Elucidating Kras-driven immune infiltration and function in pancreatic cancer

项目标题：阐明Kras在胰腺癌中的免疫渗透和功能 胰腺导管腺癌(PDA)是一种致命的恶性肿瘤，5年生存率为9%。掌上电脑 其特点是间质致密，有异质性的成纤维细胞和浸润性免疫细胞。 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。 微环境。然而，人们对串扰背后的机制知之甚少。基因突变 KRAS癌基因是PDA的主要驱动因素，90%以上的人胰腺肿瘤标本中存在KRAS癌基因。 KRAS激活不同的下游信号通路，包括MAPK、PI3K/AKT和JAK/STAT信号通路。 我的初步数据显示，Kras调节免疫细胞的招募和功能。这样做的目的是 建议确定致癌的Kras表达的上皮细胞使用的下游信号通路 将巨噬细胞极化为免疫抑制表型。此外，我生成的初步数据显示 JAK/STAT信号通路在间质中是活跃的，而MAPK和PI3K主要是上皮性的。 巨噬细胞具有活跃的JAK/STAT信号通路。因此，我将调查 JAK/STAT3信号通路在巨噬细胞免疫抑制中的作用我假设致癌因素 KRAS(Kras*)在肿瘤细胞中的表达驱动免疫抑制和靶向JAK/STAT通路 巨噬细胞中的免疫抑制表型将逆转，有助于治疗 胰腺癌。研究Kras*下游的三条信号通路及其如何参与 巨噬细胞极化I将使用体外共培养的肿瘤细胞和骨髓来源的巨噬细胞。 我将使用shRNA下调肿瘤细胞中每个途径的主要成分的表达并解剖 使用蛋白质和RNA对串扰进行分析。了解JAK/STAT在髓系极化中的作用 细胞，因此在肿瘤生长中，我将使用耗尽STAT3的基因工程小鼠模型 尤其是髓系细胞，特别是巨噬细胞。这些研究将提供关于如何 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.