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: 10889325
• 负责人: Rui Guo
• 金额: $ 24.89万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2026-07-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10889325
• 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.

项目摘要 爱泼斯坦 - 巴尔病毒（EBV）通过唾液传播，在那里建立口咽 感染，侵入扁桃体并殖民B细胞室。口服传输的EBV是 传染性单核细胞增多症以及多个B细胞和上皮癌的原因，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节使其能够将口咽和扁桃体定居至 建立持续的感染> 95％的成年人，每年引起200,000次癌症。在汤西拉 记忆B细胞，EBV表达了一种单一的免疫原性抗原。 BL，可以出现 作为颌骨的肿瘤，面部和轨道，使用类似的病毒程序来逃避抗EBV T细胞 回答。相比之下，EBV在口服毛茸茸的艾滋病患者中表达近80种病毒蛋白 舌头的白细胞，在后的八个免疫原性八延迟蛋白 移植淋巴瘤。在最初的博士后期间，申请人使用了两个CRISPR 屏幕以表征B-Cell因子对BL中EBV潜伏期重要的重要性。这些提供了见解 进入表观遗传途径，这些途径限制了B细胞中裂解周期和潜伏基因的表达 强调启动并维持高水平的DNA甲基化的表观遗传酶 EBV基因组以这种高度限制的延迟形式。 EBV DNA甲基化的扰动 抑制EBV抗原表达，并使伯基特细胞对CD8+ T细胞识别敏感。然而， 关于感染的B细胞代谢和表观遗传途径之间的串扰知之甚少， 为DNA和组蛋白修饰提供甲基必须发生。申请人 假设潜在的EBV感染的B细胞颠覆蛋氨酸和叶酸代谢 驱动EBV基因组高甲基化的途径，以使免疫原性EBV沉默所必需 基因产物和T细胞免疫驱虫。在指导的K99阶段，申请人将1） 确定蛋氨酸代谢如何有助于EBV基因组高甲基化和沉默 在潜在感染的B细胞和EBV癌蛋白中，2）表征 叶酸周期和EBV基因组甲基化在潜在感染的细胞中。在R00阶段， 申请人将进行独立研究，以确定潜在的EBV计划如何改变B细胞 在DNA和组蛋白水平上增强病毒基因组表观遗传态的代谢途径 甲基化。总的来说，这些研究将开辟EBV生物学的新领域，并承诺 支持新颖的治疗方法。职业发展计划将为申请人做好准备 作为研究者，以多学科的研究方法过渡到独立性 病毒/宿主相互作用的代谢组控制。