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The tissue is the issue: Improved methylome analysis from paraffin-embedded tissues by application of the HOPE-technique

ABSTRACT: Genome wide DNA methylation profiling of DNA samples obtained from normal lung tissue and lung carcinoma, either fixed by formalin (FFPE), HOPE (paraffin embedded) or cryo-preserved. Illumina's HumanMethylation450 BeadChip was used to obtain DNA methylation profiles across approximately 450.000 CpGs.

ORGANISM(S): Homo sapiens

PROVIDER: GSE51077 | GEO | 2016/08/22

SECONDARY ACCESSION(S): PRJNA221136

REPOSITORIES: GEO

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The tissue is the issue: Improved methylome analysis from paraffin-embedded tissues by application of the HOPE-technique

Project description:Genome wide DNA methylation profiling of DNA samples obtained from normal lung tissue and lung carcinoma, either fixed by formalin (FFPE), HOPE (paraffin embedded) or cryo-preserved. Illumina's HumanMethylation450 BeadChip was used to obtain DNA methylation profiles across approximately 450.000 CpGs. Bisulphite converted DNA from the 36 samples were hybridized to the Illumina HumanMethylation450 Beadchip.

2016-08-22 | E-GEOD-51077 | biostudies-arrayexpress

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Project description:DNA methylation is the most studied epigenetic modification due to its role in regulating gene expression and aberrations in methylation involved in the pathogenesis of cancer and several diseases. The method of choice to evaluate genome-wide methylation has been the Illumina HumanMethylation450 BeadChip (450K), but it was recently replaced with the MethylationEPIC BeadChip (EPIC). We therefore sought to validate the EPIC array in comparison to the 450K array for both fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) pediatric brain tumors. We also performed analysis on the EPIC array with paired FF and FFPE samples, to adapt to a clinical setting where FFPE is routinely used. Further, we compared two restoration methods, REPLI-g and Infinium, for FFPE-derived DNA on the EPIC array.

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Validation of the MethylationEPIC BeadChip for fresh-frozen and formalin-fixed paraffin-embedded tumours (450K)

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Epigenetic analysis of genomic DNA isolated from whole blood of normal and leukemia individuals

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A quantitative real-time PCR-based technique to determine microRNA expression in formalin-fixed paraffin-embedded samples

Project description:MicroRNAs (miRs) are non-coding RNA molecules involved in post-transcriptional regulation, with diverse functions in tissue development, differentiation, cell proliferation and apoptosis. miRs may be less prone to degradation during formalin fixation, facilitating miR expression studies in formalin-fixed paraffin-embedded (FFPE) tissue. Our study demonstrates that the TaqMan Human MicroRNA Array v1.0 (Early Access) platform is suitable for miR expression analysis in FFPE tissue with a high reproducibility (correlation coefficients of 0.95 between duplicates, p<0.00001) and outlines the optimal performance conditions of this platform using clinical FFPE samples. We outline a method of data analysis looking at differences in miR abundance between FFPE and fresh-frozen samples. By dividing the profiled miR into abundance strata of high (Ct<30), medium (30<Ct<35), and low (Ct>35), we show that reproducibility between technical replicates, equivalent dilutions, and FFPE vs. frozen samples is best in the high abundance stratum. We also demonstrate that the miR expression profiles of FFPE samples are comparable to those of fresh-frozen samples, with a correlation of up to 0.87 (p<0.001), when examining all miRs, regardless of RNA extraction method used. Examining correlation coefficients between FFPE and fresh-frozen samples in terms of miR abundance reveals correlation coefficients of up to 0.32 (low abundance), 0.70 (medium abundance) and up to 0.97 (high abundance). Our study aims to demonstrate the utility, reproducibility, and optimization steps needed in miR expression studies using FFPE samples on a high-throughput quantitative PCR-based miR platform, opening up a realm of research possibilities for retrospective studies.

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