Systematic genome-wide RNAi screen to identify novel synthetic lethal interactions in human breast cancer cell lines: implications for the development of therapeutic molecular targets.

系统性全基因组 RNAi 筛选，以确定人类乳腺癌细胞系中新型合成致死相互作用：对治疗分子靶点开发的影响。

• 批准号: 187319425
• 负责人: Dr. Eva Maria Murga Penas, Ph.D.
• 金额: --
• 依托单位: Institute for Cancer Genetics
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/187319425?language=en
• 关键词: Systematic; genome; wide; RNAi; screen

Cancer is a heterogeneous disease that arises from an accumulation of genetic and epigenetic alterations resulting in a failure of cells to respond correctly to diverse signals, such as control of proliferation, differentiation, adhesion and apoptosis. Most chemotherapeutic treatments are based on drugs that affect unspecifically the rapidly growing tumor cells and a variety of normal cells producing, therefore, severe side effects. Novel anticancer therapies are focused on treatments that selective kill cancer cells while sparing normal cells and therefore are less toxic. The application of the RNAi technology for discovering of tumor cell specific vulnerabilities is a novel strategy that will help to the development of more rational therapies in cancer treatments. In this project, I will conduct a gene silencing strategy to introduce specific perturbations to compromise the homeostasis of breast cancer cells without affecting normal ones. For this purpose, a second-generation of shRNA library of 70,000 short hairpin RNAs will be used as a perturbation method to search the entire genome for genes that are required to maintain the viability of different breast cancer cells (basal, luminal, and non-transformed cell lines).This project exploits the RNAi technology to identify genes that, when silenced, exclusively reduce the viability of breast cancer cells that carry preexistent genetic alterations without affecting normal cells. Thus, the successful completion of this research plan will provide novel targets for more efficient and less harmful breast cancer therapies.

癌症是一种异质性疾病，由遗传和表观遗传改变的积累导致细胞无法正确响应不同的信号（例如增殖、分化、粘附和凋亡的控制）而引起。大多数化疗所采用的药物会非特异性地影响快速生长的肿瘤细胞和各种正常细胞，从而产生严重的副作用。新型抗癌疗法的重点是选择性杀死癌细胞，同时不伤害正常细胞，因此毒性较小。应用RNAi技术发现肿瘤细胞特异性脆弱性是一种新策略，将有助于在癌症治疗中开发更合理的疗法。在这个项目中，我将实施一种基因沉默策略，引入特定的扰动来破坏乳腺癌细胞的稳态，而不影响正常细胞。为此，将使用包含 70,000 个短发夹 RNA 的第二代 shRNA 文库作为扰动方法，在整个基因组中搜索维持不同乳腺癌细胞（基底细胞系、管腔细胞系和非转化细胞系）活力所需的基因。该项目利用 RNAi 技术来识别基因，这些基因在沉默时会专门降低携带先前存在遗传基因的乳腺癌细胞的活力。 改变而不影响正常细胞。因此，该研究计划的成功完成将为更有效、危害更小的乳腺癌治疗提供新的靶点。