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IGF2BP1 regulates the cargo of extracellular vesicles and promotes neuroblastoma metastasis

Oncogene volume 42, pages 1558–1571 (2023)Cite this article

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Abstract

Neuroblastoma is a highly metastatic cancer, and thus is one of the leading causes of cancer-related mortalities in pediatric patients. More than 50% of NB cases exhibit 17q21-ter partial chromosomal gain, which is independently associated with poor survival, suggesting the clinical importance of genes at this locus in NB. IGF2BP1 is one such proto-oncogene located at 17q locus, and was found to be upregulated in patients with metastatic NBs. Here, utilizing multiple immunocompetent mouse models, along with our newly developed highly metastatic NB cell line, we demonstrate the role of IGF2BP1 in promoting NB metastasis. Importantly, we show the significance of small extracellular vesicles (EVs) in NB progression, and determine the pro-metastatic function of IGF2BP1 by regulating the NB-EV-protein cargo. Through unbiased proteomic analysis of EVs, we discovered two novel targets (SEMA3A and SHMT2) of IGF2BP1, and reveal the mechanism of IGF2BP1 in NB metastasis. We demonstrate that IGF2BP1 directly binds and governs the expression of SEMA3A/SHMT2 in NB cells, thereby modulating their protein levels in NB-EVs. IGF2BP1-affected levels of SEMA3A and SHMT2 in the EVs, regulate the formation of pro-metastatic microenvironment at potential metastatic organs. Finally, higher levels of SEMA3A/SHMT2 proteins in the EVs derived from NB-PDX models indicate the clinical significance of the two proteins and IGF2BP1-SEMA3A/SHMT2 axis in NB metastasis.

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Fig. 1: Generation and characterization of a novel highly metastatic transplantable mouse NB cell line - M1.
Fig. 2: IGF2BP1 promotes metastasis of NB and reduces survival in mouse models.
Fig. 3: IGF2BP1 does not impact the pro-metastatic properties of highly metastatic M1 cells in vitro.
Fig. 4: IGF2BP1 promotes metastasis in NB via small extracellular vesicles (EVs).
Fig. 5: IGF2BP1-regulated proteins SEMA3A and SHMT2 induce pre-metastatic niche in an EV-mediated manner.
Fig. 6: IGF2BP1 increases the expression of SEMA3A and SHMT2 by stabilizing their mRNAs.

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Data availability

Data generated in the study are available within the article and its Supplementary data files. Raw EV-mass spectrometry data are available upon request from the corresponding author.

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Acknowledgements

The authors are thankful to Dr. Paul Sondel’s lab for the generous gift of reagents, Venkatesha Basrur and University of Michigan mass-spectrometry core for help with EV-mass spec. We also thank Joe Bednarczyk and Flow-Cytometry Core, Han Chen and TEM Facility, and Molecular & Histopathology Core services at Penn State College of Medicine for help with cell-sorting, TEM and tissue mounting, respectively.

Funding

This study was supported by the NIH grant R01 CA243167 (VSS), and Four Diamonds Fund (VSS).

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Authors and Affiliations

  1. Division of Pediatric Hematology and Oncology, Department of Pediatrics, The Pennsylvania State University College of Medicine, Hershey, PA, USA

    Mayura R. Dhamdhere, Chethana P. Gowda, Vikash Singh, Arati Sharma, Sinisa Dovat, Hong-Gang Wang & Vladimir S. Spiegelman

  2. Department of Public Health Sciences, The Pennsylvania State University College of Medicine, Penn State Cancer Institute, Hershey, PA, USA

    Zhenqiu Liu

  3. Bioinformatics Core, University of Michigan Medical School, Ann Arbor, MI, USA

    Nicholas Carruthers

  4. Division of Pediatric Surgery, Department of Surgery, The Pennsylvania State University College of Medicine, Hershey, PA, USA

    Christa N. Grant

  5. Department of Ophthalmology, The Pennsylvania State University College of Medicine, Hershey, PA, USA

    Jeffrey M. Sundstrom

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  1. Mayura R. Dhamdhere
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Contributions

MRD: study design, data curation, methodology, formal analysis, validation, investigation, visualization, writing-original draft, revision; CPG, VS: methodology, data curation; NC, ZL: data curation, bioinformatic analysis; CNG: manuscript reviewing, data curation; JMS, AS, SD: resources, manuscript reviewing; HGW: manuscript reviewing; VSS: conception and design, study supervision, funding acquisition, writing-review & revision, project administration.

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Correspondence to Vladimir S. Spiegelman.

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Dhamdhere, M.R., Gowda, C.P., Singh, V. et al. IGF2BP1 regulates the cargo of extracellular vesicles and promotes neuroblastoma metastasis. Oncogene 42, 1558–1571 (2023). https://doi.org/10.1038/s41388-023-02671-0

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