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.

项目摘要 爱泼斯坦-巴尔病毒(EBV)会导致先天性或获得性免疫缺陷患者的重大疾病。 阐明早期癌症的标志物和了解肿瘤成分是提高诊断水平的关键 以及EBV疾病的治疗方法。EB病毒感染B细胞体外诱导淋巴母细胞系的形成 (LCLS)模拟淋巴增生性疾病的特性并作为研究B细胞的模型 转型过程。多种病毒基因产品极大地改变了B细胞环境， 已经确定了几个对维持LCL增殖至关重要的宿主因素；然而，宿主因素导致 至LCL生长以及建立持续感染所需的不同细胞亚阶段和 人们对B细胞转化知之甚少。破译这些过程中涉及的分子机制 是一个持续的挑战，因为原始B细胞对基因操作的难治性，以及观察到的 通过标准批量转录组分析检测到病毒和细胞基因表达的异质性。至 解决我们目前知识中的空白，我们将使用多组单细胞方法来定义B细胞分子 描述早期病毒介导性转化的签名和细胞标记。对于这些研究，我们将 研究临床前EB病毒模型、恒河猴(RM)细胞和肿瘤组织感染恒河猴 淋巴腺病毒(RLCV)。通过同时测量免疫表型和基因表达，我们的目标是 识别B细胞状态，成功地将感染引导至LCL轨迹。利用这些优势 为了进一步分析rLCV+淋巴组织，我们的目标是定义癌前病变的生物学特性 以及单细胞水平上的癌变状态。