Smooth Modelling of Covariate Effects in Bisulfite Sequencing-derived Measures of DNA Methylation

Kaiqiong Zhao, Karim Oualkacha, Lajmi Lakhal-Chaieb, Kathleen Klein, Marie Hudson, Celia Greenwood

Identifying disease-associated changes in DNA methylation can help us gain a better understanding of disease etiology. Bisulfite sequencing allows the generation of high-throughput methylation profiles at single-base resolution. However, optimally modeling and analyzing these sparse and discrete sequencing data is still very challenging due to variable read depth, missing data patterns, long-range correlations, data errors, and confounding from cell type mixtures. We propose a regression-based hierarchical model that allows covariate effects to vary smoothly along genomic positions. We build a specialized EM algorithm which explicitly allows for experimental errors and cell type mixtures, to make inference about smooth covariate effects in the model. Simulations show that the proposed method provides accurate estimates of covariate effects and captures the major underlying methylation patterns. We also apply our method to analyze data from rheumatoid arthritis patients and controls and find regions with altered methylation. We anticipate the results will lead to important improvements in understanding methylation patterns in rheumatoid arthritis.