The Dynamic Use of <i>EGFR</i> Mutation Analysis in Cell-Free DNA as a Follow-Up Biomarker during Different Treatment Lines in Non-Small-Cell Lung Cancer Patients

Lim

Asia-Pac J Clncl Oncology

et al. 2022

Aim Epidermal growth factor receptor (EGFR) mutations are detected in non‐small cell lung cancer (NSCLC) and associated with responses to therapy with tyrosine kinase inhibitors (TKIs). We compared the analytical performances of two real‐time PCRs and droplet digital PCR (ddPCR) to detect EGFR mutations using plasma. Methods Plasma EGFR tests were performed using 86 plasma samples from 75 prospectively enrolled NSCLC patients with early and advanced stages. Analytical performances of plasma‐using two real‐time PCR, Cobas EGFR mutation v2 and PANAMutyper, EGFR kit, and ddPCR were evaluated based on the tissue EGFR test results. The frequencies of EGFR mutations and acquired T790M mutation after TKI therapy were also assessed. Results The incidence of all EGFR mutations was 52.3% (23/44) in tissue and was up to 43.2% (19/44) in plasma. The Cobas detection rates of three EGFR mutations (exon 19 deletions, L858R, and T790M) in plasma were similar to those in tissue. The Cobas showed a higher detection rate (76.7%) than that by the PANAMutyper (60.5%). Sensitivity for T790M mutation was lower than the sensitivity for the exon 19 deletions or L858R in both tests. Mutant allele frequency measured by ddPCR was significantly correlated with the semi‐quantitative values of the Cobas. Conclusions 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.

Section: Discussionmentioning

confidence: 91%

Evaluation of molecular methods for plasma detection of EGFR mutations in non‐small cell lung cancer

Lim

Asia-Pac J Clncl Oncology

et al. 2022

“…Two of the most important advancements in personalized medicine, especially in the field of lung cancer, include the use of cfDNA as a diagnostic and prognostic biomarker and next-generation sequencing (NGS) for the mutational analysis of lung tumors. For instance, EGFR and ALK have been identified as key biomarkers in lung cancer, and molecular tests for EGFR and ALK have become common in lung cancer treatment [24] . Circulating DNA as a biomarker is easily accessible, reliable, and reproducible and reduces the suffering and cost to society associated with the disease.…”

Section: Discussionmentioning

confidence: 99%

“…This difference is more remarkable when using silica-gel columns as small DNA fragments (<100 bp) are loosed during isolation. The use of an isolation kit that captures all DNA fragments is of importance for the reliability of all downstream experiments, espe¬cially when the small DNA fractions (<100 bp) are considered the most informative as they rep¬resent characteristic qualitative changes in the primary tumor such as mutations [24] .…”

Section: Discussionmentioning

confidence: 99%

Comparative Evaluation of Diagnostic Performance of Circulating free DNA by Conventional vs. Real Time PCR in Gallbladder Cancer

Kumari

Mishra

Agarwal

et al. 2022

ijirms

Background: Circulating free DNA (cfDNA) in serum/plasma has been studied as a promising biomarker in various pathologies, including cancer. However, there is no standardized method for the isolation and quantification of serum cfDNA. An effective and reliable method for isolation and quantification is of utmost importance before any clinical decision. The current study compares the conventional and real-time PCR methods to find any differences and concordance in cfDNA levels between the two methods and the diagnostic accuracy of cfDNA by each method. Methods: Serum sample was collected from 67 subjects, including 17 normal healthy individuals (control, n=17), 19 disease controls (cholecystitis, n=19), and 31 Gallbladder cancer patients (cancer, n=31) before any treatment for cfDNA quantification. Results: The cfDNA level did not differ significantly between two methods in both control and cholecystitis groups. In cancer group, cfDNA level was significantly (P < 0.001) different and higher in real time PCR as compared to conventional PCR. There was no significant correlation between two methods in control (r=0.02, P = 0.937), cholecystitis (r=0.10, P = 0.697), cancer (r=-0.08, P = 0.657) and total cases (control + cholecystitis + cancer) (r=0.06, P = 0.622). The diagnostic accuracy of two methods was found similar (P > 0.05) when assessed between control vs. cholecystitis (Z=0.85, P = 0.397), and control vs. total cases (Z=1.35, P = 0.177). However, the diagnostic accuracy of real time PCR was found significantly different and higher as compared to conventional PCR when assessed between control vs. cancer (Z=2.98, P = 0.003), and cholecystitis vs. cancer (Z=4.41, P < 0.001). Conclusion: Quantitative real-time PCR method is of high accuracy, reproducibility, and time-effectiveness. The diagnostic accuracy of real-time PCR was higher compared to conventional PCR.

“…In addition, exosome analysis could help in the screening and early detection of lung cancer, when patients have better prognosis [113,124], and some clinical trials are already addressing this issue (www.clinicaltrials.gov). However, exosome analysis has not been included in clinical guides yet, contrary to ctDNA where there are already clinical indications for its use as biomarker in lung cancer and some commercial kits are already available for mutations' assessment [8,155]. One of the reasons is the lack of standardized protocols for exosome isolation and analysis, which impairs the implementation of their analysis in a routine clinical laboratory.…”

Section: Discussionmentioning

confidence: 99%

Exosomes in Lung Cancer: Actors and Heralds of Tumor Development

Sandúa

Alegre

González

2021

Cancers

Self Cite

Lung cancer is a leading cause of cancer-related death worldwide and in most cases, diagnosis is reached when the tumor has already spread and prognosis is quite poor. For that reason, the research for new biomarkers that could improve early diagnosis and its management is essential. Exosomes are microvesicles actively secreted by cells, especially by tumor cells, hauling molecules that mimic molecules of the producing cells. There are multiple methods for exosome isolation and analysis, although not standardized, and cancer exosomes from biological fluids are especially difficult to study. Exosomes’ cargo proteins, RNA, and DNA participate in the communication between cells, favoring lung cancer development by delivering signals for growth, metastasis, epithelial mesenchymal transition, angiogenesis, immunosuppression and even drug resistance. Exosome analysis can be useful as a type of liquid biopsy in the diagnosis, prognosis and follow-up of lung cancer. In this review, we will discuss recent advances in the role of exosomes in lung cancer and their utility as liquid biopsy, with special attention to isolating methods.