A large animal model to elucidate the role of LGR5 stem cells in cancer

项目摘要 在美国，结直肠癌（CRC）是女性癌症死亡的第三大原因， 每年，每20人中就有1人被诊断出患有CRC。然而，理解因素 控制疾病进展和转移由于缺乏合适的动物模型而受到阻碍。 《儿童权利公约》。目前，有相当多的关于在CRC中突变的候选基因的信息， 有助于肿瘤位置、肿瘤表型和患者对治疗的反应1。虽然候选基因 可以通过使用携带功能获得和功能丧失遗传的转基因系在小鼠中进行研究 修饰，候选基因如Apc的突变，导致小鼠的病变与那些似乎不同， 在人类身上。因此，需要对传统的转基因模型进行修改，以推进我们的研究。 理解CRC。相比之下，类似的APC-/-猪模型主要显示结肠肿瘤， 与人类疾病非常相似。 我们建议将现有基因编辑的LGR 5-H2 B-GFP猪系的优势与基因编辑的能力联合收割机结合。 Cre诱导的Cas9系统产生新的细胞系，其将允许在体内和体外仔细解剖 与结肠癌中LGR 5细胞行为相关的分子途径。该提案将允许 用于模型的生成和初始体外和体内验证。 这种高度相关的大型动物模型将极大地提高我们对LGR 5细胞行为的理解， 在致癌转化过程中在分子水平上控制，将对开发新的 治疗人类结肠直肠癌以及其他癌症的策略。

Project Summary In the United States, colorectal cancer (CRC) is the 3rd leading cause of cancer death among women and men combined, with of 1 in 20 people being diagnosed with CRC each year. Yet, understanding factors controlling disease progression and metastasis has been hampered by the lack of a suitable animal model of CRC. At present, there is considerable information regarding candidate genes that are mutated in CRC and contribute to tumor locations, tumor phenotype and patient response to treatment1. While candidate genes can be studied in mice by use of transgenic lines carrying gain-of-function and loss-of function genetic modifications, mutations in candidate genes such as Apc, result in lesions in mice that differ from those seem in humans. As a result, modifications of the traditional transgenic model have been required to advance our understanding of CRC. In contrast, an analogous APC-/- pig model demonstrates predominantly colon tumors, which are remarkably similar to the human disease. We propose to combine the strengths of an existing gene edited LGR5-H2B-GFP pig line with the power of a Cre-inducible Cas9 system to generate a novel line that will allow a careful dissection both in vivo and in vitro of the molecular pathways associated with the behavior of LGR5 cells in colon cancer. This proposal will allow for the generation and the initial in vitro and in vivo validation of the model. This highly relevant large animal model will greatly improve our understanding of how LGR5 cells behavior is controlled at the molecular level during oncogenic transformation and will be of great use to develop novel strategies to treat human colorectal cancer, as well as other cancers.