PMC7734064_01
Chest computed tomography (CT) for the nodules in the lung lobes and the hematoxylin-eosin staining revealed the lung adenocarcinoma cells in the three primary sites (100microm). Three light red circles show the schematic location of the nodules. (A) and (B) for the nidus in the right upper lobe carrying EGFR 19del, (C) and (D) for the nidus in the right middle lobe carrying EGFR 20ins, and (E) and (F) for the nidus in the right middle lobe carrying ROS1 fusion.
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Chest computed tomography (CT) revealed one lesion in the right upper lobe and two lesions in the right middle lobe on June 16, 2019 (Figure 1A, C and E). Immunohistochemical staining in the post-operative pathology indicated that three nodules of the patient all was clinically diagnosed with stage IA minimally invasive lung adenocarcinoma, T1N0M0 (Figure 1B, D and F). EGFR exon 19 deletion (c.2240-2260delinsCAAGAG) (Figure 2A) was detected at a mutant allele frequency (MAF) of 1.13% in the nidus (0.5 cm in diameter) of the right upper lobe (Figure 1A). EGFR exon 20 insertion (c.2311-2312delinsGGGTT) (Figure 2A) was identified at a MAF of 5.13% in one nodule (0.4 cm in diameter) in the right middle lobe (Figure 1C). And EZR exon 10-ROS1 exon 34 (EZR-ROS1) rearrangement (Figure 2B) was detected at DNA level in the other nodule (0.6 cm in diameter) in the right middle lobe (Figure 1E) and confirmed by amplification RNA sequencing with 7574 mutation fragments (Figure 2C).
NGS targeting gene test on the trilateral surgical tumor tissue. (A) Schematic diagram of EGFR 19del and 20ins. (B) Schematic diagram of EZR-ROS1 fusion. (C) Integrative Genomics Viewer (IGV) snapshot of EZR-ROS1 fusion regions, which was detected by NGS sequencing reads.
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The surgical tumor tissues were subjected to target capture-based clinical next-generation sequencing (NGS) test targeting 31 cancer-relevant genes (Berry Oncology), and three independent driver mutations had been found in three nidi, respectively (Figure 2). EGFR exon 19 deletion (c.2240-2260delinsCAAGAG) (Figure 2A) was detected at a mutant allele frequency (MAF) of 1.13% in the nidus (0.5 cm in diameter) of the right upper lobe (Figure 1A). EGFR exon 20 insertion (c.2311-2312delinsGGGTT) (Figure 2A) was identified at a MAF of 5.13% in one nodule (0.4 cm in diameter) in the right middle lobe (Figure 1C). And EZR exon 10-ROS1 exon 34 (EZR-ROS1) rearrangement (Figure 2B) was detected at DNA level in the other nodule (0.6 cm in diameter) in the right middle lobe (Figure 1E) and confirmed by amplification RNA sequencing with 7574 mutation fragments (Figure 2C).