M3D: a kernel-based test for shape changes in methylation profiles

Abstract: DNA methylation is an intensely studied epigenetic mark implicated in many biological processes of direct clinical relevance. While sequencing based technologies are increasingly allowing high resolution measurements of DNA methylation, statistical modelling of such data is still challenging. In particular, statistical identification of differentially methylated regions across different conditions poses unresolved challenges in accounting for spatial correlations within the statistical testing procedure. We propose a non-parametric, kernel-based method, M3D, to detect higher-order changes in methylation profiles, such as shape, across pre-defined regions. The test statistic explicitly accounts for differences in coverage levels between samples, thus handling in a principled way a major confounder in the analysis of methylation data. Empirical tests on real and simulated data sets show an increased power compared to established methods, as well as considerable robustness with respect to coverage and replication levels. Code is implemented in R, and is freely available via Bioconductor package M3D.