Abstract 4795: A novel gene fusion in glioblastoma and a radiation response methylation signature identified by genomic characterization of glioma sphere-forming cells
Authors: Wang Qianghu , Ezhilarasan Ravesanker , Goodman Lindsey D. , Gumin Joy , Zheng Siyuan , Yoshihara Kosuke , Sun Peng , Yang Jie , Heffernan Tim , Draetta Giulio , Aldape Kenneth D. , Lang Frederick F. , Verhaak Roel G.W. , Sulman Erik P. 
Published: August 2015

Abstract
Purpose: High fidelity models of the lethal primary brain tumor glioblastoma (GBM) are essential to develop new therapies. Glioma sphere-forming cells (GSCs) are derived from surgical specimens and are thought to play important roles in tumor maintenance and treatment resistance. We performed genomic characterization of the largest reported panel of GSCs. We hypothesized that GSCs would recapitulate the genomic alterations of their GBMs of origin while identifying novel changes identifiable only in a pure tumor cell population.
Methods: All GSCs were obtained at the time of surgical resection and all analyses were conducted at early passage. We performed exome and transcriptome sequencing, DNA methylation profiling (Illumina Infinium 450K Bead Arrays) and DNA copy number determination (Affymetrix OncoScan). Radiation (RT) sensitivity was determined by clonogenic survival and in vivo survival by orthotopic xenograft.
Results: We analyzed 43 GSCs, 40 of which had tissue available from their tumors of origin. Somatically mutated genes previously described in GBM, such as TP53, EGFR, PTEN, NF1, PIK3CA and RB1, were found at similar mutation frequencies. Likewise, DNA copy number variations were similar to their matched tumor and those reported by the TCGA, with novel or more pronounced alterations, such as MYC application and QKI deletion, identified in the GSCs. GSCs were classified into TCGA GBM subtypes by expression signatures, identifying a subset of GSCs with a subtype differing from their matched tumors that correlated to decreased stromal enrichment. GSCs exhibited upregulation of self-renewal pathways, such as MYC, WNT, and NOTCH, and of stem-cell factors, such as MSI1, NESTIN, OLIG2, and SOX2, consistent with the stem-like phenotype attributed to GSCs. Transcript analyses identified the previously reported FGFR3-TACC3 and EGFR-SEPT14 gene fusions as well as a novel KIF1B-KMT2A (MLL) fusion, which was found to have been retained in the matching recurrent GBM as well as the GSC derived from the recurrence. A signature derived by the differential methylation pattern of RT sensitive vs. resistant GSCs was applied to the subset of TCGA cases that received upfront RT. Survival by methylation class in this subset was significantly different (median survival 84 vs. 61 weeks; HR 1.64 adjusting for patient age, p-value&lt;0.008), suggesting this signature is predictive of clinical RT response.
Conclusions: Based on genomic analyses, GSCs are robust models of GBM which can be used for therapeutic development. We have identified a novel gene fusion involving MLL with a predicted driving role suggesting a new mode of gliomagenesis. A methylation signature predictive of RT response may have potential for personalizing RT treatment of GBM patients and provides insights into RT sensitivity phenotypes.
Citation Format: Qianghu Wang, Ravesanker Ezhilarasan, Lindsey D. Goodman, Joy Gumin, Siyuan Zheng, Kosuke Yoshihara, Peng Sun, Jie Yang, Tim Heffernan, Giulio Draetta, Kenneth D. Aldape, Frederick F. Lang, Roel G.W. Verhaak, Erik P. Sulman. A novel gene fusion in glioblastoma and a radiation response methylation signature identified by genomic characterization of glioma sphere-forming cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4795. doi:10.1158/1538-7445.AM2015-4795