Characterization of Pathways Controlling Cancer at the Level of Gene Regulation

基因调控水平控制癌症途径的表征

• 批准号: 7913508
• 负责人: Phillip A Sharp
• 金额: $ 27.62万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7913508
• 关键词: Characterization; Pathways; Controlling; Cancer; Level

Cancer is a major disease burden and one hope of changing this is to develop new treatments through a better understanding of the disease. Recent discoveries concerning the activities of short RNAs in mammals may provide both new insights and new treatments of cancer. A common goal of this Program is to investigate the roles of short RNAs, such as microRNAs, in regulation of genes in normal and cancer cells. There is strong and rapidly growing evidence suggesting that changes in miRNA regulation are related to malignant transformation and in fact could be a critical event in oncogenic transformation. The function of the mir-17-92-1 cluster which is frequently overexpressed/amplified in a subset of human cancers will be investigated by creation of specific mutations of these microRNAs in the context of mouse models of cancer. Changes in microRNA populations in normal cells and tumor cells of the same developmental state will be analyzed using both bead-array technology as well as new cloning technology. Vectors with regulated expression of a short hairpin RNA which generates a specific siRNA for silencing a gene will be developed for transgenic analysis of pathways. Additionally, methods will be tested for screening of small libraries of shRNA-lentiviral vectors to identify genes which, when silenced, either inhibit or stimulate tumor development. Furthermore, libraries of retro viral vectors expressing shRNAs will be used in screens to identify (a) genes that modulate the proliferation and/or survival of pRB-deficient cells , (b) genes that modulate the rate of development of a K-ras-driven lung cancer model, and (c) genes important for the differentiation of ES cells. The potential role of short RNAs in transcriptional silencing will be investigated in embryonic stem cells. These processes could be important for epigenetic silencing and genomic stability of cancer cells. ES cells will also be studied for the role of miRNAs in development and proliferation. Changes in the spectrum of microRNAs and siRNAs during T-cell development will be characterized using a cloning technology which requires small amounts of RNAi. Activation of the Arf promoter is an early signal in oncogenic transformation. This promoter is silenced under normal conditions by the E2F3B protein, linking the p19Arf-mdm2-p53 pathway to the p16INK4a-cycD/cdk4-pRB-EdF pathway. The role of E2F3B complexes and other E2F factors in regulation of the Arf promoter will be studied.

癌症是一种主要的疾病负担，改变这种状况的一个希望是通过一种新的治疗方法来开发新的治疗方法。 更好地了解疾病。哺乳动物短RNA活性的新发现 可能为癌症提供新的见解和新的治疗方法。该计划的一个共同目标是 研究短RNA（如microRNA）在正常和癌细胞基因调控中的作用。 有强有力的和迅速增长的证据表明，miRNA调控的变化与以下因素有关： 恶性转化，事实上可能是致癌转化中的关键事件。的功能 在人类癌症亚组中经常过表达/扩增的miR-17-92-1簇将被 通过在小鼠癌症模型的背景下创建这些microRNA的特定突变来研究。 在相同发育状态的正常细胞和肿瘤细胞中，microRNA群体的变化将是 使用珠阵列技术以及新的克隆技术进行分析。具有调节的载体 将开发产生用于沉默基因的特异性siRNA的短发夹RNA的表达 用于转基因途径分析。此外，还将测试用于筛选以下小文库的方法： shRNA-慢病毒载体用于鉴定沉默时抑制或刺激肿瘤的基因 发展此外，表达shRNA的逆转录病毒载体文库将用于筛选， 鉴定（a）调节pRB缺陷型细胞增殖和/或存活的基因，（B） 调节K-ras驱动的肺癌模型的发展速率，和（c）对K-ras驱动的肺癌模型的发展重要的基因。 ES细胞的分化。短RNA在转录沉默中的潜在作用将在 胚胎干细胞这些过程可能对表观遗传沉默和基因组稳定性很重要。 癌细胞还将研究ES细胞中miRNA在发育和增殖中的作用。变化 在T细胞发育过程中的microRNA和siRNA谱中， 这项技术需要少量的RNAi。Arf启动子的激活是一个早期信号， 致癌转化该启动子在正常条件下被E2 F3 B蛋白沉默，连接 p19 Arf-mdm 2-p53通路到p16 INK 4a-cycD/cdk 4-pRB-EdF通路。E2 F3 B复合物的作用 以及其他E2 F因子对Arf启动子的调控作用。