FFPE breast tumour blocks provide reliable sources of both germline and malignant DNA for investigation of genetic determinants of individual tumour responses to treatment.
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Date
2018-08-01ICR Author
Author
Wilkins, A
Chauhan, R
Rust, A
Pearson, A
Daley, F
Manodoro, F
Fenwick, K
Bliss, J
Yarnold, J
Somaiah, N
Type
Journal Article
Metadata
Show full item recordAbstract
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 Retrieved samples had been archived at different centres for at least 13 years.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.
Subject
Germ Cells
Humans
Breast Neoplasms
Genetic Predisposition to Disease
DNA, Neoplasm
Treatment Outcome
Combined Modality Therapy
Reproducibility of Results
Gene Expression Profiling
Female
Genetic Testing
DNA Copy Number Variations
High-Throughput Nucleotide Sequencing
Exome
Research team
Molecular Oncology
Clinical Trials & Statistics Unit
Targeted Therapy
Translational Breast Radiobiology
Language
eng
Date accepted
2018-04-19
License start date
2018-08
Citation
Breast cancer research and treatment, 2018, 170 (3), pp. 573 - 581
Publisher
SPRINGER