Combined targeted resequencing of cytosine DNA methylation and mutations of DNA repair genes with potential use for poly(ADP-ribose) polymerase 1 inhibitor sensitivity testing

Item Type:	Article
Title:	Combined targeted resequencing of cytosine DNA methylation and mutations of DNA repair genes with potential use for poly(ADP-ribose) polymerase 1 inhibitor sensitivity testing
Creators Name:	Grimm, C. and Fischer, A. and Farrelly, A.M. and Kalachand, R. and Castiglione, R. and Wasserburger, E. and Hussong, M. and Schultheis, Anne M and Altmüller, J. and Thiele, H. and Reinhardt, H.C. and Hauschulz, K. and Hennessy, B.T. and Herwig, R. and Lienhard, M. and Buettner, R. and Schweiger, M.R.
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.
Keywords:	BRCA1 Protein, Tumor Cell Line, Cell Survival, DNA Methylation, DNA Mutational Analysis, E1A-Associated p300 Protein, Ovarian Neoplasms, Poly (ADP-Ribose) Polymerase-1, Poly(ADP-ribose) Polymerase Inhibitors, Genetic Promoter Regions, Temozolomide / Pharmacology
Source:	Journal of Molecular Diagnostics
ISSN:	1525-1578
Publisher:	Elsevier
Volume:	21
Number:	2
Page Range:	198-213
Date:	March 2019
Official Publication:	https://doi.org/10.1016/j.jmoldx.2018.10.007
PubMed:	View item in PubMed

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