The softness of tumour-cell-derived microparticles regulates their drug-delivery efficiency

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Extracellular microparticles (MPs) can function as drug-delivery vehicles for anticancer drugs. Here, we show that the softness of MPs derived from tumour-repopulating cells (TRCs) isolated from three-dimensional fibrin gels enhances the MPs’ drug-delivery efficiency. We found that, compared with MPs derived from tumour cells cultured in conventional tissue-culture plastic, TRC-derived MPs intravenously injected in tumour-xenograft-bearing mice showed enhanced accumulation in tumour tissues, enhanced blood-vessel crossing and penetration into tumour parenchyma, and preferential uptake by highly tumorigenic TRCs. We also show that the cytoskeleton-related protein cytospin-A plays a critical role in the regulation of TRC-derived MP softness. The modulation of the mechanical properties of TRC-derived MPs could aid the efficiency of delivery of anticancer drugs.

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Fig. 1: Characterization of DOX@3D-MPs.
Fig. 2: Antitumour activity of drug-packaging 3D-MPs.
Fig. 3: Efficient tumour accumulation, extravasation and penetration of DOX@3D-MPs.
Fig. 4: Role of softness on the in vivo transport process of 3D-MPs.
Fig. 5: Involvement of cytospin-A in the regulation of softness and in vivo transport process of 3D-MPs.
Fig. 6: Involvement of cytospin-A in regulation of the anticancer activity of DOX-loaded MPs.

Data availability

The data that support the findings of this study are available within the paper and its Supplementary Information files. Raw data are available from the corresponding authors upon reasonable request.


  1. 1.

    Guillemard, V. & Saragovi, H. U. Novel approaches for targeted cancer therapy. Curr. Cancer Drug Targets 4, 313–326 (2004).

  2. 2.

    Wang, M. & Thanou, M. Targeting nanoparticles to cancer. Pharmacol. Res. 62, 90–99 (2010).

  3. 3.

    Peer, D. et al. Nanocarriers as an emerging platform for cancer therapy. Nat. Nanotech. 2, 751–760 (2007).

  4. 4.

    Maeda, H., Nakamura, H. & Fang, J. The EPR effect for macromolecular drug delivery to solid tumors: improvement of tumor uptake, lowering of systemic toxicity, and distinct tumor imaging in vivo. Adv. Drug Deliv. Rev. 65, 71–79 (2013).

  5. 5.

    Wang, H., Yu, J., Lu, X. & He, X. Nanoparticle systems reduce systemic toxicity in cancer treatment. Nanomedicine 11, 103–106 (2016).

  6. 6.

    Zuo, Z. Q. et al. Promoting tumor penetration of nanoparticles for cancer stem cell therapy by TGF-β signaling pathway inhibition. Biomaterials 82, 48–59 (2016).

  7. 7.

    Axelson, H., Fredlund, E., Ovenberger, M., Landberg, G. & Pahlman, S. Hypoxia induced dedifferentiation of tumor cells—a mechanism behind heterogeneity and aggressiveness of solid tumors. Semin. Cell Dev. Biol. 16, 554–563 (2005).

  8. 8.

    Li, Z. et al. Hypoxia inducible factors regulate tumorigenic capacity of glioma stem cells. Cancer Cell 15, 501–513 (2009).

  9. 9.

    Vader, P., Mol, E. A., Pasterkamp, G. & Schiffelers, R. M. Extracellular vesicles for drug delivery. Adv. Drug Deliv. Rev. 106, 148–156 (2016).

  10. 10.

    Tominaga, N., Yoshioka, Y. & Ochiya, T. A novel platform for cancer therapy using extracellular vesicles. Adv. Drug Deliv. Rev. 95, 50–55 (2015).

  11. 11.

    Erkan, E. P. & Saydam, O. Extracellular vesicles as novel delivery tools for cancer treatment. Curr. Cancer Drug Targets 16, 34–42 (2016).

  12. 12.

    György, B. et al. Detection and isolation of cell-derived microparticles are compromised by protein complexes resulting from shared biophysical parameters. Blood 117, 39–48 (2011).

  13. 13.

    Ratajczak, J. et al. Membrane-derived microvesicles: important and underappreciated mediators of cell-to-cell communication. Leukemia 20, 1487–1495 (2006).

  14. 14.

    Tang, K. et al. Delivery of chemotherapeutic drugs in tumour cell-derived microparticles. Nat. Commun. 3, 1282 (2012).

  15. 15.

    Ma, J. et al. Reversing drug resistance of soft tumor-repopulating cells by tumor cell-derived chemotherapeutic microparticles. Cell Res. 26, 713–727 (2016).

  16. 16.

    Hashizume, H. et al. Openings between defective endothelial cells explain tumor vessel leakiness. Am. J. Pathol. 156, 1363–1380 (2000).

  17. 17.

    Hida, K., Maishi, N., Sakurai, Y., Hida, Y. & Harashima, H. Heterogeneity of tumor endothelial cells and drug delivery. Adv. Drug Deliv. Rev. 99, 140–147 (2016).

  18. 18.

    Pries, A. R., Höpfner, M., le Noble, F., Dewhirst, M. W. & Secomb, T. W. The shunt problem: control of functional shunting in normal and tumour vasculature. Nat. Rev. Cancer 10, 587–593 (2010).

  19. 19.

    Maeda, H., Wu, J., Sawa, T., Matsumura, Y. & Hori, K. Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. J. Control. Release 65, 271–284 (2000).

  20. 20.

    Kobayashi, H., Turkbey, B., Watanabe, R. & Choyke, P. L. Cancer drug delivery: considerations in the rational design of nanosized bioconjugates. Bioconjug. Chem. 25, 2093–2100 (2014).

  21. 21.

    Wang, J. et al. The role of micelle size in tumor accumulation, penetration, and treatment. ACS Nano 9, 7195–7206 (2015).

  22. 22.

    Cabral, H. et al. Accumulation of sub-100 nm polymeric micelles in poorly permeable tumours depends on size. Nat. Nanotechnol. 6, 815–823 (2011).

  23. 23.

    Tang, L. et al. Investigating the optimal size of anticancer nanomedicine. Proc. Natl Acad. Sci. USA 11, 15344–15349 (2014).

  24. 24.

    Anselmo, A. C. & Mitragotri, S. Impact of particle elasticity on particle-based drug delivery systems. Adv. Drug Deliv. Rev. 108, 51–67 (2017).

  25. 25.

    Anselmo, A. C. et al. Elasticity of nanoparticles influences their blood circulation, phagocytosis, endocytosis, and targeting. ACS Nano 9, 3169–3177 (2015).

  26. 26.

    Key, J. et al. Soft discoidal polymeric nanoconstructs resist macrophage uptake and enhance vascular targeting in tumors. ACS Nano 9, 11628–11641 (2015).

  27. 27.

    Hartmann, R., Weidenbach, M., Neubauer, M., Fery, A. & Parak, W. J. Stiffness-dependent in vitro uptake and lysosomal acidification of colloidal particles. Angew. Chem. Int. Ed. Engl. 54, 1365–1368 (2015).

  28. 28.

    Sun, H. et al. The role of capsule stiffness on cellular processing. Chem. Sci. 6, 3505–3514 (2015).

  29. 29.

    Liu, J. et al. Soft fibrin gels promote selection and growth of tumorigenic cells. Nat. Mater. 11, 734–741 (2012).

  30. 30.

    Tan, Y. et al. Matrix softness regulates plasticity of tumour-repopulating cells via H3K9 demethylation and Sox2 expression. Nat. Commun. 5, 4619 (2014).

  31. 31.

    Discher, D. E., Mooney, D. J. & Zandstra, P. W. Growth factors, matrices, and forces combine and control stem cells. Science 324, 1673–1677 (2009).

  32. 32.

    Saadi, I. et al. Deficiency of the cytoskeletal protein SPECC1L leads to oblique facial clefting. Am. J. Hum. Genet. 89, 44–55 (2011).

  33. 33.

    Raggi, C. et al. Cholangiocarcinoma stem-like subset shapes tumor-initiating niche by educating associated macrophages. J. Hepatol. 66, 102–115 (2017).

  34. 34.

    Chen, J. et al. Efficient extravasation of tumor-repopulating cells depends on cell deformability. Sci. Rep. 6, 19304 (2016).

  35. 35.

    Lei, Z. et al. Regulation of HIF-1α and VEGF by miR-20b tunes tumor cells to adapt to the alteration of oxygen concentration. PLoS ONE 4, e7629 (2009).

  36. 36.

    Ho, M. M., Ng, A. V., Lam, S. & Hung, J. Y. Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res. 67, 4827–4833 (2007).

  37. 37.

    Nakanishi, T. et al. Side-population cells in luminal-type breast cancer have tumour-initiating cell properties, and are regulated by HER2 expression and signalling. Br. J. Cancer 102, 815–826 (2010).

  38. 38.

    Wang, H. X. et al. Surface charge critically affects tumor penetration and therapeutic efficacy of cancer nanomedicines. Nano Today 11, 133–144 (2016).

  39. 39.

    Zhou, M. et al. Shape regulated anticancer activities and systematic toxicities of drug nanocrystals in vivo. Nanomedicine 12, 181–189 (2016).

  40. 40.

    Holzinger, A. Jasplakinolide: an actin-specific reagent that promotes actin polymerization. Methods Mol. Biol. 586, 71–87 (2009).

  41. 41.

    Coué, M., Brenner, S. L., Spector, I. & Korn, E. D. Inhibition of actin polymerization by latrunculin A. FEBS Lett. 213, 316–318 (1987).

  42. 42.

    Sun, Z., Guo, S. S. & Fässler, R. Integrin-mediated mechanotransduction. J. Cell Biol. 215, 445–456 (2016).

  43. 43.

    Ohashi, K., Fujiwara, S. & Mizuno, K. Roles of the cytoskeleton, cell adhesion and rho signalling in mechanosensing and mechanotransduction. J. Biochem. 161, 245–254 (2017).

  44. 44.

    Keller, M., Rüegg, A., Werner, S. & Beer, H. D. Active caspase-1 is a regulator of unconventional protein secretion. Cell 132, 818–831 (2008).

  45. 45.

    Palmer, G. M. et al. In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent reporters. Nat. Protoc. 6, 1355–1366 (2011).

  46. 46.

    Wang, X. et al. Epirubicin-adsorbed nanodiamonds kill chemoresistant hepatic cancer stem cells. ACS Nano 8, 12151–12166 (2014).

  47. 47.

    Li, M. et al. Nanoscale imaging and mechanical analysis of Fc receptor-mediated macrophage phagocytosis against cancer cells. Langmuir 30, 1609–1621 (2014).

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We thank N. Wang from the Huazhong University of Science and Technology and University of Illinois at Urbana-Champaign for help in guiding and revising this manuscript. We thank Z. Zhang’s group from the Huazhong University of Science and Technology for help with the dorsal window chamber model. We thank the Analytical and Testing Center of the Huazhong University of Science and Technology and the Research Core Facilities for Life Science (HUST) for related analysis. This work was supported by the National Basic Research Program of China (2018YFA0208900 and 2015CB931802), National Natural Science Foundation of China (81627901, 81773653, 81672937, 81530080, 81788101 and 61572213) and Chinese Academy of Medical Sciences Initiative for Innovative Medicine (2016-I2M-1–007).

Author information

Q.L., N.B., L.G., B.H. and X.Y. conceived and designed the experiments. Q.L., N.B., T.Y., K.T., Z.W., X.Z., H.Z. and W.H. performed the experiments. Q.L., N.B., T.Y., X.S., H.J., L.G., B.H. and X.Y. collected and analysed the data. L.G., B.H. and X.Y. supervised the project. L.G., B.H., X.Y. Q.L., N.B. and T.Y. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Correspondence to Lu Gan or Bo Huang or Xiangliang Yang.

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Supplementary information

Supplementary Information

Supplementary figures and tables.

Reporting Summary

Supplementary Dataset 1

The upregulated proteins in 3D-MPs versus 2D-MPs.

Supplementary Dataset 2

The downregulated proteins in 3D-MPs versus 2D-MPs.

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