Colorectal Cancer: Characterization of a new Cre-LoxP Model

结直肠癌(CRC)是美国癌症相关死亡的主要原因之一。在……里面 我们最近的报告，我们证明了Sprouty2的促肿瘤作用，Sprouty2是一种内源性的抑制因子 受体酪氨酸激酶(RTK)/丝裂原活化蛋白激酶(MAPK)信号转导通路 结直肠癌(CRC)为了确定Sprouty2在癌症中的体内作用，我们最近建立了 我们实验室有一只2F/F小鼠。这个R03应用程序的目标是描述 胚胎(Sprouty2F/F-Villin Cre)和出生后(Sprouty2F/F-Villin)的模型及作用研究 CreERT2)删除关于CRC进展的芽胞2。我们进行这些研究的理由是，建立 Sprouty2基因在结直肠癌中的体内作用将使我们能够提交机械性RO1申请。 我们实验室利用细胞培养模型、原位和转移小鼠进行的体外研究 模型显示了Sprouty2在结直肠癌中的致癌潜力。在初步研究中，我们还 证实依赖TAT的Sprouty2蛋白转导或Sprouty2稳定转染有利于 癌症表型。相反，抑制Sprouty2支持癌细胞的上皮特征 和从Sprouty2F/F-Villin CreERT2小鼠获得的小鼠胚胎成纤维细胞(MEF)。然而， Sprouty2在体内的结直肠癌进展中的要求尚未确定。胚胎和胚胎的影响 Sprouty2F/F-Villin CRE(胚胎缺失)和Sprouty2F/F-Villin CreERT2中Sprouty2的出生后缺失 小鼠(三苯氧胺依赖的出生后缺失)将分别在AOM/DSS诱导期间进行研究 结肠癌的发生。我们还将检测体内Sprouty2缺失对肠道细胞的影响 利用两组小鼠进行分化和谱系鉴定。 据我们所知，这是第一个证明体内SPRY2缺失对结直肠癌影响的研究 进展和肠道细胞谱系。我们的假设是建立在我们的过去和 最近的出版物。拟议的研究是一个多管齐下的方法，让我们为RO1的申请做好准备。

Colorectal cancer (CRC) is one of the leading causes of cancer-related mortality in the United States. In our recent reports, we demonstrated tumor promoting role of Sprouty2, an endogenous suppressor of receptor tyrosine kinase (RTK)/ Mitogen Activated Protein Kinase (MAPK) signaling pathways in colorectal cancer (CRC). In order to establish in vivo role of Sprouty2 in cancer we recently generated Sprouty2F/F mouse in our laboratory. The objectives of this RO3 application are to characterize the model and study the effect of embryonic (Sprouty2F/F-Villin Cre) and postnatal (Sprouty2F/F-Villin CreERT2) deletion of Sprouty2 on CRC progression. Our rationale for these studies is that establishing in vivo role of Sprouty2 gene in CRC would allow us to submit a mechanistic RO1 application. In vitro studies from our laboratory utilizing cell culture models, orthotopic and metastasis mouse models demonstrated oncogenic potential of Sprouty2 in CRC. In preliminary studies, we also established that TAT dependent Sprouty2 protein transduction or Sprouty2 stable transfection favors cancer phenotype. On the contrary suppression of Sprouty2 supports epithelial features in cancer cells and murine embryonic fibroblasts (MEFs) obtained from Sprouty2F/F-Villin CreERT2 mouse. However, requirement of Sprouty2 in CRC progression in vivo is not established. Effect of embryonic and postnatal Sprouty2 deletion in Sprouty2F/F-Villin Cre (embryonic deletion) and Sprouty2F/F-Villin CreERT2 mouse (tamoxifen dependent postnatal deletion), respectively, will be studied during AOM/DSS induced colonic carcinogenesis. We will also examine effect of Sprouty2 deletion in vivo on intestinal cell differentiation and lineage by utilizing both mice groups. To our knowledge, this is the first study to demonstrate the effect of in vivo deletion of SPRY2 on CRC progression and intestinal cell lineage. We have established our hypothesis based on our past and recent publications. The proposed study is a multi-prong approach to prepare us for a RO1 application.