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Sunitinib and Axitinib increase secretion and glycolytic activity of small extracellular vesicles in renal cell carcinoma

Cancer Gene Therapy volume 29, pages 683–696 (2022)Cite this article

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Abstract

Extracellular vesicles (EVs) encompass a wide range of vesicles that are released by all cell types. They package protein, nucleic acids, metabolites, and other cargo that can be delivered to recipient cells and affect their phenotypes. However, little is known about how pharmaceutical agents can alter EV secretion, protein and metabolic cargo, and the active biological processes taking place in these vesicles. In this study, we isolated EVs from human renal cell carcinoma (RCC) cells treated with tyrosine kinase inhibitors (TKIs) Sunitinib and Axitinib. We found these TKIs increase the number of large (lEVs) and small extracellular vesicles (sEVs) secreted from RCC cells in a dose-dependent manner. In addition, quantitative proteomics revealed that metabolic proteins are enriched in sEVs secreted from Sunitinib-treated cells. In particular, the glucose transporter GLUT1 was enriched in sEVs purified from TKI-treated cells. These sEVs displayed increased glucose uptake and glycolytic metabolism compared to sEVs released from vehicle-treated cells. Overexpression of GLUT1 in RCC cells augmented GLUT1 levels in sEVs, which subsequently displayed higher glucose uptake and glycolytic activity. Together, these findings suggest that these TKIs alter metabolic cargo and activity in RCC sEVs.

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Fig. 1: Tyrosine kinase inhibitors Sunitinib and Axitinib augment EV secretion.
Fig. 2: Sunitinib increases GLUT1 packaging into RCC sEVs.
Fig. 3: GLUT1 expression is higher and correlates with poor patient survival in RCC.
Fig. 4: sEVs secreted from tyrosine kinase inhibitor-treated cells have increased glucose uptake and glycolytic activity.
Fig. 5: sEVs from cells overexpressing GLUT1 have increased glycolytic activity.

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Acknowledgements

We thank the members of the Rathmell and Weaver Laboratories for their support and technical expertise. We also thank the Vanderbilt Cell Imaging Shared Resource (CISR) for their technical expertise on electron microscopy and the Vanderbilt proteomics core for their technical expertise on proteomics. Finally, we thank the patients who agreed to participate in the neoadjuvant Pazopanib study.

Funding

This work was supported by the AACR (WKR, ARL), the Medical Scientist Training Program Grant T32GM007347 (ARL), P01CA229123 and R01CA249424 (AMW), and R01CA217987 (WKR). The Vanderbilt CISR and Mass Spectrometry Research Center receive support from the National Institutes of Health (P30CA068485).

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Authors and Affiliations

  1. Division of Hematology and Oncology, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA

    Aaron R. Lim & W. Kimryn Rathmell

  2. Program in Cancer Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    Aaron R. Lim & Alissa M. Weaver

  3. Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Benjamin G. Vincent

  4. Division of Hematology and Oncology, Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Benjamin G. Vincent

  5. Department of Microbiology and Immunology, Curriculum in Bioinformatics and Computational Biology, Computational Medicine Program, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Benjamin G. Vincent

  6. Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN, USA

    Alissa M. Weaver

  7. Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN, USA

    Alissa M. Weaver

  8. Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, TN, USA

    W. Kimryn Rathmell

  9. Vanderbilt Center for Immunobiology, Vanderbilt University Medical Center, Nashville, TN, USA

    W. Kimryn Rathmell

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  1. Aaron R. Lim
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Contributions

ARL performed the experiments, analyzed data, and wrote the manuscript. BGV provided RNAseq data from the neoadjuvant Pazopanib trial. AMW and WKR provided guidance on experimental design. BGV, AMW, and WKR edited the manuscript.

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Correspondence to W. Kimryn Rathmell.

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ARL, BGV, and AMW declare no direct conflict of interest with the contents of this manuscript. Within the past 2 years, WKR has received unrelated clinical research support Bristol-Meyers Squib, Merck, Pfizer, Peloton, Calithera, and Incyte.

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Lim, A.R., Vincent, B.G., Weaver, A.M. et al. Sunitinib and Axitinib increase secretion and glycolytic activity of small extracellular vesicles in renal cell carcinoma. Cancer Gene Ther 29, 683–696 (2022). https://doi.org/10.1038/s41417-021-00345-1

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