Colorectal Cancer: Characterization of a new Cre-LoxP Model

结直肠癌：新 Cre-LoxP 模型的表征

• 批准号: 9307305
• 负责人: SHARAD KHARE
• 金额: $ 6.08万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-15 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9307305
• 关键词: Award; Budgets; Cancer Etiology; Cancer Model; Cancer Patient; Carcinoma; Cell Culture Techniques; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cessation of life; Colorectal Cancer; Cre-LoxP; Data; Development; Embryo; Epithelial; Epithelial Cells; Fibroblasts; Foundations; Funding; Gene Deletion; Genes; Goals; Homeostasis; Human; Immunocompromised Host; In Vitro; Individual; International; Intestines; Investigation; Laboratories; LoxP-flanked allele; Malignant Neoplasms; Mitogen-Activated Protein Kinase Kinases; Modeling; Mus; Mutant Strains Mice; Neoplasm Metastasis; Non-Small-Cell Lung Carcinoma; Oncogenes; Oncogenic; Pathway interactions; Patient risk; Phenotype; Press Releases; Preventive; Proteins; Publications; Receptor Protein-Tyrosine Kinases; Regulation; Reporting; Research Personnel; Resources; Role; Signal Pathway; Signal Transduction; Tamoxifen; Testing; Therapeutic; Time; Transfection; United States; base; cancer cell; carcinogenesis; career; epithelial to mesenchymal transition; evidence base; high risk; in vitro Model; in vivo; intestinal homeostasis; migration; mortality; mouse model; novel; postnatal; small molecule inhibitor; tumor; tumor progression; tumorigenesis; villin

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.

结直肠癌（CRC）是美国癌症相关死亡率的主要原因之一。在 我们最近的报道，我们证明了Sputy 2的肿瘤促进作用，Sputy 2是一种内源性抑制剂， 受体酪氨酸激酶（RTK）/丝裂原活化蛋白激酶（MAPK）信号通路在 结直肠癌（CRC）。为了确定Sprouty 2在癌症中的体内作用，我们最近产生了 Sprouty 2F/F小鼠。本RO 3应用程序的目标是表征 研究了胚胎期（Sprouty 2F/F-Villin Cre）和出生后（Sprouty 2F/F-Villin Cre）的作用 CreERT 2）Sprouty 2缺失对CRC进展的影响。我们进行这些研究的理由是， Sprouty 2基因在CRC中体内作用将允许我们提交机械RO 1应用。 我们实验室利用细胞培养模型进行的体外研究，原位和转移小鼠 模型证实Sprouty 2在CRC中的致癌潜力。在初步研究中，我们还 证实达特依赖性Sputy 2蛋白转导或Sputy 2稳定转染有利于 癌症表型。相反，Sprouty 2的抑制支持癌细胞中的上皮特征 和从Sprouty 2F/F-Villin CreERT 2小鼠获得的鼠胚胎成纤维细胞（MEF）。然而，在这方面， Sprouty 2在体内CRC进展中的需求尚未建立。胚胎和 Sputy 2F/F-Villin Cre（胚胎缺失）和Sputy 2F/F-Villin CreERT 2中的出生后Sputy 2缺失 小鼠（他莫昔芬依赖性出生后缺失）的研究将分别在AOM/DSS诱导期间进行 结肠癌发生我们还将研究Sprouty 2缺失对体内肠细胞的影响， 分化和谱系。 据我们所知，这是第一个证明体内缺失SPRY 2对CRC影响的研究 进展和肠细胞谱系。我们根据过去的经验建立了我们的假设， 最近的出版物。拟议的研究是一个多管齐下的方法，为我们准备的RO 1应用程序。