Combined Targeted Re-Sequencing of Cytosine DNA Methylation and Mutations of 
DNA Repair Genes with Potential Use for PARP1 Inhibitor Sensitivity Testing

Grimm, Christina; Fischer, Axel; Farrelly, Angela M.; Kalachand, Roshni; Castiglione, Roberta; Wasserburger, Elena; Hussong, Michelle; Schultheis, Anne M.; Altmüller, Janine; Thiele, Holger; Reinhardt, Hans C.; Hauschulz, Kai; Hennessy, Bryan T.; Herwig, Ralf; Lienhard, Matthias; Buettner, Reinhard; Schweiger, Michal R.

doi:10.1016/j.jmoldx.2018.10.007

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Combined Targeted Re-Sequencing of Cytosine DNA Methylation and Mutations of DNA Repair Genes with Potential Use for PARP1 Inhibitor Sensitivity Testing

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Herwig, Ralf
Bioinformatics (Ralf Herwig), Dept. of Computational Molecular Biology (Head: Martin Vingron), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Lienhard, Matthias
Bioinformatics (Ralf Herwig), Dept. of Computational Molecular Biology (Head: Martin Vingron), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Citation

Grimm, C., Fischer, A., Farrelly, A. M., Kalachand, R., Castiglione, R., Wasserburger, E., et al. (2019). Combined Targeted Re-Sequencing of Cytosine DNA Methylation and Mutations of DNA Repair Genes with Potential Use for PARP1 Inhibitor Sensitivity Testing. The Journal of Molecular Diagnostics, 21(2), 198-213. doi:10.1016/j.jmoldx.2018.10.007.

Cite as: https://hdl.handle.net/21.11116/0000-0003-1503-2

Abstract

Current molecular tumor diagnostics encompass panel sequencing to detect mutations, copy number alterations, and rearrangements. However, tumor suppressor genes can also be inactivated by methylation within their promoter region. These epigenetic alterations are so far rarely assessed in the clinical setting. Therefore, we established the AllCap protocol facilitating the combined detection of mutations and DNA methylation at the coding and promoter regions of 342 DNA repair genes in one experiment. We demonstrate the use of the protocol by applying it to ovarian cancer cell lines with different responsiveness to poly(ADP-ribose) polymerase inhibition. BRCA1, ATM, ATR, and EP300 mutations and methylation of the BRCA1 promoter were detected as potential predictors for therapy response. The required amount of input DNA was optimized, and the application to formalin-fixed, paraffin-embedded tissue samples was verified to improve the clinical applicability. Thus, by adding DNA methylation values to panel resequencings, the AllCap assay will add another important level of information to clinical tests and will improve stratification of patients for systemic therapies.