Elucidating KRAS-specific vulnerabilities in pancreatic cancer

阐明胰腺癌中 KRAS 特异性的脆弱性

• 批准号: 9164819
• 负责人: Mandar Deepak Muzumdar
• 金额: $ 17.71万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9164819
• 关键词: Adenocarcinoma Cell; Adverse effects; Advisory Committees; Animal Model; Apoptosis; Award; Basic Science; Biochemical; Biological Assay; Biology; CRISPR screen; Cancer Biology; Cancer Center; Cancer Etiology; Cancer Patient; Caring; Cell Line; Cell Proliferation; Cells; Cessation of life; Clinical; Clinical Sciences; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Colon; Combination Drug Therapy; Communities; Complementary DNA; Dana-Farber Cancer Institute; Dependency; Development; Environment; Essential Genes; Evaluation; Fellowship; Fostering; Funding; Gene Expression; Gene Targeting; Genes; Genetic; Genetic study; Goals; Growth; Guide RNA; High Prevalence; Human; In Vitro; Institutes; International; KRAS2 gene; Knock-out; Laboratories; Libraries; Maintenance; Malignant Neoplasms; Malignant neoplasm of gastrointestinal tract; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Measures; Mediating; Mediator of activation protein; Medical Oncologist; Mentors; Mentorship; Methodology; Molecular Biology; Morbidity - disease rate; Mus; Mutate; Mutation; Neoplasm Transplantation; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Physicians; Play; Postdoctoral Fellow; Proteins; Proto-Oncogenes; Reagent; Research; Research Personnel; Resources; Role; Scientist; Signal Pathway; Signal Transduction; Technology; Therapeutic; Time; Training Programs; Translational Research; Translations; Transplantation; United States; Validation; Variant; Work; anticancer research; base; cDNA Library; cancer cell; cancer genetics; cancer therapy; catalyst; design; drug sensitivity; genome editing; genome-wide; genome-wide analysis; improved; in vivo; in vivo Model; inhibitor/antagonist; knock-down; medical schools; mortality; mouse model; mutant; novel; novel strategies; novel therapeutics; pancreatic neoplasm; pre-clinical; programs; research study; response; screening; targeted treatment; tool; transcriptome sequencing; tumor

PROJECT SUMMARY/ABSTRACT Despite significant therapeutic advances in other cancers, treatment of pancreatic ductal adenocarcinoma (PDAC) has improved only modestly over the past several decades. Consequently, pancreatic cancer remains the fourth leading cause of cancer death in the United States and a major cause of morbidity and mortality worldwide. Mutation of the proto-oncogene KRAS is a hallmark of PDAC, occurring in >90% of cases. Studies in animal models and cell lines have suggested that mutant KRAS is important for both the initiation and maintenance of PDAC, making KRAS an attractive target for therapy. Unfortunately, efforts to directly target KRAS have been thwarted by several unique features of the mutant protein, making KRAS “undruggable” to date. Furthermore, current strategies to inhibit putative downstream signaling pathways have been largely unsuccessful. Given the high prevalence of KRAS mutations in not only PDAC but also cancers of the lung and colon, the development of new approaches to target KRAS mutant cancers remains a significant unmet need. The long-term goal of the proposed research is to improve our understanding of KRAS biology to facilitate the development of novel therapeutic strategies in PDAC and other KRAS mutant cancers. Our preliminary studies revealed that partial or complete inhibition of mutant KRAS induced marked and reversible changes in gene expression, cell signaling, and drug sensitivity, suggesting that KRAS fosters a specific cell state, potentially amenable to targeted inhibition. In this proposal, we leverage novel PDAC cell lines in which KRAS has been inactivated by CRISPR/Cas-mediated genome editing, a lentiviral cDNA library permitting expression of a diverse array of KRAS mutant variants, and CRISPR-based genome-wide screening libraries to interrogate key KRAS-relevant functional pathways and elucidate synthetic lethal interactions with KRAS mutation. Specifically, we propose to re-express different KRAS mutant variants to identify variant-specific effectors through transcriptional and biochemical profiling, offering an unprecedented evaluation of KRAS mutant variants in cancer cells. In parallel, we plan to elucidate unique genetic dependencies in cells harboring or lacking KRAS function using CRISPR-based screening. Together, these approaches are likely to reveal novel targetable pathways and proteins in KRAS mutant cells, which we will validate in additional cell lines in vitro and transplant and autochthonous models in vivo. The proposed research strategy is part of an extensive training program designed to facilitate my transition to becoming an independent investigator in basic and translational cancer biology. The main experiments will be conducted in the Jacks laboratory at the Koch Institute for Integrative Cancer Research at MIT under the guidance of Dr. Tyler Jacks, an international leader in KRAS biology and mouse models of cancer. Co- mentorship on clinical and translational science will be provided by Dr. Charles Fuchs, a leading clinical researcher and Director of the GI SPORE at the Dana-Farber Cancer Institute and Harvard Cancer Center (DF/HCC). The scientific community, encompassing the DF/HCC, Koch Institute at MIT, and Harvard Medical School, offers an outstanding environment for scientific discussion and collaboration. In addition, I have assembled a scientific advisory committee, comprised of Drs. William Hahn, Matthew Vander Heiden, and Alec Kimmelman, who collectively have a wealth of information and expertise in basic and translational research. In addition to presenting my work locally at seminars within the Harvard and MIT community, I have also proposed a comprehensive set of didactic coursework through the Harvard Catalyst program to enhance my understanding of translational research with a focus on target identification, preclinical validation, and T1 translation. As a medical oncologist and physician-scientist, I am committed to improving the care of cancer patients, with a focus on pancreatic cancer and other gastrointestinal malignancies. As a postdoctoral fellow in the Jacks lab, I have led the development of several projects, building novel tools and reagents that I will utilize in the proposed experiments. Results from these studies will form the basis for developing an independently-funded laboratory. While I have been successful in acquiring fellowship funding to support my preliminary studies, the K08 award will provide critical funding for continued mentored research and transition to independence.

项目摘要/摘要 尽管在其他癌症的治疗方面取得了重大进展，但胰腺导管腺癌的治疗 在过去的几十年里，(PDAC)只有轻微的改善。因此，胰腺癌仍然存在。 美国癌症死亡的第四大原因，也是发病率和死亡率的主要原因 全世界。原癌基因KRAS的突变是PDAC的一个特征，发生在90%的病例中。研究 在动物模型和细胞系中，已经表明突变的KRAS对启动和 维持PDAC，使KRAS成为有吸引力的治疗靶点。不幸的是，直接针对 KRAS已经被突变蛋白的几个独特特征所阻挠，使得KRAS对 约会。此外，目前抑制下游信号通路的策略在很大程度上是 不成功。鉴于KRAS突变不仅在PDAC而且在肺癌和肺癌中的高流行率 在结肠，针对KRAS突变癌症的新方法的开发仍然是一个重要的未得到满足的需求。 拟议研究的长期目标是提高我们对KRAS生物学的理解，以促进 PDAC和其他KRAS突变癌症的新治疗策略的开发。我们的初步研究 发现部分或完全抑制突变的KRAS会导致基因的显著和可逆的变化 表达、细胞信号和药物敏感性，表明KRAS潜在地促进了特定的细胞状态 易受定向抑制的。在这个方案中，我们利用了KRAS已经在其中的新型PDAC细胞系 通过CRISPR/Cas介导的基因组编辑失活的慢病毒cDNA文库允许表达 多种KRAS突变体和基于CRISPR的全基因组筛选文库，以询问关键字 KRAS相关的功能通路，并阐明与KRAS突变的合成致死相互作用。 具体地说，我们建议重新表达不同的KRAS突变变体以识别特定于变体的效应器 通过转录和生化图谱，提供前所未有的KRAS突变评估 癌细胞中的变种。同时，我们计划阐明细胞中存在的独特的遗传依赖关系 CRISPR筛查缺乏KRAS功能。总而言之，这些方法可能会揭示出新的 KRAS突变细胞中的靶向通路和蛋白质，我们将在体外额外的细胞系中进行验证 体内移植和自体模型。 拟议的研究战略是一项广泛培训计划的一部分，旨在促进我向 成为癌症生物学基础和转译领域的独立研究员。主要的实验将是 在麻省理工学院科赫癌症综合研究所的贾克斯实验室进行 在KRAS生物学和小鼠癌症模型方面的国际领先者泰勒·杰克斯博士的指导下。共同-- 临床和翻译科学的导师将由领先的临床专家Charles Fuchs博士提供 达纳-法伯癌症研究所和哈佛癌症中心GI孢子研究员兼主任 (DF/肝细胞癌)。科学界，包括DF/HCC、麻省理工学院科赫研究所和哈佛医学院 学校，为科学讨论和合作提供了一个出色的环境。此外，我还有 组建了一个科学咨询委员会，由威廉·哈恩博士、马修·范德·海登博士和亚历克博士组成 他们在基础研究和翻译研究方面拥有丰富的信息和专业知识。在……里面 除了在哈佛和麻省理工学院的研讨会上展示我的工作外，我还 通过哈佛催化剂计划提出了一套全面的教学课程，以增强我的 了解转化性研究，重点关注目标识别、临床前验证和T1 翻译。 作为一名内科肿瘤学家和内科科学家，我致力于改善癌症患者的护理， 重点关注胰腺癌和其他胃肠道恶性肿瘤。作为贾克斯实验室的博士后研究员， 我领导了几个项目的开发，构建了我将在 拟议的实验。这些研究的结果将成为开发独立资助的 实验室。虽然我已经成功地获得了奖学金来支持我的初步学习， K08奖将为继续指导研究和向独立过渡提供关键资金。