Epigenomics of Atherosclerosis

描述（由申请人提供）：随着我们对基因调控的复杂机制的理解的增加，很明显，识别与复杂疾病相关的异常遗传机制的传统方法，如连锁和关联研究，将是不够的。此外，需要改变导致疾病的变异必须存在于基因编码区域内的传统范式。更好地建模疾病风险和理解疾病变异的一步在于扩大复杂疾病研究的范式，包括导致疾病的表观遗传影响。我们假设内皮细胞（ECs）和平滑肌细胞（SMCs）中基因DNA甲基化状态的变化可能在动脉粥样硬化的发生和进展中发挥作用。本提案特别寻求在(1)疾病和(2)非疾病状态下的EC和SMC中创建胞嘧啶甲基化基因组的综合图谱，使用一种新的和经过验证的人类替代血管共培养模型，该模型可以将EC和SMC表型重新校准为人体外的健康或动脉粥样硬化表型，以及(3)患病和非患病的人类主动脉组织。我们处理三种范式来演示如何评估健康细胞和组织、疾病细胞和组织以及早期（流动表型）和晚期（脂质和斑块组织）动脉粥样硬化中的胞嘧啶甲基化变化。此外，将利用这些数据开发高通量测序方法和工具，并提供给所有科学家。公共卫生相关性：越来越清楚的是，遗传和表观遗传变化的疾病模型无疑将解释大多数复杂的疾病表型。这里提出的实验的短期影响将是揭示另一层潜在的遗传基因调控，这将使我们更接近了解早期动脉粥样硬化，并可能为我们提供干预的候选途径。我们的长期影响将来自于我们识别的正常和疾病标记的解剖，以更好地了解细胞及其在非疾病和病变（动脉粥样硬化）状态之间的平衡。

DESCRIPTION (provided by applicant): As our understanding of the intricate mechanisms of gene regulation increases, it is apparent that traditional methods of identifying aberrant genetic mechanisms associated with complex disease, such as linkage and association studies, will not be sufficient. Additionally, the traditional paradigm that variants that lead to disease must exist within the coding region of a gene needs to be changed. One step in better modeling of disease risk and understanding disease variants lies in expanding the paradigm of complex disease study to include epigenetic influences that contribute to diseases. We hypothesize that changes in DNA methylation status of genes in endothelial cells (ECs) and smooth muscle cells (SMCs) undergoing phenotypic switching, a hallmark of atherosclerosis formation, could play a role in atherosclerosis initiation and progression. This proposal specifically seeks to create comprehensive maps of the cytosine methylated genome in (1) ECs and SMCs under a disease and (2) non-disease state influenced using a novel and validated human surrogate vascular co-culture model that can recalibrate the EC and SMC phenotype into a healthy or atheroprone phenotype outside the human body and (3) diseased and non-diseased human aorta tissue. We tackle three paradigms to demonstrate how to evaluate cytosine methylation changes, in healthy cells and tissues, in disease cells and tissues, and in early (flow phenotype) and late (lipid and plaque laden tissue) atherosclerosis. In addition, highthroughput sequencing methods and tools will be developed using these data and be made available to all scientists. Public Health Relevance: It is becoming clear that disease models of genetic and epigenetic changes will no doubt account for a majority of the complex disease phenotype. The short range impact of the experiments proposed here will be to uncover another entire layer of potentially heritable gene regulation that will bring us closer to understanding early atherosclerosis and may provide us with candidate pathways for intervention. Our long range impacts will come from the dissection of the normal and disease marks we identify to better understand the cell and its balance between a non-disease and diseased (atherosclerosis) state.