DNA METHYLATION AND GENE EXPRESSION PROFILES IN MONOCYTES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 动脉粥样硬化性血管疾病仍然是世界范围内发病率和死亡率的主要原因之一。人们普遍认为，心血管疾病风险的个体差异是环境和遗传因素共同作用的结果。在确定具体环境因素方面取得了相当大的进展。相反，只有少数DNA序列变异已被确定与动脉粥样硬化性血管疾病的风险明显相关，这些变异的明显影响是适度的。表观遗传现象如DNA甲基化代表了另一类遗传因子，其可以被环境修饰，具有影响临床表型如动脉粥样硬化的能力。我们推测单核细胞DNA甲基化改变通过改变基因表达谱介导动脉粥样硬化形成。我们推测异常的DNA甲基化可能有助于动脉粥样硬化的发病机制。我们计划开发一种可靠分离单核细胞DNA和RNA的方案，以鉴定DNA甲基化和基因表达的变化。简言之，我们计划1）从常规抽血中分离外周血单核细胞（PBMC），2）从PBMC中分离单核细胞，3）从单核细胞中同时提取DNA和RNA，以及4）定量单核细胞的DNA甲基化和基因表达水平。这项试点研究的总体目标是生成初步数据，用于额外的NIH校外R 01应用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Atherosclerotic vascular disease remains one of the leading causes of morbidity and mortality worldwide. It is widely accepted that individual variation in the risk for cardiovascular disease results from the joint effects of both environmental and genetic factors. Considerable progress has been made in identifying specific environmental factors. In contrast, only a few DNA sequence variants have been identified that are clearly associated with risk for atherosclerotic vascular disease, and the apparent effects of these variants are modest. Epigenetic phenomenon such as DNA methylation represent another class of genetic factors that may be modified by the environment which have the ability to influence clinical phenotypes such as atherosclerosis. We hypothesize that alterations in DNA methylation profiles in monocytes play a causative role in atherogenesis, mediated through altering gene expression profiles. We postulate that aberrant DNA methylation may contribute to the pathogenesis of atherosclerosis. We plan to develop a protocol to reliably isolate monocyte DNA and RNA in order to identify variations of DNA methylation and gene expression. Briefly, we plan to 1) isolate Peripheral Blood Mononuclear Cell (PBMCs) from a routine blood draw, 2) isolate monocytes from the PBMCs, 3) extract DNA and RNA simultaneously from the monocytes, and 4) quantify DNA methylation and gene expression levels from monocytes. The overall goal of this pilot study is to generate preliminary data to be used for an additional NIH extramural R01 application.