2015;33:4106C4111

2015;33:4106C4111. or real and combined types. In conclusion, although patients with SCLC and LCNEC may benefit from target therapy, they were not identifiable by clinicopathologic background. And there was not significant genetic difference between SCLC and LCNEC, including between real and combined types. Classifying SCLC and LCNEC in same category is usually affordable. However, distinguishing the real type from combined type was not validated. Comprehensive genetic analysis should be performed to detect targetable variants in any type of SCLC and LCNEC. and were frequently found. As therapeutically targetable variants, mutations in (L858R), (G12D, G12A, G12V), and (E545K) were detected in 5 cases. The mutation (E545K) was not detected by NGS in 1 case (case 28), but was found by droplet digital polymerase chain reaction (ddPCR). One case (case 22) harbored both G12V and E545K mutations (Physique ?(Figure11). Open in a separate window Physique 1 Result of next-generation sequencing and immunohistochemistry analysisHistological subtype is usually around the horizontal axis. Around the vertical axis, accompanied component, sex, smoking history, case number, examined gene by next generation sequencing, and examined proteins by immunohistochemistry are shown. According to the histological difference (SCLC or LCNEC) or intra-histological heterogeneity (real type or combined type), cases can be divided into 4 subgroups. However, the frequency of genetic variants was not difference between any subtypes. There was also no statistically significant difference in the frequency of any genetic variance between SCLC and LCNEC, including when comparing the real and combined types and cases with and without targetable variants. The statistical non-significance of the comparisons of SCLC and LCNEC did not switch when the analysis was limited to missense mutations, which was the most common type of variant (Physique ?(Figure2).2). Immunohistochemical status did not differ significantly in frequencies of genetic variants (Physique ?(Figure3).3). Immunohistochemical profiling also did not show distinguishing features between SCLC and LCNEC, including when comparing the real and combined types and cases with and without targetable variants (Physique ?(Figure44). Open in a separate window Physique 2 Comparison of genetic variant number concerning histological and/or component type or therapeutic-targetThe frequency of all genetic variants was compared among real small cell lung malignancy (SCLC), real large cell neuroendrocrine carcinoma (LCNEC), combined SCLC, and combined LCNEC (A). Frequency of genetic variants between p/c SCLC and p/c LCNEC (B), real S/L and combined S/L (C), and cases with and without therapeutic targets (D). The frequency of missense mutations was also compared among real SCLC, combined SCLC, real LCNEC, and combined LCNEC (E). Frequency of missense mutations between p/c SCLC and p/c LCNEC (F), real S/L and combined S/L (G), and cases with and without therapeutic targets (H). Data are shown as the boundaries of the 10th, 25th, 50th, 75th, and 90th percentiles. Abbreviations: p, real; c, combined; S, small cell lung malignancy; L, large cell neuroendocrine carcinoma; *NS, no significance between any subtypes; NS, no significance. Open in a separate window Physique 3 Quantity of genetic variants according to immunohistochemical profileData are shown as the boundaries of the 10th, 25th, 50th, 75th, and 90th percentiles. Abbreviations: CgA, chromogranin A; Syp, synaptophysin. Open in a separate window Physique 4 Percentage of positive cases by immunohistochemistry grouped by histological and/or component type or therapeutic-targetThe ratio of positive staining of the indicated antibodies was compared among real small cell lung malignancy (SCLC), combined SCLC, real large cell PNRI-299 neuroendocrine carcinoma (LCNEC), and combined LCNEC (A). The ratio of positive staining between p/c SCLC and p/c LCNEC (B), real S/L and combined S/L (C), and cases with and without therapeutic targets (D). For each antibody, there was no significant difference in positive frequency in any of the comparisons. Abbreviations: p, real; c, combined; S, small cell lung malignancy; L, large cell neuroendocrine carcinoma. Apart from the L858R mutation, PNRI-299 which was confirmed by an external examining body during a patients clinical course, therapeutically targetable variants detected by NGS were validated by ddPCR (Figure ?(Figure5).5). The mutations G12D, G12A, and G12V were confirmed by the ddPCR screening multiplex kit (186-3506, BIO-RAD). The E545K mutation was confirmed by ddPCR with locked nucleic acid probes. Polymerase chain reaction (PCR) conditions and the sequence of each primer and probe are shown in Supplementary Table 1. Open in a separate window Figure 5 Results of droplet digital polymerase chain reaction of each variantEach amplified mutation of G12D (A), G12A (B), G12V (C),.Sequist LV, Waltman BA, Dias-Santagata D, Digumarthy S, Turke AB, Fidias P, Bergethon K, Shaw AT, Gettinger S, Cosper AK, Akhavanfard S, Heist RS, Temel J, et al. combined types. In conclusion, although patients with SCLC and LCNEC may benefit from target therapy, they were not identifiable by clinicopathologic background. And there was not significant genetic difference between SCLC and LCNEC, including between pure and combined types. Classifying SCLC and LCNEC in same category is reasonable. However, distinguishing the pure type from combined type was not validated. Comprehensive genetic analysis should be performed to detect targetable variants in any type of SCLC and LCNEC. and were frequently found. As therapeutically targetable variants, mutations in (L858R), (G12D, G12A, G12V), and (E545K) were detected in 5 cases. The mutation (E545K) was not detected by NGS in 1 case (case 28), but was found by droplet digital polymerase chain reaction (ddPCR). One case (case 22) harbored both G12V and E545K mutations (Figure ?(Figure11). Open in a separate window Figure 1 Result of next-generation sequencing and immunohistochemistry analysisHistological subtype is on the horizontal axis. On the vertical axis, accompanied component, sex, smoking history, case number, examined gene by next generation sequencing, and examined proteins by immunohistochemistry are shown. According to the histological difference (SCLC or LCNEC) or intra-histological heterogeneity (pure type or combined type), cases can be divided into 4 subgroups. However, the frequency of genetic variants was not difference between any subtypes. There was also no statistically significant difference in the frequency of any genetic variation between SCLC and LCNEC, including PNRI-299 when comparing the pure and combined types and cases with and without targetable variants. The statistical non-significance of the comparisons of SCLC and LCNEC did not change when the analysis was limited to missense mutations, which was the most common type of variant (Figure ?(Figure2).2). Immunohistochemical status did not differ significantly in frequencies of genetic variants (Figure ?(Figure3).3). Immunohistochemical profiling also did not show distinguishing features between SCLC and LCNEC, including when comparing the pure and combined types and cases with and without targetable variants (Figure ?(Figure44). Open in a separate window Figure 2 Comparison of genetic variant number concerning histological and/or component type or therapeutic-targetThe MCMT frequency of all genetic variants was compared among pure small cell lung cancer (SCLC), pure large cell neuroendrocrine carcinoma (LCNEC), combined SCLC, and combined LCNEC (A). Frequency of genetic variants between p/c SCLC and p/c LCNEC (B), pure S/L and combined S/L (C), and cases with and without therapeutic targets (D). The frequency of missense mutations was also compared among pure SCLC, combined SCLC, pure LCNEC, and combined LCNEC (E). Frequency of missense mutations between p/c SCLC and p/c LCNEC (F), pure S/L and combined S/L (G), and cases with and without therapeutic targets (H). Data are shown as the boundaries of the 10th, 25th, 50th, 75th, and 90th percentiles. Abbreviations: p, pure; c, combined; S, small cell lung cancer; L, large cell neuroendocrine carcinoma; *NS, no significance between any subtypes; NS, no significance. Open in a separate window Figure 3 Number of genetic variants according to immunohistochemical profileData are shown as the boundaries of the 10th, 25th, 50th, 75th, and 90th percentiles. Abbreviations: CgA, chromogranin A; Syp, synaptophysin. Open in a separate window Figure 4 Percentage of positive cases by immunohistochemistry grouped by histological and/or component type or therapeutic-targetThe ratio of positive staining of the indicated antibodies was compared among pure small cell lung cancer (SCLC), combined SCLC, pure large cell neuroendocrine carcinoma (LCNEC), and combined LCNEC (A). The ratio of positive staining between p/c SCLC and p/c LCNEC (B), pure S/L and combined S/L (C), and cases with and without therapeutic targets (D). For each antibody, there was no significant difference in positive frequency in any of the comparisons. Abbreviations: p, pure; c, combined; S, small cell lung cancer; L, large cell neuroendocrine carcinoma. Apart from the L858R mutation, which was confirmed by an external examining body during a patients clinical course, therapeutically targetable variants detected by NGS were validated by ddPCR (Figure ?(Figure5).5). The mutations G12D, G12A, and G12V were confirmed by the.