Ryer Evan J, Ronning Kaitryn E, Erdman Robert, Schworer Charles M, Elmore James R, Peeler Thomas C, Nevius Christopher D, Lillvis John H, Garvin Robert P, Franklin David P, Kuivaniemi Helena, Tromp Gerard
Department of Vascular and Endovascular Surgery, Geisinger Health System, Danville, PA 17822, USA.
Sigfried and Janet Weis Center for Research, Geisinger Health System, Danville, PA 17822, USA.
Int J Mol Sci. 2015 May 18;16(5):11259-75. doi: 10.3390/ijms160511259.
Abdominal aortic aneurysm (AAA) is a complex disorder that has a significant impact on the aging population. While both genetic and environmental risk factors have been implicated in AAA formation, the precise genetic markers involved and the factors influencing their expression remain an area of ongoing investigation. DNA methylation has been previously used to study gene silencing in other inflammatory disorders and since AAA has an extensive inflammatory component, we sought to examine the genome-wide DNA methylation profiles in mononuclear blood cells of AAA cases and matched non-AAA controls. To this end, we collected blood samples and isolated mononuclear cells for DNA and RNA extraction from four all male groups: AAA smokers (n = 11), AAA non-smokers (n = 9), control smokers (n = 10) and control non-smokers (n = 11). Methylation data were obtained using the Illumina 450k Human Methylation Bead Chip and analyzed using the R language and multiple Bioconductor packages. Principal component analysis and linear analysis of CpG island subsets identified four regions with significant differences in methylation with respect to AAA: kelch-like family member 35 (KLHL35), calponin 2 (CNN2), serpin peptidase inhibitor clade B (ovalbumin) member 9 (SERPINB9), and adenylate cyclase 10 pseudogene 1 (ADCY10P1). Follow-up studies included RT-PCR and immunostaining for CNN2 and SERPINB9. These findings are novel and suggest DNA methylation may play a role in AAA pathobiology.
腹主动脉瘤（AAA）是一种复杂的疾病，对老年人群有重大影响。虽然遗传和环境风险因素都与AAA的形成有关，但所涉及的精确遗传标记以及影响其表达的因素仍是一个正在进行研究的领域。DNA甲基化先前已被用于研究其他炎症性疾病中的基因沉默，由于AAA具有广泛的炎症成分，我们试图检查AAA病例和匹配的非AAA对照的单核血细胞中的全基因组DNA甲基化谱。为此，我们从四个全男性组中收集血样并分离单核细胞以提取DNA和RNA：AAA吸烟者（n = 11）、AAA非吸烟者（n = 9）、对照吸烟者（n = 10）和对照非吸烟者（n = 11）。使用Illumina 450k人类甲基化芯片获得甲基化数据，并使用R语言和多个生物导体软件包进行分析。对CpG岛子集进行主成分分析和线性分析，确定了与AAA在甲基化方面有显著差异的四个区域：kelch样家族成员35（KLHL35）、钙调蛋白2（CNN2）、丝氨酸蛋白酶抑制剂B族（卵清蛋白）成员9（SERPINB9）和腺苷酸环化酶10假基因1（ADCY10P1）。后续研究包括对CNN2和SERPINB9进行RT-PCR和免疫染色。这些发现是新颖的，表明DNA甲基化可能在AAA病理生物学中起作用。
