Tumor Host Range Mutants of Polyoma and Their Targets

多瘤的肿瘤宿主范围突变体及其靶标

• 批准号: 7471532
• 负责人: THOMAS Livingston BENJAMIN
• 金额: $ 61.8万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7471532
• 关键词: Accounting; Affinity Chromatography; Antibodies; Apoptosis; Automobile Driving; Binding; Binding Sites; Biological Assay; C-terminal; Cell physiology; Cells; Chromatin; Co-Immunoprecipitations; Consensus; DNA Footprint; DNA Methylation; Development; Embryonal Carcinoma Cell; Epithelial; Epithelial Cells; Exons; Expression Library; Family; Frequencies; Gel; Gene Targeting; Genetic Transcription; Growth; Human; Hybrids; In Situ; In Vitro; Investigation; Large T Antigen; Libraries; Link; Luciferases; Malignant Neoplasms; Malignant neoplasm of ovary; Methylation; Molecular; Molecular Profiling; Mouse Strains; Mus; Normal Cell; Oligonucleotides; Ovarian; Ovarian Carcinoma; Ovary; Ovulation; Pathway interactions; Pattern; Phage Display; Polyomavirus; Procedures; Promoter Regions; Property; Protein Microchips; Proteins; Radiation; Range; Reporter; Research Personnel; Surface; T Virus; TP53 gene; Testing; Trans-Activators; Transgenic Mice; Tumor Antigens; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumor-Derived; Viral; Viral Tumor Antigens; Virus; Virus Replication; Yeasts; Zinc Fingers; base; cDNA Expression; cell growth; cellular targeting; chromatin immunoprecipitation; cis acting element; in vivo; mouse model; mutant; neoplastic cell; programs; promoter; sialosyl-T antigen; transcription factor; tumor

DESCRIPTION (provided by applicant): We propose to continue to isolate and characterize "tumor host range" (THR) mutants of polyoma virus in efforts to identify and characterize the cellular targets of the viral T (tumor) antigens. The rationale behind this selection is that tumor cells may have undergone the loss of particular function(s) which the wild type virus normally targets. These functions may represent tumor suppressor genes or other regulators of replication and survival which the virus needs to inactivate or alter in some manner in order to replicate efficiently. THR mutants are selected to be able to grow on certain tumor cells but not on normal cells and are presumed to have lost the targeting function. One THR mutants has been used to identify the multi-zinc finger homeotic transcription factor Sal2 as a target of the polyoma large T antigen. We have shown that Sal2 acts in some important respects like p53 in terms of its growth arrest and apoptosis inducing properties. In this application we focus on Sal2 with efforts to determine its downstream cellular gene targets, upstream regulators and protein partners. Expression profiling analysis and chromatin immunoprecipitation (ChIP) will be used in these investigations. We will also seek to determine the mutual effects which large T and Sal2 exert on each other based on their interaction. Sal2 is highly expressed in ovarian surface epithelial cells. Preliminary evidence indicates that Sal2 expression is lost in human ovarian carcinomas. We propose to examine further if such loss occurs and at what frequency in various forms of ovarian carcinoma. We will test the hypothesis that promoter methylation in one of the two alternative Sal2 promoters may account for Sal2 silencing in some cases of ovarian carcinoma. Finally, we will take several approaches to developing a mouse model of ovarian cancer based on recent findings on the large T-Sal2 interaction and on the isolation of a new strain of polyoma which efficiently induces ovarian surface epithelial tumors in certain strains of mice.

描述（由申请方提供）：我们建议继续分离和表征多瘤病毒的“肿瘤宿主范围”（THR）突变体，以鉴定和表征病毒T（肿瘤）抗原的细胞靶点。这种选择背后的基本原理是肿瘤细胞可能已经经历了野生型病毒通常靶向的特定功能的丧失。这些功能可能代表肿瘤抑制基因或其他复制和存活的调节因子，病毒需要以某种方式复制或改变这些基因或调节因子以有效复制。THR突变体被选择为能够在某些肿瘤细胞上生长，但不能在正常细胞上生长，并且推测已经失去了靶向功能。一个THR突变体已被用于鉴定多锌指同源异型转录因子Sal 2作为多瘤大T抗原的靶标。我们已经表明，Sal 2的行为在一些重要的方面像p53在其生长停滞和凋亡诱导性能。在本申请中，我们专注于Sal 2，努力确定其下游细胞基因靶点，上游调控因子和蛋白质伴侣。在这些研究中将使用表达谱分析和染色质免疫沉淀（ChIP）。我们还将寻求确定大T和Sal 2基于它们的相互作用而彼此施加的相互影响。Sal 2在卵巢表面上皮细胞中高度表达。初步证据表明Sal 2表达在人卵巢癌中丢失。我们建议进一步研究这种损失是否发生，以及在各种形式的卵巢癌中发生的频率。我们将测试的假设，在两个替代Sal 2启动子之一的启动子甲基化可能占Sal 2沉默在某些情况下，卵巢癌。最后，我们将采取几种方法来开发一种小鼠卵巢癌模型的基础上，最近的研究结果大T-Sal 2的相互作用和分离的一种新的菌株的多瘤，有效地诱导卵巢表面上皮肿瘤在某些品系的小鼠。