Epigenomics of Atherosclerosis

摘要 随着我们对基因调控复杂机制的了解加深，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，传统的范式，即导致疾病的变异必须存在于基因的编码区，需要改变。更好地模拟疾病风险和了解疾病变异的一个步骤是扩大复杂疾病研究的范式，以包括导致疾病的表观遗传影响。我们推测，发生表型转换的内皮细胞(ECs)和平滑肌细胞(SMC)中基因甲基化状态的变化可能在动脉粥样硬化的启动过程中起作用。 和进步。这一建议特别寻求创建在(1)疾病和(2)非疾病状态下的内皮细胞和平滑肌细胞中胞嘧啶甲基化基因组的全面图谱，该图谱使用一种新的和验证的人类替代血管共培养模型，该模型可以将EC和SMC的表型重新校准为健康的或体外的动脉粥样硬化表型，以及(3)患病和未患病的人类主动脉组织。我们解决了三个范例来演示如何评估胞嘧啶甲基化变化，在健康细胞和组织中，在疾病细胞和组织中，以及在早期(流动表型)和晚期(脂质和斑块组织)动脉粥样硬化中。此外，还将使用这些数据和工具开发高通量测序方法和工具 向所有科学家提供。

Abstract 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.