Elucidating Kras-driven immune infiltration and function in pancreatic cancer

项目名称：阐明胰腺癌中Kras驱动的免疫浸润和功能 胰腺导管腺癌（PDA）是一种致命的恶性肿瘤，5年生存率为9%。PDA 其特征在于致密的基质，具有成纤维细胞和浸润免疫细胞的异质群体。 肿瘤细胞、成纤维细胞和免疫细胞之间的相互作用导致高度免疫抑制的免疫抑制。 微环境然而，对串扰的潜在机制知之甚少。突变 Kras癌基因是PDA的主要驱动因素，并且在超过90%的人类胰腺肿瘤样品中发现。 Kras激活不同的下游信号通路，包括MAPK、PI 3 K/AKT和JAK/STAT信号通路。 我的初步数据表明，Kras调节免疫细胞的募集和功能。这个目标 一项提案是确定致癌性Kras表达上皮细胞所使用的下游信号通路， 将巨噬细胞转化为免疫抑制表型。另外，我得到的初步数据显示 JAK/STAT信号通路在间质中是活跃的，而MAPK和PI 3 K主要在上皮中。 具体而言，巨噬细胞具有活跃的JAK/STAT信号通路。因此，我将研究 JAK/STAT 3信号通路在巨噬细胞中驱动免疫抑制。我假设致癌基因 肿瘤细胞中的Kras（Kras*）表达驱动免疫抑制和靶向JAK/STAT途径 将导致免疫抑制表型的逆转，有助于治疗 胰腺癌为了研究三种Kras* 下游信号通路以及它们如何参与 巨噬细胞极化I将使用肿瘤细胞和骨髓来源的巨噬细胞的体外共培养物。 我将使用shRNA下调肿瘤细胞中每个通路的主要成分的表达，并解剖肿瘤细胞。 使用蛋白质和RNA分析的串扰。了解JAK/STAT在骨髓细胞分化中的作用 因此，在肿瘤生长中，我将使用一种基因工程小鼠模型， 特别是在骨髓细胞中，特别是巨噬细胞。这些研究将提供一个全面的看法， Kras* 调节肿瘤微环境。

Project Title: Elucidating Kras-driven immune infiltration and function in pancreatic cancer Pancreatic Ductal Adenocarcinoma (PDA) is a lethal malignancy with a 5-year survival rate of 9%. PDA is characterized by a dense stroma with a heterogeneous population of fibroblasts and infiltrating immune cells. The cross-talk between tumor cells, fibroblasts, and immune cells leads to a highly immunosuppressive microenvironment. However, the mechanisms underlying the cross-talk is poorly understood. Mutations in the Kras oncogene are the main drivers of PDA and are found in more than 90% of human pancreatic tumor samples. Kras activates different downstream signaling pathways, including MAPK, PI3K/AKT and JAK/STAT signaling. My preliminary data shows that Kras modulates the immune cells recruitment and function. The goal of this proposal is to identify the downstream signaling pathways used by oncogenic Kras-expressing epithelial cells to polarize macrophages to an immunosuppressive phenotype. Further, preliminary data that I generated shows that the JAK/STAT signaling pathway is active in the stroma, while MAPK and PI3K are predominantly epithelial. Specifically, macrophages have active JAK/STAT signaling pathway. Thus, I will investigate the role of JAK/STAT3 signaling pathway in driving immunosuppression in macrophages. I hypothesize that oncogenic Kras (Kras*) expression in tumor cells drives immunosuppression and that targeting JAK/STAT pathway in macrophages will lead to reversion of the immunosuppressive phenotype aiding in the treatment of pancreatic cancer. To study the three Kras* downstream signaling pathways and how they are involved in macrophage polarization I will use an in vitro co-culture of tumor cells and bone marrow-derived macrophages. I will use shRNA to downregulate expression of the main component of each pathway in tumor cells and dissect the crosstalk using protein and RNA analysis. To understand the role of JAK/STAT in the polarization of myeloid cells and consequently in tumor growth, I will use a genetically engineered mouse model that depletes STAT3 specifically in myeloid cells, specially macrophages. These studies will provide a comprehensive view on how Kras* regulates the tumor microenvironment.