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.

胰腺癌的特征在于“不可能”的KRAS原始癌基因的突变激活， 大约95％的胰腺导管腺癌（PDAC）患者含有Kras 突变。五年生存率糟糕的五年生存率为10％，制定有效的治疗策略 仍然是高度优先级。 KRASG12D突变代表PDAC中最常见的KRAS突变 其次是Krasg12v和Krasg12r。但是，其他突变，例如KRASQ61L，是 在PDAC目的地中不存在模型系统中的高度致癌性。那不是Kras突变 相等，每个突变体可以调节不同的和重叠的信号通路。我们将映射 使用2D和3D细胞模型的特定KRAS突变体的蛋白质相互作用。在AIM 1中，我们将 员工是一种新版本的生物素识别（生化）接近连接测定，称为涡轮 突变特异性蛋白相互作用。我在Krasg12r上的工作提供了一个很好的例子 KRAS突变特异性信号传导。我们假设突变特异性效应子相互作用 KRAS发射的PDAC将阐明哪种途径最容易受到治疗破坏。目标 2，我们建议扩展CDLD的细胞模型核心，以生成小鼠和患者的衍生 胰腺器官在我们的研究中以及其他MUSC中使用。 KRAS信号各种取决于 细胞是否在塑料（2D）与3D环境上生长。为了解决这种复杂性，我们将扩展 我们对正常小鼠胰腺发展开发的3D器官的生物形式研究。这种方法将允许 使用“正常”类器官线来定义KRAS之间的突变特异性信号传导差异 突变体。使用正常小鼠类器官将允许同等基因的“正常”环境 代表KRAS启动胰腺早期阶段的天然胰腺环境 癌症。在早期癌症中验证突变选择性效应途径是否需要 模型系统，我们将建立来自KRASLSL-G12D/+和KRASLSL-G12R/+的肿瘤前类器官 小鼠模型，其中Kras突变在天然基因基因座中表达以验证电势 目标。这种方法将有助于定义胰腺癌中新的突变特异性疗法。