IGF1 Is a Common Target Gene of Ewing's Sarcoma Fusion Proteins in Mesenchymal Progenitor Cells

Background The EWS-FLI-1 fusion protein is associated with 85–90% of Ewing's sarcoma family tumors (ESFT), the remaining 10–15% of cases expressing chimeric genes encoding EWS or FUS fused to one of several ets transcription factor family members, including ERG-1, FEV, ETV1 and ETV6. ESFT are dependent on insulin-like growth factor-1 (IGF-1) for growth and survival and recent evidence suggests that mesenchymal progenitor/stem cells constitute a candidate ESFT origin. Methodology/Principal Findings To address the functional relatedness between ESFT-associated fusion proteins, we compared mouse progenitor cell (MPC) permissiveness for EWS-FLI-1, EWS-ERG and FUS-ERG expression and assessed the corresponding expression profile changes. Whereas all MPC isolates tested could stably express EWS-FLI-1, only some sustained stable EWS-ERG expression and none could express FUS-ERG for more than 3–5 days. Only 14% and 4% of the total number of genes that were respectively induced and repressed in MPCs by the three fusion proteins were shared. However, all three fusion proteins, but neither FLI-1 nor ERG-1 alone, activated the IGF1 promoter and induced IGF1 expression. Conclusion/Significance Whereas expression of different ESFT-associated fusion proteins may require distinct cellular microenvironments and induce transcriptome changes of limited similarity, IGF1 induction may provide one common mechanism for their implication in ESFT pathogenesis.

[1]  G. Thomas,et al.  Gene fusion with an ETS DNA-binding domain caused by chromosome translocation in human tumours , 1992, Nature.

[2]  C. Denny,et al.  Ewing sarcoma 11;22 translocation produces a chimeric transcription factor that requires the DNA-binding domain encoded by FLI1 for transformation. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[3]  C. Denny,et al.  Multiple domains mediate transformation by the Ewing's sarcoma EWS/FLI-1 fusion gene. , 1995, Oncogene.

[4]  Marcienne M Wright,et al.  EWS/FLI1-induced manic fringe renders NIH 3T3 cells tumorigenic , 1997, Nature Genetics.

[5]  D. Leroith,et al.  The Insulin-like Growth Factor-I Receptor Is Required for EWS/FLI-1 Transformation of Fibroblasts* , 1997, The Journal of Biological Chemistry.

[6]  P. Rotwein,et al.  Essential promoter elements are located within the 5′ untranslated region of human insulin-like growth factor-I exon I , 1997, Molecular and Cellular Endocrinology.

[7]  P. Lollini,et al.  Blockage of insulin-like growth factor-I receptor inhibits the growth of Ewing's sarcoma in athymic mice. , 1998, Cancer research.

[8]  G. Childs,et al.  The Transcriptional Repressor ZFM1 Interacts with and Modulates the Ability of EWS to Activate Transcription* , 1998, The Journal of Biological Chemistry.

[9]  C. Denny,et al.  Divergent Ewing's sarcoma EWS/ETS fusions confer a common tumorigenic phenotype on NIH3T3 cells , 1999, Oncogene.

[10]  H. Chansky,et al.  EWS·Fli-1 Fusion Protein Interacts with Hyperphosphorylated RNA Polymerase II and Interferes with Serine-Arginine Protein-mediated RNA Splicing* , 2000, The Journal of Biological Chemistry.

[11]  T. Werner,et al.  Highly specific localization of promoter regions in large genomic sequences by PromoterInspector: a novel context analysis approach. , 2000, Journal of molecular biology.

[12]  S. Baker,et al.  The Splicing Factor U1C Represses EWS/FLI-mediated Transactivation* , 2000, The Journal of Biological Chemistry.

[13]  S. Welford,et al.  DNA Binding Domain-independent Pathways Are Involved in EWS/FLI1-mediated Oncogenesis* , 2001, The Journal of Biological Chemistry.

[14]  Christopher T Denny,et al.  Biology of EWS/ETS fusions in Ewing's family tumors , 2001, Oncogene.

[15]  C. Denny,et al.  Loss of p16 pathways stabilizes EWS/FLI1 expression and complements EWS/FLI1 mediated transformation , 2001, Oncogene.

[16]  T. Golub,et al.  Supplemental Information for , 2002 .

[17]  P. Lollini,et al.  Expression of an IGF‐I receptor dominant negative mutant induces apoptosis, inhibits tumorigenesis and enhances chemosensitivity in Ewing's sarcoma cells , 2002, International journal of cancer.

[18]  P. Lollini,et al.  Effectiveness of insulin-like growth factor I receptor antisense strategy against Ewing's sarcoma cells , 2002, Cancer Gene Therapy.

[19]  A. Fukamizu,et al.  Cooperative Interaction of EWS with CREB-binding Protein Selectively Activates Hepatocyte Nuclear Factor 4-mediated Transcription* , 2003, The Journal of Biological Chemistry.

[20]  P. Meltzer,et al.  Mechanisms of sarcoma development , 2003, Nature Reviews Cancer.

[21]  S. Chin,et al.  FUS/ERG gene fusions in Ewing's tumors. , 2003, Cancer research.

[22]  W. May,et al.  EWS/FLI function varies in different cellular backgrounds. , 2003, Experimental cell research.

[23]  Rainer Breitling,et al.  Rank products: a simple, yet powerful, new method to detect differentially regulated genes in replicated microarray experiments , 2004, FEBS letters.

[24]  O. Delattre,et al.  EWS/FLI-1 Silencing and Gene Profiling of Ewing Cells Reveal Downstream Oncogenic Pathways and a Crucial Role for Repression of Insulin-Like Growth Factor Binding Protein 3 , 2004, Molecular and Cellular Biology.

[25]  D. Latchman,et al.  The effects of Brn-3a on neuronal differentiation and apoptosis are differentially modulated by EWS and its oncogenic derivative EWS/Fli-1 , 2004, Oncogene.

[26]  D. Suvà,et al.  Non‐hematopoietic human bone marrow contains long‐lasting, pluripotential mesenchymal stem cells , 2004, Journal of cellular physiology.

[27]  P. Meltzer,et al.  Gene expression profiling of human sarcomas: insights into sarcoma biology. , 2005, Cancer research.

[28]  M. Wynes,et al.  Transcription of macrophage IGF-I exon 1 is positively regulated by the 5'-untranslated region and negatively regulated by the 5'-flanking region. , 2005, American journal of physiology. Lung cellular and molecular physiology.

[29]  W. Gerald,et al.  The EWS-WT1 gene fusion in desmoplastic small round cell tumor. , 2005, Seminars in cancer biology.

[30]  A. Trumpp,et al.  Development of Ewing's sarcoma from primary bone marrow-derived mesenchymal progenitor cells. , 2005, Cancer research.

[31]  J. Richardson,et al.  Expression of the EWS/FLI-1 oncogene in murine primary bone-derived cells Results in EWS/FLI-1-dependent, ewing sarcoma-like tumors. , 2005, Cancer research.

[32]  P. Lollini,et al.  Antitumor activity of the insulin-like growth factor-I receptor kinase inhibitor NVP-AEW541 in musculoskeletal tumors. , 2005, Cancer research.

[33]  Rainer Breitling,et al.  RankProd: a bioconductor package for detecting differentially expressed genes in meta-analysis , 2006, Bioinform..

[34]  O. Delattre,et al.  Mesenchymal stem cell features of Ewing tumors. , 2007, Cancer cell.

[35]  K. Yutzey,et al.  Developmental regulation of the mouse IGF-I exon 1 promoter region by calcineurin activation of NFAT in skeletal muscle. , 2007, American journal of physiology. Cell physiology.

[36]  Nicolò Riggi,et al.  Sarcomas: genetics, signalling, and cellular origins. Part 1: The fellowship of TET , 2007, The Journal of pathology.

[37]  Catherine M. Verfaillie,et al.  Pluripotency of mesenchymal stem cells derived from adult marrow , 2007, Nature.

[38]  M. Suvà,et al.  EWS-FLI-1 expression triggers a Ewing's sarcoma initiation program in primary human mesenchymal stem cells. , 2008, Cancer research.

[39]  D. Suvà,et al.  In vitro activated human T lymphocytes very efficiently attach to allogenic multipotent mesenchymal stromal cells and transmigrate under them , 2008, Journal of cellular physiology.