Core Essential Genes in Primary Effusion Lymphoma Cell Lines

总结 卡波西肉瘤相关疱疹病毒（KSHV）是一种人类疱疹病毒， 原发性渗出性B细胞淋巴瘤（PEL），特别是在获得性 免疫缺陷综合征（艾滋病）。PEL肿瘤发生在很大程度上由 病毒癌基因对正常细胞生物学的干扰。因此，KSHV阳性PEL 衍生的细胞系需要持续的病毒感染以使它们在细胞中存活和增殖。 文化KSHV所需的特定细胞基因和途径导致 然而，淋巴瘤并不完全清楚。在过去的两年里， 人类细胞系中的基因编辑已经变得可用，并且已经适应于 基因组规模的敲除筛选。我们进行了全基因组CRISPR/Cas9- 介导的敲除筛选，以全面鉴定细胞基因和途径 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 鉴定和验证PEL细胞系存活所需的核心基因组。在aim中 2，我们建议专注于一组功能相关基因的生物学，这些基因是 在我们的初步研究中， CRISPR/Cas9筛选。目标2的拟议工作包括深入验证 这些基因对于PEL细胞存活和增殖的需要，以及实验 来研究这些基因如何在基因表达级联中协作。在一起， 这些目标预计将导致对KSHV关键参与者的全面概述- 介导的B细胞转化，并可能确定新的药物靶点。

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.