodgkin-like Lymphomas in Humanized Mice

EBV LMP1/LMP2A 蛋白促进人源化小鼠霍奇金样淋巴瘤的发生

• 批准号: 10403940
• 负责人: Shannon Celeste Kenney
• 金额: $ 37.39万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-13 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10403940
• 关键词: AIDS-Related Hodgkin' s Lymphoma; AIDS-Related Lymphoma; Acquired Immunodeficiency Syndrome; Attenuated; Autocrine Communication; Automobile Driving; B-Cell Antigen Receptor; B-Cell Lymphomas; B-Lymphocytes; CD4 Positive T Lymphocytes; Cell Line; Cells; Collagen; DDR1 gene; Development; EBNA2 protein; Environment; Epstein-Barr Virus Infections; Epstein-Barr Virus latency; Goals; Growth; HIV Infections; Hodgkin Disease; Human; Human Herpesvirus 4; Immunocompetence; Immunocompromised Host; Immunosuppression; In Vitro; Inflammatory; LMP1; Laboratories; Lead; Lymphoma; Lymphoma cell; Malignant Neoplasms; Membrane Proteins; Modeling; PDGFA gene; PDGFRA gene; Paracrine Communication; Pathway interactions; Patients; Phenotype; Proteins; Receptor Protein-Tyrosine Kinases; Research; Role; Signal Pathway; Signal Transduction; T-Lymphocyte; TNFRSF5 gene; Transcription Coactivator; Tumor-Derived; Umbilical Cord Blood; Viral Proteins; Virus; Virus Latency; cell growth; humanized mouse; immunogenic; improved; in vitro Model; in vivo; in vivo Model; insight; mouse model; mutant; neoplastic cell; notch protein; novel; novel therapeutics; overexpression; public health relevance; tumor; tumor growth; tumor microenvironment

PROJECT SUMMARY/ABSTRACT Epstein-Barr virus is an important cause of AIDS-related lymphomas (ARLs) world-wide, including Hodgkin lymphomas (HLs) that are driven by the EBV latent membrane proteins LMP1 (a CD40 mimic) and LMP2A (a BCR mimic), in conjunction with a supportive tumor microenvironment. Although the EBV protein EBNA2 (which mimics notch signaling) is required for EBV-induced transformation of B cells in vitro, cells that express EBNA2 have the “type III” form of viral latency, which is highly immunogenic. Thus, most EBV-infected human lymphomas (including ARLs in cART-treated patients) do not express EBNA2. However, there is currently no in vivo or in vitro model available to study how EBV infection can cause lymphomas in the absence of EBNA2 expression, and the only EBV+ HL cell line, L591, has switched to type III latency. Increasing evidence suggests that HIV infection collaborates with EBV to induce lymphomas not only by inducing immunosuppression, but by creating a pro-inflammatory microenvironment that promotes tumors with more stringent forms of EBV latency (such as the “type II” form that occurs in HLs) that do not express EBNA2. Our lab has developed a novel cord blood-humanized mouse model which provides a highly supportive CD4 T cell-rich environment that allows certain EBV mutants previously considered “non-transforming” in vitro to induce lymphomas in vivo. Our new exciting preliminary results reveal that a naturally occurring EBNA2-deleted EBV strain (P3HR1) that is completely non-transforming in vitro causes Hodgkin-like lymphomas that express LMP1 and LMP2A in this model. Furthermore, our preliminary results suggest that similar to human HLs, lymphomas induced by EBNA2- deleted EBV are heavily infiltrated with collagen, express the pro-survival collagen-stimulated receptor tyrosine kinase DDR1, and have activated PDGFRA and notch-1 signaling. Thus, we believe we have created the first in vivo model for EBV-induced Hodgkin lymphoma. In this proposal, we will dissect the roles of the EBV proteins LMP1 and LMP2A, as well as the tumor microenvironment, in driving lymphomas induced by EBNA2-deleted EBV in humanized mice. We hypothesize that both LMP1 and LMP2A, as well as CD4 T cell- and collagen- derived signals in the microenvironment, are required for the growth of these lymphomas. In Aim 1, we will compare the phenotypes of wild-type (WT) versus EBNA2-deleted (Akata strain) EBV in humanized mouse models, and identify paracrine and/or autocrine signaling pathways (derived from T cells, B cells and/or collagen) that compensate for loss of EBNA2 expression both in vivo and in L591 HL cells in vitro. In Aim 2, we will define the roles of LMP1 and LMP2A on tumor cell growth, and in regulating the tumor microenvironment, in both the CBH model in vivo and in L591 cells in vitro. In Aim 3, we will determine whether inhibiting notch, PDGFRA and/or DDR1 signaling attenuates the growth of lymphomas induced by WT virus and/or EBNA2-deleted virus in humanized mice or in L591 cells in vitro. The results of these studies should provide key insights into the mechanism(s) by which LMP1 and LMP2A promote AIDS-related Hodgkin lymphomas, and elucidate how the tumor microenvironment cooperates with type II latent EBV infection to induce human lymphomas in vivo when EBNA2 expression is shut off.

项目摘要/摘要 爱泼斯坦 - 巴尔病毒是全球艾滋病相关淋巴瘤（ARLS）的重要原因，包括霍奇金 由EBV潜在膜蛋白LMP1（A CD40 MIMIC）和LMP2A（a）驱动的淋巴瘤（HLS）（A BCR模拟），与支持性肿瘤微环境结合使用。虽然EBV蛋白EBNA2（它 模仿Notch信号传导是EBV诱导的B细胞体外转化所必需的，即表达EBNA2的细胞 具有高度免疫原性的病毒潜伏期的“ II型”形式。那，大多数EBV感染的人 淋巴瘤（包括手推车治疗的患者中的ARL）不表达EBNA2。但是，目前没有 可用于研究在没有EBNA2的情况下EBV感染如何引起淋巴瘤的体内或体外模型 表达，唯一的EBV+ HL细胞系L591已切换到III型潜伏期。越来越多的证据表明 艾滋病毒感染与EBV合作，不仅是通过诱导的免疫抑制，而且是通过 创建促炎性微环境，该环境促进具有更严格形式的EBV潜伏期的肿瘤 （例如在HLS中发生的“ II型”形式）不表达EBNA2。我们的实验室已经开发了一条新颖的绳子 血液人类化的小鼠模型提供了高度支持的CD4 T细胞环境，可以 某些EBV突变体以前在体外考虑过“非转化”以在体内诱导淋巴瘤。我们的新 令人兴奋的初步结果表明，天然存在的EBNA2删除EBV菌株（P3HR1）是 完全不转化的体外导致表达LMP1和LMP2A的霍奇金样淋巴瘤 模型。此外，我们的初步结果表明，与人HLS相似，EBNA2-诱导的淋巴瘤 删除的EBV用胶原蛋白大量渗入，表达促生物胶原蛋白刺激的受体酪氨酸 激酶DDR1，并激活了PDGFRA和Notch-1信号传导。那就是我们相信我们创造了第一个 EBV诱导的霍奇金淋巴瘤的体内模型。在此提案中，我们将剖析EBV蛋白的作用 LMP1和LMP2A以及肿瘤微环境，在驱动由EBNA2骨骼诱导的淋巴瘤中 人源化小鼠的EBV。我们假设LMP1和LMP2A，以及CD4 T细胞和胶原蛋白 这些淋巴瘤的生长需要微环境中的派生信号。在AIM 1中，我们将 比较人源化小鼠中野生型（WT）与EBNA2删除（Akata菌株）的表型 模型，并识别旁分泌和/或自分泌信号通路（源自T细胞，B细胞和/或胶原蛋白） 在体外和L591 HL细胞中，EBNA2表达的损失均在体外。在AIM 2中，我们将定义 LMP1和LMP2A在肿瘤细胞生长以及控制肿瘤微环境中的作用 体内和L591细胞的CBH模型在体外。在AIM 3中，我们将确定是否抑制缺口，PDGFRA 和/或DDR1信号传导减弱了由WT病毒和/或EBNA2骨骼病毒诱导的淋巴瘤的生长 在人性化小鼠或L591细胞中，体外。这些研究的结果应提供对 LMP1和LMP2A促进与艾滋病相关的霍奇金淋巴瘤的机制，并阐明如何阐明 肿瘤微环境与II型潜在EBV感染合作，在体内诱导人类淋巴瘤 EBNA2表达被关闭。