It was recently reported that a functional single nucleotide
It was recently reported that a functional single nucleotide polymorphism (rs75932628) within Triggering receptor expressed on myeloid ProteOrange 2 (TREM2) is associated with AD (Guerreiro et al., 2013). Homozygous loss of function mutations in TREM2 are also associated with an autosomal recessive form of early onset dementia, presenting with bone cysts and consequent fractures called polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy, or Nasu-Hakola disease (Paloneva et al., 2003). TREM2 activates the innate immune response in macrophages and dendritic cells (N\'Diaye et al., 2009). In the brain, TREM2 is expressed in microglia and promotes phagocytosis of apoptotic neurons, cellular debris, and misfolded proteins by the recognition of specific endogenous ligands on the surface of apoptotic cells (Jonsson and Stefansson, 2013). Subsequently, TREM2 delays the inflammatory response by suppressing microglial cytokine production (Hsieh et al., 2009). TREM2 deficiency augmented Aβ accumulation and neuronal loss in a mouse model of AD (Wang et al., 2015). In addition, higher TREM2 mRNA expression was predominantly associated with the increase of phosphorylated tau in AD patient brains (Lue et al., 2015).
Recently, peripheral leukocytes were reported to be indicated in AD pathogenesis (Zenaro et al., 2015). Gene expression and DNA methylation changes in leukocytes have been observed in neuropsychiatric conditions including AD (Yamazaki et al., 2016, Yoshino et al., 2016a, Yoshino et al., 2016b, Yoshino et al., 2016c, Funahashi et al., 2016). DNA methylation, a type of epigenetic modification, plays a key role in regulating gene expression (Abdolmaleky et al., 2004). Although we reported higher TREM2 mRNA expression in the leukocytes of AD subjects (Mori et al., 2015), methylation rates of TREM2 have not yet been examined.
In the present study, we examined the methylation rates of the TREM2 intron 1 in leukocytes of AD subjects and investigated the relationship between these methylation rates and TREM2 mRNA expression.
Materials and methods
Discussion This is the first study showing TREM2 DNA hypomethylation in AD subjects and significant correlation between TREM2 mRNA expression and TREM2 DNA methylation rates of intron 1 in peripheral leukocytes of AD subjects. Furthermore, we could replicate our preceding study with larger sample showing the elevated TREM2 mRNA expression in AD subjects compared to control subjects (Mori et al., 2015). In the present study, there are three major findings. First, we confirmed that TREM2 mRNA expression in the leukocytes of AD subjects is significantly higher than that in control subjects. Our results are also consistent with another study that reported that TREM2 mRNA expression in monocytes was elevated in AD (Hu et al., 2014). Although our preceding report revealed that TREM2 mRNA expression was significantly higher in schizophrenia subjects than control subjects (Mori et al., 2015), there was no association between TREM2 expression or methylation and use of antipsychotic drugs or psychiatric symptoms in NPI sub-items. In the brain, TREM2 is primarily expressed in microglia and it is absent or expressed at low levels in granulocytes, monocytes, and macrophages (Neumann and Takahashi, 2007). The microglial expression of TREM2 was reported to be high in cultured mouse neuronal cells (Wang et al., 2015). Although there was no significant correlation between TREM2 mRNA expression and clinical characteristics of AD in our study, elevated TREM2 expression in leukocytes can be utilized as a novel biological marker for AD. Second, we found significantly lower methylation rates of three CpG sites in intron 1 in TREM2 of AD patients versus healthy controls. These CpG sites are the sites to which the largest number of transcription factors are predicted to bind. The methylation rates at CpG1, 2 and 3, and the average methylation rate at these sites in AD subjects were significantly lower than those in control subjects even after Bonferroni correction. Discriminant analysis demonstrated good sensitivity and specificity of 78.0% and 84.0%, respectively, of this assay.