DNA Methylase Covalent Complexes in Cancer

描述(申请人提供)：DNA甲基化是一种表观遗传加密系统，对生命至关重要。DNA甲基化在人类中的重要性被其错误调控的后果所突显，其中包括严重的遗传缺陷和癌症。这个过程很复杂，因为甲基化是由许多不同的酶执行的；然而，人们对它们在体内的作用知之甚少。提出了一种全球、全基因组的方法来了解所有DNA甲基转移酶的作用，使用了允许在体内任何给定细胞中的结合事件的快照视图的新技术。这种方法被称为体内甲基酶复合体(ICM)，将使研究人员能够在染色体环境中识别几乎所有催化活性的内源性甲基酶的基因和同源DNA结合位点。由于ICM检测是以抗体为基础的，所以可以很容易地区分甲基转移酶亚型。这种方法是高度量化和易于处理的。基于DNA甲基化反应机制的现有生物化学和分子生物学工具，提出了一系列实验，以检验甲基酶的微妙之处：活细胞中的基因组相互作用。鉴于甲基化作为分子靶标的强烈兴趣，这些实验也与使用DNA低甲基化试剂开发有效的抗癌治疗有关。最后，ICM方法代表了一种明确定义的新的生物相关性，这可能会在DNA低甲基化药物的临床试验中带来显著的好处。

DESCRIPTION (provided by applicant): DNA methylation is an epigenetic encryption system that is essential for life. The importance of DNA methylation in man is underscored by the consequences of its misregulation, which include and serious genetics defects and cancer. The process is complicated by the fact that methylation is carried out by a number of different enzymes; however their in vivo roles are poorly understood. A global, genome wide approach to understanding the action of all DNA methyltransferases is proposed using novel technology that allows a snapshot view of the binding event in any given cell in vivo. This method, called the In vivo Complex of Methylase (ICM), will allow empower researchers with the ability to identify genes and cognate DNA binding sites for virtually all catalytically active, endogenous methylases in a chromosomal setting. Because the ICM assay is antibody based, one can readily differentiate methyltransferase isoforms. The approach is highly quantitative and tractable. A series of experiments are proposed, based on current tools of biochemistry and molecular biology of DNA methylation reaction mechanism, to examine subtleties of methylase:genome interactions in living cells. Given the acute interest of methylation as a molecular target, these experiments are also relevant to the development of effective anti-cancer therapy using DNA hypomethylating agents. Finally, the ICM method represents a clearly defined and novel biological correlate that could be a significant benefit in clinical trials with DNA hypomethylating drugs.