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Widschwendter, M.

Publications and source records attributed to Widschwendter, M..

3 recordsLinked to original sources

Devising reliable and accurate epigenetic clocks:choosing the optimal computational solution

Illumina DNA methylation arrays are frequently used for the discovery of methylation signatures associated with aging and disease. One of the major hurdles to overcome when training trait prediction models is the high dimensionality of the data, with the number of features (CpGs) greatly exceeding the typical number of samples assessed. In addition, most large-scale DNA methylation-based studies do not include replicate measurements for a given sample, making it impossible to estimate the degree of measurement uncertainty or the reliability of the prediction models. A recent study proposed that training penalized regression models on derived principal components (PCs) rather than on the original features (CpGs) results in more reliable age predictions, as estimated from technical replication. Moreover, the same method could be applied for predicting other phenotypes more reliably. Here, we aimed at validating the proposed PC method. We found that although dimension reduction with PCA consistently led to small improvements in the reliability of age prediction models, it severely compromised their accuracy. PC-based models needed far larger training set sizes to be similarly accurate as CpG-based models, whereas reliability did not depend on the sample size of the training set data for either approach. Finally, the PC version of a novel multiclass predictor for breast, ovarian and endometrial cancer we trained using weighted ensembles of deep-learning models also had a markedly lower predictive accuracy compared to a CpG version, suggesting limited applicability of the proposed PC method for predicting phenotypes beyond age.

genomics↗

Technical and biological sources of unreliability of Infinium type II probes of the Illumina MethylationEPIC BeadChip microarray

The Illumina Methylation array platform has facilitated countless epigenetic studies on DNA methylation (DNAme) in health and disease, yet relatively few studies have so studied its reliability, i.e., the consistency of repeated measures. Here we focus on the reliability of both type I and type II Infinium probes. We propose a method for excluding unreliable probes based on dynamic thresholds for mean intensity (MI) and unreliability, estimated by probe-level simulation of the influence of technical noise on methylation {beta}-values using the background intensities of negative control probes. We validate our method in several datasets, including Illumina MethylationEPIC BeadChip v1.0 data from paired whole blood samples taken six weeks apart. Our analysis revealed that specifically probes with low MI exhibit higher {beta}-value variability between repeated samples. MI was associated with the number of C-bases in the respective probe sequence and correlated negatively with unreliability scores. The unreliability scores were substantiated through validation in a new EPIC v1.0 (blood and cervix) and a publicly available 450k (blood) dataset, as they effectively captured the variability observed in {beta}-values between technical replicates. Finally, despite promising higher robustness, the newer version v2.0 of the MethylationEPIC BeadChip retained a substantial number of probes with poor unreliability scores. To enhance current pre-processing pipelines, we developed an R package to calculate MI and unreliability scores and provide guidance on establishing optimal dynamic score thresholds for a given data set.

genomics↗

Correcting For Cell-Type Heterogeneity In Epigenome-Wide Association Studies: Premature Analyses And Conclusions

Recently, a study by Rahmani et al [1] claimed that a reference-free cell-type deconvolution method, called ReFACTor, leads to improved power and improved estimates of cell-type composition compared to competing reference-free and reference-based methods in the context of Epigenome-Wide Association Studies (EWAS). However, we identified many critical flaws (both conceptual and statistical in nature), which seriously question the validity of their claims. We outlined constructive criticism in a recent correspondence letter, Zheng et al [2]. The purpose of this letter is two-fold. First, to present additional analyses, which demonstrate that our original criticism is statistically sound. Second, to highlight additional serious concerns, which Rahmani et al have not yet addressed. In summary, we find that ReFACTor has not been demonstrated to outperform state-of-the-art reference-free methods such as SVA or RefFreeEWAS, nor state-of-the-art reference-based methods. Thus, the claim by Rahmani et al (a claim reiterated in their recent response letter [3]) that ReFACT or represents an advance over the state-of-the-art is not supported by an objective and rigorous statistical analysis of the data.

bioinformatics↗