Dynamics of Plasma EGFR T790M Mutation in Advanced NSCLC: A Multicenter Study

Abstract

Background

Droplet digital polymerase chain reaction (ddPCR) is an emerging technology for quantitative cell-free DNA oncology applications. However, a ddPCR assay for the epidermal growth factor receptor (EGFR) p.Thr790Met (T790M) mutation suitable for clinical use remains to be established with analytical and clinical validations.

Objective

We aimed to develop and validate a new ddPCR assay to quantify the T790M mutation in plasma for monitoring and predicting the progression of advanced non-small-cell lung cancer (NSCLC).

Methods

Specificity of the ddPCR assay was evaluated with genomic DNA samples from healthy individuals. The inter- and intraday variations of the assay were evaluated using mixtures of plasmid DNA containing wild-type EGFR and T790M mutation sequences. We assessed the clinical utility of the T790M assay in a multicenter prospective study in patients with advanced NSCLC receiving tyrosine kinase inhibitor (TKI) treatment by analyzing longitudinal plasma DNA samples.

Results

We set the criteria for a positive call when the following conditions were satisfied: (1) T790M mutation frequency > 0.098% (3 standard deviations above the background signal); (2) at least two positive droplets in duplicate ddPCR reactions. Among the 62 patients with advanced NSCLC exhibiting resistance to TKI treatment, 15 had one or more serial plasma samples that tested positive for T790M. T790M mutation was detected in the plasma as early as 205 days (median 95 days) before disease progression, determined by imaging analysis. Plasma T790M concentrations also correlated with intervention after disease progression.

Conclusions

We developed a ddPCR assay to quantify the T790M mutation in plasma. Quantification of longitudinal plasma T790M mutation may allow noninvasive assessment of drug resistance and guide follow-up treatment in TKI-treated patients with NSCLC.

Trial Registration

Clinical Trials.gov identifier: NCT02804100.

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Acknowledgements

The authors thank all the patients who participated in the study and all the support from the doctors and nurses for the collection of samples from 16 centers throughout Zhejiang Province.

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Affiliations

Authors

Contributions

Zhengquan Yang, Jialu Li, Yujie Hu, Meihua Chen, Danli Peng, Dan Zong, Qingjuan Shang, Lianqin Tao, Yanling Zhao, Yiyun Ni, Jinyan Ye, Yupeng Xie, Li Yang, Quan Lin, Chang Cai, Ning Xu, Xiaoping Huang, Xiaoting Dong, Zhonghui Zhou, Yali Yu, Zongxiao Shangguan, Yangyang Xu, Weiping Ying, Meiling Weng, Zuguo Yuan, Zhijun Dong, Jifa Li, Zhe Zheng, Jiongwei Pan, Lu Liu, Junhui Ye, Zhan Zhang, Wenfeng Li, Junfei Zhu, and Yuping Li enrolled patients into the study. Zhengquan Yang and Jialu Li contributed to the protocol design. Zhengquan Yang and Jialu Li interpreted the data. Zhengquan Yang, Jialu Li, SJ, and CD performed the statistical analyses; YL and JZ were the clinical leads for the study; SJ and CD conceived and designed the study. All authors participated in the writing and/or critical review of the manuscript and agreed on the final version.

Corresponding authors

Correspondence to Junfei Zhu or Shengnan Jin or Yuping Li or Chunming Ding.

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Funding

The study was funded by Innovation Team in Zhejiang Province Universities, Innovation Discipline of Zhejiang Province in Nucleic Acid Molecular Diagnostics (437201702G), Key Discipline of Zhejiang Province in Medical Technology (First Class, Category A) (No. 437601607) and Zhejiang Provincial Natural Science Foundation (No. LY18H200007).

Conflict of Interest

Zhengquan Yang, Jialu Li, Yujie Hu, Meihua Chen, Danli Peng, Dan Zong, Qingjuan Shang, Lianqin Tao, Yanling Zhao, Yiyun Ni, Jinyan Ye, Yupeng Xie, Li Yang, Quan Lin, Chang Cai, Ning Xu, Xiaoping Huang, Xiaoting Dong, Zhonghui Zhou, Yali Yu, Zongxiao Shangguan, Yangyang Xu, Weiping Ying, Meiling Weng, Zuguo Yuan, Zhijun Dong, Jifa Li, Zhe Zheng, Jiongwei Pan, Lu Liu, Junhui Ye, Zhan Zhang, Wenfeng Li, Junfei Zhu, Shengnan Jin, Yuping Li and Chunming Ding have no conflicts of interest that might be relevant to the contents of this manuscript.

Ethics Approval and Consent to Participate

The study was approved by the Clinical Research Ethics Committee of Wenzhou Medical University. Written informed consent was obtained from participants.

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Yang, Z., Li, J., Hu, Y. et al. Dynamics of Plasma EGFR T790M Mutation in Advanced NSCLC: A Multicenter Study. Targ Oncol 14, 719–728 (2019). https://doi.org/10.1007/s11523-019-00682-0

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