Wheeler Lindsay J, Watson Zachary L, Qamar Lubna, Yamamoto Tomomi M, Sawyer Brandon T, Sullivan Kelly D, Khanal Santosh, Joshi Molishree, Ferchaud-Roucher Veronique, Smith Harry, Vanderlinden Lauren A, Brubaker Sky W, Caino Cecilia M, Kim Hyunmin, Espinosa Joaquin M, Richer Jennifer K, Bitler Benjamin G
Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Colorado School of Medicine, Aurora, CO 80045, USA.
Division of Reproductive Sciences, Department of Obstetrics and Gynecology, University of Colorado School of Medicine, 12700 E. 19(th) Avenue, MS 8613, Aurora, CO 80045, USA.
iScience. 2019 Sep 27;19:474-491. doi: 10.1016/j.isci.2019.07.049. Epub 2019 Aug 6.
High-grade serous ovarian cancers (HGSOCs) arise from exfoliation of transformed cells from the fallopian tube, indicating that survival in suspension, and potentially escape from anoikis, is required for dissemination. We report here the results of a multi-omic study to identify drivers of anoikis escape, including transcriptomic analysis, global non-targeted metabolomics, and a genome-wide CRISPR/Cas9 knockout (GeCKO) screen of HGSOC cells cultured in adherent and suspension settings. Our combined approach identified known pathways, including NOTCH signaling, as well as novel regulators of anoikis escape. Newly identified genes include effectors of fatty acid metabolism, ACADVL and ECHDC2, and an autophagy regulator, ULK1. Knockdown of these genes significantly inhibited suspension growth of HGSOC cells, and the metabolic profile confirmed the role of fatty acid metabolism in survival in suspension. Integration of our datasets identified an anoikis-escape gene signature that predicts overall survival in many carcinomas.
高级别浆液性卵巢癌(HGSOCs)起源于输卵管中转化细胞的脱落,这表明细胞在悬浮状态下存活并可能逃避失巢凋亡是其扩散所必需的。我们在此报告一项多组学研究的结果,以确定失巢凋亡逃逸的驱动因素,包括转录组分析、全局非靶向代谢组学,以及对在贴壁和悬浮条件下培养的HGSOC细胞进行全基因组CRISPR/Cas9基因敲除(GeCKO)筛选。我们的综合方法确定了包括NOTCH信号通路在内的已知途径,以及失巢凋亡逃逸的新调节因子。新鉴定的基因包括脂肪酸代谢效应器ACADVL和ECHDC2,以及自噬调节因子ULK1。敲低这些基因显著抑制了HGSOC细胞的悬浮生长,代谢谱证实了脂肪酸代谢在悬浮存活中的作用。我们数据集的整合确定了一个失巢凋亡逃逸基因特征,该特征可预测许多癌症的总生存期。