Sasaki Aya, Kim Bona, Murphy Kellie E, Matthews Stephen G
Department of Physiology, University of Toronto, Toronto, ON, Canada.
Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON, Canada.
Front Genet. 2020 Mar 24;11:224. doi: 10.3389/fgene.2020.00224. eCollection 2020.
The profiling of DNA methylation modifications in peripheral blood has significant potential to determine risk factors for human disease. Little is known concerning the sensitivity of DNA methylation profiles to sample handling. Here, we studied typical conditions prior to sample storage associated with cord blood samples obtained from clinical investigations using reduced representation bisulfite sequencing. We examined both whole blood collected shortly after birth and dried blood spots, a potentially important source of neonatal blood for investigation of the DNA methylome and the Developmental Origins of Health and Disease in human cohorts because they are routinely collected during clinical care. Samples were matched across different time conditions, as they were from the same cord blood samples obtained from the same individuals. Maintaining whole blood up to 24 h (4°C) or dried blood spots up to 7 days (room temp.) had little effect on DNA methylation profiles. Minimal differences were detected between cord blood immediately frozen and dried blood spots. Our results indicate that DNA methylation profiles are resilient to sample handling conditions prior to storage. These data will help guide future human studies focused toward determination of DNA methylation modifications in whole blood.
对外周血中DNA甲基化修饰进行分析，在确定人类疾病风险因素方面具有巨大潜力。关于DNA甲基化谱对样本处理的敏感性，目前所知甚少。在这里，我们使用简化代表性亚硫酸氢盐测序技术，研究了临床研究中获得的脐带血样本在储存前的典型条件。我们检查了出生后不久采集的全血和干血斑，干血斑是新生儿血液的一个潜在重要来源，可用于研究人类队列中的DNA甲基化组以及健康与疾病的发育起源，因为它们是在临床护理过程中常规采集的。样本在不同时间条件下进行了匹配，因为它们来自同一脐带血样本，而这些样本又来自同一个体。全血保存长达24小时（4°C）或干血斑保存长达7天（室温）对DNA甲基化谱影响很小。立即冷冻的脐带血和干血斑之间检测到的差异极小。我们的结果表明，DNA甲基化谱对储存前的样本处理条件具有弹性。这些数据将有助于指导未来专注于确定全血中DNA甲基化修饰的人体研究。
