Linnertz Colton, Anderson Lauren, Gottschalk William, Crenshaw Donna, Lutz Michael W, Allen Jawara, Saith Sunita, Mihovilovic Mirta, Burke James R, Welsh-Bohmer Kathleen A, Roses Allen D, Chiba-Falek Ornit
Division of Neurology at the Department of Medicine, Duke University Medical Center, Durham, NC, USA.
Division of Neurology at the Department of Medicine, Duke University Medical Center, Durham, NC, USA; Division of Neurology, Joseph and Kathleen Bryan Alzheimer's Disease Research Center, Duke University Medical Center, Durham, NC, USA.
Alzheimers Dement. 2014 Sep;10(5):541-51. doi: 10.1016/j.jalz.2013.08.280. Epub 2014 Jan 15.
We investigated the genomic region spanning the Translocase of the Outer Mitochondrial Membrane 40-kD (TOMM40) and Apolipoprotein E (APOE) genes, that has been associated with the risk and age of onset of late-onset Alzheimer's disease (LOAD) to determine whether a highly polymorphic, intronic poly-T within this region (rs10524523; hereafter, 523) affects expression of the APOE and TOMM40 genes. Alleles of this locus are classified as S, short; L, long; and VL, very long based on the number of T residues.
We evaluated differences in APOE messenger RNA (mRNA) and TOMM40 mRNA levels as a function of the 523 genotype in two brain regions from APOE ε3/ε3 white autopsy-confirmed LOAD cases and normal controls. We further investigated the effect of the 523 locus in its native genomic context using a luciferase expression system.
The expression of both genes was significantly increased with disease. Mean expression of APOE and TOMM40 mRNA levels were higher in VL homozygotes compared with S homozygotes in the temporal and occipital cortexes from normal and LOAD cases. Results of a luciferase reporter system were consistent with the human brain mRNA analysis; the 523 VL poly-T resulted in significantly higher expression than the S poly-T. Although the effect of poly-T length on reporter expression was the same in HepG2 hepatoma and SH-SY5Y neuroblastoma cells, the magnitude of the effect was greater in the neuroblastoma than in the hepatoma cells, which implies tissue-specific modulation of the 523 poly-T.
These results suggest that the 523 locus may contribute to LOAD susceptibility by modulating the expression of TOMM40 and/or APOE transcription.
我们研究了跨越线粒体外膜转位酶40-kD(TOMM40)和载脂蛋白E(APOE)基因的基因组区域,该区域与晚发性阿尔茨海默病(LOAD)的发病风险和发病年龄相关,以确定该区域内一个高度多态的内含子多聚T(rs10524523;以下简称523)是否影响APOE和TOMM40基因的表达。基于T残基的数量,该位点的等位基因分为S(短)、L(长)和VL(非常长)。
我们评估了来自APOE ε3/ε3白种人尸检确诊的LOAD病例和正常对照的两个脑区中,APOE信使核糖核酸(mRNA)和TOMM40 mRNA水平随523基因型的差异。我们使用荧光素酶表达系统进一步研究了523位点在其天然基因组背景下的作用。
随着疾病的发展,这两个基因的表达均显著增加。在正常和LOAD病例的颞叶和枕叶皮质中,VL纯合子中APOE和TOMM40 mRNA水平的平均表达高于S纯合子。荧光素酶报告系统的结果与人类脑mRNA分析一致;523 VL多聚T导致的表达显著高于S多聚T。尽管多聚T长度对报告基因表达的影响在HepG2肝癌细胞和SH-SY5Y神经母细胞瘤细胞中相同,但在神经母细胞瘤中的影响程度大于肝癌细胞,这意味着523多聚T存在组织特异性调节。
这些结果表明,523位点可能通过调节TOMM40和/或APOE转录的表达来影响LOAD易感性。