A multi-faceted approach to identifying K-Ras synthetic lethal relationships

识别 K-Ras 合成致死关系的多方面方法

• 批准号: 10224565
• 负责人: STEPHEN J ELLEDGE
• 金额: $ 15万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-25 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224565
• 关键词: multi; faceted; approach; identifying; Ras

 DESCRIPTION (provided by applicant): More than 200,000 Americans die as a result of lung and colorectal cancer each year. Decreasing the frequency of deaths due to these cancers will undoubtedly require tailoring an individual's treatment to the specific mutations that have occurred in their cancer. Activating mutations in the K-Ras oncoprotein are common in lung and colorectal cancers and are associated with particularly poor response to both conventional and targeted therapies. Our overarching goal is to understand the mechanisms underlying the oncogenic properties of mutant K-Ras in order to develop targeted therapeutic strategies. This project includes three phases. In the first phase of our project, we will use CRISPR technology to generate K-Ras wild-type derivatives of lung and colorectal cancer cells expressing endogenous mutant K-Ras. We will then comprehensively characterize the cellular and molecular phenotypes associated with loss of mutant K-Ras, for example by combining multiplexed mass spectrometry with computational modeling to identify the signal transduction network utilized by mutant K-Ras to transform cells. This study will also include an analysis of radiation response in wild-type and mutant cells, as activated K-Ras is known to confer resistance to ionizing radiation. In the second phase of our project, we will perform a variety of genome-wide and targeted screens for genes that when knocked down or over-expressed cause lethality in the context of mutant KRas in vitro and in vivo. These studies will utilize stat-of-the-art high-throughput screening technologies, including doxycycline-inducible shRNAs and open reading frames, that we have perfected over the past decade. In the final phase of the project, we will identify K-Ras synthetic lethals that are therapeutically targetable and then perform preclinical studies in genetically engineered mouse models of lung and colon cancer. The utilization of genetically controlled mouse and human experimental systems will allow us to identify gene products that are truly selectively required in cancer cells expressing mutant K-Ras. In the end, this work will have a major impact for patients who develop K-Ras mutant cancer.

 描述（由适用提供）：每年肺和有色癌症导致20万多名美国人死亡。降低由于这些癌症而导致的死亡频率无疑将需要根据其癌症发生的特定突变来调整个人的治疗。 K-RAS癌蛋白中的激活突变在肺和有色癌症中很常见，并且对常规疗法和靶向疗法的反应特别差。我们的总体目标是了解突变K-RAS致癌特性的基础机制，以制定有针对性的治疗策略。该项目包括三个阶段。在我们项目的第一阶段，我们将使用CRISPR技术来生成表达内源性突变体K-RAS的肺和有色癌细胞的K-RAS野生型衍生物。然后，我们将全面地表征与突变K-RAS丢失相关的细胞和分子表型，例如，通过将多路复用质谱与计算模型相结合，以识别突变k-Ras使用的信号翻译网络来转化细胞。这项研究还将包括对野生型和突变细胞中辐射反应的分析，因为已知活化的K-RAS赋予电离辐射的耐药性。在我们项目的第二阶段中，我们将对基因的各种基因组和靶向筛选执行各种基因筛选，这些筛选被敲低或表达过表达时会在突变体KRAS体外和体内引起致死性。这些研究将利用我们在过去十年中完善的高通量筛查技术，包括强力霉素诱导的shrnas和开放式阅读框架。在该项目的最后阶段，我们将确定具有热靶向靶向的K-RAS合成致死剂，然后在肺和颜色癌的基因工程小鼠模型中进行临床前研究。一般控制的小鼠和人类实验系统的利用将使我们能够鉴定出表达突变K-RAS的癌细胞中真正需要的基因产物。最后，这项工作将对患有K-RAS突变癌的患者产生重大影响。

项目成果

KRAS Alleles: The Devil Is in the Detail.

• DOI: 10.1016/j.trecan.2017.08.006
• 发表时间: 2017-10
• 期刊: Trends in cancer
• 影响因子: 18.4
• 作者: Haigis KM

A Role for Mitochondrial Translation in Promotion of Viability in K-Ras Mutant Cells.

• DOI: 10.1016/j.celrep.2017.06.061
• 发表时间: 2017-07-11
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Martin TD;Cook DR;Choi MY;Li MZ;Haigis KM;Elledge SJ
• 通讯作者: Elledge SJ

No back seat for a progression event-K-RAS as a therapeutic target in CRC.

• DOI: 10.1101/gad.297630.117
• 发表时间: 2017-02-15
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Poulin EJ;Haigis KM
• 通讯作者: Haigis KM

Functional genomics reveals that tumors with activating phosphoinositide 3-kinase mutations are dependent on accelerated protein turnover.

• DOI: 10.1101/gad.290122.116
• 发表时间: 2016-12-15
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Davoli T;Mengwasser KE;Duan J;Chen T;Christensen C;Wooten EC;Anselmo AN;Li MZ;Wong KK;Kahle KT;Elledge SJ
• 通讯作者: Elledge SJ