Defining the KRAS mutation specific interactions in pancreas cancer

胰腺癌的特征在于“不可治疗的”KRAS原癌基因的突变激活， 大约95%的胰腺导管腺癌（PDAC）患者具有KRAS 突变五年生存率只有10%，发展有效的治疗策略 仍然是一个高度优先事项。KRASG 12 D突变代表PDAC中最常见的KRAS突变 其次是KRASG 12 V和KRASG 12 R。然而，其他突变，如KRASQ 61 L， 在PDAC中不存在，尽管在模型系统中是高度致癌的。因此，KRAS突变不是 每个突变体都可以调节不同的以及重叠的信号传导途径。我们将绘制 利用2D和3D细胞模型的特定KRAS突变体的蛋白质相互作用。在目标1中，我们 采用新版本的生物素鉴定（BioID）邻近连接测定法，称为TurboID，以确定 突变特异性蛋白质相互作用我在KRASG 12 R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，在细胞中， KRAS启动的PDAC将阐明哪些途径最容易受到治疗中断的影响。在Aim中 2，我们建议扩展CDLD的细胞模型核心，以产生小鼠和患者来源的 胰腺类器官用于我们的研究和MUSC的其他研究。KRAS信号传导取决于 细胞是否生长在塑料（2D）和3D环境中。为了解决这一复杂性，我们将 我们的BioID研究是从正常小鼠胰腺发育的3D类器官。这种方法将允许 使用“正常”类器官系来定义KRAS-1和KRAS-2之间的突变特异性信号传导差异， 变种人正常小鼠类器官的使用将允许一个等基因的“正常”环境， 代表KRAS启动胰腺炎早期阶段的天然胰腺环境， 癌验证突变选择性效应通路是否在早期癌症中是必需的 模型系统，我们将从KrasLSL-G12 D/+和KrasLSL-G12 R/+建立肿瘤前类器官 小鼠模型，其中KRAS突变在天然基因座中表达，以验证 目标的这种方法将有助于定义一类新的胰腺癌突变特异性疗法。

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.