TRANSFORMATION BY EBV LATENT MEMBRANE PROTEINS 1 AND 2 (LMP1 AND LMP2)

EBV 潜伏膜蛋白 1 和 2（LMP1 和 LMP2）的转化

基本信息

批准号：
7617146
负责人：
NANCY JOAN RAAB-TRAUB
金额：
$ 35.03万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7617146
关键词：
Affect Animals B-Cell Lymphomas B-Lymphocytes Biochemical Biological Assay Biological Models Breeding Carcinogens Carcinoma Cells Complement Data Development Embryo Engineering Enhancers Epithelial Cells Epstein-Barr Virus Infections Epstein-Barr Virus-Related Malignant Neoplasm Exposure to Fibroblasts Funding Gene Expression Genes Genetic Goals Growth Growth and Development function Heavy-Chain Immunoglobulins Hodgkin Disease Human Development Human Herpesvirus 4 Hypersensitivity skin testing In Vitro Keratin Lymphocyte Lymphoma Malignant - descriptor Malignant Neoplasms Membrane Membrane Proteins Modeling Mus Mutation Nasopharynx Carcinoma Oncogenic Pathway interactions Process Property Regulation Research Rodent Role Signal Pathway Skin Carcinogenesis TP53 gene Testing Time Transgenic Mice Transgenic Organisms Transplantation Tumor Promoters Tumor Suppressor Genes Tumor Suppressor Proteins Viral Proteins cell growth complement pathway in vitro Assay in vivo in vivo Model keratin 14, K14 p19ARF promoter tumor tumor initiation tumor progression

项目摘要

The viral proteins, latent membrane protein 1 (LMP1) and latent membrane protein 2 (LMP2), are expressed in many of the malignancies associated with EBV. We have previously produced and characterized transgenic mice that express LMP1 in B-lymphocytes using the immunoglobulin heavy chain promoter/enhancer (Ig-LMPl). These mice develop clonal B-cell lymphomas that express LMP1 at high levels. The clonal development of these cancers indicates that additional genetic changes must occur that complement the cellular pathways activated by LMP1. During this last period of funding, we have identified remarkable oncogenic synergy between the loss of the p16INK4/p19ARF locus and expression of LMP1 in B-lymphocytes. To evaluate the effects of LMP1 on epithelial cell growth, we have produced transgenic mice that express LMP1 under the control of the keratin 14 (K14) promoter. In these mice using classical tumor initiation and promotion analyses, LMP1 functions largely as a tumor promoter with a possible role in tumor progression. Transgenic mice that express LMP2 in B-lymphocytes using the Ig promoter and in epithelial cells from the K14 promoter have been obtained from Dr. Richard Longnecker. The K14-LMP2 transgenic mice will be tested to determine if LMP2 affects the oncogenic process through initiation, promotion, or progression in skin tests. Possible synergistic effects of LMP1 and LMP2 expression on B-cell and epithelial cell growth will be analyzed in dually transgenic mice and in transformation assays in vitro. Our specific aims are: 1) The transgenic LMP1+ lymphocytes, LMP1+ lymphomas, LMP1+ p16 null lymphomas, and LMP1+ p53 heterozygous lymphomas will be further characterized to identify activated signaling pathways and effects on cellular gene expression. The growth properties of the LMP1+ lymphocytes and lymphoma cells will be analyzed in vitro. 2) LMP1 and LMP2 affect and activate distinct cellular signaling pathways. To test the hypothesis that coordinate expression of LMP1 and LMP2 activates complementing pathways that synergistically alter growth regulation, Ig-LMP1/LMP2 transgenic mice will be produced by cross-breeding. The time to tumor development and the growth and biochemical properties of the cells in vitro will be determined. 3) To determine the effects of expression of LMP1 and LMP2 in normal epithelial cells, K14 promoter transgenic mice that express LMP1 and/or LMP2 in epithelial cells will be tested in classic skin carcinogenesis assays in combination with exposure to carcinogens and tumor promoters. 4) Characterize LMP1 transformation of rodent fibroblasts. The essential domains of LMP1 will be identified and the signaling pathways that are activated will be determined. The effects on rodent fibroblast growth properties of LMP1 alone and in the absence of the p16 and p19 tumor suppressor genes will be determined.

病毒蛋白，潜在的膜蛋白1（LMP1）和潜在膜蛋白2（LMP2）在与EBV相关的许多恶性肿瘤中表达。我们先前使用免疫球蛋白重链启动子/增强子（IG-LMPL）在B淋巴细胞中表达LMP1的转基因小鼠（IG-LMPL）。这些小鼠会出现克隆B细胞淋巴瘤高水平表达LMP1。这些癌症的克隆发育表明，必须发生其他遗传变化，以补充LMP1激活的细胞途径。在这最后的资金阶段，我们已经确定了p16INK4/p19arf基因座的丧失与B-淋巴细胞中LMP1的表达之间的显着致癌协同作用。为了评估LMP1对上皮细胞生长的影响，我们产生了在角蛋白14（K14）启动子控制下表达LMP1的转基因小鼠。在这些小鼠使用经典肿瘤的开始和促进分析中，LMP1在很大程度上起着在肿瘤进展中可能作用的肿瘤启动子。使用IG启动子和来自K14启动子的上皮细胞中B淋巴细胞中表达LMP2的转基因小鼠已从Richard Longnecker博士中获得。将测试K14-LMP2转基因小鼠，以确定LMP2是否通过皮肤测试中的启动，促进或进展影响致癌过程。 LMP1和LMP2表达对B细胞和上皮细胞生长的可能协同作用将在双重转基因小鼠和体外转化分析中分析。我们的具体目的是：1）转基因LMP1+淋巴细胞，LMP1+淋巴瘤，LMP1+ P16无效淋巴瘤和 LMP1+ p53杂合淋巴瘤将进一步表征，以鉴定活化的信号通路和对细胞基因表达的影响。 LMP1+淋巴细胞和淋巴瘤细胞的生长特性将在体外分析。 2）LMP1和LMP2影响并激活不同的细胞信号通路。为了测试LMP1和LMP2坐标表达的假设激活协同改变生长调节的补充途径，IG-LMP1/LMP2转基因小鼠将通过交叉繁殖产生。将确定肿瘤发育的时间以及体外细胞的生长和生化特性。 3）为了确定LMP1和LMP2在正常上皮细胞中的表达，K14启动子转基因小鼠在上皮细胞中表达LMP1和/或LMP2的转基因小鼠将在经典的皮肤致癌法中进行测试，并结合暴露于癌症和肿瘤启动子。 4）表征啮齿动物成纤维细胞的LMP1转化。将确定LMP1的基本域，并确定激活的信号通路。将确定对LMP1的啮齿动物成纤维细胞生长特性的影响，并且在没有p16和p19肿瘤抑制基因的情况下。