Novel Methodology for Quantitative High-throughput Cancer Epigenetics

描述(由申请人提供) 摘要：在没有基因序列改变的情况下发生的基因表达的可遗传变化被称为表观遗传，这种表观遗传信号的失调是人类一些生物学过程和疾病(如癌症)的基础。表观遗传机制的中心是关键的染色质调控网络，如DNA甲基化和组蛋白翻译后修饰(PTM)。组蛋白PTMS与基因激活和沉默都有关联，这取决于特定的单一修饰位点。然而，同时发生的组蛋白修饰(组蛋白密码)的不同组合对细胞事件的影响还知之甚少。这种缺乏知识的主要原因是不存在用于通过任何生物学手段对组合组蛋白编码进行定量表征甚至定性鉴定的健壮的高通量方法。我们的工作解决了这一不足，开发了一个新的基于质谱学的蛋白质组学平台，用于组合组蛋白密码信号的定量分子水平描述。我们的初步数据显示，数百种不同的组蛋白密码存在于人类基因组中，我们计划应用这一方法来发现动态组蛋白密码如何影响几种类型的癌症的基因表达。我们将努力实现这样的目标：在疾病状态下获取基因组的任何定义部分，并准确量化组蛋白密码，检测任何非组蛋白蛋白质，然后将这些蛋白质映射回特定的基因组位置，从而拍摄癌症进展过程中染色质景观的快照。目前人们认识到，癌症是一种既有遗传转化又有表观遗传转化的疾病。虽然遗传因素与几种类型的癌症有关，但与组蛋白修饰相关的表观遗传学方面仍然难以捉摸。我们相信，在这些过程中，在组蛋白编码水平上理解这些改变的组蛋白PTM机制可能会导致对人类健康有益的诊断或表观遗传疗法的发展。 公共卫生相关性：组蛋白修饰的异常变化与癌症等疾病有关，但目前在组合多位点水平上检测这些变化的技术严重受限。我们计划使用新技术来解决这个问题，以发现与几种类型癌症的疾病进展相关的组蛋白修饰模式，从而创建可能有助于疾病诊断或治疗的初步知识。

DESCRIPTION (Provided by the applicant) Abstract: Heritable changes in gene expression occurring without alterations in gene sequence are referred to as "epigenetic", and dysregulation of this epigenetic signaling underlies several human biological processes and diseases such as cancer. At the center of epigenetic mechanisms are critical chromatin regulatory networks such as DNA methylation and histone post-translational modifications (PTMs). Histone PTMs have been linked to both gene activation and silencing depending on the specific single modification site. Nevertheless, what type of effect distinct combinations of simultaneously occurring histone modifications (Histone Codes) have upon cellular events is poorly understood. The main reason for this lack of knowledge is that robust high- throughput methods for quantitative characterization or even qualitative identification of combinatorial Histone Codes by any biological means do not exist. Our work has addressed this deficiency by developing a novel mass spectrometry based proteomic platform for quantitative molecular level descriptions of combinatorial Histone Code signaling. Our preliminary data reveal that hundreds of distinct Histone Codes exist throughout the human genome, and we plan to apply this approach to discover how dynamic Histone Codes influence gene expression in several types of cancers. We will work towards the goal of taking any defined part of the genome during disease state and accurately quantifying the Histone Codes, detecting any non-histone proteins, and then mapping these back to the specific genomic locations, thereby taking a snapshot of the chromatin landscape during cancer progression. It is currently appreciated that cancer is a disease of both genetic and epigenetic transformations. While genetic causes have been associated with several types of cancers, epigenetic aspects related to histone modifications remain elusive. We believe that comprehension of these altered histone PTM mechanisms at the Histone Code level in these processes may lead to enhanced diagnostics or development of epigenetic therapy beneficial for human health. Public Health Relevance: Aberrant changes in histone modifications have been linked to diseases such as cancer, but currently the technology to detect these changes on a combinatorial multisite level is severely limited. We plan to use novel technology to address this problem to discover histone modification patterns associated with disease progression in several types of cancers, thereby creating initial knowledge that may be beneficial for disease diagnostics or treatment.