Nostalgia in the WNT signaling pathway; fatty acids, epigenetics and leukemia

描述（由申请人提供）：研究表明，基因调控的表观遗传改变允许癌前细胞存活，饮食影响表观遗传调控。通过了解影响基因表达的表观遗传调控的饮食因素，我们可能能够设计有效的癌症预防策略。这项探索性研究将调查的假设，ω 3/ω 6脂肪酸比例的操纵可以影响WNT通路信号通过表观遗传调节SFRP水平的髓性白血病。将使用两个实验模型。为了测定癌症进展、ω脂肪酸和SFRP甲基化之间的联系，将使用小鼠BCR-ABL慢性髓性白血病（CML）原始细胞危象模型。在第一个具体目标中，我们将确定高欧米伽3菜籽油与低欧米伽3玉米油对急性CML发病率的影响。这将与使用甲基化特异性PCR的五个SFRP基因的启动子甲基化状态相关联。还将通过流式细胞术和共聚焦显微镜在该动物模型中测定WNT途径的活性。在特定目标2中，造血干细胞分化的第二个体外模型将用于omega 3/ omega 6脂肪酸对分化和DNA甲基化的影响的更多操纵性研究。EML-克隆1细胞分化系统将用于表征ω 3和ω 6脂肪酸的比例对骨髓细胞分化为巨噬细胞和粒细胞谱系的影响。将采用流式细胞术进行免疫表型分析以跟踪细胞分化。本提案中的研究将在体内和体外系统中检验我们的中心假设，并为使用膳食脂肪酸的预防性治疗方法提供初步证据。 项目健康相关性：饮食因素会影响我们患癌症等疾病的风险，这一点越来越明显。该提案将在接触特定类型的脂肪酸（如鱼油中存在的脂肪酸）与预防癌症之间建立联系。已经提出了一种机制作为这种相互作用的基础，并将对其进行测试。

DESCRIPTION (provided by applicant): Research indicates that epigenetic alteration of gene regulation allows survival of premalignant cells and that diet influences epigenetic regulation. By understanding dietary factors that influence epigenetic regulation of gene expression, we may be able to design effective cancer preventive strategies. This exploratory study will investigate the hypothesis that manipulation of omega 3/omega 6 fatty acid ratios can affect WNT pathway signaling through epigenetic regulation of SFRP levels in myeloid leukemias. Two experimental models will be used. To assay the connection between cancer progression, omega fatty acids, and SFRP methylation a mouse BCR-ABL chronic myelogenous leukemia (CML) blast crisis model will be used. In the first Specific Aim, we will determine the effect of high omega 3 canola oil vs. low omega 3 corn oil on the incidence of blast crisis CML. This will be correlated with the promoter methylation status of five SFRP genes using methylation specific PCR. The activity of the WNT pathway will also be determined in this animal model through flow cytometry and confocal microscopy. In Specific Aim 2, a second in vitro model of hematopoietic stem cell differentiation will be used for more manipulative studies of the effect of omega 3/ omega 6 fatty acids on differentiation and DNA methylation. The EML-clone1 cell differentiation system will be used to characterize the effect of ratios of omega 3 and omega 6 fatty acids on myeloid cell differentiation into the macrophage and granulocyte lineages. Immunophenotyping with flow cytometry will be employed to follow cellular differentiation. The studies in this proposal will test our central hypothesis in both in vivo and in vitro systems and provide preliminary evidence for preventive therapeutic approaches using dietary fatty acids. PROJECT HEALTH RELEVANCE: It has become increasingly apparent that dietary factors can affect our risk to diseases such as cancer. This proposal will draw connections between exposure to specific types of fatty acids, such as those present in fish oils, to protection from cancer. A mechanism has been proposed for the basis of this interaction and will be tested.