Exploration of resistance mechanisms for epidermal growth factor receptor-tyrosine kinase inhibitors based on plasma analysis by digital polymerase chain reaction and next-generation sequencing

Eiji Iwama, Kazuko Sakai, Koichi Azuma, Daijiro Harada, Kaname Nosaki, Katsuyuki Hotta, Makoto Nishio, Takayasu Kurata, Tatsuro Fukuhara, Hiroaki Akamatsu, Koichi Goto, Takayuki Shimose, Junji Kishimoto, Yoichi Nakanishi, Kazuto Nishio, Isamu Okamoto

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Liquid biopsy offers a potential alternative to tissue biopsy for detection of genetic alterations in cancer, and it has been introduced into clinical practice to detect the tyrosine kinase inhibitor (TKI) resistance-conferring T790M mutation of epidermal growth factor receptor (EGFR) in patients with non-small-cell lung cancer (NSCLC). We prospectively collected tumor and plasma samples from 25 NSCLC patients who harbored activating mutations of EGFR and experienced failure of treatment with afatinib. The samples were analyzed by digital PCR (dPCR) and next-generation sequencing (NGS). T790M was detected in plasma with a respective sensitivity and specificity of 83.3% and 70.0% by dPCR and 50.0% and 70.0% by NGS relative to analysis of corresponding tumor samples. Quantitation of T790M based on the ratio of the number of T790M alleles to that of activating mutation alleles (T/A ratio) improved the specificity of plasma analysis to 100% for both dPCR and NGS without a reduction in sensitivity. Although several afatinib resistance mechanisms other than T790M—including copy number gain of NRAS or MET—were identified in tumor samples, the corresponding genetic alterations were not detected in plasma. TP53 mutations were frequently identified in plasma and tumor samples, with most such mutations also having been detected before afatinib treatment. The presence of de novo TP53 mutations was associated with reduced progression-free survival. Quantitation of T790M in plasma is thus a clinically relevant approach to determine the T790M status of tumors. In addition, genetic alterations coexisting with EGFR mutations can affect the efficacy of EGFR-TKI treatment.

Original languageEnglish
Pages (from-to)3921-3933
Number of pages13
JournalCancer Science
Volume109
Issue number12
DOIs
Publication statusPublished - Dec 1 2018

Fingerprint

Epidermal Growth Factor Receptor
Protein-Tyrosine Kinases
Polymerase Chain Reaction
Mutation
Neoplasms
Non-Small Cell Lung Carcinoma
Alleles
Biopsy
Treatment Failure
Disease-Free Survival
Sensitivity and Specificity
Therapeutics
BIBW 2992

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Exploration of resistance mechanisms for epidermal growth factor receptor-tyrosine kinase inhibitors based on plasma analysis by digital polymerase chain reaction and next-generation sequencing. / Iwama, Eiji; Sakai, Kazuko; Azuma, Koichi; Harada, Daijiro; Nosaki, Kaname; Hotta, Katsuyuki; Nishio, Makoto; Kurata, Takayasu; Fukuhara, Tatsuro; Akamatsu, Hiroaki; Goto, Koichi; Shimose, Takayuki; Kishimoto, Junji; Nakanishi, Yoichi; Nishio, Kazuto; Okamoto, Isamu.

In: Cancer Science, Vol. 109, No. 12, 01.12.2018, p. 3921-3933.

Research output: Contribution to journalArticle

Iwama, E, Sakai, K, Azuma, K, Harada, D, Nosaki, K, Hotta, K, Nishio, M, Kurata, T, Fukuhara, T, Akamatsu, H, Goto, K, Shimose, T, Kishimoto, J, Nakanishi, Y, Nishio, K & Okamoto, I 2018, 'Exploration of resistance mechanisms for epidermal growth factor receptor-tyrosine kinase inhibitors based on plasma analysis by digital polymerase chain reaction and next-generation sequencing', Cancer Science, vol. 109, no. 12, pp. 3921-3933. https://doi.org/10.1111/cas.13820
Iwama, Eiji ; Sakai, Kazuko ; Azuma, Koichi ; Harada, Daijiro ; Nosaki, Kaname ; Hotta, Katsuyuki ; Nishio, Makoto ; Kurata, Takayasu ; Fukuhara, Tatsuro ; Akamatsu, Hiroaki ; Goto, Koichi ; Shimose, Takayuki ; Kishimoto, Junji ; Nakanishi, Yoichi ; Nishio, Kazuto ; Okamoto, Isamu. / Exploration of resistance mechanisms for epidermal growth factor receptor-tyrosine kinase inhibitors based on plasma analysis by digital polymerase chain reaction and next-generation sequencing. In: Cancer Science. 2018 ; Vol. 109, No. 12. pp. 3921-3933.
@article{120d35a459ca4f438111b645470925f3,
title = "Exploration of resistance mechanisms for epidermal growth factor receptor-tyrosine kinase inhibitors based on plasma analysis by digital polymerase chain reaction and next-generation sequencing",
abstract = "Liquid biopsy offers a potential alternative to tissue biopsy for detection of genetic alterations in cancer, and it has been introduced into clinical practice to detect the tyrosine kinase inhibitor (TKI) resistance-conferring T790M mutation of epidermal growth factor receptor (EGFR) in patients with non-small-cell lung cancer (NSCLC). We prospectively collected tumor and plasma samples from 25 NSCLC patients who harbored activating mutations of EGFR and experienced failure of treatment with afatinib. The samples were analyzed by digital PCR (dPCR) and next-generation sequencing (NGS). T790M was detected in plasma with a respective sensitivity and specificity of 83.3{\%} and 70.0{\%} by dPCR and 50.0{\%} and 70.0{\%} by NGS relative to analysis of corresponding tumor samples. Quantitation of T790M based on the ratio of the number of T790M alleles to that of activating mutation alleles (T/A ratio) improved the specificity of plasma analysis to 100{\%} for both dPCR and NGS without a reduction in sensitivity. Although several afatinib resistance mechanisms other than T790M—including copy number gain of NRAS or MET—were identified in tumor samples, the corresponding genetic alterations were not detected in plasma. TP53 mutations were frequently identified in plasma and tumor samples, with most such mutations also having been detected before afatinib treatment. The presence of de novo TP53 mutations was associated with reduced progression-free survival. Quantitation of T790M in plasma is thus a clinically relevant approach to determine the T790M status of tumors. In addition, genetic alterations coexisting with EGFR mutations can affect the efficacy of EGFR-TKI treatment.",
author = "Eiji Iwama and Kazuko Sakai and Koichi Azuma and Daijiro Harada and Kaname Nosaki and Katsuyuki Hotta and Makoto Nishio and Takayasu Kurata and Tatsuro Fukuhara and Hiroaki Akamatsu and Koichi Goto and Takayuki Shimose and Junji Kishimoto and Yoichi Nakanishi and Kazuto Nishio and Isamu Okamoto",
year = "2018",
month = "12",
day = "1",
doi = "10.1111/cas.13820",
language = "English",
volume = "109",
pages = "3921--3933",
journal = "Cancer Science",
issn = "1347-9032",
publisher = "Wiley-Blackwell",
number = "12",

}

TY - JOUR

T1 - Exploration of resistance mechanisms for epidermal growth factor receptor-tyrosine kinase inhibitors based on plasma analysis by digital polymerase chain reaction and next-generation sequencing

AU - Iwama, Eiji

AU - Sakai, Kazuko

AU - Azuma, Koichi

AU - Harada, Daijiro

AU - Nosaki, Kaname

AU - Hotta, Katsuyuki

AU - Nishio, Makoto

AU - Kurata, Takayasu

AU - Fukuhara, Tatsuro

AU - Akamatsu, Hiroaki

AU - Goto, Koichi

AU - Shimose, Takayuki

AU - Kishimoto, Junji

AU - Nakanishi, Yoichi

AU - Nishio, Kazuto

AU - Okamoto, Isamu

PY - 2018/12/1

Y1 - 2018/12/1

N2 - Liquid biopsy offers a potential alternative to tissue biopsy for detection of genetic alterations in cancer, and it has been introduced into clinical practice to detect the tyrosine kinase inhibitor (TKI) resistance-conferring T790M mutation of epidermal growth factor receptor (EGFR) in patients with non-small-cell lung cancer (NSCLC). We prospectively collected tumor and plasma samples from 25 NSCLC patients who harbored activating mutations of EGFR and experienced failure of treatment with afatinib. The samples were analyzed by digital PCR (dPCR) and next-generation sequencing (NGS). T790M was detected in plasma with a respective sensitivity and specificity of 83.3% and 70.0% by dPCR and 50.0% and 70.0% by NGS relative to analysis of corresponding tumor samples. Quantitation of T790M based on the ratio of the number of T790M alleles to that of activating mutation alleles (T/A ratio) improved the specificity of plasma analysis to 100% for both dPCR and NGS without a reduction in sensitivity. Although several afatinib resistance mechanisms other than T790M—including copy number gain of NRAS or MET—were identified in tumor samples, the corresponding genetic alterations were not detected in plasma. TP53 mutations were frequently identified in plasma and tumor samples, with most such mutations also having been detected before afatinib treatment. The presence of de novo TP53 mutations was associated with reduced progression-free survival. Quantitation of T790M in plasma is thus a clinically relevant approach to determine the T790M status of tumors. In addition, genetic alterations coexisting with EGFR mutations can affect the efficacy of EGFR-TKI treatment.

AB - Liquid biopsy offers a potential alternative to tissue biopsy for detection of genetic alterations in cancer, and it has been introduced into clinical practice to detect the tyrosine kinase inhibitor (TKI) resistance-conferring T790M mutation of epidermal growth factor receptor (EGFR) in patients with non-small-cell lung cancer (NSCLC). We prospectively collected tumor and plasma samples from 25 NSCLC patients who harbored activating mutations of EGFR and experienced failure of treatment with afatinib. The samples were analyzed by digital PCR (dPCR) and next-generation sequencing (NGS). T790M was detected in plasma with a respective sensitivity and specificity of 83.3% and 70.0% by dPCR and 50.0% and 70.0% by NGS relative to analysis of corresponding tumor samples. Quantitation of T790M based on the ratio of the number of T790M alleles to that of activating mutation alleles (T/A ratio) improved the specificity of plasma analysis to 100% for both dPCR and NGS without a reduction in sensitivity. Although several afatinib resistance mechanisms other than T790M—including copy number gain of NRAS or MET—were identified in tumor samples, the corresponding genetic alterations were not detected in plasma. TP53 mutations were frequently identified in plasma and tumor samples, with most such mutations also having been detected before afatinib treatment. The presence of de novo TP53 mutations was associated with reduced progression-free survival. Quantitation of T790M in plasma is thus a clinically relevant approach to determine the T790M status of tumors. In addition, genetic alterations coexisting with EGFR mutations can affect the efficacy of EGFR-TKI treatment.

UR - http://www.scopus.com/inward/record.url?scp=85056453575&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85056453575&partnerID=8YFLogxK

U2 - 10.1111/cas.13820

DO - 10.1111/cas.13820

M3 - Article

VL - 109

SP - 3921

EP - 3933

JO - Cancer Science

JF - Cancer Science

SN - 1347-9032

IS - 12

ER -