Characterization of Target Gene Regulation by the Two Epstein-Barr Virus Oncogene LMP1 Domains Essential for B-cell Transformation.
Characterization of Target Gene Regulation by the Two Epstein-Barr Virus Oncogene LMP1 Domains Essential for B-cell Transformation.
复制标题
B 细胞转化所必需的两个 Epstein-Barr 病毒癌基因 LMP1 结构域对靶基因调控的表征。
DOI：
10.1101/2023.04.10.536234
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Gewurz,BenjaminE
中科院分区：
文献类型：
--
作者：
Mitra,Bidisha;Beri,NinaRose;Guo,Rui;Burton,EricM;Murray-Nerger,LauraA;Gewurz,BenjaminE
EB病毒（EBV）致癌基因潜伏膜蛋白1（LMP 1）模拟CD 40信号传导，并在多种恶性肿瘤中表达。两个LMP 1 C-末端胞质尾区，称为转化必需位点（TES）1和2，对于EBV将B淋巴细胞转化为永生化淋巴母细胞样细胞系（LCL）至关重要。然而，TES 1与TES 2 B细胞靶基因的特征仍然不完全，两者是否都是LCL生存所必需的仍然未知。为了定义LCL LMP 1靶基因，我们分析了细胞死亡前急性LMP 1 CRISPR敲除（KO）的全转录组效应。为了表征特定的LCL TES 1和TES 2作用，我们在内源性LMP 1 KO后条件性表达野生型、TES 1无效、TES 2无效或双TES 1/TES 2无效LMP 1等位基因。出乎意料的是，TES 1而不是TES 2信号传导对LCL存活至关重要。LCL依赖性因子cFLIP在LCL凋亡阻断中起着强制性作用，其通过TES 1信号传导的丧失而高度下调。为了进一步表征TES 1与TES 2的作用，我们在两个Burkitt模型中条件性表达野生型、TES 1和/或TES 2无效LMP 1等位基因。系统的RNAseq分析揭示了对TES 1和TES 2反应更强烈的基因簇，对两者都有强烈反应的基因簇或相反调节的基因簇。再次注意到TES 1对cFLIP诱导的稳健作用。确定了TES 1和2对其他LCL依赖性因子（包括BATF和IRF 4）表达以及对EBV超级增强子的影响。总的来说，这些研究提出了一个模型，通过该模型，LMP 1 TES 1和TES 2共同重塑B细胞转录组，并突出TES 1作为一个关键的治疗靶点。重要的是，EB病毒（EBV）导致多种人类癌症，包括B细胞淋巴瘤。在细胞培养中，EBV将健康的人类B细胞转化为不断生长的永生化细胞，这是移植后淋巴瘤的模型。EBV癌基因潜伏膜蛋白1（LMP 1）的两个胞质尾域的组成性信号传导是这种转化所需的，但尚未对其宿主基因靶点进行系统分析。我们发现，只有来自膜近端结构域的信号传导是这些EBV永生化细胞存活所必需的，并且其丢失触发细胞凋亡。我们确定了关键的LMP 1靶基因，其丰度随着LMP 1信号的丢失而显著变化，或者在EBV未感染的人类B细胞模型中，响应于一个或两个LMP 1结构域的信号转导而上调。这些包括EB病毒感染的B细胞生存所需的主要抗凋亡因子。生物信息学分析确定了B细胞基因簇，它们对任一或两个结构域的信号传导反应不同。
The Epstein-Barr virus (EBV) oncogene latent membrane protein 1 (LMP1) mimics CD40 signaling and is expressed by multiple malignancies. Two LMP1 C-terminal cytoplasmic tail regions, termed transformation essential sites (TES) 1 and 2, are critical for EBV transformation of B lymphocytes into immortalized lymphoblastoid cell lines (LCL). However, TES1 versus TES2 B-cell target genes have remained incompletely characterized, and whether both are required for LCL survival has remained unknown. To define LCL LMP1 target genes, we profiled transcriptome-wide effects of acute LMP1 CRISPR knockout (KO) prior to cell death. To then characterize specific LCL TES1 and TES2 roles, we conditionally expressed wildtype, TES1 null, TES2 null, or double TES1/TES2 null LMP1 alleles upon endogenous LMP1 KO. Unexpectedly, TES1 but not TES2 signaling was critical for LCL survival. The LCL dependency factor cFLIP, which plays obligatory roles in blockade of LCL apoptosis, was highly downmodulated by loss of TES1 signaling. To further characterize TES1 vs TES2 roles, we conditionally expressed wildtype, TES1, and/or TES2 null LMP1 alleles in two Burkitt models. Systematic RNAseq analyses revealed gene clusters that responded more strongly to TES1 vs TES2, that respond strongly to both or that are oppositely regulated. Robust TES1 effects on cFLIP induction were again noted. TES1 and 2 effects on expression of additional LCL dependency factors, including BATF and IRF4, and on EBV super-enhancers were identified. Collectively, these studies suggest a model by which LMP1 TES1 and TES2 jointly remodel the B-cell transcriptome and highlight TES1 as a key therapeutic target.IMPORTANCEEpstein-Barr virus (EBV) causes multiple human cancers, including B-cell lymphomas. In cell culture, EBV converts healthy human B-cells into immortalized ones that grow continuously, which model post-transplant lymphomas. Constitutive signaling from two cytoplasmic tail domains of the EBV oncogene latent membrane protein 1 (LMP1) is required for this transformation, yet there has not been systematic analysis of their host gene targets. We identified that only signaling from the membrane proximal domain is required for survival of these EBV-immortalized cells and that its loss triggers apoptosis. We identified key LMP1 target genes, whose abundance changed significantly with loss of LMP1 signals, or that were instead upregulated in response to switching on signaling by one or both LMP1 domains in an EBV-uninfected human B-cell model. These included major anti-apoptotic factors necessary for EBV-infected B-cell survival. Bioinformatics analyses identified clusters of B-cell genes that respond differently to signaling by either or both domains.
影响因子：
5.1
作者：
GIVNER, LB;ABRAMSON, JS;WASILAUSKAS, B
通讯作者：
WASILAUSKAS, B
与中毒性休克生命综合症和猩红热毒素相关的严重 A 组链球菌感染
影响因子：
3.6
作者：
A. D. L. Stevens
通讯作者：
A. D. L. Stevens
化脓性链球菌引起的感染模式的变化
影响因子：
4.2
作者：
E. Gaworzewska;G. Colman
通讯作者：
G. Colman