TEST:

cobas® EGFR Mutation Test v2

Overall, test performance was equally robust with either blood collection tube, eg, regarding limit of detection, linearity, and reproducibility, making Roche cfDNA tubes suitable for routine clinical laboratory use in this setting. Importantly, the Roche cfDNA tubes provided more flexibility for specimen handling versus K2EDTA tubes, eg, in terms of tube mixing, plasma separation, and sample stability, and do not require processing of blood within 8 hours thereby increasing the reach of plasma biopsies in NSCLC.

Confirmation of EGFR mutation is important step in the diagnosis and selection of optimal drug for cancer patients. However, tissue biopsy has several limitations such as difficulties to get tissue samples and inaccurate information due to invasive method and tumor heterogeneity. Detection of EGFR mutation in CTC might overcome and make up for limitations of tissue biopsy.

In 2017, the Roche Cobas® EGFR Mutation Test v2 for the detection of the EGFR c.2369C>T p.(T790M) mutation in plasma-derived ctDNA was FDA-approved to identify patients eligible for osimertinib treatment...This implies that centralization of cfDNA-testing is necessary to optimize cost-efficiency. With the advent of ctDNA analysis beyond EGFR in the context of targeted therapies for lung cancer, multigene detection assays will become more cost-effective with high throughput.

It has been observed that new mutations can be detected not only at the time of diagnosis, but also in LB samples taken during the follow-up period, leading to the determination of targeted therapy. The results showed that "liquid biopsy" is a successful and alternative non-invasive method for detecting cancer-causing executive mutations, given the limitations of conventional biopsies.

The liquid-first approach enhances the yield of rare mutations by tissue NGS which is resource-dependent. It benefits regions where tissue biopsy is a rate limiting factor and EGFR mutations are prevalent.

P3; Trial completion date: Mar 2029 --> Jul 2029 | Trial primary completion date: Mar 2024 --> Jun 2024

In our pilot study, tumor microenvironment can be analyzed by multiplex IHC staining. TLS can be identified and calculated by our working definition. TLS-High predicts longer DFS regardless EGFR mutation types, stages, gender and PD-L1 TPS.

The patient initially received 1st line Erlotinib with remission maintained more than 4 years, despite an early dose reduction due to skin toxicity. Despite loss of the original compound EGFR mutation, the rebiopsy revealed a new pathogenic and druggable driver (BRAF p.V600E), which gave the patient a new possibility of further treatment. Yet, replacing Osimertinib with a BRAF/MEK-Is combination resulted only in mixed response, suggesting that some lesions might have remained, at least in part, dependent on the original EGFR mutations. Indeed, re-challenge with Osimertinib in reduced dose together with continuation of Dabrafenib and Trametinib resulted in SD and reflected spatial and temporal heterogeneity of NSCLC.

ctDNA monitoring during Osimertinib treatment helps identify a subgroup of patients with poor prognosis. A lack of cfDNA negativization could identify a subgroup of patients requiring adaptive therapy

This study investigated the distribution of EGFR E20ins in the Asian population, with p.A767_V769dup and p.S768_D770dup being the most frequently observed mutations. Since some studies have reported single EGFR E20ins sequences using different names, it is important to be cautious when describing EGFR E20ins. To ensure comprehensive coverage in real-world settings, each method used for the detection of EGFR E20ins must be annotated according to the HGVS nomenclature.

Further study is suggested to review other parameters in characterizing the insufficient sample.

CTC were isolated by our unique blood-filtration device. And it could be useful to investigate EGFR status. However, more experiments are required to clarify the performance.

P3; Trial primary completion date: Jun 2023 --> Nov 2023

Results Guide sheath flush-based genotyping yielded a mutation detection rate of 61.8% (17/24 EGFR, 1/2 ERBB2, 1/1 KRAS, 1/1 MAP2K, 1/4 MET, and 0/1 STK11), compared to 33% in plasma-based genotyping (p = 0.0151). Conclusion The detection of tumor DNA in the supernatant of guide sheath flush fluid collected during bronchoscopy represents a feasible and more sensitive alternative to circulating tumor DNA genotyping in non-small cell lung cancer.

If a negative result was obtained from methods with lower sensitivity (e.g., Cobas), repeated liquid biopsy testing and/or tissue biopsy analysis should be performed whenever possible, to identify T790M-positive patients to allow them to receive the optimal second-line treatment with a third-generation EGFR TKI.

The Oncomine Lung cfDNA Assay can be used to identify plasma EGFR mutations in patients with lung cancer, although further large-scale studies are required to evaluate the analytical validity for other types of aberrations and genes using clinical samples.

ctDNA analysis is a critical challenge that often leads to false negative results due to biological variables such as stage of the disease, tumor volume, low DNA-shedding, and preanalytical variables. The current study demonstrates the utility of ctDNA as a non-invasive promising tool to detect targetable alterations in situations where tissue biopsy is inaccessible or tumor tissue is depleted. Real-time PCR- based platform is the most preferred option for molecular analysis of ctDNA in clinical settings over NGS, considering factors like cost, turnaround time, and availability of resources.

The Idylla EGFR testing is an accurate and simple tool useful for the early screening of EGFR mutations. The Idylla GeneFusion is a potential screening panel with short turnaround time using a minimal amount of tissue and might be considered as a relevant complementary testing to IHC in diagnostic molecular laboratories.

"Based on the EGFR genotyping by ADPS, the treatment options could be switched in five patients. The highly sensitive and specific ADPS™ EGFR Mutation Test Kit would be useful in detecting the patients who have lung cancer with EGFR mutation, and can benefit from the EGFR targeted therapy."

False positives for exon 20 insertion may occur when using cobas EGFR, and false negatives for exon 19 deletion and L858R mutations may occur when using ODxTT.

We demonstrated the accuracy and potential clinical utility of the Idylla EGFR Mutation Test as a molecular screening platform in terms of turnaround time and molecular testing cost if applied to a cohort with a high EGFR mutation incidence (>17.9%).

P2; Trial completion date: Mar 2024 --> Sep 2024 | Trial primary completion date: Mar 2024 --> Sep 2024

The cobas EGFR Mutation Test, ddPCR, and NGS appear to be feasible methods for analyzing CSF ctDNA in patients with NSCLC and LM. In addition, NGS may provide comprehensive information regarding the mechanisms underlying osimertinib resistance.

NGS ctDNA analysis provided a more comprehensive genetic testing than conventional single gene testing kits. The lower stage which could imply lower or lack of ctDNA shedding into the blood was associated with a discordant result of the blood-based NGS ctDNA assay. Co-occurring mutations might have an impact on the treatment duration of EGFR-TKI.

P2; Trial completion date: Mar 2023 --> Mar 2024 | Trial primary completion date: Mar 2023 --> Mar 2024

The serial monitoring of ct-DNA T790M-status in advanced EGFR-mutant NSCLC during treatment with first-generation EGFR inhibitors was feasible, and a molecular progression before RECIST PD led to an earlier switch to osimertinib in 17% of patients with satisfactory PFS and OS outcomes.

P2; Recruiting --> Active, not recruiting | N=80 --> 116

P2; N=36 --> 0 | Trial completion date: Dec 2024 --> Feb 2023 | Not yet recruiting --> Withdrawn | Trial primary completion date: Dec 2024 --> Feb 2023

Plasma EGFR tests showed similar detection rates for common EGFR mutations compared to the tissue EGFR tests. Cobas showed higher sensitivity in detection of EGFR mutations in body fluids than the PANAMutyper. Real-time PCR using plasma or body fluids could be a suitable first test for the detection of EGFR mutations.

"We described a patient with lung adenocarcinoma harboring METex14del as a potential acquired resistance to osimertinib, who responded to subsequent tepotinib therapy. Re-biopsy and re-analysis of genetic profiles should be considered in NSCLC patients who develop osimertinib resistance."

Although non-plasma samples provided a superior T790M mutation detection rate, we found that liquid biopsy can offer a valuable tool for T790M mutation detection, when a tissue biopsy is not available. Alternatively, a liquid biopsy can be used as a screening test, when re-biopsy should be considered in case of wild-type results.

P2; Trial primary completion date: Sep 2022 --> May 2022

Liquid biopsy can be a prognostic factor in EGFR-tyrosine kinase inhibitor treatments at diagnosis. Tumor re-biopsy can be omitted in patients with positive EGFR mutations by liquid biopsy at PD.

The success rate of ODxTT is slightly inferior to that of cobas EGFR. ODxTT shared a high concordance rate and k-coefficient with cobas EGFR in detecting EGFR mutations, but discordant results between the two tests were observed in a few cases, mainly due to the difference of detectable EGFR variants.

P3; Trial primary completion date: Jan 2023 --> Jul 2023

The most overlooked mutations were exon 20 insertions. A comprehensive EGFR mutation assay can provide significant benefits to patients with NSCLC.

In 3 samples (50 %), the T790M mutation was detected while the corresponding blood sample showed only the presence of the driver mutation. Conclusions EGFR mutation testing from pleural effusion has proven valuable as an alternative liquid biopsy sample to blood, for detecting the T790M acquired resistance mutation in patients who progressed on first-line EGFR-TKIs in Serbia.

Background APPLE is a randomized, non-comparative, open-label, 3-arm, phase II study in patients with common EGFR-mutant, treatment-naïve NSCLC, aimed to evaluate the feasibility of longitudinal plasma EGFR T790M monitoring and the best sequencing strategy from gefitinib to osimertinib. Toxicity was as expected for both drugs. Conclusions Serial monitoring of ctDNA T790M status in patients with advanced EGFR mutant-NSCLC by Cobas v2.0 PCR test treated with first-generation EGFR inhibitors is feasible, and a molecular progression before RECIST PD led to earlier switch to osimertinib in 17% of patients with satisfactory PFS and OS outcomes.

P2; Initiation date: Feb 2022 --> Feb 2023

"Lucence will present new data at the upcoming IASLC 2022 World Conference on Lung Cancer on August 6-9, 2022, highlighting novel applications of its ultrasensitive amplicon-based next generation sequencing technology in the detection and treatment of lungLucence recently announced the publication of a validation study for its flagship LiquidHALLMARK ctDNA Liquid Biopsy Assay. The study, published in PLOS ONE, establishes LiquidHALLMARK’s performance and identifies actionable biomarkers in 70% of lung cancer patients. In addition to showing high sensitivity of both LiquidHALLMARK and Lucence’s AmpliMark amplicon next-generation sequencing (NGS) platform, external validation with cobas® EGFR Mutation Test v2 for lung cancer specimens demonstrated an overall concordance of 84.00% with a 100% concordance rate for EGFR variants above 0.4% VAF. cancer."

P2; Active, not recruiting --> Completed | Trial completion date: Dec 2022 --> Jul 2022 | Trial primary completion date: Dec 2022 --> Jul 2022

P2; Trial primary completion date: Mar 2022 --> Mar 2023