VAMP2–NRG1 Fusion Gene is a Novel Oncogenic Driver of Non–Small-Cell Lung Adenocarcinoma Yeonjoo Jung, PhD, Seunghui Yong, BS, Pora Kim, PhD, Hee-Young.

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VAMP2–NRG1 Fusion Gene is a Novel Oncogenic Driver of Non–Small-Cell Lung Adenocarcinoma Yeonjoo Jung, PhD, Seunghui Yong, BS, Pora Kim, PhD, Hee-Young Lee, MS, Yeonhwa Jung, BS, Juhee Keum, MS, Sanghyuk Lee, PhD, Jhingook Kim, MD, Jaesang Kim, PhD Journal of Thoracic Oncology Volume 10, Issue 7, Pages 1107-1111 (July 2015) DOI: 10.1097/JTO.0000000000000544 Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 A, Alignment of sequence reads mapping to vesicle-associated membrane protein 2 (VAMP2) and Neuregulin 1 (NRG1) from RNA-seq data. The sequence of the assembled fusion transcript appears at the top, and individual reads are below. The vertical line indicates the fusion point. Sequences to the left matches Exon 4 of VAMP2 and to the right matches Exon 5 of NRG1. B, A schematic of fusion genes with exons from VAMP2 and NRG1. Expected RT-PCR product is seen only from tumor (T) but not adjacent normal (N) tissues. Sequence traces from Sanger sequencing show the nucleotides around the exon–exon boundary. Journal of Thoracic Oncology 2015 10, 1107-1111DOI: (10.1097/JTO.0000000000000544) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 A, Schematic illustrations of vesicle-associated membrane protein 2 (VAMP2)–Neuregulin 1-I-β3 (NRG1-I-β3) (left) and VAMP2–NRG1-I-γ (right) proteins. Note that the trans-membrane is provided by VAMP2 and that V5 epitope is in the C-terminus of each protein. B, Live V5 staining images of H1568 cells after retroviral transduction. Top row panels show control virus-infected cells with only green fluorescent protein (GFP) signal. Middle row panels and bottom row panels show cells infected, respectively, with VAMP2–NRG1-I-β3 and VAMP2–NRG1-I-γ. Note V5 staining in GFP positive cells. Journal of Thoracic Oncology 2015 10, 1107-1111DOI: (10.1097/JTO.0000000000000544) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 A, Immunoblotting for ERBB2/ERBB3 signaling cascade proteins. Note that phosphorylation levels of ERBB2, ERBB3, AKT, and ERK increased only with VAMP2–NRG1-I-β3 virus. B, Representative images from soft agar assays. Note the increase in the number of colonies with VAMP2–NRG1-I-β3 virus. C, Results from soft agar assay are quantitated. Bars represent the average number of colonies, and error bars represent SD of three independent experiments. * represents p value less than 0.05. Journal of Thoracic Oncology 2015 10, 1107-1111DOI: (10.1097/JTO.0000000000000544) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions