EGFR-mutation detection in VTX-1-enriched CTCs, ctDNA, and comparison to tumor in patients with non-small cell lung cancer (NSCLC).

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e20525-e20525
Author(s):  
Jonathan Wade Goldman ◽  
Haiyan Liu ◽  
Meghah Vuppalapaty ◽  
Charles Wilkerson ◽  
Clementine Lemaire ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Kinase Inhibitors ◽  
Low Cost ◽  
Growth Factor Receptor ◽  
High Sensitivity ◽  
Circulating Tumor Dna ◽  
Egfr Mutations ◽  
Label Free ◽  
Enhanced Sensitivity ◽  
Nsclc Patients

e20525 Background: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI's) have shown dramatic clinical benefits when EGFR mutations are detected. These assays are mainly performed on tumor biopsies, which carry risks, are expensive and are not always successful. Moreover, secondary mutations, causing resistance to 1st & 2nd generation TKI’s develop during treatment and therefore mutations require ongoing monitoring. Circulating tumor cells (CTCs) or circulating tumor DNA (ctDNA) are promising for monitoring the patient over time and show enhanced sensitivity when combined1. The VTX-1 Liquid Biopsy System enables label-free capture of CTCs from blood and genomic assays downstream2. In this study, we demonstrate the sensitivity of a combined EGFR assay on VTX-1-enriched CTCs and ctDNA, using blood spiked with cancer cells & DNA and then with NSCLC patients. Methods: NSCLC cells (A549: wt, H1975: L858R+ & T790M+, HCC827: 19del+) were used to validate the assay. H1975 cells and HCC827 DNA were spiked in blood. Plasma was extracted and the plasma depleted blood was processed through VTX-1. The collected plasma and enriched cells were subjected to the detection of 19del, L858R and T790M EGFR mutations using the EntroGen ctEGFR kit. Later, blood and tumor were collected from NSCLC patients and analyzed for EGFR mutations on biopsies, ctDNA and CTCs. Results: Mutant DNA was detected for an input as low as 0.5 ng (~83 cells), with a sensitivity from 0.1% to 2% for a total DNA of 25ng (~4 cancer cells & 4000 WBCs) to 1ng (~4 cancer cells & 200 WBCs). Processing of plasma-depleted-blood with Vortex showed the same efficiency compared to whole blood. Mutations were detected from enriched cells and plasma respectively for 100 H1975 cells and 0.5ng of HCC827 DNA spiked in 2mL blood. NSCLC patients are being enrolled and results will be presented at the conference. Conclusions: The ctEGFR assay performed well on VTX-1 enriched cells and ctDNA, enabling a low cost approach to analyze EGFR mutations from a single blood tube with high sensitivity, potentially being a useful tool for guiding treatment of NSCLC patients. 1.Sundaresan, et al. Clin Cancer Res 2016. 2. Kidess-Sigal, et al. Oncotarget 2016.

scholarly journals Neoadjuvant EGFR-TKI Therapy for EGFR-Mutant NSCLC in Traditionally Unresectable Patients

2021 ◽  
Author(s):  
Ke-Cheng Chen ◽  
Shuenn-Wen Kuo ◽  
Chen-Hao Hsiao ◽  
Jing-Shing Chen
Keyword(s):  
Thoracic Surgery ◽  
Lung Adenocarcinoma ◽  
Surgical Resection ◽  
Kinase Inhibitors ◽  
Growth Factor Receptor ◽  
Progression Free Survival ◽  
Egfr Mutations ◽  
Multidisciplinary Management ◽  
Advanced Stage ◽  
Nsclc Patients

Abstract IntroductionAdvanced stage non-small cell lung cancer (NSCLC) patients harboring epidermal growth factor receptor (EGFR) mutations may have benefit from tyrosine kinase inhibitors (TKI). However, the role of multidisciplinary management including neoadjuvant TKI therapy and thoracic surgery is uncertain. This study assessed the possible impact of neoadjuvant TKI therapy and thoracic surgery in selected advanced stage patients.MethodsAdvanced stage of IIIB and IVA NSCLC patients were retrospectively reviewed from 2010 to 2013. Patients with EGFR mutations who received neoadjuvant TKI followed by surgical resection were included. All patients were followed up for 5 years or until death.ResultsThere were total 15 advanced stage lung adenocarcinoma patients in the study. 8 patients were stage IIIB and 7 were stage IVA. All tumor sizes significantly decreased after neoadjuvant TKI therapy (p value = 0.0002). 11 patients received adjuvant TKI therapy after surgical resection and others received adjuvant cisplatin-based chemotherapy. Progression-free survival was superior in the group of adjuvant TKI therapy than in the group of adjuvant chemotherapy (median 14 months versus 5.9 months, p value = 0.016). Overall survival (OS) was not different between two groups (p value = 0.755). In the group of adjuvant TKI therapy, median OS in patients harboring exon 19 deletion and exon 21 L858R was 60 months and 44.9 months, respectively (p value = 0.078). ConclusionTKI may decrease the size of EGFR mutation lung adenocarcinoma. A multidisciplinary management including neoadjuvant TKI therapy and thoracic surgery may be discussed in selected advanced stage lung adenocarcinoma.

Routine detection of EGFR mutations in patients with NSCLC: A comparative study of three alternative methods in 105 patients

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 7184-7184
Author(s):  
G. Zalcman ◽  
N. Richard ◽  
A. Hardouin ◽  
R. Gervais ◽  
M. Antoine ◽  
...  
Keyword(s):  
High Performance ◽  
Kinase Inhibitors ◽  
Direct Sequencing ◽  
Tumor Resection ◽  
Growth Factor Receptor ◽  
Alternative Methods ◽  
Egfr Mutations ◽  
Genomic Sequencing ◽  
Surgical Biopsy ◽  
Nsclc Patients

7184 Background: The efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) has been reported in lung adenocarcinoma patients with tumoral EGFR mutations. Those mutations were found mainly by direct genomic sequencing on snap-frozen surgical specimens. Conversely, TKIs are used in metastatic patients who do not undergo tumor resection. In these patients, there is a need for routine sensitive molecular procedures, to overcome the small size of non-surgical bronchoscopy paraffin-embedded biopsy samples. Methods: Patients were selected on clinical and pathological characteristics: never (n = 43) or former smokers, patients with non-squamous NSCLC (n = 105) or patients with bronchioloalveolar adenocarcinoma (n = 34). Direct genomic sequencing assay was performed as reported elsewhere. Denaturing, high-performance, liquid chromatography (DHPLC) assay was performed with EGFR-specific primers that amplify exons 18, 19, and 21. A multiplex, allele-specific, oligonucleotide PCR (MASO-PCR) assay was carried out with a set of primers that identify the 14 most frequent molecular events described for the EGFR gene, which covers 90% of EGFR gain-of-function mutations described to date. Results: 123 samples were screened from 105 non-squamous NSCLC patients (female/male ratio = 0.84). Non-surgical biopsy specimens were available in 38 patients. EGFR mutations were detected by at least two of three procedures in 18/105 patients (17%). In paraffin-embedded specimens with low tumor content, EGFR heterozygous mutations were found either by MASO-PCR alone (n = 2, confirmed in the matched surgical sample by another procedure), or both by MASO-PCR and DHPLC (n = 16); they were missed by nucleotidic sequencing in 6 samples. 18 patients received TKI. 6 dramatic responses were achieved in patients with EGFR mutation, while no mutation could be found in non-responsive patients. Overall disease control was obtained in 8/18 patients (44%). Conclusions: MASO-PCR and DHPLC appear more sensitive than direct sequencing in non-surgical paraffin-embedded biopsies, which represent the bulk of samples in lung cancer patients. We propose that the cost-effective MASO-PCR be used for routine screening of EGFR mutations in NSCLC patients. No significant financial relationships to disclose.

scholarly journals Detection of EGFR Mutations in Plasma Cell-Free Tumor DNA of TKI-Treated Advanced-NSCLC Patients by Three Methodologies: Scorpion-ARMS, PNAClamp, and Digital PCR

Diagnostics ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1062
Author(s):  
Annamaria Siggillino ◽  
Paola Ulivi ◽  
Luigi Pasini ◽  
Maria Sole Reda ◽  
Elisa Chiadini ◽  
...  
Keyword(s):  
Kinase Inhibitors ◽  
Growth Factor Receptor ◽  
Digital Pcr ◽  
Egfr Mutations ◽  
Amplification Refractory Mutation System ◽  
Clinical Method ◽  
Thermo Fisher Scientific ◽  
Exon 19 Deletion ◽  
Nsclc Patients ◽  
Tumor Dna

Analysis of circulating cell-free tumor DNA (cftDNA) has emerged as a specific and sensitive blood-based approach to detect epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) patients. Still, there is some debate on what should be the preferential clinical method for plasma-derived cftDNA analysis. We tested 31 NSCLC patients treated with anti-EGFR tyrosine kinase inhibitors (TKIs), at baseline and serially during therapy, by comparing three methodologies in detecting EGFR mutations (L858R, exon 19 deletion, and T790M) from plasma: scorpions-amplification refractory mutation system (ARMS) methodology by using EGFR Plasma RGQ PCR Kit-QIAGEN, peptide nucleic acid (PNA) clamp and PANA RealTyper integration by using PNAClamp EGFR-PANAGENE, and digital real time PCR by using QuantStudio 3D Digital PCR System-Thermo Fisher Scientific. Specificity was 100% for all three mutations, independently from the platform used. The sensitivity for L858R (42.86%) and T790M (100%) did not change based on the method, while the sensitivity for Del 19 differed markedly (Scorpion-ARMS 45%, PNAClamp 75%, and Digital PCR 85%). The detection rate was also higher (94.23%) as measured by Digital PCR, and when we monitored the evolution of EGFR mutations over time, it evidenced the extreme inter-patient heterogeneity in terms of levels of circulating mutated copies. In our study, Digital PCR showed the best correlation with tissue biopsy and the highest sensitivity to attain the potential clinical utility of monitoring plasma levels of EGFR mutations.

scholarly journals FBXL2 counteracts Grp94 to destabilize EGFR and inhibit EGFR-driven NSCLC growth

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Mengmeng Niu ◽  
Jing Xu ◽  
Yang Liu ◽  
Yuhuang Li ◽  
Tao He ◽  
...  
Keyword(s):  
Kinase Inhibitors ◽  
Critical Role ◽  
Growth Factor Receptor ◽  
Egfr Mutations ◽  
Inhibitory Effects ◽  
Molecule Inhibitor ◽  
Epidermal Growth ◽  
F Box Protein ◽  
Nsclc Patients ◽  
Egfr Tki

AbstractAbnormal activation of epidermal growth factor receptor (EGFR) drives non-small cell lung cancer (NSCLC) development. EGFR mutations-mediated resistance to tyrosine-kinase inhibitors (TKIs) is a major hurdle for NSCLC treatment. Here, we show that F-box protein FBXL2 targets EGFR and EGFR TKI-resistant mutants for proteasome-mediated degradation, resulting in suppression of EGFR-driven NSCLC growth. Reduced FBXL2 expression is associated with poor clinical outcomes of NSCLC patients. Furthermore, we show that glucose-regulated protein 94 (Grp94) protects EGFR from degradation via blockage of FBXL2 binding to EGFR. Moreover, we have identified nebivolol, a clinically used small molecule inhibitor, that can upregulate FBXL2 expression to inhibit EGFR-driven NSCLC growth. Nebivolol in combination with osimertinib or Grp94-inhibitor-1 exhibits strong inhibitory effects on osimertinib-resistant NSCLC. Together, this study demonstrates that the FBXL2-Grp94-EGFR axis plays a critical role in NSCLC development and suggests that targeting FBXL2-Grp94 to destabilize EGFR may represent a putative therapeutic strategy for TKI-resistant NSCLC.

Sensitivity of TargetSelector in clinical experience in ctDNA profiling of NSCLC 2000 cases.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e23033-e23033
Author(s):  
Veena M. Singh ◽  
Anthony J. Daher ◽  
Jeffery J. Chen ◽  
Lyle Arnold ◽  
Cecile Rose T. Vibat
Keyword(s):  
Kinase Inhibitors ◽  
Acquired Resistance ◽  
Growth Factor Receptor ◽  
Egfr Mutations ◽  
Wild Type ◽  
T790m Mutation ◽  
Blood Samples ◽  
Copy Numbers ◽  
Activating Mutation ◽  
Nsclc Patients

e23033 Background: Targeted cancer therapy relies on identifying specific DNA mutations from a patient’s tumor. Tyrosine kinase inhibitors (TKIs) tend to be effective for non-small cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) activating mutations, of which exon 19 deletions (Del19) and L858R are most common. Acquired resistance to TKI therapy is associated with a T790M mutation. Standard biomarker analyses may not reflect tumor heterogeneity; they entail tissue biopsies often with surgical complications. To address these limitations, Biocept developed a minimally invasive method to characterize cancer biomarkers in blood. Biocept's proprietary TargetSelector assays selectively amplify relevant mutations from circulating tumor DNA (ctDNA). Clinical validations demonstrated high concordances between molecular tests in blood vs tissue. As further validation, EGFR mutation detection frequencies were compared to US averages (mycancergenome.org). Here we analyze 2000 blood samples received at Biocept from 1Mar 2016 to 4Jan 2017 from late stage NSCLC patients. Methods: Blood was collected in Biocept OncoCEE BCT validated to preserve DNA ≤ 8 days. TargetSelector was used to detect ctDNA L858R, Del19 and T790M.EGFR allele copy numbers for wild type and each mutant were calculated. The prevalence of each mutation was compared to US averages. Results: Del19, L858R, and T790M mutations were detected in 12.9%, 8.5%, and 9.9% of the analyzed blood samples, respectively. This is concordant with US averages, which are 10% for each mutation. Median copy numbers/ml of blood were 30 for Del19 (range: 1 – 91974), 15 for L858R (range: 1 – 91200), and 10 for T790M (range: 1 – 137360). The median wild type EGFR copy number detected/ml blood was 2304 (range: 8 – 2498725). In ~80% of T790M cases, ≥ 1 concomitant activating mutation was detected. Conclusions: Biocept's TargetSelector detects EGFR mutations (Del19, L585R, and T790M) at a very high level of sensitivity down to 1 mutant copy/ml in advanced NSCLC patients at frequencies consistent with cited US rates. Moreover, the underlying activating mutation was detected in ~80% of T790M cases.

scholarly journals Iterative Upgrading of Small Molecular Tyrosine Kinase Inhibitors for EGFR Mutation in NSCLC: Necessity and Perspective

Pharmaceutics ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 1500
Author(s):  
Jing Zhu ◽  
Qian Yang ◽  
Weiguo Xu
Keyword(s):  
Lung Cancer ◽  
Kinase Inhibitors ◽  
Egfr Mutation ◽  
Growth Factor Receptor ◽  
Gene Mutations ◽  
Egfr Mutations ◽  
Convenient Route ◽  
Epidermal Growth ◽  
Nsclc Patients ◽  
Genome Information

Molecular targeted therapy has been reported to have fewer adverse effects, and offer a more convenient route of administration, compared with conventional chemotherapy. With the development of sequencing technology, and research on the molecular biology of lung cancer, especially whole-genome information on non-small cell lung cancer (NSCLC), various therapeutic targets have been unveiled. Among the NSCLC-driving gene mutations, epidermal growth factor receptor (EGFR) mutations are the most common, and approximately 10% of Caucasian, and more than 50% of Asian, NSCLC patients have been found to have sensitive EGFR mutations. A variety of targeted therapeutic agents for EGFR mutations have been approved for clinical applications, or are undergoing clinical trials around the world. This review focuses on: the indications of approved small molecular kinase inhibitors for EGFR mutation-positive NSCLC; the mechanisms of drug resistance and the corresponding therapeutic strategies; the principles of reasonable and precision molecular structure; and the drug development discoveries of next-generation inhibitors for EGFR.

SHP2 Inhibition Benefits Epidermal Growth Factor Receptor mutated Non-Small Cell Lung Cancer Therapy

2020 ◽  
Vol 20 ◽  
Author(s):  
Leiming Xia ◽  
Lu Wen ◽  
Siying Wang
Keyword(s):  
Stem Cells ◽  
Synergistic Effects ◽  
Growth Factor Receptor ◽  
Egfr Mutations ◽  
Combination Strategies ◽  
Lung Cancer Therapy ◽  
Src Homology ◽  
Nsclc Patients ◽  
Egfr Mutated ◽  
Egfr Tkis

: EGFR-TKIs are facing a big challenge of everlasting activated EGFR mutations which lack of effective binding sites, this barrier confers the dark sides that largely limited the outcome of NSCLC patients in clinic. Combination strategies show impressive anti-tumor efficacy comparing with EGFR-TKI mono-treatment, especially targeting both stem cells and non-stem cells. SHP2 (Src homology 2-containing phosphotyrosine phosphatase 2) plays an important role in regulating various malignant biology through hyper-activating intracellular pathways due to either over expression or catalytical mutation. Some pathways that SHP2 involved in were overlaps with EGFR downstream, and others were not subject to EGFR. Interestingly, SHP2 suppression was reported that can destroy the stemness of cancer. Therefore, we hypothesize SHP2 inhibitor might be an promising drug that could synergistically enhance or sensitize the anti-tumor efficacy of EGFR-TKIs in EGFR mutated NSCLC patients. Here, we summarized the mechanisms of SHP2 in regulating EGFR mutated NSCLC patients, attempted to reveal the potential synergistic effects of SHP2 inhibitor combined with EGFR-TKIs.

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