FFPE breast tumour blocks provide reliable sources of both germline and malignant DNA for investigation of genetic determinants of individual tumour responses to treatment
| Autor: | Wilkins, Anna, Chauhan, Ritika, Rust, Alistair, Pearson, Alex, Daley, Frances, Manodoro, Floriana, Fenwick, Kerry, Bliss, Judith, Yarnold, John, Somaiah, Navita |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Germline determinants
DNA Copy Number Variations Gene Expression Profiling Whole exome sequencing High-Throughput Nucleotide Sequencing Reproducibility of Results Breast Neoplasms DNA Neoplasm Clinical Trial Combined Modality Therapy Breast cancer Germ Cells Treatment Outcome Humans Exome Female Genetic Predisposition to Disease Genetic Testing Archival formalin fixed paraffin embedded tissue |
| Zdroj: | Breast Cancer Research and Treatment |
| ISSN: | 1573-7217 0167-6806 |
| Popis: | Background Bio-banked formalin-fixed paraffin-embedded (FFPE) tissues provide an excellent opportunity for translational genomic research. Historically matched blood has not always been collected as a source of germline DNA. This project aimed to establish if normal FFPE breast tissue could be used as an alternative to blood. Methods Exome sequencing was carried out on matched tumour tissue, normal breast tissue and blood on five patients in the START trial. Retrieved samples had been archived at different centres for at least 13 years. Following tissue macro-dissection and DNA extraction, targeted exome capture was performed using SureSelect Human All Exome v5 reagents (Agilent). Illumina paired-end libraries were prepared from the captured target regions and sequenced on a HiSeq2500 (Illumina) acquiring 2 × 75 bp reads. Somatic variants were called using the MuTect software analysis tool and copy number abnormalities (CNA) were identified using CNVkit. Targeted sequencing and droplet digital PCR were used to validate somatic variants and CNA, respectively. Results Overlap of somatic variants and CNA called on tumour versus blood and tumour versus normal breast tissue was good. Agreement in somatic variant calling ranged from 76.9 to 93.6%. Variants with an allele frequency lower than 10% were more difficult to validate irrespective of the type of germline DNA used. Pearson’s correlation coefficients for paired comparisons of CNA using blood or normal tissue as reference ranged from 0.70 to 0.94. Conclusions There is good correlation between the somatic mutations and CNA called using archived blood or normal breast tissue as germline reference material. Electronic supplementary material The online version of this article (10.1007/s10549-018-4798-7) contains supplementary material, which is available to authorised users. |
| Databáze: | OpenAIRE |
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