Table 2.

Associations between HSD11β2/NR3C1 methylation and cardiovascular diseases.

Authors	Study design	Gene	Method	Tissue type	Exposure/Outcome	Sample	Results
Friso et al. (2008)	Pro-spective	HSD11β2	Methylation-specific PCR	Peripheral blood (PBMC)	Glucocorticoid treatment/hypertension	N = 57 Italian subjects. 32 glucocorticoid-treated patients normotensive at baseline (mean age: 54.56 years; 4 men). 25 essential hypertensive patients (EH) (mean age: 51.92 years; 14 men)	Glucocorticoid-treated patients who developed hypertension had: •Greater HSD11β2 promoter methylation •Higher urinary THFs/THE ratio, which indicates lower HSD11β2 protein activity
Pizzolo et al. (2015)	Cross-sectional	HSD11β2	Pyrosequencing	Peripheral blood (PBMC)	AME/hypertension	N = 12 members of the same Italian family. 2 proband brothers (7 and 13 years old) with apparent mineralocorticoid excess (AME; A221G missense mutation) and 10 relatives (age range: 46–71 years), 6 of whom had the A221G mutation	Among the 6 family members heterozygous for 221AG: •a higher methylation index was documented in the 4 hypertensive subjects compared to the 2 normotensive subjects •Statistical analysis was not permitted due to the small sample size
Zhao et al. (2015)	Cross-sectional	NR3C1	Pyrosequencing	Peripheral blood (leukocytes)	Combat-related PTSD/Subclinical atherosclerosis	N = 168 US veterans (84 monozygotic [MZ] twin pairs) (mean age: 55.1 years). The majority of twins were Caucasian (94%) and free of history of cardiovascular disease at enrollment	•Except for LDL, there was no significant association between NR3C1 1F promoter methylation and standard coronary risk factors (e.g., triglycerides, blood pressure) •Intra-pair difference in methylation was significantly and positively associated with intra-pair difference in flow-mediated dilation (FMD; determined using bi-mode ultrasound) at 50% (12/22) of studied CpG sites, and with mean DNA methylation across all studied CpG sites in the 1F promoter •On average, 1% increase in the intra-pair difference in mean DNA methylation was associated with 2.83% increase in the intra-pair difference in FMD •Methylation variation at each individual CpG site only conferred a small risk to disease, but the authors identified three methylation blocks (− 3365 to − 3371, − 3418 to − 3445, and − 3456 to − 3470 bp from ATG) that were significantly associated with FMD in their sample •FMD-related methylation changes may therefore be systematically organized, rather than randomly distributed