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| Status |
Public on Aug 03, 2020 |
| Title |
Insulin-like growth factor 2 mRNA-binding protein 3 modulates aggressiveness of Ewing sarcoma by regulating the CD164-CXCR4 axis |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Ewing sarcoma (EWS) is the second most common bone and soft tissue-associated malignancy in children and young adults. It is driven by the fusion oncogene EWS/FLI1 and characterized by rapid growth and early metastasis. We have previously discovered that the mRNA binding protein IGF2BP3 constitutes an important biomarker for EWS as high expression of IGF2BP3 in primary tumors predicts poor prognosis of EWS patients. We additionally demonstrated that IGF2BP3 enhances anchorage-independent growth and migration of Ewing sarcoma cells suggesting that IGF2BP3 might work as molecular driver and predictor of EWS progression.
The aim of this study was to further define the role of IGF2BP3 in EWS progression. We demonstrated that high IGF2BP3 mRNA expression levels correlated with EWS metastasis and disease progression in well-characterized EWS tumor specimens. EWS tumors with high IGF2BP3 levels were characterized by a specific gene signature enriched in chemokine-mediated signaling pathways. We also discovered that IGF2BP3 regulated the expression of CXCR4 through CD164.
We identified for the first time a tumorigenic axis consisting of IGF2BP3/CD164 and CXCR4, which confers migratory advantage to EWS cells, particularly under stress-adaptive conditions.
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| Overall design |
Specimens from 48 primary localized EWS tumors and 51 EWS metastatic lesions were analyzed by quantitative RT-PCR (qRT-PCR) to establish expression levels and correlations between IGF2BP3, CD164 and CXCR4. 50 paraffin-embedded primary localized EWS specimens were mounted as tissue microarrays and processed for immunohistochemical evaluation of IGF2BP3, CD164 and CXCR4 for validation of gene expression results. 6 primary localized EWS specimens characterized by high (3 samples) or low (3 samples) expression of IGF2BP3 were profiled by RNAseq. Enrichment analysis of differentially expressed genes was performed using MetaCore software (GeneGo, Thomson Reuters). CXCR4 expression was investigated in IGF2BP3-depleted or empty vector-transfected EWS cells using RT2 Profiler Cancer Inflammation and Immunity Crosstalk PCR Array, qRT-PCR and western blotting for validation of RNAseq results. CD164 expression was assessed by western blot in stable IGF2BP3 knockdown EWS cellular models compared to controls while ribo-immunoprecipitation assay and qRT-PCR were used to test IGF2BP3-CD164 interaction. We used confocal microscopy analysis to asses colocalization between CD164 and CXCR4 in response to CXCL12 stimulation. Cell migration in response to CXCL12 stimulation was investigated after treatment with anti-CD164 siRNA or controls and in stable IGF2BP3 knockdown EWS cellular models in hypoxic and normal conditions to asses functional relevance of IGF2BP3/CD164/CXCR4 axis.
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| Contributor(s) |
Mancarella C, Caldoni G, Ribolsi I, Parra A, Manara M, Mercurio AM, Morrione A, Scotlandi K |
| Citation(s) |
32719743 |
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| Submission date |
May 17, 2020 |
| Last update date |
Aug 03, 2020 |
| Contact name |
Katia Scotlandi |
| E-mail(s) |
katia.scotlandi@ior.it
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| Organization name |
IRCCS Istituto Ortopedico Rizzoli
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| Lab |
Laboratory of Experimental Oncology
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| Street address |
via di Barbiano 1/10
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| City |
Bologna |
| ZIP/Postal code |
40136 |
| Country |
Italy |
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| Platforms (1) |
| GPL15456 |
Illumina HiScanSQ (Homo sapiens) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA633384 |
| SRA |
SRP262048 |
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