Phosphoproteomic Profiling Reveals IL6-Mediated Paracrine Signaling within the Ewing Sarcoma Family of Tumors

被引:20
作者
Anderson, Jennifer L. [1 ,2 ]
Titz, Bjoern [3 ,4 ,5 ]
Akiyama, Ryan [2 ]
Komisopoulou, Evangelia [3 ,4 ,5 ]
Park, Ann [2 ]
Tap, William D. [6 ,7 ]
Graeber, Thomas G. [3 ,4 ,5 ,8 ,9 ]
Denny, Christopher T. [1 ,2 ,4 ,8 ]
机构
[1] Univ Calif Los Angeles, Inst Mol Biol, Los Angeles, CA 90095 USA
[2] Univ Calif Los Angeles, Dept Pediat, Div Hematol Oncol, Gwynne Hazen Cherry Mem Labs, Los Angeles, CA 90095 USA
[3] Univ Calif Los Angeles, Crump Inst Mol Imaging, Los Angeles, CA 90095 USA
[4] Univ Calif Los Angeles, Jonsson Comprehens Canc Ctr, Los Angeles, CA 90095 USA
[5] Univ Calif Los Angeles, David Geffen Sch Med, Dept Mol & Med Pharmacol, Los Angeles, CA 90095 USA
[6] Mem Sloan Kettering Canc Ctr, Dept Med, Div Solid Tumors, Sarcoma Med Oncol Serv, New York, NY 10021 USA
[7] Weill Cornell Med Coll, Dept Med, New York, NY USA
[8] Univ Calif Los Angeles, Calif NanoSyst Inst, Los Angeles, CA 90095 USA
[9] Univ Calif Los Angeles, Metabol Ctr, Los Angeles, CA 90095 USA
关键词
CHILDRENS ONCOLOGY GROUP; LARGE GENE LISTS; STAT3; ACTIVATION; EWS/ETS FUSIONS; RAF INHIBITORS; TARGET GENE; BCR-ABL; CELLS; CANCER; RESISTANCE;
D O I
10.1158/1541-7786.MCR-14-0159
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Members of the Ewing sarcoma family of tumors (ESFT) contain tumor-associated translocations that give rise to oncogenic transcription factors, most commonly EWS/FLI1. EWS/FLI1 plays a dominant role in tumor progression by modulating the expression of hundreds of target genes. Here, the impact of EWS/FLI1 inhibition, by RNAi-mediated knockdown, on cellular signaling was investigated using mass spectrometry-based phospho-proteomics to quantify global changes in phosphorylation. This unbiased approach identified hundreds of unique phosphopeptides enriched in processes such as regulation of cell cycle and cytoskeleton organization. In particular, phosphotyrosine profiling revealed a large upregulation of STAT3 phosphorylation upon EWS/FLI1 knockdown. However, single-cell analysis demonstrated that this was not a cell-autonomous effect of EWS/FLI1 deficiency, but rather a signaling effect occurring in cells in which knockdown does not occur. Conditioned media from knockdown cells were sufficient to induce STAT3 phosphorylation in control cells, verifying the presence of a soluble factor that can activate STAT3. Cytokine analysis and ligand/receptor inhibition experiments determined that this activation occurred, in part, through an IL6-dependent mechanism. Taken together, the data support a model in which EWS/FLI1 deficiency results in the secretion of soluble factors, such as IL6, which activate STAT signaling in bystander cells that maintain EWS/FLI1 expression. Furthermore, these soluble factors were shown to protect against apoptosis. (C)2014 AACR.
引用
收藏
页码:1740 / 1754
页数:15
相关论文
共 50 条
[1]   Critical Role of STAT3 in IL-6-Mediated Drug Resistance in Human Neuroblastoma [J].
Ara, Tasnim ;
Nakata, Rie ;
Sheard, Michael A. ;
Shimada, Hiroyuki ;
Buettner, Ralf ;
Groshen, Susan G. ;
Ji, Lingyun ;
Yu, Hua ;
Jove, Richard ;
Seeger, Robert C. ;
DeClerck, Yves A. .
CANCER RESEARCH, 2013, 73 (13) :3852-3864
[2]   Biology of EWS/ETS fusions in Ewing's family tumors [J].
Arvand, A ;
Denny, CT .
ONCOGENE, 2001, 20 (40) :5747-5754
[3]   A phase 2 trial of trabectedin in children with recurrent rhabdomyosarcoma, Ewing sarcoma and non-rhab domyosarcoma soft tissue sarcomas: A report from the Children's Oncology Group [J].
Baruchel, Sylvain ;
Pappo, Alberto ;
Krailo, Mark ;
Baker, K. Scott ;
Wu, Bing ;
Villaluna, Doojduen ;
Lee-Scott, Michelle ;
Adamson, Peter C. ;
Blaney, Susan M. .
EUROPEAN JOURNAL OF CANCER, 2012, 48 (04) :579-585
[4]  
Behjati S, 2012, SARCOMA, V2012
[5]   Small-molecule screen identifies modulators of EWS/FLI1 target gene expression and cell survival in Ewing's sarcoma [J].
Boro, Aleksandar ;
Pretre, Kathya ;
Rechfeld, Florian ;
Thalhammer, Verena ;
Oesch, Susanne ;
Wachtel, Marco ;
Schaefer, Beat W. ;
Niggli, Felix K. .
INTERNATIONAL JOURNAL OF CANCER, 2012, 131 (09) :2153-2164
[6]   Stat3 activation is required for cellular transformation by v-src [J].
Bromberg, JF ;
Horvath, CM ;
Besser, D ;
Lathem, WW ;
Darnell, JE .
MOLECULAR AND CELLULAR BIOLOGY, 1998, 18 (05) :2553-2558
[7]  
Chaturvedi Aashi, 2012, Genes Cancer, V3, P102, DOI 10.1177/1947601912457024
[8]   Differential Disruption of EWS-FLI1 Binding by DNA-Binding Agents [J].
Chen, Changmin ;
Wonsey, Diane R. ;
Lemieux, Madeleine E. ;
Kung, Andrew L. .
PLOS ONE, 2013, 8 (07)
[9]   The quest to overcome resistance to EGFR-targeted therapies in cancer [J].
Chong, Curtis R. ;
Jaenne, Pasi A. .
NATURE MEDICINE, 2013, 19 (11) :1389-1400
[10]   QUANTITATIVE-ANALYSIS OF DOSE-EFFECT RELATIONSHIPS - THE COMBINED EFFECTS OF MULTIPLE-DRUGS OR ENZYME-INHIBITORS [J].
CHOU, TC ;
TALALAY, P .
ADVANCES IN ENZYME REGULATION, 1984, 22 :27-55