Radial porous SiO₂ nanoflowers potentiate the effect of antigen/adjuvant in antitumor immunotherapy

Abstract

Here, we reported a cancer nanovaccine based on SiO₂ nanoflowers with a special radial pore structure, which greatly enhanced cross-presentation and induced the production of cytotoxic T lymphocyte cells secreting granzymes B and interferon-γ. The antigen ovalbumin was covalently conjugated onto the as-synthesized hierarchical SiO₂ nanoflowers, and the adjuvant cytosine-phosphate-guanine was electrostatically adsorbed into their radial pore by simple mixing before use. The nanovaccine exhibited excellent storage stability without antigen release after 27 days of incubation, negligible cytotoxicity to dendritic cells, and a high antigen loading capacity of 430 ± 66 mg·g⁻¹ support. Besides, the nanovaccine could be internalized by dendritic cells via multiple pathways. And the enhancement of antigen/adjuvant uptake and lysosome escape of antigen were observed. Noteworthy, in vitro culture of bone marrow-derived dendritic cells in the presence of nanovaccine proved the activation of dendritic cells and antigen cross-presentation as well as secretion of proinflammatory cytokines. Besides, in vivo study verified the targeting of nanovaccine to draining lymph nodes, the complete suppression of tumor in six out of ten mice, and the triggering of notable tumor growth delay. Overall, the present results indicated that the nanovaccine can be served as a potential therapeutic vaccine to treat cancer.

Key words： silica nanoflower; antigen delivery; cancer immunotherapy; nanovaccine

Acknowledgements

The authors thank for the National Natural Science Foundation of China (Grant No. 81972899), Natural Science Foundation of Tianjin City (Grant No. 18JCQNJC14500), CAMS Innovation Fund for Medical Sciences (Grant No. 2017-I2M-3-022), Specific Program for High-Tech Leader&Team of Tianjin Government, Tianjin innovation and promotion plan key innovation team of immunoreactive biomaterials. The authors declare that they have no conflict of interest.

Electronic Supplementary Material

Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s11705-020-2034-6 and is accessible for authorized users.

References

1 Kelly P N. The cancer immunotherapy revolution. Science, 2018, 359(6382): 1344–1345

2 Sahin U, Türeci Ö. Personalized vaccines for cancer immunotherapy. Science, 2018, 359(6382): 1355–1360

3 Nam J, Son S, Park K S, Zou W, Shea L D, Moon J J. Cancer nanomedicine for combination cancer immunotherapy. Nature Reviews. Materials, 2019, 4(6): 398–414

4 Hollingsworth R E, Jansen K. Turning the corner on therapeutic cancer vaccines. NPJ Vaccines, 2019, 4(1): 7

5 Wculek S K, Cueto F J, Mujal A M, Melero I, Krummel M F, Sancho D. Dendritic cells in cancer immunology and immunotherapy. Nature Reviews. Immunology, 2020, 20(1): 1–18

6 Hu Z, Ott P A, Wu C J. Towards personalized, tumour-specific, therapeutic vaccines for cancer. Nature Reviews. Immunology, 2017, 18(3): 168–182

7 Riley R S, June C H, Langer R, Mitchell M J. Delivery technologies for cancer immunotherapy. Nature Reviews. Drug Discovery, 2019, 18(3): 175–196

8 Huang P, Wang X, Liang X, Yang J, Zhang C, Kong D, Wang W. Nano-, micro-, and macroscale drug delivery systems for cancer immunotherapy. Acta Biomaterialia, 2019, 85: 1–26

9 Newton J M, Sikora A G, Young S. Chapter 41—Materials-Based Cancer Immunotherapies: Principles of Regenerative Medicine. 3rd ed. Boston: Academic Press, 2019, 715–739

10 Gause K T, Wheatley A K, Cui J, Yan Y, Kent S J, Caruso F. Immunological principles guiding the rational design of particles for vaccine delivery. ACS Nano, 2017, 11(1): 54–68

11 Gu L. Tailored silica nanomaterials for immunotherapy. ACS Central Science, 2018, 4(5): 527–529

12 Giglio V, Varelaaramburu S, Travaglini L, Fiorini F, Seeberger P H, Maggini L, de Cola L. Reshaping silica particles: mesoporous nanodiscs for bimodal delivery and improved cellular uptake. Chemical Engineering Journal, 2018, 340: 148–154

13 Yang Y, Lu Y, Abbaraju P L, Zhang J, Zhang M, Xiang G, Yu C. Multi-shelled dendritic mesoporous organosilica hollow spheres: roles of composition and architecture in cancer immunotherapy. Angewandte Chemie International Edition, 2017, 56(29): 8446–8450

14 Abbaraju P L, Meka A K, Song H, Yang Y, Jambhrunkar M, Zhang J, Xu C, Yu M, Yu C. Asymmetric silica nanoparticles with tunable headtail structures enhance hemocompatibility and maturation of immune cells. Journal of the American Chemical Society, 2017, 139(18): 6321–6328

15 Wang X, Li X, Ito A, Watanabe Y, Sogo Y, Tsuji N M, Ohno T. Stimulation of in vivo antitumor immunity with hollow mesoporous silica nanospheres. Angewandte Chemie International Edition, 2016, 55(5): 1899–1903

16 Li X, Wang X, Sogo Y, Ohno T, Onuma K, Ito A. Mesoporous silicacalcium phosphate-tuberculin purified protein derivative composites as an effective adjuvant for cancer immunotherapy. Advanced Healthcare Materials, 2013, 2(6): 863–871

17 Li W A, Lu B Y, Gu L, Choi Y, Kim J, Mooney D J. The effect of surface modification of mesoporous silica micro-rod scaffold on immune cell activation and infiltration. Biomaterials, 2016, 83: 249–256

18 Wang X, Li X, Yoshiyuki K, Watanabe Y, Sogo Y, Ohno T, Tsuji N M, Ito A. Comprehensive mechanism analysis of mesoporous-silica-nanoparticle-induced cancer immunotherapy. Advanced Healthcare Materials, 2016, 5(10): 1169–1176

19 An M, Li M, Xi J, Liu H. Silica nanoparticle as a lymph node targeting platform for vaccine delivery. ACS Applied Materials & Interfaces, 2017, 9(28): 23466–23475

20 Cha B G, Jeong J H, Kim J. Extra-large pore mesoporous silica nanoparticles enabling co-delivery of high amounts of protein antigen and toll-like receptor 9 agonist for enhanced cancer vaccine efficacy. ACS Central Science, 2018, 4(4): 484–492

21 Moon D S, Lee J K. Tunable synthesis of hierarchical mesoporous silica nanoparticles with radial wrinkle structure. Langmuir, 2012, 28(33): 12341–12347

22 Du X, Qiao S. Dendritic silica particles with center-radial pore channels: promising platforms for catalysis and biomedical applications. Small, 2015, 11(4): 392–413

23 Shen Z, Li Y, Wen H, Ren X, Liu J, Yang L. Investigation on the role of surfactants in bubble-algae interaction in flotation harvesting of Chlorella vulgaris. Scientific Reports, 2018, 8(1): 3303

24 Kuai R, Ochyl L J, Bahjat K S, Schwendeman A, Moon J J. Designer vaccine nanodiscs for personalized cancer immunotherapy. Nature Materials, 2016, 16(4): 489–496

25 Bowen W S, Svrivastava A K, Batra L, Barsoumian H, Shirwan H. Current challenges for cancer vaccine adjuvant development. Expert Review of Vaccines, 2018, 17(3): 207–215

26 Warrier V U, Makandar A I, Garg M, Sethi G, Kant R, Pal J K, Yuba E, Gupta R K. Engineering anti-cancer nanovaccine based on antigen cross-presentation. Bioscience Reports, 2019, 39(10): BSR20193220

27 Casaravilla C, Pittini Á, Rückerl D, Seoane P I, Jenkins S J, MacDonald A S, Ferreira A M, Allen J E, Díaz Á. Unconventional maturation of dendritic cells induced by particles from the laminated layer of larval Echinococcus granulosus. Infection and Immunity, 2014, 82(8): 3164–3176

28 Yoshida S, Gaeta I, Pacitto R, Krienke L, Alge O, Gregorka B, Swanson J A. Differential signaling during macropinocytosis in response to M-CSF and PMA in macrophages. Frontiers in Physiology, 2015, 6: 8

29 Kasai H, Inoue K, Imamura K, Yuvienco C, Montclare J K, Yamano S. Efficient siRNA delivery and gene silencing using a lipopolypeptide hybrid vector mediated by a caveolae-mediated and temperature-dependent endocytic pathway. Journal of Nanobiotechnology, 2019, 17(1): 11

30 El-Sayed A, Harashima H. Endocytosis of gene delivery vectors: from clathrin-dependent to lipid raft-mediated endocytosis. Molecular Therapy, 2013, 21(6): 1118–1130

31 Kubiak Ossowska K, Jachimska B, Al Qaraghuli M, Mulheran P A. Protein interactions with negatively charged inorganic surfaces. Current Opinion in Colloid & Interface Science, 2019, 41: 104–117

32 Slowing I, Trewyn B G, Lin V S Y. Effect of surface functionalization of MCM-41-type mesoporous silica nanoparticles on the endocytosis by human cancer cells. Journal of the American Chemical Society, 2006, 128(46): 14792–14793

33 Latz E, Schoenemeyer A, Visintin A, Fitzgerald K A, Monks B G, Knetter C F, Lien E, Nilsen N J, Espevik T, Golenbock D T. TLR9 signals after translocating from the ER to CpG DNA in the lysosome. Nature Immunology, 2004, 5(2): 190–198

34 Hjortø G M, Larsen O, Steen A, Daugvilaite V, Berg C, Fares S, Hansen M, Ali S, Rosenkilde M M. Differential CCR7 targeting in dendritic cells by three naturally occurring CC-chemokines. Frontiers in Immunology, 2016, 7: 568

35 Gerlach C, Moseman E A, Loughhead S M, Alvarez D, Zwijnenburg A J, Waanders L, Garg R, de la Torre J C, von Andrian U H. The chemokine receptor CX3CR1 defines three antigen-experienced CD8⁺ T cell subsets with distinct roles in immune surveillance and homeostasis. Immunity, 2016, 45(6): 1270–1284

36 Lutz M B, Schuler G. Immature, semi-mature and fully mature dendritic cells: Which signals induce tolerance or immunity? Trends in Immunology, 2002, 23(9): 445–449

37 Tugues S, Burkhard S H, Ohs I, Vrohlings M, Nussbaum K, Vom Berg J, Kulig P, Becher B. New insights into IL-12-mediated tumor suppression. Cell Death and Differentiation, 2015, 22(2): 237–246

38 Kaplanski G, Marin V, Montero Julian F, Mantovani A, Farnarier C. IL-6: a regulator of the transition from neutrophil to monocyte recruitment during inflammation. Trends in Immunology, 2003, 24(1): 25–29

39 Sixt M, Kanazawa N, Selg M, Samson T, Roos G, Reinhardt D P, Pabst R, Lutz M B, Sorokin L. The conduit system transports soluble antigens from the afferent lymph to resident dendritic cells in the T cell area of the lymph node. Immunity, 2005, 22(1): 19–29

40 Sallusto F, Geginat J, Lanzavecchia A. Central memory and effector memory T cell subsets: function, generation, and maintenance. Annual Review of Immunology, 2004, 22(1): 745–763

41 Nimmerjahn F, Bruhns P, Horiuchi K, Ravetch J V. FcgRIV: a novel FcR with distinct IgG subclass specificity. Immunity, 2005, 23(1): 41–51