Institute of Legal Medicine, University Hospital Düsseldorf, 40225, Düsseldorf, Germany.
Department of General Paediatrics, University Hospital Düsseldorf, 40225, Düsseldorf, Germany.
Int J Legal Med. 2022 Jul;136(4):987-996. doi: 10.1007/s00414-022-02826-w. Epub 2022 May 12.
Age estimation based on DNA methylation (DNAm) can be applied to children, adolescents and adults, but many CG dinucleotides (CpGs) exhibit different kinetics of age-associated DNAm across these age ranges. Furthermore, it is still unclear how growth disorders impact epigenetic age predictions, and this may be particularly relevant for a forensic application. In this study, we analyzed buccal mucosa samples from 95 healthy children and 104 children with different growth disorders. DNAm was analysed by pyrosequencing for 22 CpGs in the genes PDE4C, ELOVL2, RPA2, EDARADD and DDO. The relationship between DNAm and age in healthy children was tested by Spearman's rank correlation. Differences in DNAm between the groups "healthy children" and the (sub-)groups of children with growth disorders were tested by ANCOVA. Models for age estimation were trained (1) based on the data from 11 CpGs with a close correlation between DNAm and age (R ≥ 0.75) and (2) on five CpGs that also did not present significant differences in DNAm between healthy and diseased children. Statistical analysis revealed significant differences between the healthy group and the group with growth disorders (11 CpGs), the subgroup with a short stature (12 CpGs) and the non-short stature subgroup (three CpGs). The results are in line with the assumption of an epigenetic regulation of height-influencing genes. Age predictors trained on 11 CpGs with high correlations between DNAm and age revealed higher mean absolute errors (MAEs) in the group of growth disorders (mean MAE 2.21 years versus MAE 1.79 in the healthy group) as well as in the short stature (sub-)groups; furthermore, there was a clear tendency for overestimation of ages in all growth disorder groups (mean age deviations: total growth disorder group 1.85 years, short stature group 1.99 years). Age estimates on samples from children with growth disorders were more precise when using a model containing only the five CpGs that did not present significant differences in DNAm between healthy and diseased children (mean age deviations: total growth disorder group 1.45 years, short stature group 1.66 years). The results suggest that CpGs in genes involved in processes relevant for growth and development should be avoided in age prediction models for children since they may be sensitive for alterations in the DNAm pattern in cases of growth disorders.
基于 DNA 甲基化(DNAm)的年龄估计可应用于儿童、青少年和成年人,但许多 CG 二核苷酸(CpG)在这些年龄范围内表现出与年龄相关的 DNAm 不同的动力学。此外,目前尚不清楚生长障碍如何影响表观遗传年龄预测,这对于法医学应用可能尤为重要。在这项研究中,我们分析了 95 名健康儿童和 104 名患有不同生长障碍的儿童的口腔颊黏膜样本。通过焦磷酸测序分析了 PDE4C、ELOVL2、RPA2、EDARADD 和 DDO 基因中 22 个 CpG 的 DNAm。通过 Spearman 等级相关检验了健康儿童中 DNAm 与年龄的关系。通过 ANCOVA 检验了“健康儿童”组和生长障碍(亚)组之间的 DNAm 差异。(1)基于与 DNAm 和年龄密切相关的 11 个 CpG(R≥0.75)的数据和(2)基于在健康和患病儿童之间 DNAm 没有显著差异的 5 个 CpG 建立年龄估计模型。统计分析显示,健康组与生长障碍组(11 个 CpG)、身材矮小组(12 个 CpG)和非身材矮小组(3 个 CpG)之间存在显著差异。结果与身高相关基因的表观遗传调控假设一致。在与 DNAm 和年龄相关性高的 11 个 CpG 上训练的年龄预测器在生长障碍组中显示出更高的平均绝对误差(MAE)(生长障碍组的平均 MAE 为 2.21 岁,健康组的 MAE 为 1.79 岁),以及身材矮小(亚)组;此外,所有生长障碍组都存在年龄高估的明显趋势(平均年龄偏差:总生长障碍组 1.85 岁,身材矮小组 1.99 岁)。在使用仅包含在健康和患病儿童之间 DNAm 无显著差异的 5 个 CpG 的模型时,对生长障碍儿童样本的年龄估计更精确(总生长障碍组的平均年龄偏差为 1.45 岁,身材矮小组的平均年龄偏差为 1.66 岁)。结果表明,在生长障碍病例中,DNAm 模式发生改变时,与生长和发育相关的基因中的 CpG 可能会变得敏感,因此在儿童的年龄预测模型中应避免使用这些 CpG。