Exploiting Metabolism to Uncloak Epstein-Barr Virus Immunogens in Latently Infected B-cells

利用代谢揭示潜伏感染 B 细胞中的 Epstein-Barr 病毒免疫原

• 批准号: 10589798
• 负责人: Rui Guo
• 金额: $ 11.58万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-11 至 2024-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10589798
• 关键词: Acquired Immunodeficiency Syndrome; Address; Adult; Amino Acids; Antigens; Area; B-Lymphocytes; Biology; Burkitt Lymphoma; CD8-Positive T-Lymphocytes; CRISPR screen; Carcinoma; Cell Compartmentation; Cells; Cellular Metabolic Process; Clustered Regularly Interspaced Short Palindromic Repeats; Code; DNA; DNA Methylation; DNA Modification Process; Data; Detection; Development; Development Plans; Disease; Double Stranded DNA Virus; Enzyme Inhibitor Drugs; Enzymes; Epigenetic Process; Episome; Epithelium; Epstein-Barr Virus latency; Face; Folic Acid; Genes; Genetic Transcription; Genome; Hairy Leukoplakia; Human; Human Genome; Human Herpesvirus 4; Hypermethylation; Immune; Immunotherapy; Infection; Infectious Mononucleosis; Intervention; Invaded; Jaw; Learning; Lymphocyte; Lymphoma; Lymphoma cell; Lymphoproliferative Disorders; Lytic Phase; Malignant Neoplasms; Membrane Proteins; Memory B-Lymphocyte; Mentors; Metabolic; Metabolic Control; Metabolic Pathway; Metabolism; Methionine; Methionine Metabolism Pathway; Methylation; Nasopharynx Carcinoma; Nuclear Antigens; Ocular orbit; Oncoproteins; Oral; Oropharyngeal; Pathway interactions; Patients; Phase; Postdoctoral Fellow; Proteins; Regimen; Research; Research Personnel; SA01; SA02; SA03; Saliva; T cell response; T-Lymphocyte; Tongue; Tonsil; Tumor Escape; Viral; Viral Antigens; Viral Genome; Viral Proteins; Virus; career development; chemical genetics; chronic infection; derepression; dietary restriction; epigenetic regulation; epigenome; experimental study; folic acid metabolism; gene product; genetic approach; genome-wide; histone methylation; histone modification; immunogenic; infected B cell; insight; interdisciplinary approach; metabolomics; methyl group; neoplastic cell; novel therapeutic intervention; pathogen; post-transplant; programs; small molecule; transmission process; tumor

Project Summary Epstein-Barr virus (EBV) is spread through the saliva, where it establishes oropharyngeal infection, invades the tonsils and colonizes the B-cell compartment. Orally transmitted EBV is the cause of infectious mononucleosis and of multiple B-cell and epithelial cancers, including Burkitt lymphoma (BL) and nasopharyngeal carcinoma. Much remains to be learned about how epigenetic regulation of the viral genome enables it to colonize the oropharynx and tonsils to establish persistent infection of >95% adults and to cause 200,000 cancers per year. In tonsillar memory B-cells, EBV expresses a single, poorly immunogenic antigen. BL, which can present as tumors of the jaws, face and orbit, use a similar viral program to evade anti-EBV T-cell responses. By contrast, EBV expresses nearly 80 viral proteins in AIDS patients with oral hairy leukoplakia of the tongue, and as many as immunogenic eight latency proteins in post- transplant lymphoma. During the initial post-doctoral period, the applicant used two CRISPR screens to characterize B-cell factors important for EBV latency in BL. These provided insights into epigenetic pathways that restrict expression of lytic cycle and latency genes in B-cells and highlighted epigenetic enzymes that initiate and maintain a high level of DNA methylation of the EBV genome in this highly restricted form of latency. Perturbation of EBV DNA methylation de- represses EBV antigen expression and sensitizes Burkitt cells to CD8+ T-cell recognition. Yet, little is known about cross-talk between infected B-cell metabolism and epigenetic pathways, which must occur to supply methyl groups for DNA and histone modification. The applicant hypothesizes that latently EBV-infected B cells subvert methionine and folate metabolism pathways to drive EBV genome hypermethylation necessary for silencing of immunogenic EBV gene products and T-cell immunoevasion. During the mentored K99 phase, the applicant will 1) identify how methionine metabolism contributes to EBV genome hypermethylation and silencing in latently infected B-cells and EBV oncoproteins, and 2) characterize the interplay between the folate cycle and EBV genome methylation in latently-infected cells. During the R00 phase, the applicant will perform independent research to identify how latent EBV programs alter B-cell metabolic pathways to reinforce viral genome epigenetic states at the level of DNA and histone methylation. Collectively, these studies will open a new area of EBV biology and promise to support novel therapeutic approaches. The career development plan will prepare the applicant for transition to independence as an investigator with a multi-disciplinary approach to study metabolomic control of virus/host interactions.

项目概要 EB 病毒 (EBV) 通过唾液传播，并在口咽部建立 感染，侵入扁桃体并定植于 B 细胞区室。口服传播的 EBV 是 传染性单核细胞增多症以及多种 B 细胞癌和上皮癌的病因，包括 伯基特淋巴瘤（BL）和鼻咽癌。关于如何 病毒基因组的表观遗传调控使其能够定植于口咽和扁桃体， 造成 >95% 的成年人持续感染，每年导致 200,000 例癌症。在扁桃体 记忆 B 细胞中，EBV 表达单一的、免疫原性较差的抗原。 BL，可以呈现 作为颌骨、面部和眼眶的肿瘤，使用类似的病毒程序来逃避抗 EBV T 细胞 回应。相比之下，口腔毛茸茸的艾滋病患者中的 EBV 表达近 80 种病毒蛋白。 舌头白斑，以及多达八种免疫原性潜伏蛋白 移植淋巴瘤。在最初的博士后期间，申请人使用了两次CRISPR 筛选对 BL 中 EBV 潜伏期重要的 B 细胞因子进行表征。这些提供了见解 进入限制 B 细胞中裂解周期和潜伏基因表达的表观遗传途径， 强调了启动并维持高水平 DNA 甲基化的表观遗传酶 EBV 基因组处于这种高度受限的潜伏期。 EBV DNA 甲基化去扰动 抑制 EBV 抗原表达并使 Burkitt 细胞对 CD8+ T 细胞识别敏感。然而， 对于受感染的 B 细胞代谢和表观遗传途径之间的相互作用知之甚少， 必须发生这种情况才能为 DNA 和组蛋白修饰提供甲基。申请人 假设潜伏感染 EBV 的 B 细胞破坏蛋氨酸和叶酸代谢 驱动 EBV 基因组高甲基化的途径是沉默免疫原性 EBV 所必需的 基因产物和T细胞免疫逃避。在指导的 K99 阶段，申请人将 1) 确定蛋氨酸代谢如何导致 EBV 基因组高甲基化和沉默 在潜伏感染的 B 细胞和 EBV 癌蛋白中，2) 表征了 潜伏感染细胞中的叶酸循环和 EBV 基因组甲基化。在 R00 阶段， 申请人将进行独立研究，以确定潜在 EBV 程序如何改变 B 细胞 在 DNA 和组蛋白水平上强化病毒基因组表观遗传状态的代谢途径 甲基化。总的来说，这些研究将开辟 EBV 生物学的新领域，并有望 支持新的治疗方法。职业发展计划将为申请人做好准备 过渡为独立的研究者，采用多学科方法进行研究 病毒/宿主相互作用的代谢组学控制。