Coordinated Splicing of Regulatory Detained Introns within Oncogenic Transcripts Creates an Exploitable Vulnerability in Malignant Glioma

October 9, 2017

... rug DMSO control point) was plated in a 96-well plate 2-fold dilution steps, starting at 500 mM. 100 nl of the compound solutions were pin-transferred (V&P Scientific, CA, pin tool mounted onto Tecan Freedom Evo 150 MCA96 head, Tecan, CA) into 96-well plates and incubated for 120h. Cells were seeded in 96-plates on the day prior to drug treatment. Seeding cell numbers were adjusted for different c ... e no-drug DMSO control point) was plated in a 96-well plate 2-fold dilution steps, starting at 500 mM. 100 nl of the compound solutions were pin-transferred (V&P Scientific, CA, pin tool mounted onto Tecan Freedom Evo 150 MCA96 head, Tecan, CA) into 96-well plates and incubated for 120h. Cells were seeded in 96-plates on the day prior to drug treatment. Seeding cell numbers were adjusted for diffe

Glioblastoma (GBM) is a devastating malignancy with few therapeutic options. We identify PRMT5 in an in vivo GBM shRNA screen and show that PRMT5 knockdown or inhibition potently suppresses in vivo GBM tumors, including patient-derived xenografts. Pathway analysis implicates splicing in cellular PRMT5 dependency, and we identify a biomarker that predicts sensitivity to PRMT5 inhibition. We find that PRMT5 deficiency primarily disrupts the removal of detained introns (DIs). This impaired DI splicing affects proliferation genes, whose downregulation coincides with cell cycle defects, senescence and/or apoptosis. We further show that DI programs are evolutionarily conserved and operate during neurogenesis, suggesting that they represent a physiological regulatory mechanism. Collectively, these findings reveal a PRMT5-regulated DI-splicing program as an exploitable cancer vulnerability. Copyright © 2017 Elsevier Inc. All rights reserved.

Braun, CJ; Stanciu, M; Boutz, PL; Patterson, JC; Calligaris, D; Higuchi, F; Neupane, R; Fenoglio, S; Cahill, DP; Wakimoto, H; Agar, NYR; Yaffe, MB; Sharp, PA; Hemann, MT; Lees, JA;

Journal: Cancer Cell Pages: 411-426.e11

Original article (28966034)