Making hypomorphic tumor supressors in vivo using RNAi

利用 RNAi 在体内制造亚型肿瘤抑制因子

• 批准号: 7392158
• 负责人: SCOTT W. LOWE
• 金额: $ 89.31万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7392158
• 关键词: Animals; Behavior; Breast; Cancer Model; Carcinoma; Complex; Cultured Cells; Development; Disabled Persons; Epigenetic Process; Gene Expression; Gene Silencing; Genes; Genetic; Genetic Screening; Genome; Libraries; Maintenance; Malignant Neoplasms; Mediating; Methods; Modeling; Molecular Profiling; Mus; Neoplasms; Pathology; Phenotype; Pre-Clinical Model; RNA Interference; Range; Research Personnel; Series; System; Technology; Testing; Therapeutic Intervention; Tumor Suppressor Genes; Tumor Suppressor Proteins; Tumor-Suppressor Gene Inactivation; cancer therapy; carcinogenesis; gene function; human cancer mouse model; in vivo; insight; mouse genome; neoplastic cell; novel; rapid technique; response; small hairpin RNA; success; tumor; tumor progression

DESCRIPTION (provided by applicant): Tumor suppressors act as components of complex networks whose overall function can be impaired by many different genetic or epigenetic alterations. While many of these alterations have been elegantly recapitulated in mouse, existing technology has been limited in its ability to model the significant range and complexity of gene suppression that occurs during neoplastic progression. The recent development of retroviral systems capable of mediating stable gene silencing has vastly increased our capacity to recreate the precise gene expression profiles seen in tumor cells. We propose to exploit the emerging power of RNA interference (RNAi) to study tumor suppressor gene networks in vivo, in particular, the impact of tumor suppressor hypomorphs on tumor development and responses to cancer therapy. Our team includes investigators that have been pioneers in establishing methods for using short hairpin RNAs (shRNAs) to stably suppress gene function in culture cells and in animals. As well as investigators with substantial expertise in modeling cancer and cancer therapy in the mouse who have successfully demonstrated that shRNAs can create 'epi-allelic' series of hypomorphs that produce distinct tumor phenotypes in vivo. Our experimental approach will be to: 1) develop RNAi technology for suppressing gene function in both chimeric (genetic mosaic) and germline settings; 2) use conditional systems to determine the extent to which tumor suppressor gene inactivation is required for tumor maintenance, and the consequences of gene reactivation on tumor behavior; 3) use this technology to produce an 'epi-allelic' series of tumor suppressor hypomorphs that may produce different pathologies depending on the strength of suppression; 4) take advantage of new shRNA libraries presently capable of targeting cancer relevant genes in the mouse genome (and likely to expand genome-wide) to conduct unbiased genetic screens for modulators of tumor phenotypes; 5) build upon our previous success with the hematopoeitic system to model carcinomas, with a particular emphasis on developing rapid methods to evaluate genetic interactions during breast carcinogenesis and therapy. We expect that these studies will provide new insights into how tumor suppressor networks are disabled during the development of particular neoplasias, and ultimately identify key nodes in these networks that may be sensitive to therapeutic intervention. Moreover, they will produce new mouse models of human cancer that can be used to understand treatment responses and as preclinical models for testing novel therapies.

描述(由申请人提供)： 肿瘤抑制因子作为复杂网络的组成部分，其整体功能可因许多不同的遗传或表观遗传改变而受损。虽然许多这些改变已经在小鼠身上得到了优雅的概括，但现有技术在模拟肿瘤进展过程中发生的基因抑制的显著范围和复杂性方面的能力一直受到限制。最近发展的逆转录病毒系统能够介导稳定的基因沉默，极大地提高了我们重建在肿瘤细胞中看到的精确基因表达谱的能力。我们建议利用RNA干扰(RNAi)的新兴力量来研究体内的抑癌基因网络，特别是抑癌基因亚型对肿瘤发生和癌症治疗反应的影响。我们的团队包括在建立使用短发夹状RNA(ShRNAs)稳定抑制培养细胞和动物中基因功能的方法方面具有开拓性的研究人员。以及在小鼠癌症建模和癌症治疗方面拥有丰富专业知识的研究人员，他们成功地证明shRNAs可以在体内产生一系列产生不同肿瘤表型的表观等位基因亚型。我们的实验方法将是：1)开发在嵌合(遗传镶嵌)和生殖系环境中抑制基因功能的RNAi技术；2)使用条件系统来确定肿瘤维持所需的肿瘤抑制基因失活的程度，以及基因重新激活对肿瘤行为的影响；3)使用该技术产生一系列可能产生不同病理变化的表观等位基因；4)利用目前能够针对小鼠基因组中的癌症相关基因(并可能在全基因组范围内扩展)的新的shRNA文库，对肿瘤表型的调节器进行无偏见的遗传筛选；5)在我们之前成功利用造血系统建立癌症模型的基础上，特别强调开发快速方法来评估乳腺癌发生和治疗过程中的遗传交互作用。我们希望这些研究将提供新的见解，了解肿瘤抑制网络在特定肿瘤的发展过程中是如何被禁用的，并最终确定这些网络中可能对治疗干预敏感的关键节点。此外，他们还将生产新的人类癌症小鼠模型，可以用来了解治疗反应，并作为测试新疗法的临床前模型。