Department of Psychiatry, Social Psychiatry and Psychotherapy, Hannover Medical School, Carl-Neuberg-Str. 1, Hannover, 30625, Germany.
Laboratory for Molecular Neuroscience, Feodor-Lynen-Str. 35, Hannover, 30625, Germany.
BMC Med Genomics. 2024 Nov 29;17(1):279. doi: 10.1186/s12920-024-02053-9.
MAGEL2 is an autism susceptibility gene whose deficiency has been associated with autism-related behaviors in animal models and in syndromic human autism spectrum disorders (ASDs) such as Schaaf-Yang syndrome, but has not been studied in the broader autism spectrum. Given the capabilities of long-read sequencing technologies, this pilot study used a targeted nanopore sequencing approach to simultaneously examine MAGEL2 DNA sequence and methylation in adults with high-functioning autism (HFA) compared to neurotypical controls (NC).
Using DNA extracted from peripheral blood, Cas9-targeted nanopore DNA sequencing was used to analyze MAGEL2, including its entire regulatory construct (chr15:23639316-23651466), for sequence variation and 5-methyl-cytosine (5mC) modification in a cohort of adults with HFA compared to sex- and age-matched NC. Given the known sex differences in ASD and MAGEL2 KO animal models, results were further analyzed by sex.
20 adults with HFA (10 males, 10 females) and 20 NC were included. While there were no overall differences in MAGEL2 DNA sequence and 5mC modification between HFA and NC, we found a significant difference in MAGEL2 gene promoter methylation between males and females with HFA and NC of both sexes, with HFA males tending to show hypomethylation in a 300 bp long differentially methylated region (chr15:23647640-23647939) around the MAGEL2 transcription start site.
In this pilot study utilizing nanopore Cas9 targeted DNA sequencing, significant sex-specific differences in MAGEL2 gene promoter methylation were identified in male adults with HFA in comparison to control groups, suggesting the potential for sex-specific epigenetic differences. However, further replication in larger cohorts is required to validate these findings.
MAGEL2 是一个自闭症易感基因，其缺陷与动物模型中的自闭症相关行为以及综合征性人类自闭症谱系障碍（ASD）有关，如 Schaaf-Yang 综合征，但在更广泛的自闭症谱系中尚未研究过。鉴于长读测序技术的能力，这项初步研究使用靶向纳米孔测序方法，比较了高功能自闭症（HFA）成人与神经典型对照（NC）之间 MAGEL2 的 DNA 序列和甲基化。
使用从外周血中提取的 DNA，使用 Cas9 靶向纳米孔 DNA 测序分析 MAGEL2，包括其整个调控结构（chr15:23639316-23651466），以分析与性别和年龄匹配的 NC 相比，HFA 成人的序列变异和 5-甲基胞嘧啶（5mC）修饰。鉴于 ASD 和 MAGEL2 KO 动物模型中的已知性别差异，进一步按性别分析结果。
纳入了 20 名 HFA 成人（10 名男性，10 名女性）和 20 名 NC。尽管 HFA 和 NC 之间的 MAGEL2 DNA 序列和 5mC 修饰没有总体差异，但我们发现 HFA 和 NC 中男性和女性的 MAGEL2 基因启动子甲基化存在显着差异，HFA 男性在 MAGEL2 转录起始位点周围的一个 300bp 长的差异甲基化区域（chr15:23647640-23647939）中表现出低甲基化。
在这项利用纳米孔 Cas9 靶向 DNA 测序的初步研究中，与对照组相比，HFA 男性成人的 MAGEL2 基因启动子甲基化存在显著的性别特异性差异，表明存在潜在的性别特异性表观遗传差异。但是，需要在更大的队列中进行进一步复制以验证这些发现。
