American Association for Cancer Research
10780432ccr151627t-sup-152484_1_supp_3149951_n993k9.xlsx (23.62 kB)

Supplemental Table 1 from Bias-Corrected Targeted Next-Generation Sequencing for Rapid, Multiplexed Detection of Actionable Alterations in Cell-Free DNA from Advanced Lung Cancer Patients

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posted on 2023-03-31, 19:08 authored by Cloud P. Paweletz, Adrian G. Sacher, Chris K. Raymond, Ryan S. Alden, Allison O'Connell, Stacy L. Mach, Yanan Kuang, Leena Gandhi, Paul Kirschmeier, Jessie M. English, Lee P. Lim, Pasi A. Jänne, Geoffrey R. Oxnard

Supplemental Table 1: Summary of probe sequences


Department of Defense, Conquer Cancer Foundation of ASCO, Phi Beta Psi Sorority, Stading-Younger Cancer Research Foundation, Expect Miracles Foundation, Harold and Gail Kirstein Lung Cancer Research Fund, and US National Institutes of Health



Purpose: Tumor genotyping is a powerful tool for guiding non–small cell lung cancer (NSCLC) care; however, comprehensive tumor genotyping can be logistically cumbersome. To facilitate genotyping, we developed a next-generation sequencing (NGS) assay using a desktop sequencer to detect actionable mutations and rearrangements in cell-free plasma DNA (cfDNA).Experimental Design: An NGS panel was developed targeting 11 driver oncogenes found in NSCLC. Targeted NGS was performed using a novel methodology that maximizes on-target reads, and minimizes artifact, and was validated on DNA dilutions derived from cell lines. Plasma NGS was then blindly performed on 48 patients with advanced, progressive NSCLC and a known tumor genotype, and explored in two patients with incomplete tumor genotyping.Results: NGS could identify mutations present in DNA dilutions at ≥0.4% allelic frequency with 100% sensitivity/specificity. Plasma NGS detected a broad range of driver and resistance mutations, including ALK, ROS1, and RET rearrangements, HER2 insertions, and MET amplification, with 100% specificity. Sensitivity was 77% across 62 known driver and resistance mutations from the 48 cases; in 29 cases with common EGFR and KRAS mutations, sensitivity was similar to droplet digital PCR. In two cases with incomplete tumor genotyping, plasma NGS rapidly identified a novel EGFR exon 19 deletion and a missed case of MET amplification.Conclusions: Blinded to tumor genotype, this plasma NGS approach detected a broad range of targetable genomic alterations in NSCLC with no false positives including complex mutations like rearrangements and unexpected resistance mutations such as EGFR C797S. Through use of widely available vacutainers and a desktop sequencing platform, this assay has the potential to be implemented broadly for patient care and translational research. Clin Cancer Res; 22(4); 915–22. ©2015 AACR.See related commentary by Tsui and Berger, p. 790

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