Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Delivery of nitric oxide with a nanocarrier promotes tumour vessel normalization and potentiates anti-cancer therapies

Nature Nanotechnology volume 14, pages1160–1169(2019)Cite this article

Subjects

Abstract

Abnormal tumour vasculature has a significant impact on tumour progression and response to therapy. Nitric oxide (NO) regulates angiogenesis and maintains vascular homeostasis and, thus, can be delivered to normalize tumour vasculature. However, a NO-delivery system with a prolonged half-life and a sustained release mechanism is currently lacking. Here we report the development of NanoNO, a nanoscale carrier that enables sustained NO release to efficiently deliver NO into hepatocellular carcinoma. Low-dose NanoNO normalizes tumour vessels and improves the delivery and effectiveness of chemotherapeutics and tumour necrosis factor-related, apoptosis-inducing, ligand-based therapy in both primary tumours and metastases. Furthermore, low-dose NanoNO reprogrammes the immunosuppressive tumour microenvironment toward an immunostimulatory phenotype, thereby improving the efficacy of cancer vaccine immunotherapy. Our findings demonstrate the ability of nanoscale NO delivery to efficiently reprogramme tumour vasculature and immune microenvironments to overcome resistance to cancer therapy, resulting in a therapeutic benefit.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

Rent or Buy

All prices are NET prices.

Fig. 1: Schematic showing the mechanism by which NanoNO suppresses HCC progression in mice.
Fig. 2: NanoNO accumulates in tumours, releases NO and mediates anti-tumour effects.
Fig. 3: Low-dose NanoNO normalizes tumour vasculature in HCC.
Fig. 4: Low-dose NanoNO improves drug delivery efficiency and enhances anti-cancer efficacy.
Fig. 5: Low-dose NanoNO reprogrammes immunosuppressive TAMs towards an immunostimulatory phenotype, increases tumour-infiltrating T cells and achieves synergistic anti-cancer effects when combined with a vaccine in orthotopic HCC models.
Fig. 6: Low-dose NanoNO modulates TME in metastatic lesions and suppresses metastatic progression of HCC.

Data availability

The data supporting the findings of this study are available within the paper and its Supplementary Information. All relevant data are available from the authors upon reasonable request.

References

  1. 1.

    Jain, R. K., Munn, L. L. & Fukumura, D. Dissecting tumour pathophysiology using intravital microscopy. Nat. Rev. Cancer 2, 266–276 (2002).

    CAS  Article  Google Scholar 

  2. 2.

    Fukumura, D. & Jain, R. K. Tumor microenvironment abnormalities: causes, consequences, and strategies to normalize. J. Cell Biochem. 101, 937–949 (2007).

    CAS  Article  Google Scholar 

  3. 3.

    Schaaf, M. B., Garg, A. D. & Agostinis, P. Defining the role of the tumor vasculature in antitumor immunity and immunotherapy. Cell Death Dis. 9, 115 (2018).

    Article  CAS  Google Scholar 

  4. 4.

    Gilkes, D. M., Semenza, G. L. & Wirtz, D. Hypoxia and the extracellular matrix: drivers of tumour metastasis. Nat. Rev. Cancer 14, 430–439 (2014).

    CAS  Article  Google Scholar 

  5. 5.

    Tredan, O., Galmarini, C. M., Patel, K. & Tannock, I. F. Drug resistance and the solid tumor microenvironment. J. Natl Cancer Inst. 99, 1441–1454 (2007).

    CAS  Article  Google Scholar 

  6. 6.

    Jain, R. K. Normalizing tumor microenvironment to treat cancer: bench to bedside to biomarkers. J. Clin. Oncol. 31, 2205–2218 (2013).

    CAS  Article  Google Scholar 

  7. 7.

    Jain, R. K. Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science 307, 58–62 (2005).

    CAS  Article  Google Scholar 

  8. 8.

    Carmeliet, P. & Jain, R. K. Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases. Nat. Rev. Drug Discov. 10, 417–427 (2011).

    CAS  Article  Google Scholar 

  9. 9.

    Huang, Y., Stylianopoulos, T., Duda, D. G., Fukumura, D. & Jain, R. K. Benefits of vascular normalization are dose and time dependent—letter. Cancer Res. 73, 7144–7146 (2013).

    CAS  Article  Google Scholar 

  10. 10.

    Huang, Z., Fu, J. & Zhang, Y. Nitric oxide donor-based cancer therapy: advances and prospects. J. Med. Chem. 60, 7617–7635 (2017).

    CAS  Article  Google Scholar 

  11. 11.

    Cheng, H. et al. Nitric oxide in cancer metastasis. Cancer Lett. 353, 1–7 (2014).

    CAS  Article  Google Scholar 

  12. 12.

    Fukumura, D., Kashiwagi, S. & Jain, R. K. The role of nitric oxide in tumour progression. Nat. Rev. Cancer 6, 521–534 (2006).

    CAS  Article  Google Scholar 

  13. 13.

    Wu, S. C. et al. Water-soluble dinitrosyl iron complex (DNIC): a nitric oxide vehicle triggering cancer cell death via apoptosis. Inorg. Chem. 55, 9383–9392 (2016).

    CAS  Article  Google Scholar 

  14. 14.

    Kashiwagi, S. et al. NO mediates mural cell recruitment and vessel morphogenesis in murine melanomas and tissue-engineered blood vessels. J. Clin. Invest. 115, 1816–1827 (2005).

    CAS  Article  Google Scholar 

  15. 15.

    Kashiwagi, S. et al. Perivascular nitric oxide gradients normalize tumor vasculature. Nat. Med. 14, 255–257 (2008).

    CAS  Article  Google Scholar 

  16. 16.

    Zhu, A. X., Duda, D. G., Sahani, D. V. & Jain, R. K. HCC and angiogenesis: possible targets and future directions. Nat. Rev. Clin. Oncol. 8, 292–301 (2011).

    CAS  Article  Google Scholar 

  17. 17.

    Llovet, J. M. et al. Hepatocellular carcinoma. Nat. Rev. Dis. Primers 2, 16018 (2016).

    Article  Google Scholar 

  18. 18.

    Reiberger, T. et al. An orthotopic mouse model of hepatocellular carcinoma with underlying liver cirrhosis. Nat. Protoc. 10, 1264–1274 (2015).

    CAS  Article  Google Scholar 

  19. 19.

    Powell, J. A., Mohamed, S. N., Kerr, J. S. & Mousa, S. A. Antiangiogenesis efficacy of nitric oxide donors. J. Cell Biochem. 80, 104–114 (2000).

    CAS  Article  Google Scholar 

  20. 20.

    Ziche, M. & Morbidelli, L. Nitric oxide and angiogenesis. J. Neurooncol. 50, 139–148 (2000).

    CAS  Article  Google Scholar 

  21. 21.

    Chauhan, V. P. et al. Normalization of tumour blood vessels improves the delivery of nanomedicines in a size-dependent manner. Nat. Nanotechnol. 7, 383–388 (2012).

    CAS  Article  Google Scholar 

  22. 22.

    Stylianopoulos, T., Munn, L. L. & Jain, R. K. Reengineering the tumor vasculature: improving drug delivery and efficacy. Trends Cancer 4, 258–259 (2018).

    CAS  Article  Google Scholar 

  23. 23.

    Wahl, K. et al. Increased apoptosis induction in hepatocellular carcinoma by a novel tumor-targeted TRAIL fusion protein combined with bortezomib. Hepatology 57, 625–636 (2013).

    CAS  Article  Google Scholar 

  24. 24.

    von Karstedt, S., Montinaro, A. & Walczak, H. Exploring the TRAILs less travelled: TRAIL in cancer biology and therapy. Nat. Rev. Cancer 17, 352–366 (2017).

    Article  CAS  Google Scholar 

  25. 25.

    Liu, C. H. et al. A multifunctional nanocarrier for efficient TRAIL-based gene therapy against hepatocellular carcinoma with desmoplasia in mice. Hepatology 67, 899–913 (2018).

    CAS  Article  Google Scholar 

  26. 26.

    Huang, Y., Goel, S., Duda, D. G., Fukumura, D. & Jain, R. K. Vascular normalization as an emerging strategy to enhance cancer immunotherapy. Cancer Res. 73, 2943–2948 (2013).

    CAS  Article  Google Scholar 

  27. 27.

    Huang, Y., Snuderl, M. & Jain, R. K. Polarization of tumor-associated macrophages: a novel strategy for vascular normalization and antitumor immunity. Cancer Cell 19, 1–2 (2011).

    Article  CAS  Google Scholar 

  28. 28.

    Peterson, T. E. et al. Dual inhibition of Ang-2 and VEGF receptors normalizes tumor vasculature and prolongs survival in glioblastoma by altering macrophages. Proc. Natl Acad. Sci. USA 113, 4470–4475 (2016).

    CAS  Article  Google Scholar 

  29. 29.

    Kloepper, J. et al. Ang-2/VEGF bispecific antibody reprograms macrophages and resident microglia to anti-tumor phenotype and prolongs glioblastoma survival. Proc. Natl Acad. Sci. USA 113, 4476–4481 (2016).

    CAS  Article  Google Scholar 

  30. 30.

    Jiang, Y., Li, Y. & Zhu, B. T-cell exhaustion in the tumor microenvironment. Cell Death Dis. 6, e1792 (2015).

    CAS  Article  Google Scholar 

  31. 31.

    Ribas, A. & Wolchok, J. D. Cancer immunotherapy using checkpoint blockade. Science 359, 1350–1355 (2018).

    CAS  Article  Google Scholar 

  32. 32.

    Calderaro, J. et al. Programmed death ligand 1 expression in hepatocellular carcinoma: relationship with clinical and pathological features. Hepatology 64, 2038–2046 (2016).

    CAS  Article  Google Scholar 

  33. 33.

    Chen, L. & Han, X. Anti-PD-1/PD-L1 therapy of human cancer: past, present, and future. J. Clin. Invest. 125, 3384–3391 (2015).

    Article  Google Scholar 

  34. 34.

    Sun, C., Mezzadra, R. & Schumacher, T. N. Regulation and function of the PD-L1 checkpoint. Immunity 48, 434–452 (2018).

    CAS  Article  Google Scholar 

  35. 35.

    Tao, L. H. et al. A polymorphism in the promoter region of PD-L1 serves as a binding-site for SP1 and is associated with PD-L1 overexpression and increased occurrence of gastric cancer. Cancer Immunol. Immunother. 66, 309–318 (2017).

    CAS  Article  Google Scholar 

  36. 36.

    Lin, C. C. et al. The efficacy of a novel vaccine approach using tumor cells that ectopically express a codon-optimized murine GM-CSF in a murine tumor model. Vaccine 34, 134–141 (2016).

    CAS  Article  Google Scholar 

  37. 37.

    Yu, T. W., Chueh, H. Y., Tsai, C. C., Lin, C. T. & Qiu, J. T. Novel GM-CSF-based vaccines: one small step in GM-CSF gene optimization, one giant leap for human vaccines. Hum. Vaccin. Immunother. 12, 3020–3028 (2016).

    Article  Google Scholar 

  38. 38.

    Lu, T. T. et al. Anionic Roussin's red esters (RREs) syn-/anti-[Fe(mu-SEt)(NO)2]2(-): the critical role of thiolate ligands in regulating the transformation of RREs into dinitrosyl iron complexes and the anionic RREs. Inorg. Chem. 47, 6040–6050 (2008)..

    CAS  Article  Google Scholar 

  39. 39.

    Cerami, E. et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2, 401–404 (2012).

    Article  Google Scholar 

  40. 40.

    Chen, Y. et al. Differential effects of sorafenib on liver versus tumor fibrosis mediated by stromal-derived factor 1 alpha/C-X-C receptor type 4 axis and myeloid differentiation antigen-positive myeloid cell infiltration in mice. Hepatology 59, 1435–1447 (2014).

    CAS  Article  Google Scholar 

  41. 41.

    Ueno, T., Suzuki, Y., Fujii, S., Vanin, A. F. & Yoshimura, T. In vivo distribution and behavior of paramagnetic dinitrosyl dithiolato iron complex in the abdomen of mouse. Free Radic. Res. 31, 525–534 (1999).

    CAS  Article  Google Scholar 

  42. 42.

    Kim, S. H. et al. Death induction by recombinant native TRAIL and its prevention by a caspase 9 inhibitor in primary human esophageal epithelial cells. J. Biol. Chem. 279, 40044–40052 (2004).

    CAS  Article  Google Scholar 

  43. 43.

    Schaub, F. X. et al. Fluorophore-NanoLuc BRET reporters enable sensitive in vivo optical imaging and flow cytometry for monitoring tumorigenesis. Cancer Res. 75, 5023–5033 (2015).

    CAS  Article  Google Scholar 

  44. 44.

    Campeau, E. et al. A versatile viral system for expression and depletion of proteins in mammalian cells. PLoS ONE 4, e6529 (2009).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by the Ministry of Science and Technology (nos. 104-2628-B-007-001-MY3, 105-2628-E-007-007-MY3, 108-2321-B-009-004 and 108-2221-E-007-104-MY5 to Y.C. and 103-2632-M-033-001-MY3, 104-2113-M-033-005-MY2, 106-2113-M-007-028-MY2 and 106-2113-M-033-009-MY2 to T.-T.L.); by the Chang Gung Memorial Hospital-National Tsinghua University Joint Research Grant (no. 108Q2508E1); by the National Health Research Institutes (no. NHRI-EX108-10609BC); by the ‘Frontier Research Center on Fundamental and Applied Sciences of Matters’ from the Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (no. 108QR001I5); and by the Ministry of Science and Technology (no. 108-3017-F-007-003). We are also grateful to H.-Y. Tang at College of Biomedical Science and Engineering Center, NTHU for assistance with flow cytometry and confocal laser microscopy and to C.-Y.S. Lai for assistance with pharmacokinetics analysis. We thank the National Laboratory Animal Center, NARLabs, Taiwan for technical support with measurement of blood pressure in mice. We acknowledge Prof. C.-H. Hung, W.-M. Ching, and K.-J. Hsing at the Institute of Chemistry, Academia Sinica and Instrumentation Center, National Taiwan Normal University for EPR measurements.

Author information

Affiliations

  1. Institute of Biomedical Engineering and Frontier Research Center on Fundamental and Applied Sciences of Matters, National Tsing Hua University, Hsinchu, Taiwan

    Yun-Chieh Sung, Pei-Ru Jin, Chih-Chun Chang, Dong-Yu Gao, John Jun-Sheng Ko, Tsai-Te Lu & Yunching Chen

  2. Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan

    Yun-Chieh Sung & Jane Wang

  3. Brain Research Center, National Tsing Hua University, Hsinchu, Taiwan

    Li-An Chu & Ann-Shyn Chiang

  4. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    Fu-Fei Hsu

  5. Department of Chemistry, Chung Yuan Christian University, Taoyuan, Taiwan

    Mei-Ren Wang & Tsai-Te Lu

  6. Department of Applied Chemistry, National Chiayi University, Chiayi, Taiwan

    Show-Jen Chiou

  7. School of Medicine, Chang Gung University, Taoyuan, Taiwan

    Jiantai Timothy Qiu

  8. Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Taoyuan, Taiwan

    Jiantai Timothy Qiu

  9. Department of Biomedical Sciences, School of Medicine, Chang Gung University, Taoyuan, Taiwan

    Jiantai Timothy Qiu, Chu-Chi Lin & Yu-Sing Chen

  10. Department of Biomedical Sciences and Engineering, National Central University, Taoyuan, Taiwan

    Yi-Chiung Hsu

  11. Department of Biomedical Engineering and Environmental Sciences, ​National Tsing Hua University, Hsinchu, Taiwan

    Fu-Nien Wang & Pei-Lun Yu

  12. Graduate Institute of Clinical Medical Science, China Medical University, Taichung, Taiwan

    Ann-Shyn Chiang

  13. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan

    Ann-Shyn Chiang

  14. Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli, Taiwan

    Ann-Shyn Chiang

  15. Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan

    Anthony Yan-Tang Wu, John Jun-Sheng Ko & Charles Pin-Kuang Lai

  16. Chemical Biology and Molecular Biophysics Program, Taiwan International Graduate Program, Academia Sinica, Taipei, Taiwan

    Anthony Yan-Tang Wu & Charles Pin-Kuang Lai

  17. Department and Graduate Institute of Pharmacology, National Taiwan University, Taipei, Taiwan

    Anthony Yan-Tang Wu

  18. Genome and Systems Biology Degree Program, National Taiwan University and Academia Sinica, Taipei, Taiwan

    Charles Pin-Kuang Lai

Authors
  1. Yun-Chieh Sung
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pei-Ru Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Li-An Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Fu-Fei Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mei-Ren Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chih-Chun Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Show-Jen Chiou
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jiantai Timothy Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Dong-Yu Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Chu-Chi Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Yu-Sing Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yi-Chiung Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Jane Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Fu-Nien Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Pei-Lun Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Ann-Shyn Chiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. Anthony Yan-Tang Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. John Jun-Sheng Ko
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. Charles Pin-Kuang Lai
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. Tsai-Te Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  21. Yunching Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Y.-C.S. and Y.C. conceived and designed the experiments and analysed the data. Y.-C.S., P.-R.J., F.-F.H., L.-A.C., C.-C.C., D.-Y.G., J.T.Q., C.-C.L., Y.-S.C., Y.-C.H., J.W., F.-N.W., P.-L.Y., T.-T.L. and Y.C. performed the experiments. F.-F.H., M.-R.W., S.-J.C., J.T.Q., C.-C.L., Y.-S.C., A.-S.C., A.Y-.T.W., J.J.-S.K., C.P.-K.L. and T.-T.L. contributed materials and analysis tools. Y.-C.S., T.-T.L. and Y.C. co-wrote the paper. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Tsai-Te Lu or Yunching Chen.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Tables 1–6 and Figs. 1–16.

Reporting Summary

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Sung, YC., Jin, PR., Chu, LA. et al. Delivery of nitric oxide with a nanocarrier promotes tumour vessel normalization and potentiates anti-cancer therapies. Nat. Nanotechnol. 14, 1160–1169 (2019). https://doi.org/10.1038/s41565-019-0570-3

Download citation

Further reading

Search

Quick links

Find nanotechnology articles, nanomaterial data and patents all in one place. Visit Nano by Nature Research