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The exit of nanoparticles from solid tumours

Abstract

Nanoparticles enter tumours through endothelial cells, gaps or other mechanisms, but how they exit is unclear. The current paradigm states that collapsed tumour lymphatic vessels impair the exit of nanoparticles and lead to enhanced retention. Here we show that nanoparticles exit the tumour through the lymphatic vessels within or surrounding the tumour. The dominant lymphatic exit mechanism depends on the nanoparticle size. Nanoparticles that exit the tumour through the lymphatics are returned to the blood system, allowing them to recirculate and interact with the tumour in another pass. Our results enable us to define a mechanism of nanoparticle delivery to solid tumours alternative to the enhanced permeability and retention effect. We call this mechanism the active transport and retention principle. This delivery principle provides a new framework to engineer nanomedicines for cancer treatment and detection.

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Fig. 1: Nanoparticles drain into intratumoural lymphatic vessels.
Fig. 2: Nanoparticles drain into peritumoural lymphatic vessels.
Fig. 3: Nanoparticles exit the tumour through the blood vessels.
Fig. 4: The dominant exit mechanism is a function of the nanoparticle size.
Fig. 5: Nanoparticles exiting the tumour through lymphatic vessels are returned to the blood circulation.
Fig. 6: The active transport and retention (ATR) principle.

Data availability

All data supporting the findings in this study are available within the Article and Supplementary Figs. 139. Raw data and videos are available via Figshare at

https://figshare.com/projects/Supporting_Information_-_The_exit_of_nanoparticles_from_solid_tumours/167957

Additional data are available from the corresponding author (W.C.W.C, warren.chan@utoronto.ca) upon reasonable request.

Code availability

The accompanying code for Figs. 1b–d and 2b–d and Supplementary Fig. 16 can be found at the GitHub repository: https://github.com/luan-matthew/Nanoparticle-and-Tumour-Lymphatic-3D-Image-Analysis. The accompanying code and data for Supplementary Figs. 2729 can be found at the GitHub repository: https://github.com/luan-matthew/Nanoparticle-Exit-Compartment-Model.

References

  1. Wilhelm, S. et al. Analysis of nanoparticle delivery to tumours. Nat. Rev. Mater. 1, 16014 (2016).

    CAS  Google Scholar 

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

    CAS  Google Scholar 

  3. Nakamura, H., Fang, J., Jun, F. & Maeda, H. Development of next-generation macromolecular drugs based on the EPR effect: challenges and pitfalls. Expert Opin. Drug Del. 12, 53–64 (2014).

    Google Scholar 

  4. Jain, R. K. & Stylianopoulos, T. Delivering nanomedicine to solid tumors. Nat. Rev. Clin. Oncol. 7, 653–664 (2010).

    CAS  Google Scholar 

  5. Chauhan, V. P. & Jain, R. K. Strategies for advancing cancer nanomedicine. Nat. Mater. 12, 958–962 (2013).

    CAS  Google Scholar 

  6. Cancer and nanotechnology. National Cancer Institute (accessed 1 November 2022); https://www.cancer.gov/nano/cancer-nanotechnology

  7. Sindhwani, S. et al. The entry of nanoparticles into solid tumours. Nat. Mater. 19, 566–575 (2020).

    CAS  Google Scholar 

  8. Kingston, B. R. et al. Specific endothelial cells govern nanoparticle entry into solid tumors. ACS Nano 15, 14080–14094 (2021).

    CAS  Google Scholar 

  9. Padera, T. P. et al. Lymphatic metastasis in the absence of functional intratumor lymphatics. Science 296, 1883–1886 (2002).

    CAS  Google Scholar 

  10. Leu, A. J., Berk, D. A., Lymboussaki, A., Alitalo, K. & Jain, R. K. Absence of functional lymphatics within a murine sarcoma: a molecular and functional evaluation. Cancer Res. 60, 4324–4327 (2000).

    CAS  Google Scholar 

  11. Padera, T. P. et al. Cancer cells compress intratumour vessels. Nature 427, 695–695 (2004).

    CAS  Google Scholar 

  12. Cheng, Y.-H., He, C., Riviere, J. E., Monteiro-Riviere, N. A. & Lin, Z. Meta-analysis of nanoparticle delivery to tumors using a physiologically based pharmacokinetic modeling and simulation approach. ACS Nano 14, 3075–3095 (2020).

    CAS  Google Scholar 

  13. Ballou, B. et al. Sentinel lymph node imaging using quantum dots in mouse tumor models. Bioconjugate Chem. 18, 389–396 (2007).

    CAS  Google Scholar 

  14. Liang, C. et al. Tumor metastasis inhibition by imaging‐guided photothermal therapy with single‐walled carbon nanotubes. Adv. Mater. 26, 5646–5652 (2014).

    CAS  Google Scholar 

  15. Liu, J. et al. Enhanced primary tumor penetration facilitates nanoparticle draining into lymph nodes after systemic injection for tumor metastasis inhibition. ACS Nano 13, 8648–8658 (2019).

    CAS  Google Scholar 

  16. Valdés-Olmos, R. A. et al. Evaluation of mammary lymphoscintigraphy by a single intratumoral injection for sentinel node identification. J. Nucl. Med. 41, 1500–1506 (2000).

    Google Scholar 

  17. Jiang, X. et al. Intratumoral administration of STING-activating nanovaccine enhances T cell immunotherapy. J. Immunother. Cancer 10, e003960 (2022).

    Google Scholar 

  18. Kwong, B., Gai, S. A., Elkhader, J., Wittrup, K. D. & Irvine, D. J. Localized immunotherapy via liposome-anchored anti-CD137 + IL-2 prevents lethal toxicity and elicits local and systemic antitumor immunity. Cancer Res. 73, 1547–1558 (2013).

    CAS  Google Scholar 

  19. Vaahtomeri, K. & Alitalo, K. Lymphatic vessels in tumor dissemination versus immunotherapy. Cancer Res. 80, 3463–3465 (2020).

    CAS  Google Scholar 

  20. Stacker, S. A. et al. Lymphangiogenesis and lymphatic vessel remodelling in cancer. Nat. Rev. Cancer 14, nrc3677 (2014).

    Google Scholar 

  21. Hu, X. & Luo, J. Heterogeneity of tumor lymphangiogenesis: progress and prospects. Cancer Sci. 109, 3005–3012 (2018).

    CAS  Google Scholar 

  22. Dai, Q. et al. Quantifying the ligand-coated nanoparticle delivery to cancer cells in solid tumors. ACS Nano 12, 8423–8435 (2018).

    CAS  Google Scholar 

  23. Ouyang, B. et al. The dose threshold for nanoparticle tumour delivery. Nat. Mater. 19, 1362–1371 (2020).

    CAS  Google Scholar 

  24. Leak, L. V. Studies on the permeability of lymphatic capillaries. J. Cell Biol. 50, 300–323 (1971).

    CAS  Google Scholar 

  25. Feng, D., Nagy, J. A., Dvorak, H. F. & Dvorak, A. M. Ultrastructural studies define soluble macromolecular, particulate, and cellular transendothelial cell pathways in venules, lymphatic vessels, and tumor‐associated microvessels in man and animals. Microsc. Res. Tech. 57, 289–326 (2002).

    CAS  Google Scholar 

  26. Kobayashi, H. et al. Delivery of gadolinium-labeled nanoparticles to the sentinel lymph node: comparison of the sentinel node visualization and estimations of intra-nodal gadolinium concentration by the magnetic resonance imaging. J. Control. Release 111, 343–351 (2006).

    CAS  Google Scholar 

  27. Benezra, M. et al. Multimodal silica nanoparticles are effective cancer-targeted probes in a model of human melanoma. J. Clin. Invest. 121, 2768–2780 (2011).

    CAS  Google Scholar 

  28. Zhang, X., Shen, Y.-P., Li, J.-G. & Chen, G. Clinical feasibility of imaging with indocyanine green combined with carbon nanoparticles for sentinel lymph node identification in papillary thyroid microcarcinoma. Medicine 98, e16935 (2019).

    CAS  Google Scholar 

  29. Liu, R. et al. Prevention of nodal metastases in breast cancer following the lymphatic migration of paclitaxel-loaded expansile nanoparticles. Biomaterials 34, 1810–1819 (2013).

    CAS  Google Scholar 

  30. Thomas, S. N., Vokali, E., Lund, A. W., Hubbell, J. A. & Swartz, M. A. Targeting the tumor-draining lymph node with adjuvanted nanoparticles reshapes the anti-tumor immune response. Biomaterials 35, 814–824 (2014).

    CAS  Google Scholar 

  31. Kirkin, V. et al. Characterization of indolinones which preferentially inhibit VEGF‐C‐ and VEGF‐D‐induced activation of VEGFR‐3 rather than VEGFR‐2. Eur. J. Biochem. 268, 5530–5540 (2001).

    CAS  Google Scholar 

  32. Aspelund, A., Robciuc, M. R., Karaman, S., Makinen, T. & Alitalo, K. Lymphatic system in cardiovascular medicine. Circ. Res. 118, 515–530 (2016).

    CAS  Google Scholar 

  33. Kim, K. S., Suzuki, K., Cho, H., Youn, Y. S. & Bae, Y. H. Oral nanoparticles exhibit specific high-efficiency intestinal uptake and lymphatic transport. ACS Nano 12, 8893–8900 (2018).

    CAS  Google Scholar 

  34. Kobayashi, H. et al. Comparison of dendrimer‐based macromolecular contrast agents for dynamic micro‐magnetic resonance lymphangiography. Magn. Reson. Med. 50, 758–766 (2003).

    CAS  Google Scholar 

  35. Matsumura, Y. & Maeda, H. A new concept for macromolecular therapeutics in cancer chemotherapy: mechanism of tumoritropic accumulation of proteins and the antitumor agent smancs. Cancer Res. 46, 6387–6392 (1986).

    CAS  Google Scholar 

  36. Nichols, J. W. & Bae, Y. H. EPR: evidence and fallacy. J. Control. Release 190, 451–464 (2014).

    CAS  Google Scholar 

  37. Danhier, F. To exploit the tumor microenvironment: since the EPR effect fails in the clinic, what is the future of nanomedicine? J. Control. Release 244, 108–121 (2016).

    CAS  Google Scholar 

  38. Nakamura, Y., Mochida, A., Choyke, P. L. & Kobayashi, H. Nanodrug delivery: is the enhanced permeability and retention effect sufficient for curing cancer? Bioconjugate Chem. 27, 2225–2238 (2016).

    CAS  Google Scholar 

  39. Park, K. The drug delivery field at the inflection point: time to fight its way out of the egg. J. Control. Release 267, 2–14 (2017).

    CAS  Google Scholar 

  40. Sun, D., Zhou, S. & Gao, W. What went wrong with anticancer nanomedicine design and how to make it right. ACS Nano 14, 12281–12290 (2020).

    CAS  Google Scholar 

  41. Zi, Y. et al. Strategies to enhance drug delivery to solid tumors by harnessing the EPR effects and alternative targeting mechanisms. Adv. Drug Deliv. Rev. 188, 114449 (2022).

    CAS  Google Scholar 

  42. Tran, V. et al. Quantitative tissue pharmacokinetics and EPR effect of AGuIX nanoparticles: a multimodal imaging study in an orthotopic glioblastoma rat model and healthy macaque. Adv. Health. Mater. 10, 2100656 (2021).

    CAS  Google Scholar 

  43. Ejigah, V. et al. Approaches to improve macromolecule and nanoparticle accumulation in the tumor microenvironment by the enhanced permeability and retention effect. Polymers 14, 2601 (2022).

    CAS  Google Scholar 

  44. Fang, J., Islam, W. & Maeda, H. Exploiting the dynamics of the EPR effect and strategies to improve the therapeutic effects of nanomedicines by using EPR effect enhancers. Adv. Drug Deliv. Rev. 157, 142–160 (2020).

    CAS  Google Scholar 

  45. Balogh, L. P. (ed.) Nano-Enabled Medical Applications (Jenny Stanford Publishing, 2020); https://doi.org/10.1201/9780429399039

  46. Harney, A. S. et al. Real-time imaging reveals local, transient vascular permeability, and tumor cell intravasation stimulated by TIE2hi macrophage–derived VEGFA. Cancer Discov. 5, 932–943 (2015).

    CAS  Google Scholar 

  47. Naumenko, V. A. et al. Extravasating neutrophils open vascular barrier and improve liposomes delivery to tumors. ACS Nano 13, 12599–12612 (2019).

    CAS  Google Scholar 

  48. Lin, Z. P. et al. Macrophages actively transport nanoparticles in tumors after extravasation. ACS Nano 16, 6080–6092 (2022).

    CAS  Google Scholar 

  49. Dewhirst, M. W. & Secomb, T. W. Transport of drugs from blood vessels to tumour tissue. Nat. Rev. Cancer 17, 738–750 (2017).

    CAS  Google Scholar 

  50. Perrault, S. D. & Chan, W. C. W. Synthesis and surface modification of highly monodispersed, spherical gold nanoparticles of 50−200 nm. J. Am. Chem. Soc. 131, 17042–17043 (2009).

    CAS  Google Scholar 

  51. Perrault, S. D., Walkey, C., Jennings, T., Fischer, H. C. & Chan, W. C. W. Mediating tumor targeting efficiency of nanoparticles through design. Nano Lett. 9, 1909–1915 (2009).

    CAS  Google Scholar 

  52. Proulx, S. T. et al. Use of a PEG-conjugated bright near-infrared dye for functional imaging of rerouting of tumor lymphatic drainage after sentinel lymph node metastasis. Biomaterials 34, 5128–5137 (2013).

    CAS  Google Scholar 

  53. Mesquita, S. D. et al. Functional aspects of meningeal lymphatics in ageing and Alzheimer’s disease. Nature 560, 185–191 (2018).

    Google Scholar 

  54. Zhang, Y.-N. et al. Nanoparticle size influences antigen retention and presentation in lymph node follicles for humoral immunity. Nano Lett. 19, 7226–7235 (2019).

    CAS  Google Scholar 

  55. Skobe, M. et al. Induction of tumor lymphangiogenesis by VEGF-C promotes breast cancer metastasis. Nat. Med. 7, 192–198 (2001).

    CAS  Google Scholar 

  56. Berg, S. et al. ilastik: interactive machine learning for (bio)image analysis. Nat. Methods 16, 1226–1232 (2019).

    CAS  Google Scholar 

  57. Lin, Y., Xue, J. & Liao, S. Blocking lymph flow by suturing afferent lymphatic vessels in mice. J. Vis. Exp. https://doi.org/10.3791/61178 (2020).

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Acknowledgements

We thank M. Ganguly, V. Bradaschia, K. Duffin and G. Ossetchkine at the Centre for Phenogenomics for histology and fluorescence imaging. We thank A. Darbandi and D. Holmyard at SickKids hospital for their help in preparing tissue samples for TEM. We thank K. Lau and P. Paroutis at the SickKids Imaging Facility for use of the light-sheet microscope. We thank G. Zheng, J. Chen and J. Bu at the Toronto Nanomedicine Fabrication Centre for use of the ICP-MS instrument, and J. Jonkman at the Advanced Optical Microscopy Facility for the help with and use of the intravital microscope and fluorescence macroscope. We thank B. Kingston, M. Osborne and Y. Zhang for editorial comments. This work was supported by the Canadian Cancer Society (grant 705285-1), the Canadian Institute of Health Research (grants FDN159932 and MOP-1301431), NanoMedicines Innovation Network (2019-T3-01) and the Canadian Research Chairs Program (grant 950-223824). We thank the Natural Sciences and Engineering Research Council of Canada (S.M.M.), Ontario Graduate Scholarship (P.M. and S.M.M.), the Walter Summer Memorial Scholarship (P.M.) and the Faculty of Applied Science & Engineering Graduate Student Endowment Fund (B.S.) for student fellowships and scholarships.

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Authors and Affiliations

Authors

Contributions

L.N.M.N., S.S. and W.C.W.C. conceptualized the project. L.N.M.N., Z.P.L., P.M. and B.S. designed and performed the nanoparticle synthesis and characterization. L.N.M.N. and Z.P.L. performed the nanoparticle biodistribution experiments. L.N.M.N., Z.P.L., P.M. and S.M.M. performed the microscopy and imaging experiments. L.N.M.N. performed the mathematical modelling. L.N.M.N., S.S. and W.C.W.C. wrote the initial manuscript draught. W.N. assisted in editing the revised document. All authors contributed to editing and revising the manuscript.

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Correspondence to Warren C. W. Chan.

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W.C.W.C consults for the Cystic Fibrosis Foundation, Foresite Capital and Merck.

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Nature Materials thanks Shuming Nie, Natalie Trevaskis, Jie Zheng and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Extended data

Extended Data Fig. 1 Quantifying nanoparticle exit from the tumour.

a, Quantification of 15 nm gold nanoparticles in B16F10 tumours at 1 hr, 4 hr, 12 hr, 24 hr, 48 hr, 72 hr, and 120 hr post-intravenous administration. The amount of nanoparticles in the tumour rises from 0.9%ID to 10.4%ID between 1 hr and 48 hr, but decreases to 5.7%ID after 120hrs post-injection. This decrease shows approximately half of accumulated nanoparticles exit the tumour. The tumour kinetic profile shows three regions: 1. nanoparticle tumour entry is dominant between 1 hr and 48 hr (labelled as Entry), 2. nanoparticle tumour exit is dominant between 48 hr and 72hrs (labelled as Exit), and 3. amount of nanoparticles retained between 72 hr and 120 hr  (labelled as Retention). Data points and error bars represent the mean ± s.e.m. Statistical significance between the 48 hr and 120 hr timepoints using a two-tailed unpaired t-test with Welch’s correction. *P < 0.05. Exact P values are as follows: P = 0.0433. NP = nanoparticle. n = 10 mice for 1 hr timepoint; n = 6 mice for 4 hr timepoint; n = 9 mice for the 12 hr timepoint; n = 10 mice for the 24 hr timepoint; n = 8 mice for the 48 hr timepoint; n = 11 mice for the 72 hr timepoint; n = 14 mice for the 120 hr timepoint. b, Three potential mechanisms responsible for nanoparticle exit: 1. Lymphatic vessels inside of the tumour (intratumoural lymphatic vessels), 2. Lymphatic vessels surrounding the tumour (peritumoural lymphatic vessels), 3. Tumour blood vessels. Extended Data Fig. 1b created with BioRender.com.

Supplementary information

Supplementary Information

Supplementary Discussions 1–9, Figs. 1–39, Tables 1–7, Methods, video captions and references.

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Nguyen, L.N.M., Lin, Z.P., Sindhwani, S. et al. The exit of nanoparticles from solid tumours. Nat. Mater. 22, 1261–1272 (2023). https://doi.org/10.1038/s41563-023-01630-0

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