How Can Better Identification of T790M Help to Inform Treatment Sequencing Decisions in EGFR Mutation-Positive Non-Small-Cell Lung Cancer?
2
Citation
20
Reference
10
Related Paper
Citation Trend
Abstract:
Optimizing the detection of T790M after progression on first-/second-generation TKIs should allow more patients to benefit from subsequent osimertinib treatment.Keywords:
T790M
Identification
The present pilot study assessed the usefulness of nanofluidic digital polymerase chain reaction (PCR) arrays in epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma after tyrosine kinase inhibitor (TKI) resistance.We enrolled 12 patients with primary lung adenocarcinoma with sensitive EGFR mutation-confirmed T790M status by re-biopsy after TKI resistance. Nanofluidic digital PCR arrays were used to quantify T790M in genomic DNA from the pre-treatment primary site and in serum cell-free DNA (cfDNA).On digital PCR, quantified T790M at the pre-treatment primary site was higher in re-biopsy-positive T790M patients (n=4) than in re-biopsy-negative patients (n=8) (0.78%±0.36% vs. 0.07%±0.09%, p<0.01). T790M at the pre-treatment primary site correlated with progression-free survival (PFS) after gefitinib therapy (r=0.67, p=0.016).Use of digital PCR to quantify T790M at the primary site of EGFR-mutant lung adenocarcinoma predicted T790M emergence in re-biopsies after TKI resistance and PFS after gefitinib therapy.
T790M
Liquid biopsy
Resistance mutation
Cite
Citations (17)
Sel-CapTM, a digital enrichment next-generation sequencing (NGS)-based cancer panel, was assessed for detection of epidermal growth factor receptor (EGFR) gene mutations in plasma for non-small cell lung cancer (NSCLC), and for application in monitoring EGFR resistance mutation T790M in plasma following first-line EGFR-tyrosine kinase inhibitor (EGFR-TKI) treatment. Using Sel-Cap, we genotyped plasma samples collected from 185 patients for mutations Ex19del, L858R, and T790M, and compared results to those of PNAclampTM tumor biopsy (reference method, a peptide nucleic acid-mediated polymerase chain reaction clamping) and two other NGS liquid biopsies. Over two-thirds of activating mutations (Ex19del and L858R), previously confirmed by PNAclamp, were detected by Sel-Cap, which is 4–5 times more sensitive than NGS liquid biopsy. Sel-Cap showed particularly high sensitivity for T790M (88%) and for early-stage plasma samples. The relationship between initial T790M detection in plasma and progression-free survival (PFS) following first-line EGFR-TKIs was evaluated in 34 patients. Patients with T790M detected at treatment initiation (±3 months) had significantly shorter PFS than patients where T790M was first detected >3 months post treatment initiation (median PFS: 5.9 vs. 26.5 months; p < 0.0001). However, time from T790M detection to disease progression was not significantly different between the two groups (median around 5 months). In conclusion, Sel-Cap is a highly sensitive platform for EGFR mutations in plasma, and the timing of the first appearance of T790M in plasma, determined via highly sensitive liquid biopsies, may be useful for prediction of disease progression of NSCLC, around 5 months in advance.
T790M
Liquid biopsy
Cite
Citations (2)
T790M
Cite
Citations (0)
Abstract A sensitive and convenient method for detecting epidermal growth factor receptor ( EGFR ) T790M mutations from circulating tumor DNA (ct DNA ) in advanced non–small cell lung cancer ( NSCLC ) patients with acquired EGFR ‐ TKI resistance would be desirable to direct patient sequential treatment strategy. A comparison of two platforms for detecting EGFR mutations in plasma ct DNA was undertaken. Plasma samples and tumor samples were collected from patients with acquired EGFR ‐ TKI resistance in Zhejiang Cancer Hospital from December 2014 to December 2015. Extracted ct DNA was analyzed using two platforms (Droplet Digital PCR and ARMS [ dPCR ]). A total of 108 patients were enrolled in this study. One hundred and eight patient plasma samples were detected by dd PCR and 75 were detected by ARMS . And 16 patients obtained tissue re‐biopsy, using ARMS assay for detecting EGFR T790M mutation. In all, 43.7% (47/108) had acquired T790M mutation by dd PCR . In 75 patient plasma samples, comparing dd PCR with ARMS , the rates of T790M mutation were 46.7% (35/75) and 25.3% (19/75) by dd PCR and ARMS , respectively. Of all, 16 patients both had tumor and plasma samples, the T790M mutation rates were 56.3% (9/16) by ARMS in tissue and 50.5% (8/16) by dd PCR in plasma ct DNA . The progression mode tended to gradual progression in T790M mutation patients (40.4%), but the T790M negative was inclined to the mode of dramatic progression (39.3%). The patients with T790M‐positive tumors had a longer time to disease progression after treatment with EGFR ‐ TKI s (median, 13.1 months vs. 10.8 months; P = 0.010) and overall survival (median, 35.3 months vs. 30.3 months; P = 0.214) compared with those with T790M‐negative patients. Our study demonstrates ddPCR assay may provide a highly sensitive method to detect EGFR T790M gene in plasma. And T790M‐positive patients have better clinical outcomes to EGFR ‐ TKI s than T790M‐negative patients.
T790M
Liquid biopsy
Resistance mutation
Cite
Citations (85)
Monitoring EGFR T790M with plasma DNA from lung cancer patients in a prospective observational study
Use of plasma DNA to detect mutations has spread widely as a form of liquid biopsy. EGFR T790M has been observed in half of lung cancer patients who have acquired resistance to EGFR tyrosine kinase inhibitors (EGFR-TKI). Effectiveness of monitoring T790M via plasma DNA during treatment with EGFR-TKI has not been established as an alternative to re-biopsy. This was a prospective multicenter observational study involving non-small cell lung cancer patients carrying EGFR L858R or exon 19 deletions, treated with EGFR-TKI. The primary objective was to determine whether T790M could be detected using plasma DNA in patients with progressive disease (PD). T790M was examined using the mutation-biased PCR and quenching probe (MBP-QP) method, a sensitive, fully-automated system developed in our laboratory. Eighty-nine non-small cell lung cancer patients were enrolled from seven hospitals in Japan. Sequential examinations revealed T790M in plasma DNA among 40% of patients who developed PD. Activating mutations, such as L858R and exon 19 deletions, were detected in 40% of patients using plasma DNA, and either T790M or activating mutations were observed in 62%. Dividing into four periods (before PD, at PD, at discontinuation of EGFR-TKI and subsequently), T790M was detected in 10, 19, 24 and 27% of patients, respectively. Smokers, males, patients having exon 19 deletions and patients who developed new lesions evidenced significantly frequent presence of T790M in plasma DNA. Monitoring T790M with plasma DNA using MBP-QP reflects the clinical course of lung cancer patients treated with EGFR-TKI. Detection of T790M with plasma DNA was correlated with EGFR mutation type, exon 19 deletions and tumor progression. Re-biopsy could be performed only in 14% of PD cases, suggesting difficulty in obtaining re-biopsy specimens in practice. Monitoring T790M with plasma DNA reflects the clinical course, and is potentially useful in designing strategies for subsequent treatment.
T790M
Liquid biopsy
Cite
Citations (67)
T790M
Osimertinib
EGFR Inhibitors
Cite
Citations (10)
11054 Background: Patients with EGFR-mutant lung adenocarcinoma develop acquired resistance to EGFR-TKI (TKI) in 10-16 months. About 50% of patients with acquired resistance to TKI have T790M mutation (T790M). However, repeat biopsy to monitor mutation status is not practical. Digital PCR arrays limiting dilution of DNA and can detect single molecules, thus enabling extremely sensitive detection and quantification. We evaluated the usefulness of nanofluidic digital PCR arrays in EGFR-mutant lung adenocarcinoma. Methods: We enrolled 12 patients with primary lung adenocarcinoma and EGFR mutation (exon 19 deletion in 8; L858R in 4) at the pretreatment primary site who acquired resistance to gefitinib. Patients were divided into 2 groups according to T790M status after TKI resistance, as confirmed by rebiopsy. Nanofluidic digital PCR arrays (BioMark HD System, Fluidigm Japan K.K. Tokyo, Japan) were used to quantify T790M in genomic DNA from the pretreatment primary site and serum cell-free DNA (cfDNA) after TKI resistance. Numbers of mutant molecules were estimated by the number of positive chambers in the digital PCR chip and were corrected using the Poisson equation. We assessed the ratio of the number of positive T790M molecules to the number of positive exon 2 molecules. Results: The digital PCR array detected and quantified T790M (0.00-1.21%) in samples from the pretreatment primary site and serum cfDNA after TKI resistance. On digital PCR, the rebiopsy-positive T790M group (n=4) had a significantly higher quantified T790M at the pretreatment primary site than did the T790M-negative group (n=8) (0.59±0.58% vs 0.07±0.09%, p < 0.001). However, the results of analysis of serum cfDNA after TKI resistance did not significantly differ (0.09±0.14% vs 0.05±0.15%, respectively, p = 0.41). T790M at the pretreatment primary site as quantified by digital PCR significantly positively correlated with progression-free survival (PFS) after gefitinib therapy (r = 0.71, p < 0.001). Conclusions: Use of digital PCR to quantify T790M at the primary site of EGFR-mutant lung adenocarcinoma was useful for predicting T790M positivity in rebiopsies after TKI resistance and PFS after gefitinib therapy.
T790M
Liquid biopsy
Resistance mutation
Acquired resistance
Cite
Citations (0)
T790M
EGFR Inhibitors
Cite
Citations (16)
Abstract BACKGROUND: Reversible EGFR TKIs, gefitinib and erlotinib, have shown antitumor efficacy in NSCLC patients with activating mutations in EGFR kinase domain. But the clinical efficacy of these agents is limited by the development of acquired drug resistance, which is most commonly caused by T790M resistance mutation in EGFR. This mutation has been detected in approximately 50% to 60% of patients. The 2nd generation irreversible EGFR inhibitors inhibit EGFR with T790M, but their clinical efficacy to NSCLC patients with T790M appears to be limited due to severe adverse effects caused by concomitant WT EGFR inhibition. Therefore, an EGFR TKI which inhibits T790M mutant EGFR selectively with less activity against WT EGFR may be beneficial. Here we report ASP8273, a novel, small molecule EGFR TKI that inhibits the kinase activity of EGFR containing the activating and T790M resistance mutations with less activity against WT EGFR. METHODS: The inhibitory effect and the selectivity of ASP8273 were evaluated against mutant EGFR (L858R, del ex19, L858R/T790M and del ex19/T790M) and WT EGFR using in vitro enzymatic and cell-based assay. Binding mode of ASP8273 to EGFR was assessed by mass spectrometry. Antitumor activity of ASP8273 was evaluated in xenograft models using PC-9 (del ex19), HCC827 (del ex19), NCI-H1975 (L858R/T790M) and PC-9ER (Erlotinib Resistant)(del ex19/T790M) NSCLC cells. RESULTS: ASP8273 inhibited mutant EGFR containing del ex19 or L858R activating mutations as well as the T790M resistance mutation with lower IC50 values than WT EGFR. Mass spectrometry analysis revealed that ASP8273 is covalently bound to a mutant EGFR(L858R/T790M) via C797 in the kinase domain of EGFR. In NCI-H1975 cells, ASP8273 induced long-lasting inhibition of EGFR phosphorylation for 24 h after washout of compound. In assays using endogenously EGFR-dependent cells, ASP8273 inhibited the growth of PC-9(del ex19), HCC827(del ex19), NCI-H1975(del ex19/T790M) and PC-9ER(del ex19/T790M) with IC50 values of 8-33 nM, more potently than that of NCI-H1666(WT) with IC50 value of 230 nM. In mouse xenograft studies, ASP8273 induced tumor regression in NCI-H1975 (L858R/T790M), HCC827 (del ex19) and PC-9 (del ex19) xenograft models by repeated oral dosing in a dose-dependent manner. Dosing schedules did not affect the efficacy of ASP8273. In an NCI-H1975 xenograft model, complete regression of tumor was achieved after 14-days of ASP8273 treatment. Complete regression was maintained in 50% of mice more than 85 days after cessation of ASP8273 treatment. CONCLUSIONS: ASP8273 inhibits the growth of NSCLC cells with EGFR activating and T790M resistance mutations with evidence of tumor regression. Therefore, ASP8273 may show therapeutic efficacy in NSCLC patients with EGFR mutations. Clinical trials of ASP8273 in NSCLC patients are planned in the US/EU and Asia. Citation Format: Hideki Sakagami, Satoshi Konagai, Hiroko Yamamoto, Hiroaki Tanaka, Takahiro Matsuya, Masamichi Mori, Hiroyuki Koshio, Masatoshi Yuri, Masaaki Hirano, Sadao Kuromitsu. ASP8273, a novel mutant-selective irreversible EGFR inhibitor, inhibits growth of non-small cell lung cancer (NSCLC) cells with EGFR activating and T790M resistance mutations. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 1728. doi:10.1158/1538-7445.AM2014-1728
T790M
EGFR Inhibitors
Erlotinib Hydrochloride
Resistance mutation
Cite
Citations (20)