Article Figures & Data
Figures
- Figure 1.
Human NR3C1 gene and the CpG region analyzed by bisulfite pyrosequencing. The 5′ region of the NR3C1 gene contains multiple first exons, corresponding to multiple transcriptional start sites and multiple mRNA splice variants. The region analyzed by pyrosequencing is illustrated by enlarged box, below the exon 1F. Numbering is relative to the alternative transcription start site in exon 2. The CpG3 and CpG4 sites are encompassed by the potential NGFI-A binding site. Adapted from Oberlander et al. (2008).
- Figure 2.
DNA methylation of NR3C1 promoter and PTSD symptoms in male genocide survivors. Association (Spearman's rank correlation, −log10p) of DNA methylation marks across eight CpG positions in the human NR3C1 gene promoter with PTSD symptoms clusters intrusions, avoidance, and hyperarousal. The association between CpG3 and intrusions reached Bonferroni-corrected statistical significance (after correcting for 8 tests; PBonferroni = 0.008). The CpGs are aligned by genomic position. The position of the potential NGFI-A binding site is shaded in purple. The horizontal dashed line indicates the p < 0.05 Bonferroni-corrected significance threshold.
- Figure 3.
Fitted values of probability for lifetime PTSD risk against DNA methylation at the NGFI-A binding site of the NR3C1 promoter for males and females. Lifetime PTSD risk was assessed against NR3C1 CpG3 DNA methylation and the number of lifetime traumatic event types via binary logistic regression (nmales = 83, pnominal_males = 0.008, βmales = −0.519; nfemales = 69, pnominal_females = 0.243, βfemales = −0.170). This graph also contains the raw CpG methylation values obtained in participants without (bottom line) and with (top line) PTSD.
- Figure 4.
Methylation-dependent differences in brain activity related to successful recognition of previously seen pictures in healthy men. Displayed are voxels with a positive correlation between methylation values (at NR3C1_CpG3) and activity, using color-coded t values. The blue circles show the activation in the pars triangularis and pars orbitalis of the inferior frontal gyrus (centered at 44, 25, −4; peak pFWE corrected < 0.05; displayed at puncorrected < 0.001; a random-effects, linear-regression model). Activations are overlaid on coronal (upper left), sagittal (upper right), and axial sections of the study-specific group template (see main text). L, Left side of the brain; R, right side of the brain.
Tables
- Table 1.
Association of PTSD symptom clusters and DNA methylation of NR3C1 gene promoter in Rwandan mena
PTSD symptom clusters (nmales = 83) NR3C1_CpG1 NR3C1_CpG2 NR3C1_CpG3 NR3C1_CpG4 NR3C1_CpG5 NR3C1_CpG6 NR3C1_CpG7 NR3C1 _CpG8 Intrusionsb Spearman's ρ −0.235 −0.039 −0.355 −0.128 −0.203 −0.059 −0.118 −0.18 p value Nominal 0.037* 0.733 0.001*** 0.261 0.073 0.606 0.3 0.112 Bonferroni correction for 8 CpGs 0.296 1 0.008 1 0.584 1 1 0.896 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 0.048 1 1 1 1 1 Avoidanceb Spearman's ρ −0.115 −0.033 −0.260 −0.033 −0.101 −0.049 −0.019 −0.074 p value Nominal 0.315 0.772 0.021* 0.77 0.376 0.666 0.87 0.517 Bonferroni correction for 8 CpGs 1 1 0.168 1 1 1 1 1 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 1 1 1 1 1 1 Hyperarousalb Spearman's ρ −0.187 −0.097 −0.346 −0.038 −0.165 −0.074 −0.08 −0.053 p value Nominal 0.099 0.396 0.009** 0.74 0.145 0.514 0.483 0.645 Bonferroni correction for 8 CpGs 0.792 1 0.072 1 1 1 1 1 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 0.432 1 1 1 1 1 PDS scoreb,c Spearman's ρ −0.207 −0.056 −0.355 −0.081 −0.172 −0.036 −0.078 −0.112 p value Nominal 0.067 0.627 0.001*** 0.476 0.131 0.756 0.496 0.324 Bonferroni correction for 8 CpGs 0.536 1 0.008 1 1 1 1 1 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 0.048 1 1 1 1 1 NR3C1 DNA methylation Mean 2.86 1.52 2.84 1.01 1.77 3.52 4.88 1.83 SD 1.32 1.55 1.56 1.45 1.58 1.81 1.82 1.56 ↵aCorrelation of NR3C1 gene promoter DNA methylation with sum of PTSD-specific symptom subscores according to PDS and DSM-IV. To account for trauma load, PTSD symptom cluster scores were divided by the sum of lifetime traumatic event types. Spearman's correlation ρs coefficients and the corresponding nominal and Bonferroni-corrected p values are shown for each symptom cluster.
↵bSum of specific symptom clusters corrected for total number of lifetime traumatic events.
↵cPDS score is not an additional symptom measure, as it represents the sum of the subscores of Intrusions, Avoidance, and Hyperarousal.
↵***Correlation is nominally significant at the 0.001 level (2-tailed, Spearman's ρ).
↵**Correlation is nominally significant at the 0.01 level (2-tailed, Spearman's ρ).
↵*Correlation is nominally significant at the 0.05 level (2-tailed, Spearman's ρ).
- Table 2.
Association of PTSD symptom clusters and DNA methylation of NR3C1 gene promoter in Rwandan womena
PTSD symptom clusters (nfemales = 69) NR3C1_CpG1 NR3C1_CpG2 NR3C1_CpG3 NR3C1_CpG4 NR3C1_CpG5 NR3C1_CpG6 NR3C1_CpG7 NR3C1_CpG8 Intrusionsb Spearman's ρ −0.166 0.013 −0.073 0.037 0.065 0.033 −0.071 0.019 p value Nominal 0.173 0.916 0.553 0.763 0.598 0.785 0.561 0.876 Bonferroni correction for 8 CpGs 1 1 1 1 1 1 1 1 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 1 1 1 1 1 1 Avoidanceb Spearman's ρ −0.116 −0.118 −0.139 0.006 −0.059 −0.058 −0.045 0.092 p value P-value (nominal) 0.344 0.333 0.256 0.963 0.633 0.635 0.714 0.45 Bonferroni correction for 8 CpGs 1 1 1 1 1 1 1 1 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 1 1 1 1 1 1 Hyperarousalb Spearman's ρ −0.003 −0.014 −0.052 0.053 0.002 −0.04 −0.057 −0.033 p value Nominal 0.983 0.912 0.672 0.667 0.99 0.746 0.642 0.79 Bonferroni correction for 8 CpGs 1 1 1 1 1 1 1 1 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 1 1 1 1 1 1 PDS scoreb,c Spearman's ρ −0.114 −0.046 −0.089 0.011 0.003 −0.006 −0.063 0.054 p value Nominal 0.352 0.706 0.469 0.928 0.982 0.959 0.605 0.657 Bonferroni correction for 8 CpGs 1 1 1 1 1 1 1 1 Bonferroni correction for 8 CpGs, sex, and 3 PTSD symptom clusters 1 1 1 1 1 1 1 1 NR3C1 DNA methylation Mean 3.23 1.90 3.55 1.10 2.42 4.16 5.43 2.29 SD 1.62 1.81 1.85 1.59 1.83 2.32 1.96 1.90 ↵aCorrelation of NR3C1 gene promoter DNA methylation with sum of PTSD-specific symptom subscores according to PDS and DSM-IV. To account for trauma load, PTSD symptom cluster scores were divided by the sum of lifetime traumatic event types. Spearman's correlation ρ coefficients and the corresponding nominal and Bonferroni-corrected p values are shown for each symptom cluster.
↵bSum of specific symptom clusters corrected for total number of traumatic events in a lifetime.
↵cPDS score is not an additional symptom measure, as it represents the sum of the subscores of Intrusions, Avoidance and Hyperarousal.
NR3C1 CpG3 Women (nfemales = 47) Men (nmales = 25) Recognition of all pictures β 0.065 −0.455 p value 0.667 0.012* Arousal for positive pictures β −0.148 0.343 p value 0.320 0.094 Arousal for negative pictures β −0.009 0.431 p value 0.954 0.031* Arousal for neutral pictures β −0.135 0.266 p value 0.366 0.199 Recognition of positive pictures corrected for arousal β −0.036 −0.447 p value 0.817 0.041* Recognition of negative pictures corrected for arousal β 0.289 −0.481 p value 0.050* 0.010** Recognition of neutral pictures corrected for arousal β −0.014 −0.643 p value 0.927 0.002** ↵aRecognition corrected for arousal. To account for methylation-dependent differences in arousal during recognition of previously seen pictures, both arousal and NR3C1 CpG3 DNA methylation were included in the linear model. We modeled valence-specific recognition performance as dependent on NR3C1 DNA methylation and valence-specific arousal. β Coefficients and the corresponding p values are shown for each recognition and arousal category (DNA methylation mean ± SD: μNR3C1 CpG3 Females = 3.20 ± 1.29, μNR3C1 CpG3 Males = 3.28 ± 1.21).
↵**Correlation is significant at the 0.01 level.
↵*Correlation is significant at the 0.05 level.
- Table 4.
Methylation-dependent differences in brain activity related to the successful recognition of previously seen pictures in healthy male individualsa
Maximum t value within cluster Regional correspondence of the maximum MNI coordinates at maximum X Y Z 6.4174 Cortex-right hemisphere-pars orbitalis (36%), cortex-right hemisphere-pars triangularis (31%) of the inferior frontal gyrus 44 24.75 −4 6.5307 Near superior temporal cortex 44 −16.5 −16 6.3278 Cortex-left hemisphere-cuneus (48%), cortex-left hemisphere-pericalcarine (10%) 0 −85.25 12 6.4978 Near superior frontal cortex 19.25 38.5 32 ↵apFWE whole-brain corrected < 0.05. Swiss sample, nmales = 24. Regions and probabilities in accordance to the in-house atlas (see Materials and Methods).
