Crystal digital droplet PCR for detection and quantification of circulating <i>EGFR</i> sensitizing and resistance mutations in advanced non-small cell lung cancer

<div><p>Over the past years, targeted therapies using tyrosine kinase inhibitors (TKI) have led to an increase in progression-free survival and response rate for a subgroup of non-small cell lung cancer (NSCLC) patients harbouring specific gene abnormalities compared with chemotherapy. However long-lasting tumor regression is rarely achieved, due to the development of resistant tumoral subclones, which requires alternative therapeutic approaches. Molecular profile at progressive disease is a challenge for making adaptive treatment decisions. The aim of this study was to monitor <i>EGFR</i>-mutant tumors over time based on the quantity of mutant DNA circulating in plasma (ctDNA), comparing two different methods, Crystal™ Digital™ PCR and Massive Parallel Sequencing (MPS). In plasma circulating cell free DNA (cfDNA) of 61 advanced NSCLC patients we found an overall correlation of 78% between mutated allelic fraction measured by Crystal Digital PCR and MPS. 7 additional samples with sensitizing mutations and 4 additional samples with the resistance mutation were detected with Crystal Digital PCR, but not with MPS. Monitoring levels of both mutation types over time showed a correlation between levels and trends of mutated ctDNA detected and clinical assessment of disease for the 6 patients tested. In conclusion, Crystal Digital PCR exhibited good performance for monitoring mutational status in plasma cfDNA, and also appeared as better suited to the detection of known mutations than MPS in terms of features such as time to results.</p></div

Patient characteristics.

<p>Patient characteristics.</p

Monitoring of targeted sensitizing and resistance EGFR mutations and circulating cell-free DNA levels over time in 6 metastatic NSCLC patients using dPCR.

<p>Coloured region indicates period of chemotherapy. Radiological assessment of patient response is indicated above the figures. Empty circles and squares indicates sensitizing and resistance mutations positives by dPCR but not detected by NGS respectively.</p

Wild-type and mutant DNA levels measured by dPCR in 87 plasma samples from 61 metastatic NSCLC patients.

<p>Greyed bar: circulating cell-free DNA (cfDNA) concentration in NSCLC patients with confirmed targetable EGFR mutations in tumor tissue. Filled bar: sensitizing (Del19, p.L858R and p.L861Q) mutations concentration. Dashed bar: wild-type DNA concentration in NSCLC patients with wild-type EGFR in tumor tissue. Grey dots: p.T790M mutation concentration. Empty circles and squares indicates sensitizing and resistance mutations positives by dPCR but not detected by NGS respectively.</p

sj-docx-1-jtt-10.1177_1357633X241229462 - Supplemental material for Evaluation of the satisfaction and experiences of oncology patients and doctors using teleconsultation during the COVID-19 pandemic

Supplemental material, sj-docx-1-jtt-10.1177_1357633X241229462 for Evaluation of the satisfaction and experiences of oncology patients and doctors using teleconsultation during the COVID-19 pandemic by Myrto Kastrisiou, Maryam Karimi, Evangelos AA Christou, Alexandra Bizot, Marie-Alix Ropers, Anne De-Jesus, Meriem Mokdad-Adi, Thi Hong Van To, Alessandro Viansone, Suzette Delaloge, Benjamin Besse, and Maria Kfoury in Journal of Telemedicine and Telecare</p

Additional file 3: Table S2. of Method for semi-automated microscopy of filtration-enriched circulating tumor cells

FISH spots per CTC of a patient with an ERG-rearranged tumor depending on the step. (DOC 63 kb

Additional file 1: Figure S1. of Method for semi-automated microscopy of filtration-enriched circulating tumor cells

Examples of gene rearrangement and gain/amplification detection in filtration enriched-cell lines by filter-adapted-FISH (FA-FISH). (A) Example of gene rearrangement detection. (B) Example of gain/amplification detection. Scale: white bars = 10 μm. (TIF 8523 kb

Additional file 2: Table S1. of Method for semi-automated microscopy of filtration-enriched circulating tumor cells

FISH spots detected per CTC in a patient with a ROS1-rearranged tumor depending on the step. (DOC 68 kb

Liquid biopsies for residual disease and recurrence

Detection of minimal residual disease in patients with cancer, who are in complete remission with no cancer cells detectable, has the potential to improve recurrence-free survival through treatment selection. Studies analyzing circulating tumor DNA (ctDNA) in patients with solid tumors suggest the potential to accurately predict and detect relapse, enabling treatment strategies that may improve clinical outcomes. Over the past decade, assays for ctDNA detection in plasma samples have steadily increased in sensitivity and specificity. These are applied for the detection of residual disease after treatment and for earlier detection of recurrence. Novel clinical trials are now assessing how assays for “residual disease and recurrence” (RDR) may influence current treatment paradigms and potentially change the landscape of risk classification for cancer recurrence. In this review, we appraise the progress of RDR detection using ctDNA and consider the emerging role of liquid biopsy in the monitoring and management of solid tumors

Additional file 1: Figure S1. of Osteopontin and thrombospondin-1 play opposite roles in promoting tumor aggressiveness of primary resected non-small cell lung cancer

(A). Graphical representation of the ELISA data for OPN serum levels in healthy donors (n = 20) and population study (n = 171). Figure S1 (B). Graphical representation of the ELISA data for TSP-1 serum levels in healthy donors (n = 20) and population study (n = 171). (DOCX 154 kb