PMID- 27940449 OWN - NLM STAT- MEDLINE DCOM- 20170522 LR - 20240210 IS - 1530-8561 (Electronic) IS - 0009-9147 (Print) IS - 0009-9147 (Linking) VI - 63 IP - 2 DP - 2017 Feb TI - Next Generation Sequencing of Circulating Cell-Free DNA for Evaluating Mutations and Gene Amplification in Metastatic Breast Cancer. PG - 532-541 LID - 10.1373/clinchem.2016.261834 [doi] AB - BACKGROUND: Breast cancer tissues are heterogeneous and show diverse somatic mutations and somatic copy number alterations (CNAs). We used a novel targeted next generation sequencing (NGS) panel to examine cell-free DNA (cfDNA) to detect somatic mutations and gene amplification in women with metastatic breast cancer (MBC). METHODS: cfDNA from pretreated patients (n = 42) and 9 healthy controls were compared with matched lymphocyte DNA by NGS, using a custom 158 amplicon panel covering hot-spot mutations and CNAs in 16 genes, with further validation of results by droplet digital PCR. RESULTS: No mutations were identified in cfDNA of healthy controls, whereas exactly half the patients with metastatic breast cancer had at least one mutation or amplification in cfDNA (mean 2, range 1-6) across a total of 13 genes. Longitudinal follow up showed dynamic changes to mutations and gene amplification in cfDNA indicating clonal and subclonal response to treatment that was more dynamic than cancer antigen 15-3 (CA15-3). Interestingly, at the time of blood sampling disease progression was occurring in 7 patients with erb-b2 receptor tyrosine kinase 2 (ERBB2) gene amplification in their cfDNA and 3 of these patients were human epidermal growth factor receptor 2 (HER2) negative at diagnosis, suggesting clonal evolution to a more aggressive phenotype. Lastly, 6 patients harbored estrogen receptor 1 (ESR1) mutations in cfDNA, suggesting resistance to endocrine therapy. Overall 9 of 42 patients (21%) had alterations in cfDNA that could herald a change in treatment. CONCLUSIONS: Targeted NGS of cfDNA has potential for monitoring response to targeted therapies through both mutations and gene amplification, for analysis of dynamic tumor heterogeneity and stratification to targeted therapy. CI - (c) 2016 American Association for Clinical Chemistry. FAU - Page, Karen AU - Page K AD - Department of Cancer Studies and Cancer Research UK Leicester Centre, University of Leicester, Leicester, UK. FAU - Guttery, David S AU - Guttery DS AD - Department of Cancer Studies and Cancer Research UK Leicester Centre, University of Leicester, Leicester, UK. FAU - Fernandez-Garcia, Daniel AU - Fernandez-Garcia D AD - Department of Cancer Studies and Cancer Research UK Leicester Centre, University of Leicester, Leicester, UK. FAU - Hills, Allison AU - Hills A AD - Department of Surgery and Cancer, Division of Cancer, and. FAU - Hastings, Robert K AU - Hastings RK AD - Department of Cancer Studies and Cancer Research UK Leicester Centre, University of Leicester, Leicester, UK. FAU - Luo, Jinli AU - Luo J AD - Department of Cancer Studies and Cancer Research UK Leicester Centre, University of Leicester, Leicester, UK. FAU - Goddard, Kate AU - Goddard K AD - Department of Medical Oncology, Imperial College London, Charing Cross Hospital, London, UK. FAU - Shahin, Vedia AU - Shahin V AD - Department of Medical Oncology, Imperial College London, Charing Cross Hospital, London, UK. FAU - Woodley-Barker, Laura AU - Woodley-Barker L AD - Department of Medical Oncology, Imperial College London, Charing Cross Hospital, London, UK. FAU - Rosales, Brenda M AU - Rosales BM AD - Department of Surgery and Cancer, Division of Cancer, and. FAU - Coombes, R Charles AU - Coombes RC AD - Department of Surgery and Cancer, Division of Cancer, and. FAU - Stebbing, Justin AU - Stebbing J AD - Department of Surgery and Cancer, Division of Cancer, and js39@le.ac.uk j.stebbing@imperial.ac.uk. FAU - Shaw, Jacqueline A AU - Shaw JA AD - Department of Cancer Studies and Cancer Research UK Leicester Centre, University of Leicester, Leicester, UK; js39@le.ac.uk j.stebbing@imperial.ac.uk. LA - eng GR - 23464/CRUK_/Cancer Research UK/United Kingdom GR - MR/M018687/1/MRC_/Medical Research Council/United Kingdom GR - 13462/CRUK_/Cancer Research UK/United Kingdom GR - G1100425/MRC_/Medical Research Council/United Kingdom GR - 2013 RIF - SHAW/PANCREATICCANUK_/Pancreatic Cancer UK/United Kingdom GR - NIHR-RP-011-053/DH_/Department of Health/United Kingdom GR - 20276/CRUK_/Cancer Research UK/United Kingdom PT - Journal Article DEP - 20161209 PL - England TA - Clin Chem JT - Clinical chemistry JID - 9421549 RN - 0 (DNA, Neoplasm) SB - IM MH - Adult MH - Aged MH - Aged, 80 and over MH - Breast Neoplasms/*genetics/*pathology MH - DNA, Neoplasm/blood/*genetics MH - Female MH - Gene Expression Profiling MH - *High-Throughput Nucleotide Sequencing MH - Humans MH - Middle Aged MH - Mutation MH - Neoplasm Metastasis/*genetics MH - Particle Size MH - Polymerase Chain Reaction MH - *Sequence Analysis, DNA PMC - PMC6241835 MID - EMS79773 COIS- Competing interests: The authors declare that they have no competing interests. EDAT- 2016/12/13 06:00 MHDA- 2017/05/23 06:00 PMCR- 2018/11/19 CRDT- 2016/12/13 06:00 PHST- 2016/06/08 00:00 [received] PHST- 2016/09/19 00:00 [accepted] PHST- 2016/12/13 06:00 [pubmed] PHST- 2017/05/23 06:00 [medline] PHST- 2016/12/13 06:00 [entrez] PHST- 2018/11/19 00:00 [pmc-release] AID - clinchem.2016.261834 [pii] AID - 10.1373/clinchem.2016.261834 [doi] PST - ppublish SO - Clin Chem. 2017 Feb;63(2):532-541. doi: 10.1373/clinchem.2016.261834. Epub 2016 Dec 9.