Molecular Dissection of Colorectal CanceR: Effects of Loss of Imprinting of IGF2

描述(由申请者提供)：该项目的总体目标是确定生长因子IGF2的印迹丢失(LOI)如何影响结直肠癌(CRC)的发展。印记缺失指的是一种表观遗传现象，即正常的单等位基因(仅从父系或母系拷贝表达)变为双等位表达。在LOI的情况下，这发生在5%-10%的美国人口中，这被发现使患CRC的概率增加了5倍。具体目标1：利用免疫荧光和荧光蛋白融合以收集高含量数据，确定LOI影响(IGF2)和结直肠癌控制(Wnt/3-catenin)通路之间的相互作用，以从我们的模型小鼠和结直肠癌细胞系来源的成纤维细胞上获得数据。具体目标2：进一步描述我们的模型鼠之间的差异，特别关注发生在结肠隐窝的差异，以确定生长、分化和凋亡平衡的变化；以及在血液中，确定更容易筛查LOI的可能靶点。具体目标3：确定LOI是否改变了CRC获得的突变类型，方法是对储存的样本进行LOI分型，然后进行分析，以确定它们在各种把关基因中的突变类型和位置。更好地了解普通人群对结直肠癌可能具有的机制和易感性是至关重要的。这不仅有助于开发新的筛查方法，还为开发针对个人患有的确切癌症类型的量身定制的个性化疗法提供了可能性，事实证明，这应该比广谱化疗更有效。

DESCRIPTION (provided by applicant): The overall goal of the project is to determine how loss of imprinting (LOI) of IGF2, a growth factor, affects the development of colorectal cancer (CRC). Loss of imprinting refers to the epigenetic phenomenon where a normally monoallelically expressed gene (expressed from paternal or maternal copy only) becomes biallelically expressed. In the case of LOI, which occurs in 5-10% of the US population, this has been found to increase the probability of developing CRC as much as 5 times. Specific Aim 1: Determine the interaction between LOI affected (IGF2) and CRC controlling (Wnt/3-catenin) pathways using immunofluorescence and fluorescent protein fusions to gather high content data, on fibroblasts derived from our model mice and CRC cell lines. Specific Aim 2: Further characterize the differences between our model mice, paying special attention to differences occurring in the colonic crypts, to determine the changes in the balance of growth, differentiation and apoptosis; and in blood to determine possible targets for easier screening of LOI. Specific Aim 3: Determine if LOI shifts the type of mutations acquired by CRC by typing banked samples for LOI, then assaying to determine the type and location of their mutations in various gatekeeper genes. A greater understanding of the mechanisms and predispositions that the general population may have towards colorectal cancer is essential. Not only does this aid in developing new methods for screening, it gives the potential to develop tailored, personal therapies to the exact kind of cancer that an individual has, which should prove more effective than broad-spectrum chemotherapies.