Ultra-sensitive detection of the pretreatment EGFR T790M mutation in non-small cell lung cancer patients with an EGFR-activating mutation using droplet digital PCR

Watanabe, Masaru; Kawaguchi, Tomoya; Isa, Shun-ichi; Ando, Masahiko; Tamiya, Akihiro; Kubo, Akihito; Saka, Hideo; Takeo, Sadanori; Adachi, Hirofumi; Tagawa, Tsutomu; Kakegawa, Seiichi; Yamashita, Motohiro; Kataoka, Kazuhiko; Ichinose, Yukito; Takeuchi, Yukiyasu; Sakamoto, Kazuhiro; Matsumura, Akihide; Koh, Yasuhiro

doi:10.1158/1078-0432.ccr-14-2151

Cited by 216Lung Cancer Treatments and MutationsCancer Genomics and DiagnosticsHER2/EGFR in Cancer Research

Ultra-sensitive detection of the pretreatment EGFR T790M mutation in non-small cell lung cancer patients with an EGFR-activating mutation using droplet digital PCR

Watanabe, Masaru, Kawaguchi, Tomoya, Isa, Shun-ichi, Ando, Masahiko, Tamiya, Akihiro, Kubo, Akihito, Saka, Hideo, Takeo, Sadanori +10 more·2015·Clinical Cancer Research

DOI 10.1158/1078-0432.ccr-14-2151

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Abstract

PURPOSE: The resistance to the EGFR tyrosine kinase inhibitors (TKI) is a major concern in non-small cell lung cancer (NSCLC) treatment. T790M mutation in EGFR accounts for nearly 50% of the acquired resistance to EGFR-TKIs. Earlier studies suggested that T790M mutation was also detected in TKI-naïve NSCLCs in a small cohort. Here, we use an ultra-sensitive droplet digital PCR (ddPCR) technique to address the incidence and clinical significance of pretreatment T790M in a larger cohort. EXPERIMENTAL DESIGN: ddPCR was established as follows: wild-type or T790M mutation-containing DNA fragments were cloned into plasmids. Candidate threshold was identified using wild-type plasmid, normal human genomic DNA, and human A549 cell line DNA, which expresses wild type. Surgically resected tumor tissues from 373 NSCLC patients with EGFR-activating mutations were then examined for the presence of T790M using ddPCR. RESULTS: Our data revealed a linear performance for this ddPCR method (R(2) = 0.998) with an analytical sensitivity of approximately 0.001%. The overall incidence of the pretreatment T790M mutation was 79.9% (298/373), and the frequency ranged from 0.009% to 26.9%. The T790M mutation was detected more frequently in patients with a larger tumor size (P = 0.019) and those with common EGFR-activating mutations (P = 0.022), as compared with the others. CONCLUSIONS: The ultra-sensitive ddPCR assay revealed that pretreatment T790M was found in the majority of NSCLC patients with EGFR-activating mutations. ddPCR should be utilized for detailed assessment of the impact of the low frequency pretreatment T790M mutation on treatment with EGFR-TKIs.

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Pretreatment T790M mutation status in formalin-fixed, paraffin-embedded tissue using standard sequencing platforms has classically been described in less than 5% of treatment-naïve tumors.
“determination of pretreatment T790M mutation status in formalin-fixed, paraffinembedded tissue by using standard sequencing platforms has been classically described in less than 5% of treatment-naïve tumors (Table 5)”
From The International Association for the Study of Lung Cancer Consensus Statement on Optimizing Management of EGFR Mutation–Positive Non–Small Cell Lung Cancer: Status in 2016 (2016) · cites this paper as [37]
With more sensitive techniques such as droplet digital PCR, T790M can be detected in up to 80% of treatment-naïve tumors.
“but with more sensitive techniques (e.g., droplet digital PCR), T790M can be detected in up to 80% of these tumors.34,37”
From The International Association for the Study of Lung Cancer Consensus Statement on Optimizing Management of EGFR Mutation–Positive Non–Small Cell Lung Cancer: Status in 2016 (2016) · cites this paper as [37]

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Abstract 617: Ultrasensitive detection of the pretreatment EGFR T790M mutation in non-small cell lung cancer patients with an EGFR-activating mutation using picodroplet digital PCR
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Ultrasensitive detection of EGFR T790M mutation by droplet digital PCR (ddPCR) in TKI naïve non-small cell lung cancer (NSCLC) harboring EGFR mutation: Results of the nationwide program Biomarkers France of the French Cooperative Thoracic Intergroup (IFCT)
Leduc, C., Pencreach, E., Merlio, J. P., Bringuier, P. P., de Fraipont, F., Escande, F., Lemoine, A., Ouafik, L., Blons, H., Denis, M. G., et al. (2017) · Annals of Oncology · doi:10.1093/annonc/mdx363.001 · Cited by 1 · see citing works
Abstract 5248: Non-invasive EGFR T790M detection using droplet digital PCR system
Hashida, S., Ohashi, K., Matsubara, T., Ohtsuka, T., Watanabe, M., Suzawa, K., Maki, Y., Yamamoto, H., Soh, J., Asano, H., et al. (2015) · Cancer Research · doi:10.1158/1538-7445.am2015-5248
Detection of Low-level EGFR c.2369 C > T (p.Thr790Met) Resistance Mutation in Pre-treatment Non-small Cell Lung Carcinomas Harboring Activating EGFR Mutations and Correlation with Clinical Outcomes
Ye, L., Ardakani, N. M., Thomas, C., Spilsbury, K., Leslie, C., Amanuel, B., & Millward, M. (2020) · Pathology & Oncology Research · doi:10.1007/s12253-020-00833-z · Cited by 10 · see citing works
Abstract 760: Detection of EGFR T790M mutation by ddPCR in untreated NSCLC patients: Correlation with clinical outcome
Ganzinelli, M., Rulli, E., Tamborini, E., Busico, A., Russo, G. L., Corrao, G., Vitali, M., Garassino, M. C., Broggini, M., & Marabese, M. (2017) · Cancer Research · doi:10.1158/1538-7445.am2017-760

Authors: Watanabe, M., Kawaguchi, T., Isa, S.-I., Ando, M., Tamiya, A., Kubo, A., Saka, H., Takeo, S., Adachi, H., Tagawa, T., Kakegawa, S., Yamashita, M., Kataoka, K., Ichinose, Y., Takeuchi, Y., Sakamoto, K., Matsumura, A., & Koh, Y.
Affiliations: Wakayama Medical University (JP)
Osaka City University (JP)
NHO Kinki Chuo Chest Medical Center (JP)
Nagoya University Hospital (JP)
Aichi Medical University (JP)
National Hospital Organization (JP)
National Kyushu Medical Center (JP)
Sapporo National Hospital (JP)
Nagasaki Medical Center (JP)
Shikoku Cancer Center (JP)
Iwakuni Medical Center (JP)
National Hospital Organization Kyushu Cancer Center (JP)
Toneyama National Hospital (JP)
National Hospital Organization Yokohama Medical Center (JP)
Corresponding author: Koh, Yasuhiro
ORCID: Watanabe, Masaru
Kawaguchi, Tomoya
Ando, Masahiko
Tamiya, Akihiro
Kubo, Akihito
Saka, Hideo
Sakamoto, Kazuhiro
Koh, Yasuhiro
Venue: Clinical Cancer Research
Publisher: American Association for Cancer Research (AACR)
Published: 2015-08-01
Volume: 21
Issue: 15
Pages: 3552-3560
ISSN: 1078-0432 (ISSN-L)
1557-3265
DOI: 10.1158/1078-0432.ccr-14-2151
OpenAlex: W2006126363
Cited by: 216
References: 42
Citations by year: 2026: 1 · 2025: 4 · 2024: 3 · 2023: 9 · 2022: 24 · 2021: 19 · 2020: 22 · 2019: 30 · 2018: 19 · 2017: 47 · 2016: 32 · 2015: 6
FWCI: 27.38
Type: journal article
Language: en
Open access: closed
Locations: Clinical Cancer Research (publishedVersion)
PubMed (publishedVersion)
Funders: Japan Society for the Promotion of Science (25460896, 23501314)
Topics: Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Lung Cancer Treatments and Mutations
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer Genomics and Diagnostics
Health Sciences › Medicine › Oncology › HER2/EGFR in Cancer Research
Keywords: T790M mutation, non-small cell lung cancer, EGFR activating mutations, droplet digital PCR, EGFR tyrosine kinase inhibitors, acquired drug resistance, tumor size, analytical sensitivity
MeSH terms: Adult, Aged, Aged, 80 and over, Carcinoma, Non-Small-Cell Lung, Female, Gene Frequency, Humans, Male, Middle Aged, Mutation, Polymerase Chain Reaction, Drug Resistance, Neoplasm, Protein Kinase Inhibitors, Precision Medicine, ErbB Receptors

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Ultra-sensitive detection of the pretreatment EGFR T790M mutation in non-small cell lung cancer patients with an EGFR-activating mutation using droplet digital PCR

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