Droplet digital polymerase chain reaction (ddPCR), which could perform thousands of
PCRs on a nanoliter scale simultaneously, would be an attractive method to massive
parallel sequencing for identifying and studying the significance of low-frequency
rare mutations. Recent evidence has shown that the key potential mechanisms of the
failure of aromatase inhibitors–based therapy involve identifying activating mutations
affecting the ligand-binding domain of the ESR1 gene. Therefore, the detection of ESR1 mutations may be useful as a biomarker predicting an effect of the treatment. We
aimed to develop a ddPCR-based method for the sensitive detection of ESR1 mutations in 325 breast cancer specimens, in which 270 primary and 55 estrogen receptor–positive
(ER+) metastatic breast cancer (MBC) specimens. Our ddPCR assay could detect the ESR1 mutant molecules with low concentration of 0.25 copies/μL. According to the selected
cutoff, ESR1 mutations occurred in 7 (2.5%) of 270 primary breast cancer specimens and in 11 (20%)
of 55 ER+ MBC specimens. Among the 11 MBC specimens, 5 specimens (45.5%) had the most
common ESR1 mutation, Y537S, 4 specimens (36.3%) each had D538G, Y537N, and Y537C. Interestingly,
2 patients had 2 ESR1 mutations, Y537N/D538G and Y537S/Y537C, and 2 patients had 3 ESR1 mutations, Y537S/Y537N/D538G. Biopsy was performed in heterochrony in 8 women twice.
In 8 women, 4 women had primary breast cancer and MBC specimens and 4 women had 2
specimens when treatment was failure. Four of these 8 women acquired ESR1 mutation, whereas no ESR1 mutation could be identified at first biopsy. ddPCR technique could be a promising
tool for the next-generation sequencing-free precise detection of ESR1 mutations in endocrine therapy resistant cases and may assist in determining the
treatment strategy.
Abbreviations:
LBD (ligand-binding domain), ERα (estrogen receptor α), NGS (next generation sequencing), ER+ (ER-positive), AIs (aromatase inhibitors), MBC (metastatic breast cancer), ddPCR (droplet digital polymerase chain reaction), FFPE (formalin-fixed paraffin embedded), PR (partial response), SD (stable disease), PD (progressive disease), gDNA (genomic DNA), LMD (laser microdissection), MGB (minor groove binding), PgR (progesterone receptor), HER2 (human epidermal growth factor receptor 2), AR (androgen receptor), IBTR (ipsilateral breast tumor recurrence), HR (hormone receptor), cfDNA (cell-free DNA), DCIS (ductal carcinoma in situ)To read this article in full you will need to make a payment
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Biography
Takashi Takeshita, PhD, is an Assistant Professor in the Department of Breast and Endocrine Surgery in the Graduate School of Medical Science at Kumamoto University. Dr. Takeshita's research interests are in the development of non-invasive biomarkers, in particular, cell-free DNA.
Article info
Publication history
Published online: September 14, 2015
Accepted:
September 5,
2015
Received in revised form:
September 3,
2015
Received:
July 15,
2015
Identification
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© 2015 Elsevier Inc. Published by Elsevier Inc. All rights reserved.
