OTHER FUNCTIONS SBIR PHASE II TOPIC 290 "RNAI SYNTHETIC LETHAL SCREEN IN CANCER

其他功能 SBIR 第二阶段主题 290“癌症中的 RNAI 合成致死筛查

• 批准号: 8565071
• 负责人: ALEXANDR CHENCHIK
• 金额: $ 99.96万
• 依托单位: CELLECTA, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-28 至 2014-09-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8565071
• 关键词: A549; Ablation; Antineoplastic Agents; Apoptosis; Breast; Breast Cancer Cell; Cancer cell line; Cell Cycle; Cell Proliferation; Cell model; Cells; Computer software; Contracts; Custom; DNA Repair; Data; Data Analyses; Data Set; Databases; Development; Documentation; Drug Delivery Systems; Funding; Gene Combinations; Genes; Genetic Screening; Goals; Individual; Lethal Genes; Libraries; Malignant Neoplasms; Malignant neoplasm of prostate; Maps; Messenger RNA; Mutate; Non-Malignant; Outcome; Phase; Plasmids; Quality Control; RNA Interference; Reporting; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Screening procedure; Services; Small Business Innovation Research Grant; Software Tools; Specificity; Technology; Time; Toxic effect; Update; Validation; base; cancer cell; combinatorial; cytotoxic; cytotoxicity; design; gene interaction; novel; small hairpin RNA

A promising direction in the development of anti-cancer drugs with high specificity and low toxicity is the possibility of inhibiting the functions of synthetically lethal (SL) (synergetic) DNA Damage and Repair (DDR) drug targets that are specifically modulated in cancer development. As outlined in the 290 SBIR contract proposals, one of the promising, although unrealized, strategies for identifying SL targets is to apply a systematic, unbiased RNAi viability screen based on the ablation of two targets from a complete set of DDR genes. As a first step for constructing a cancer-SL gene interaction map, which used Phase I SBIR contract funding, we developed the technology for the construction of novel bi-specific pooled lentiviral shRNA libraries designed for the combinatorial knockdown of any two targets in a set of rationally selected genes. Furthermore, we successfully demonstrated the application of the bi-specific shRNA library to detect SL binary interactions between 40 DDR genes in an isogenic breast cancer cell model. The ultimate scientific goal of the Phase II contract studies is to apply the developed combinatorial RNAi screening technology to the large-scale discovery of all possible SL gene pairs in a set of 400 DDR targets from a panel of three cancer cell lines. As a practical outcome of the proposed studies, we will develop, validate and commercialize a novel open-resource SL DDR RNAi screening platform for the unbiased discovery and annotation of SL interactions in cancer cell models. To provide researchers with easy access to the combinatorial RNAi screening technology, Cellecta will offer custom bi-specific shRNA libraries and custom RNAi screening services in cancer cell models.

开发具有高特异性和低毒性的抗癌药物的一个有希望的方向是抑制在癌症发展中特异性调节的合成致死（SL）（协同）DNA损伤和修复（DDR）药物靶标的功能的可能性。正如在290 SBIR合同提案中所概述的，用于鉴定SL靶标的有希望的（尽管尚未实现）策略之一是基于从一组完整的DDR基因中切除两个靶标来应用系统的、无偏见的RNAi活力筛选。作为构建癌症-SL基因相互作用图谱的第一步，使用I期SBIR合同资金，我们开发了用于构建新型双特异性合并慢病毒shRNA文库的技术，该文库设计用于组合敲除一组合理选择的基因中的任何两个靶标。此外，我们成功地证明了双特异性shRNA文库的应用，以检测在同基因乳腺癌细胞模型中的40个DDR基因之间的SL二元相互作用。第二阶段合同研究的最终科学目标是将开发的组合RNAi筛选技术应用于大规模发现来自三种癌细胞系的一组400个DDR靶标中的所有可能的SL基因对。作为拟议研究的实际成果，我们将开发，验证和商业化一种新的开放资源SL DDR RNAi筛选平台，用于在癌细胞模型中无偏见地发现和注释SL相互作用。为了使研究人员能够轻松获得组合RNAi筛选技术，Cellecta将在癌细胞模型中提供定制的双特异性shRNA文库和定制RNAi筛选服务。