A digital PCR assay development to detect EGFR T790M mutation in NSCLC patients

e19101 Background: Approximately 50% of advanced non-small cell lung cancer (A-NSCLC) patients with EGFR sensitive mutation who develop acquired resistance to EGFR-TKIs reportedly have a secondary EGFR T790M mutation. Establishing a dynamical, quantitative and noninvasive detection system of EGFR T790M mutation in process of disease therapy for NSCLC is critical to personalized targeted therapy. Methods: 135 A-NSCLC patients with EGFR mutation who received EGFR-TKIs and presented acquired resistance (PFS≥6 months) were included into this study. All patients provided the plasma samples for molecular analysis when disease progressed. 109 patients of them had matched TKI-naive plasma. T790M mutation was measured qualitatively and quantitatively by ARMS and Digital PCR (DggPCR), respectively. Association of T790M mutation with clinical charateristics were evaluated. Results: DgPCR was more sensitive than ARMs to detect T790M mutation in plasma [pre-treatment 29.4% (32/109) VS 5.5% (6/109); post-treatment: 43.0% (58/135) VS 25.2% (34/135)]. 32 patients with pre-treatment T790M mutation predicted shorter PFS and OS compared with 77 T790 M negative patients (PFS, F 12.7 VS 9.2 months, P=0.004, GOS, F 27.0 VS 18.8 months, P=0.002). Patients with or without post-treatment T790M mutation have no significantly different PFS and OS. However, quantified the ratio of copy number of mutant T790M to wild-type by DgPCR, patients were divided into high-frequency groups (≥5%), low-frequency group (0%-5%) and wild-group (0%) according to the number of positive signals observed from DgPCR results. 12 patients in high-frequency group showed shorter PFS and OS compared with wild group and low-frequency group (PFS 9.5 VS 11.9 months, P=0.033, G9.5 VS 13.6 months, P=0.028, GOS, F 18.5 VS 21.2 months, P=0.044, 18.5 VS 28.8 months, P=0.001). Conclusions: Non-invisive and quantitative detection of T790m mutation by digital PCR is feasible in clinical practice. High contents of T790M when disease progression after EGFR-TKIs therapy predicted poor prognosis.

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Detect T790M in cell free tumor DNA of Chinese advanced non-small cell lung cancer adenocarcinoma patients by different platforms and evaluate clinical outcomes of T790M positive patients with osimertinib monotherapy.

Journal of Clinical Oncology ◽

10.1200/jco.2017.35.15_suppl.tps9104 ◽

2017 ◽

Vol 35 (15_suppl) ◽

pp. TPS9104-TPS9104

Author(s):

Zhiyong Liang ◽

Ying Cheng ◽

Yuan Chen ◽

Weiping Liu ◽

You Lu ◽

...

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Digital Pcr ◽

Small Cell ◽

Small Cell Lung ◽

T790m Mutation ◽

Nsclc Patients ◽

Egfr Tki ◽

Tumor Dna ◽

Egfr T790m

TPS9104 Background: EGFR T790M mutation occurs in approximately 50-60% of non-small cell lung cancer adenocarcinoma (NSCLC) patients with acquired EGFR-TKI resistance, based on tumor re-biopsies using an invasive clinical procedure. Recently, Cell free tumor DNA (ctDNA) has emerged as a specific and sensitive blood-based biomarker and studies have demonstrated ctDNA as a feasible and minimally invasive alternative to tissue biopsy. Data on different technology platforms used for EGFR T790M detection in blood in China is limited. We aim to compare the methods currently available in hospital practise, including cobas EGFR Mutation Test (Roche Molecular Systems), super-ARMS, digital PCR and NGS, to compare each platform and clinically validate each as companion diagnostic to osimertinib. Methods: This is an open-label, multi-center study in 250 locally advanced or metastatic NSCLC patients with documented EGFR sensitizing mutation and progression on previous EGFR-TKI. T790M mutation in plasma ctDNA will be tested by four methods: cobas, super-ARMS, digital PCR and NGS in order to evaluate the concordance, sensitivity and specificity of T790M testing in plasma between the cobas test and the other platforms. T790M positive patients by any of the four platforms will receive osimertinib treatment (administered orally as one 80 mg tablet once a day in ASTRIS study, NCT02474355) and the clinical outcomes (PFS, ORR, OS) will be followed. Patients will continue to receive osimertinib until disease progression (PD), as assessed by investigators. Digital PCR and NGS will be used to monitor the molecular evolution of T790M and C797S in plasma from NSCLC patients during osimertinib treatment. NGS will also be used to explore acquired resistance mechanisms before osimertinib treatment and after PD. 23 of planned 250 patients have been enrolled in the study as of January 2017. Clinical trial information: NCT02997501.

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Diagnostic accuracy of droplet digital PCR for detection of EGFR T790M mutation in circulating tumor DNA

Cancer Management and Research ◽

10.2147/cmar.s161382 ◽

2018 ◽

Vol Volume 10 ◽

pp. 1209-1218 ◽

Cited By ~ 16

Author(s):

Rui Zhang ◽

Bojiang Chen ◽

Xiang Tong ◽

Ye Wang ◽

Chengdi Wang ◽

...

Keyword(s):

Diagnostic Accuracy ◽

Digital Pcr ◽

Circulating Tumor Dna ◽

Droplet Digital Pcr ◽

T790m Mutation ◽

Tumor Dna ◽

Egfr T790m

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Clinical implementation of plasma EGFR T790M testing using droplet digital PCR in TKI-resistant NSCLC patients

Experimental and Molecular Pathology ◽

10.1016/j.yexmp.2020.104515 ◽

2020 ◽

Vol 116 ◽

pp. 104515

Author(s):

Dana Leng Hui Chan ◽

Grace Li Xian Toh ◽

Liuh Ling Goh

Keyword(s):

Digital Pcr ◽

Droplet Digital Pcr ◽

Clinical Implementation ◽

Nsclc Patients ◽

Egfr T790m

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Plasma-based early screening and monitoring of EGFR mutations in NSCLC patients by a 3-color digital PCR assay

British Journal of Cancer ◽

10.1038/s41416-020-1024-2 ◽

2020 ◽

Vol 123 (9) ◽

pp. 1437-1444

Author(s):

Xiang Song ◽

Jian Gong ◽

Xiaoling Zhang ◽

Xiaoyan Feng ◽

Hui Huang ◽

...

Keyword(s):

Digital Pcr ◽

Egfr Mutations ◽

Pcr Assay ◽

Early Screening ◽

Nsclc Patients

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Real Clinical Practice of Using Afatinib Therapy in NSCLC Patients with an Acquired EGFR T790M Mutation

Anticancer Research ◽

10.21873/anticanres.12871 ◽

2018 ◽

Vol 38 (9) ◽

pp. 5409-5415 ◽

Cited By ~ 1

Author(s):

KUNIHIKO MIYAZAKI ◽

TOMOHIRO TAMURA ◽

TAKAYUKI KABURAGI ◽

KAZUHITO SAITO ◽

MASAHARU INAGAKI ◽

...

Keyword(s):

Clinical Practice ◽

T790m Mutation ◽

Nsclc Patients ◽

Egfr T790m ◽

Real Clinical Practice

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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

10.1158/1538-7445.am2015-617 ◽

2015 ◽

Author(s):

Yasuhiro Koh ◽

Tomoya Kawaguchi ◽

Masaru Watanabe ◽

Shun-ichi Isa ◽

Masahiko Ando ◽

...

Keyword(s):

Lung Cancer ◽

Cancer Patients ◽

Digital Pcr ◽

Small Cell ◽

Ultrasensitive Detection ◽

Small Cell Lung ◽

T790m Mutation ◽

Lung Cancer Patients ◽

Activating Mutation ◽

Egfr T790m

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Clinical Characteristics of Osimertinib Responder in Non-Small Cell Lung Cancer Patients with EGFR-T790M Mutation

Cancers ◽

10.3390/cancers11030365 ◽

2019 ◽

Vol 11 (3) ◽

pp. 365 ◽

Cited By ~ 2

Author(s):

Akihiro Yoshimura ◽

Tadaaki Yamada ◽

Naoko Okura ◽

Takayuki Takeda ◽

Kazuki Hirose ◽

...

Keyword(s):

Lung Cancer ◽

Small Cell Lung Cancer ◽

Cell Lung Cancer ◽

Kinase Inhibitors ◽

Small Cell ◽

Small Cell Lung ◽

T790m Mutation ◽

Nsclc Patients ◽

Egfr T790m ◽

Egfr Tkis

Osimertinib is a mutant-selective EGFR inhibitor that is effective against non-small cell lung cancer (NSCLC) in patients with the EGFR-T790M mutation, who are resistant to EGFR-tyrosine kinase inhibitors (EGFR-TKIs). However, the factors affecting response to osimertinib treatment are unknown. In this retrospective study, 27 NSCLC patients with the EGFR-T790M mutation were enrolled at five institutions in Japan. Among several parameters tested, the progression-free survival (PFS) associated with the initial EGFR-TKIs was positively correlated with the PFS after osimertinib treatment (p = 0.021). The median PFS following osimertinib treatment and the overall survival (OS) were longer in patients who responded to osimertinib than in those who did not (17.7 months versus 3.5 months, p = 0.009 and 24.2 months versus 13.5 months, p = 0.021, respectively). A multivariate analysis demonstrated that the PFS with initial EGFR-TKIs was significantly related to the PFS with osimertinib treatment (p = 0.035), whereas osimertinib response was significantly related to the PFS and OS with osimertinib treatment (p = 0.016 and p = 0.006, respectively). Our retrospective observations indicate that PFS following the initial EGFR-TKI treatment and the response rate to osimertinib might be promising predictors for effective osimertinib treatment in NSCLC patients with the EGFR-T790M mutation.

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