Core Essential Genes in Primary Effusion Lymphoma Cell Lines

原发性渗出性淋巴瘤细胞系的核心必需基因

基本信息

批准号：
9277430
负责人：
Eva Henriette Gottwein
金额：
$ 16.8万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9277430
关键词：
Acquired Immunodeficiency Syndrome B-Cell Lymphomas B-Lymphocytes Biology CRISPR screen CRISPR/Cas technology Cell Line Cell Proliferation Cell Survival Cells Cellular biology Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Data Data Analyses Data Set Disease Eligibility Determination Essential Genes Etiology Future Gene Expression Genes Genetic Transcription Goals Herpesviridae Human Human Cell Line Human Herpesvirus 8 IRF4 gene Individual Knock-out Laboratories Link Lymphoma Malignant Neoplasms Mediating Methods Mutation Non-Hodgkin&apos s Lymphoma Normal Cell Oncogenic Pathway interactions Patients Positioning Attribute Risk Role Solid Somatic Mutation Spermidine Synthase System TCF3 gene Technology Validation Viral Interference Viral Oncogene Virus Diseases Work addiction base cell transformation computerized tools data mining effusion experimental study follow-up genome editing genome-wide mammalian genome new therapeutic target novel primary effusion lymphoma screening tool transcription factor treatment strategy tumor tumorigenesis

Acquired Immunodeficiency Syndrome B-Cell Lymphomas B-Lymphocytes Biology CRISPR screen CRISPR/Cas technology Cell Line Cell Proliferation Cell Survival Cells Cellular biology Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Data Data Analyses Data Set Disease Eligibility Determination Essential Genes Etiology Future Gene Expression Genes Genetic Transcription Goals Herpesviridae Human Human Cell Line Human Herpesvirus 8 IRF4 gene Individual Knock-out Laboratories Link Lymphoma Malignant Neoplasms Mediating Methods Mutation Non-Hodgkin&apos s Lymphoma Normal Cell Oncogenic Pathway interactions Patients Positioning Attribute Risk Role Solid Somatic Mutation Spermidine Synthase System TCF3 gene Technology Validation Viral Interference Viral Oncogene Virus Diseases Work addiction base cell transformation computerized tools data mining effusion experimental study follow-up genome editing genome-wide mammalian genome new therapeutic target novel primary effusion lymphoma screening tool transcription factor treatment strategy tumor tumorigenesis

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项目摘要

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.

概括 Kaposi的肉瘤相关疱疹病毒（KSHV）是人类疱疹病毒一级积液B细胞淋巴瘤（PEL），特别是在获得的患者中免疫缺陷综合征（AIDS）。 PEL肿瘤发生在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PEL 衍生的细胞系需要持续的病毒感染才能生存和增殖文化。 KSHV所需的特定细胞基因和途径然而，淋巴瘤尚不清楚。在过去的两年中，有效的方法人类细胞系中的基因编辑已经可用，并且已适应基因组规模的淘汰赛屏幕。我们已经进行了全基因组CRISPR/CAS9- 介导的敲除筛网，以全面识别细胞基因和途径 KSHV诱导的PEL细胞存活和增殖所必需的。在AIM 1中，我们建议识别并验证pel细胞系生存所需的一组核心基因。目标 2，我们建议专注于一组功能相关的基因的生物学对于我们的初步中的PEL细胞存活和增殖至关重要 CRISPR/CAS9屏幕。 AIM 2的拟议工作包括深度验证这些基因对PEL细胞存活和增殖的需求以及实验研究这些基因如何在基因表达级联反应中进行协作。一起，预计这些目标将导致KSHV-的主要参与者的全面概述。介导的B细胞转化，可以鉴定新的药物靶标。