Saliva cell type DNA methylation reference panel for epidemiology studies in children
Lauren Y. M. Middleton,
Jonathan A Heiss,
Allan C. Just,
Erin B. Ware,
Justin A Colacino,
Kelly M Bakulski
Posted 15 Sep 2020
medRxiv DOI: 10.1101/2020.09.14.20191361
Posted 15 Sep 2020
Saliva is a widely used biological sample, especially in pediatric research, containing a heterogenous mixture of immune and epithelial cells. Associations of exposure or disease with saliva DNA methylation can be influenced by cell-type proportions. Here, we developed a saliva cell-type DNA methylation reference panel to estimate interindividual cell-type heterogeneity in whole saliva studies. Saliva was collected from 22 children (7-16 years) and sorted into immune and epithelial cells, using size exclusion filtration and magnetic bead sorting. DNA methylation was measured using the Illumina MethylationEPIC BeadChip. We assessed cell-type differences in DNA methylation profiles and tested for enriched biological pathways. Immune and epithelial cells differed at 164,793 (20.7%) DNA methylation sites (t-test p < 10-8). Immune cell hypomethylated sites mapped to genes enriched for immune pathways (p < 3.2 x 10-5). Epithelial cell hypomethylated sites were enriched for cornification (p = 5.2 x 10-4), a key process for hard palette formation. Saliva immune and epithelial cells have distinct DNA methylation profiles which can drive whole saliva DNA methylation measures. A primary saliva DNA methylation reference panel, easily implemented with an R package, will allow estimates of cell proportions from whole saliva samples and improve epigenetic epidemiology studies by accounting for measurement heterogeneity by cell-type proportions.
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