Characterizing single cell states of activated and transformed B cells in rhesus macaque models

恒河猴模型中活化和转化 B 细胞的单细胞状态特征

• 批准号: 10665491
• 负责人: Rebecca L Skalsky
• 金额: $ 23.62万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-02 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665491
• 关键词: Acquired Immunodeficiency Syndrome; Address; Archives; Area; B-Cell Acute Lymphoblastic Leukemia; B-Cell Development; B-Cell Lymphomas; B-Lymphocytes; B-cell receptor repertoire sequencing; Biological; CRISPR screen; Cancer Burden; Cancerous; Cell Culture Techniques; Cell Line; Cell surface; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Dependence; Development; Diagnostic; Differentiation Antigens; Dimensions; Disease; EBV-associated disease; Environment; Epstein Barr Nuclear Antigen 3; Epstein-Barr Virus Infections; Epstein-Barr Virus-Related Malignant Neoplasm; Epstein-Barr pathogenesis; Gene Expression; Genetic Transcription; Goals; Growth; Heterogeneity; Human; Human Herpesvirus 4; IRF4 gene; Immune; Immunocompromised Host; Immunophenotyping; In Vitro; Individual; Infection; Infectious Agent; Integration Host Factors; Knowledge; LMP1; Link; Longitudinal Studies; Lymph Node Tissue; Lymphocryptovirus; Lymphoid Tissue; Lymphoma; Lymphoproliferative Disorders; MS4A1 gene; Macaca mulatta; Malignant Neoplasms; Maps; Measures; Mediating; Methods; MicroRNAs; Modeling; Molecular; Molecular Profiling; Nature; Pathway interactions; Patients; Pre-Clinical Model; Predisposition; Process; Proliferating; Proliferation Marker; Property; Refractory; Resolution; Resources; Rest; Rhesus; Sampling; Side; Techniques; Therapeutic; Tumor Antigens; Tumor Tissue; Viral; Viral Cancer; Viral Proteins; Virus; Virus Diseases; cell transformation; chronic infection; experimental study; gammaherpesvirus; gene product; genetic manipulation; genetic signature; immune checkpoint; improved; in vivo; infected B cell; infectious disease model; latent infection; lymphoblastoid cell line; molecular dynamics; molecular marker; multiple omics; mutant; permissiveness; pre-clinical; premalignant; programs; single-cell RNA sequencing; time use; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; tumor; tumor heterogeneity

Project Summary Epstein-Barr virus (EBV) causes significant disease in patients with congenital or acquired immune deficiencies. Elucidating markers of early cancer and understanding tumor composition are essential in improving diagnostics and therapies for EBV diseases. In vitro, EBV infection of B cells induces formation of lymphoblastoid cell lines (LCLs) which mimic properties of lymphoproliferative disease and serve as a model to investigate B cell transformation processes. Multiple viral gene products dramatically reprogram the B cell environment, and several host factors critical for maintaining LCL proliferation have been identified; however, host factors leading to LCL outgrowth as well as the distinct cellular substages required for establishment of persistent infection and B cell transformation are less understood. Deciphering the molecular mechanisms involved in these processes is an ongoing challenge due to the refractory nature of primary B cells for genetic manipulation and the observed heterogeneity in viral and cellular gene expression detected through standard bulk transcriptome analysis. To address gaps in our current knowledge, we will use multi-omic single-cell approaches to define B cell molecular signatures and cell markers that delineate early virus-mediated transformation. For these studies, we will investigate a pre-clinical EBV model, rhesus macaque (RM) cells and tumor tissues infected with rhesus lymphocryptovirus (rLCV). By simultaneously measuring immunophenotypes and gene expression, we aim to identify B cell states that successfully navigate infection towards the LCL trajectory. Leveraging these approaches to further analyze rLCV+ lymphoid tissues, we aim to define biological properties of pre-cancerous and cancerous states at the single cell level.

项目概要 EB 病毒 (EBV) 会导致先天性或后天性免疫缺陷患者罹患严重疾病。 阐明早期癌症的标志物和了解肿瘤的组成对于改善诊断至关重要 以及 EBV 疾病的治疗。在体外，B 细胞的 EBV 感染诱导类淋巴母细胞系的形成 (LCL) 模拟淋巴增殖性疾病的特性并作为研究 B 细胞的模型 转变过程。多种病毒基因产物极大地重新编程 B 细胞环境，并且 已经确定了一些对于维持 LCL 增殖至关重要的宿主因素；然而，宿主因素导致 LCL 生长以及建立持续感染所需的不同细胞亚阶段 B细胞转化知之甚少。破译这些过程中涉及的分子机制 由于原代 B 细胞难以进行基因操作，并且观察到的结果，这是一个持续的挑战 通过标准批量转录组分析检测病毒和细胞基因表达的异质性。到 为了解决我们当前知识的空白，我们将使用多组学单细胞方法来定义 B 细胞分子 描述早期病毒介导转化的特征和细胞标记。对于这些研究，我们将 研究临床前 EBV 模型、恒河猴 (RM) 细胞和感染恒河猴的肿瘤组织 淋巴隐病毒（rLCV）。通过同时测量免疫表型和基因表达，我们的目标是 识别成功将感染引导至 LCL 轨迹的 B 细胞状态。利用这些 进一步分析 rLCV+ 淋巴组织的方法，我们的目标是定义癌前病变的生物学特性 和单细胞水平的癌态。