Cross-reactive probes on Illumina DNA methylation arrays: A large study on ALS shows that a cautionary approach is warranted in interpreting epigenome-wide association studies

Paul J. Hop, Ramona A.J. Zwamborn, Eilis J. Hannon, Annelot M. Dekker, Kristel R. van Eijk, Emma M. Walker, Alfredo Iacoangeli, Ashley R. Jones, Aleksey Shatunov, Ahmad Al Khleifat, Sarah Opie-Martin, Christopher E. Shaw, Karen E. Morrison, Pamela J. Shaw, Russell L. McLaughlin, Orla Hardiman, Ammar Al-Chalabi, Leonard H. van Den Berg, Jonathan Mill, Jan H. Veldink*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

Illumina DNA methylation arrays are a widely used tool for performing genome-wide DNA methylation analyses. However, measurements obtained from these arrays may be affected by technical artefacts that result in spurious associations if left unchecked. Cross-reactivity represents one of the major challenges, meaning that probes may map to multiple regions in the genome. Although several studies have reported on this issue, few studies have empirically examined the impact of cross-reactivity in an epigenome-wide association study (EWAS). In this paper, we report on cross-reactivity issues that we discovered in a large EWAS on the presence of the C9orf72 repeat expansion in ALS patients. Specifically, we found that that the majority of the significant probes inadvertently cross-hybridized to the C9orf72 locus. Importantly, these probes were not flagged as cross-reactive in previous studies, leading to novel insights into the extent to which cross-reactivity can impact EWAS. Our findings are particularly relevant for epigenetic studies into diseases associated with repeat expansions and other types of structural variation. More generally however, considering that most spurious associations were not excluded based on pre-defined sets of cross-reactive probes, we believe that the presented data-driven flag and consider approach is relevant for any type of EWAS.

Original languageEnglish
Article numberlqaa105
JournalNAR Genomics and Bioinformatics
Volume2
Issue number4
DOIs
Publication statusPublished - 1 Dec 2020

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