Core Essential Genes in Primary Effusion Lymphoma Cell Lines

摘要 卡波西肉瘤相关疱疹病毒是一种人类疱疹病毒，可引起 原发性渗出性B细胞淋巴瘤(PEL)，尤其是获得性 免疫缺陷综合征(艾滋病)。PEL肿瘤的发生很大程度上是由 病毒癌基因对正常细胞生物学的干扰。因此，KSHV阳性的PEL 衍生的细胞系需要持续的病毒感染才能在 文化。KSHV致病所需的特定细胞基因和途径 然而，淋巴瘤并不完全为人所知。在过去的两年里，有效的方法 人类细胞系中的基因编辑已经成为可能，并已被适应于 基因组规模的基因敲除筛选。我们已经进行了全基因组CRISPR/Cas9- 用于全面识别细胞基因和途径的介导性基因敲除筛选 是KSHV诱导的PEL细胞存活和增殖所必需的。在目标1中，我们建议 确定并验证PEL细胞系生存所需的核心基因集。在AIM 2，我们建议专注于一组与功能相关的基因的生物学 在我们的初步研究中发现对PEL细胞的存活和增殖是必不可少的 CRISPR/CAS9屏幕。目标2的拟议工作包括深入验证 这些基因对PEL细胞存活和增殖的要求及实验 来研究这些基因是如何在基因表达级联中协作的。一起， 这些目标预计将导致对KSHV主要参与者的全面概述- 介导B细胞转化，并可能识别新的药物靶点。

Summary Kaposi's sarcoma-associated herpesvirus (KSHV) is a human herpesvirus that causes primary effusion B cell lymphoma (PEL), particularly in patients with acquired immunodeficiency syndrome (AIDS). PEL tumorigenesis is largely driven by the interference of viral oncogenes with normal cell biology. Accordingly, KSHV-positive PEL derived cell lines require continued viral infection for their survival and proliferation in culture. The specific cellular genes and pathways required for KSHV to cause lymphomas, however, are not fully known. In the last two years, efficient methods for gene editing in human cell lines have become available and have been adapted for genome-scale knock-out screens. We have conducted genome-wide CRISPR/Cas9- mediated knockout screens to comprehensively identify cellular genes and pathways necessary for KSHV-induced PEL cell survival and proliferation. In Aim 1, we propose to identify and validate a core set of genes required for the survival of PEL cell lines. In Aim 2, we propose to focus on the biology of a set of functionally related genes that were identified as essential for PEL cell survival and proliferation in our preliminary CRISPR/Cas9 screen. The proposed work for Aim 2 includes the in depth validation of the requirement of these genes for PEL cell survival and proliferation, and experiments to investigate how these genes collaborate in a gene expression cascade. Together, these aims are expected to result in a comprehensive overview of key players in KSHV- mediated B cell transformation and may identify new drug targets.