Defining the KRAS mutation specific interactions in pancreas cancer

定义胰腺癌中 KRAS 突变的特异性相互作用

• 批准号: 10586129
• 负责人: Guy Hobbs
• 金额: $ 80.61万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-03 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586129
• 关键词: 3-Dimensional; Address; Biological Assay; Biological Models; Biotin; Cancer Model; Cell model; Cells; Centers of Research Excellence; Digestive System Disorders; Environment; KRAS2 gene; KRASG12D; Ligation; Liver diseases; Malignant neoplasm of pancreas; Maps; Mus; Mutation; Oncogenic; Organoids; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Predisposition; Proteins; Proto-Oncogenes; Signal Pathway; Signal Transduction; Survival Rate; Therapeutic; Work; genomic locus; mouse model; mutant; neoplastic; therapeutically effective

Pancreatic cancer is characterized by mutational activation of the “undruggable” KRAS proto-oncogene, and approximately 95% of all pancreatic ductal adenocarcinoma (PDAC) patients harbor a KRAS mutation. With an abysmal five-year survival rate of 10%, developing effective therapeutic strategies remains a high priority. The KRASG12D mutation represents the most common KRAS mutation in PDAC followed by KRASG12V and KRASG12R. However, other mutations, such as KRASQ61L, are non-existent in PDAC despite being highly oncogenic in model systems. Thus, KRAS mutations are not equal, and each mutant may regulate distinct as well as overlapping signaling pathways. We will map the protein interactions of specific KRAS mutants utilizing both 2D and 3D cell models. In Aim 1, we will employ a new version of Biotin Identification (BioID) proximity ligation assays, termed TurboID, to define the mutation-specific protein interactions. My work on KRASG12R provides an excellent example of KRAS mutation-specific signaling. We hypothesize that the mutant-specific effector interactions in KRAS-initiated PDAC will illuminate which pathways are most susceptible to therapeutic disruption. In Aim 2, we propose to expand the Cell Models Core of the CDLD to generate mouse- and patient-derived pancreatic organoids for use in our studies and others at MUSC. KRAS signaling varies depending on whether cells are grown on plastic (2D) vs 3D environments. To address this complexity, we will extend our BioID studies into 3D organoids developed from normal mouse pancreas. This approach will allow for the use of a “normal” organoid line to define the mutation-specific signaling differences between KRAS mutants. The use of normal mouse organoids will allow for an isogenic “normal” environment that is more representative of the natural pancreas environment in which KRAS initiates the early stages of pancreatic cancer. To validate whether the mutation-selective effector pathways are necessary in an early cancer model system, we will establish pre-neoplastic organoids from KrasLSL-G12D/+ and KrasLSL-G12R/+ mouse models, where the KRAS mutation is expressed in the native gene locus to validate the potential targets. This approach will help define a new class of mutation-specific therapies in pancreatic cancer.

胰腺癌的特征是“无法用药”的KRAS原癌基因的突变激活， 约95%的胰腺导管腺癌(PDAC)患者存在KRAS 突变。五年存活率高达10%，正在制定有效的治疗策略 仍然是一个高度优先的问题。KrasG12D突变是PDAC中最常见的KRAS突变 其次是KRASG12V和KRASG12R。然而，其他突变，如KRASQ61L， 在PDAC中不存在，尽管在模型系统中具有高度致癌性。因此，KRAS突变并不是 相同的情况下，每个突变体可以调节不同的和重叠的信号通路。我们将绘制 利用2D和3D细胞模型研究特定KRAS突变体的蛋白质相互作用。在目标1中，我们将 使用一种新版本的生物素鉴定(BioID)邻近连接分析，称为TurboID，以定义 突变特有的蛋白质相互作用。我在KRASG12R上的工作提供了一个很好的例子 KRAS突变特异性信号转导系统。我们假设突变体特有的效应器相互作用在 KRAS发起的PDAC将阐明哪些途径最容易受到治疗中断的影响。在AIM 2、我们建议扩展CDLD的细胞模型核心，以生成小鼠和患者派生的 在我们的研究和MUSC的其他研究中使用的胰腺有机化合物。KRAS信令因以下因素而异 细胞是在塑料(2D)环境中生长还是在3D环境中生长。为了解决这种复杂性，我们将扩展 我们对从正常小鼠胰腺发育而来的3D有机化合物进行了BioID研究。这种方法将允许 用“正常”类器官系来定义KRAS之间的突变特异性信号差异 变种人。使用正常的小鼠器官将允许一个同源的“正常”环境 代表KRAS启动胰腺早期阶段的自然胰腺环境 癌症。为了验证突变选择性效应通路在早期癌症中是否是必要的 模型系统，我们将建立KrasLSL-G12D/+和KrasLSL-G12R/+的癌前有机物 小鼠模型，其中KRAS突变在天然基因位点表达，以验证潜在的 目标。这一方法将有助于确定胰腺癌突变特异性治疗的新类别。