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

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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.

Author information

Author notes

  1. Zhengquan Yang and Jialu Li are joint first authors.

Authors and Affiliations

  1. School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, China

    Zhengquan Yang, Jialu Li, Yujie Hu, Meihua Chen, Danli Peng, Dan Zong, Qingjuan Shang, Yanling Zhao, Yiyun Ni, Shengnan Jin & Chunming Ding

  2. Key Laboratory of Laboratory Medicine, Ministry of Education, Wenzhou Medical University, Wenzhou, Zhejiang, China

    Zhengquan Yang, Jialu Li, Yujie Hu, Meihua Chen, Danli Peng, Dan Zong, Qingjuan Shang, Yanling Zhao, Yiyun Ni, Shengnan Jin & Chunming Ding

  3. Department of Pulmonary Medicine, Taizhou Central Hospital, Taizhou, Zhejiang, China

    Lianqin Tao & Junfei Zhu

  4. Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China

    Jinyan Ye, Yupeng Xie, Li Yang, Quan Lin, Chang Cai & Yuping Li

  5. Department of Respiratory Medicine, Ningbo First Hospital, Ningbo, Zhejiang, China

    Ning Xu & Xiaoping Huang

  6. Wenzhou Hospital of Traditional Chinese Medicine, Wenzhou, Zhejiang, China

    Xiaoting Dong

  7. Ningbo Hospital of Traditional Chinese Medicine, Ningbo, Zhejiang, China

    Zhonghui Zhou & Yali Yu

  8. The Third Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China

    Zongxiao Shangguan

  9. Zhejiang Jinhua Guangfu Hospital, Jinhua, Zhejiang, China

    Yangyang Xu & Weiping Ying

  10. Department of Oncology, The People’s Hospital of Quzhou, Quzhou, Zhejiang, China

    Meiling Weng

  11. Department of Radiotherapy and Chemotherapy, Yinzhou Hospital Affiliated to Medical School of Ningbo University, Ningbo, Zhejiang, China

    Zuguo Yuan

  12. ChinacoHealthcare Corporation International Hospital, Ningbo, Zhejiang, China

    Zhijun Dong

  13. Yue Qing General Hospital, Yue Qing, Zhejiang, China

    Jifa Li

  14. The Ping Yang Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China

    Zhe Zheng

  15. Department of Respiratory and Critical Care Medicine, Lishui People’s Hospital, Lishui, Zhejiang, China

    Jiongwei Pan

  16. Department of Interventional Radiology, Zhejiang University Lishui Hospital, Lishui, Zhejiang, China

    Lu Liu

  17. Department of Respiratory Medicine, Sanmen People’s Hospital of Zhejiang, Taizhou, Zhejiang, China

    Junhui Ye

  18. Huangyan Hospital of Wenzhou Medical University, Taizhou, Zhejiang, China

    Zhan Zhang

  19. Department of Radiotherapy and Chemotherapy, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China

    Wenfeng Li

Authors
  1. Zhengquan Yang
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  2. Jialu Li
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  3. Yujie Hu
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  4. Meihua Chen
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  5. Danli Peng
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  6. Dan Zong
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  7. Qingjuan Shang
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  8. Lianqin Tao
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  9. Yanling Zhao
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  10. Yiyun Ni
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  11. Jinyan Ye
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  12. Yupeng Xie
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  13. Li Yang
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  14. Quan Lin
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  15. Chang Cai
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  16. Ning Xu
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  17. Xiaoping Huang
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  18. Xiaoting Dong
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  19. Zhonghui Zhou
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  20. Yali Yu
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  21. Zongxiao Shangguan
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  22. Yangyang Xu
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  23. Weiping Ying
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  24. Meiling Weng
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  26. Zhijun Dong
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  27. Jifa Li
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  28. Zhe Zheng
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  29. Jiongwei Pan
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  30. Lu Liu
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  31. Junhui Ye
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  32. Zhan Zhang
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  33. Wenfeng Li
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  34. Junfei Zhu
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  35. Shengnan Jin
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  36. Yuping Li
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  37. Chunming Ding
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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, Shengnan Jin, Yuping Li or Chunming Ding.

Ethics declarations

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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