Oncogenes and oncogenic microRNAs in chicken tumors

鸡肿瘤中的癌基因和致癌 microRNA

基本信息

批准号：
8107864
负责人：
Karen L. Beemon
金额：
$ 32.47万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-01 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8107864
关键词：
3&apos Untranslated Regions Alternative Splicing Animal Model Apoptosis Avian Leukosis Virus B-Cell Lymphomas B-Lymphocytes Bioinformatics Biological Assay Biological Models Cell Proliferation Cells Chick Embryo Chickens Cultured Cells DNA Integration Development Disease Down-Regulation Embryo Enhancers Event Fibroblasts Fluorescent in Situ Hybridization Functional RNA Gel Gene Expression Gene Targeting Genes Genetic Genetic Transcription Genome Growth Health Human Individual Infection Insertional Activations Insertional Mutagenesis Length Luciferases Maintenance Malignant Neoplasms Measures MicroRNAs Microarray Analysis Monitor Mus Mutate Oncogene Activation Oncogenes Oncogenic Pattern Phenotype Process Promoter Regions Property Protein Analysis Proteins Proteomics Proto-Oncogenes RNA RNA Interference Relative (related person)Reporter Retroviral Vector Retroviridae Reverse Transcriptase Polymerase Chain Reaction Role Site Southern Blotting System Techniques Telomerase Testing Time Tumor Suppressor Genes Tumor Suppressor Proteins Tumor-Suppressor Gene Inactivation Virus Integration Western Blotting in vivo interest interstitial knock-down meetings mutant novel research study small hairpin RNA telomerase reverse transcriptase telomere therapeutic development tissue culture tumor tumorigenesis two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Tumors are caused by multiple genetic events, usually involving activation of oncogenes and inactivation of tumor suppressor genes. In humans, but not mice, activation of telomerase reverse transcriptase (TERT) is also observed in ~90% of tumors. Recently, activation of TERT by avian leukosis virus (ALV) integration was observed in B-cell lymphomas in chickens. Since the TERT integrations were clonal, this appears to be an early event in tumor development. In the first aim, this hypothesis will be tested by infection of 10-day chicken embryos with a retrovirus expressing the TERT gene. The rate of tumor induction will be monitored. Retroviral tagging will be employed to identify oncogenes and microRNAs that cooperate with TERT to generate tumors. Cooperativity will be studied in tissue culture systems and in tumor induction. Telomere lengths in the tumors will be analyzed in an attempt to determine whether TERT has roles in addition to telomere lengthening. Finally, telomere expression will be studied at the level of transcription and alternative splicing in the tumors. The hypothesis that retroviruses inactivate tumor suppressors by insertional activation of microRNAs will be tested in the second Aim. miR-155, processed from the non-coding bic gene, is activated in metastatic B-cell lymphomas induced by ALV. Targets of ALV will be identified by bioinformatics, microarray and proteomics analyses and validated with reporter assays. Levels of targets in tumors expressing bic will be determined. The role of individual targets in tumor induction will be studied in tissue culture assays and in vivo. PUBLIC HEALTH RELEVANCE: Since telomerase is involved in most human cancers, it is important to have an animal model system that involves telomerase to further understanding of the disease and development of therapeutics. Chicken B-cell lymphomas induced by retroviral DNA integration into the genome have been shown to involve telomerase and will be studied here. In addition, the role of microRNA-155 in tumor induction will be studied by the identification and characterization of targets important for oncogenesis.

描述（由申请人提供）：肿瘤是由多个遗传事件引起的，通常涉及癌基因的激活和肿瘤抑制基因失活。在人类而不是小鼠中，在约90％的肿瘤中也观察到端粒酶逆转录酶（TERT）的激活。最近，在鸡的B细胞淋巴瘤中观察到了禽白血病病毒（ALV）积分的TERT激活。由于TERT整合是克隆的，因此这似乎是肿瘤发育的早期事件。在第一个目标中，该假设将通过用表达TERT基因的逆转录病毒感染10天的鸡胚胎来检验。将监测肿瘤诱导的速率。逆转录病毒标记将用于识别与TERT合作以产生肿瘤的癌基因和microRNA。合作性将在组织培养系统和肿瘤诱导中进行研究。将分析肿瘤中的端粒长度，以确定除端粒延长外是否具有作用。最后，将在肿瘤中的转录水平和替代剪接水平上研究端粒表达。在第二个目标中测试了逆转录病毒通过插入microRNA的逆转录病毒抑制肿瘤的假设。从非编码BIC基因加工的miR-155在ALV诱导的转移性B细胞淋巴瘤中被激活。 ALV的靶标将通过生物信息学，微阵列和蛋白质组学分析来确定，并通过记者测定进行验证。将确定表达BIC的肿瘤中的靶标水平。将在组织培养测定法和体内研究各个靶标在肿瘤诱导中的作用。公共卫生相关性：由于端粒酶参与大多数人类癌症，因此拥有一个涉及端粒酶以进一步了解疗法和发展疗法的动物模型系统很重要。逆转录病毒DNA整合到基因组中诱导的鸡B细胞淋巴瘤已显示涉及端粒酶，并将在此处研究。另外，通过对肿瘤发生重要的靶标的鉴定和表征，将研究microRNA-155在肿瘤诱导中的作用。