Nanoparticle-Modified Apoferritin Nanotransfer for Targeted Cytostatic Transport

Česká verzia

Authors: M. Čížek ^1,2; M. Gargulák ^1,2; K. Sehnal ^1,2; D. Uhlířová ²; M. Staňková ²; M. Dočekalová ²; B. Ruttkay-Nedecký ¹; J. Zídková ³; R. Kizek ^1,2,4
Authors place of work: Ústav humánní farmakologie a toxikologie, Farmaceutická fakulta, Veterinární a farmaceutická univerzita Brno ¹; Oddělení výzkumu a vývoje, Prevention Medicals s. r. o., Studénka ²; Ústav bio chemie a mikrobio logie, Vysoká škola chemicko-technologická Praha ³; Department of Biomedical and Environmental Analyses, Wroclaw Medical University, Poland ⁴
Published in the journal: Klin Onkol 2019; 32(3): 197-200
Category: Original Articles
doi: https://doi.org/10.14735/amko2019197

Summary

Background: Ferritin is a globular intracellular protein that acts as the main reservoir for iron. Malignancies are associated with increased plasma ferritin concentrations. A number of studies show that tumor cells express high levels of transferrin receptors (TfR). Increased TfR expression was observed in prostate carcinoma. Apoferritin (APO) can be used as a protein nanotransporter into which a suitable medicinal substance can be encapsulated. Nanoparticles increase the permeability of tumor cells to nanotransporters and have a photothermal effect. The aim of this study was to encapsulate doxorubicin (DOX) into APO and to modify the resulting APO/DOX with gold (AuNPs) and silver nanoparticles prepared by green synthesis (AgNPsGS).

Methods: APO was characterized using 10% sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) – 120 V, 60 min, 24 mM Tris, 0.2 M glycine, 3 mM SDS. DOX fluorescence (Ex 480 nm; Em 650 nm) was observed, with a typical absorption maximum at 560 nm. Electrochemical measurement was performed in Brdicka solution (three-electrode setup). AgNPsGS were prepared by green synthesis using clover (Trifolium pratense L.).

Results: An electrophoretic study of APO and APO/DOX (5–100 μg/mL) was performed and the behavior of APO and APO/DOX (10 μM) as a function of pH was monitored. In an acidic environment, APO forms subunits of about 20 kDa; in an alkaline medium, it forms a globular protein of about 450 kDa. A change in APO/DOX mobility (about by 10%) was observed. A film of gold nanoparticles was applied to the APO/DOX surface. APO/DOX-AuNPs were washed with ultra-pure water. pH-dependent release of DOX a was monitored. The amount of DOX analyzed was increased by up to 50%. Furthermore, an AgNPsGS-DOX complex (1 mg AgNPsGS/100 μM DOX) was generated and prepared. Subsequently, the AgNPsGS-DOX complex was encapsulated into APO. To further improve therapeutic efficacy, the APO/AgNPsGS-DOX complex was coated with an Au layer. APO/AgNPsGS-DOX/AuNPs were stable and DOX was released from the complex after physical parameters had changed.

Conclusion: APO nanocomplexes were prepared and modified to increase therapeutic efficacy against tumors. Tumor cell targeting was achieved by binding to TfR and via increased tumor cell permeability and retention. Release of the drug was made possible due to a pH change and photothermal activation that will now be tested.

This work was supported by COST European Cholangiocarcinoma Network CA18122 and International Collaboration Project of The European Technology Platform for Nanomedicine.

The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.

The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.

Submitted: 21. 3. 2019

Accepted: 14. 5. 2019

Keywords:

apoferritin nanotransporter – transferin receptors – targeted therapy – prostate tumors – nanomedicine – silver nanoparticles – gold nanoparticles

Zdroje

1. Kawabata H. Transferrin and transferrin receptors update. Free Radic Biol Med 2019; 133: 46–54. doi: 10.1016/j.freeradbiomed.2018.06.037.

2. Heger Z, Skalickova S, Zitka O et al. Apoferritin applications in nanomedicine. Nanomedicine 2014; 9 (14): 2233–2245. doi: 10.2217/nnm.14.119.

3. Heger Z, Eckschlager T, Stiborová M et al. Modern nanomedicine in treatment of lung carcinomas. Klin Onkol 2015; 28 (4): 245–250. doi: 10.14735/amko2015245.

4. Liang M, Fan K, Zhou M et al. H-ferritin-nanocaged doxorubicin nanoparticles specifically target and kill tumors with a single-dose injection. Proc Natl Acad Sci USA 2014; 111 (41): 14900–14905. doi: 10.1073/pnas.1407808 111.

5. Li AP, Yuchi QX, Zhang LB. Ferritin: a powerful platform for nanozymes. Prog Biochem Biophys 2018; 45 (2): 193–203.

6. Keer HN, Kozlowski JM, Tsai YC et al. Elevated transferrin receptor content in human prostate cancer cell lines assessed in vitro and in vivo. J Urol 1990; 143 (2): 381–385.

7. Niitsu Y, Kohgo Y, Nishisato T et al. Transferrin receptors in human cancerous tissues. Tohoku J Exp Med 1987; 153 (3): 239–243.

8. Maeda H, Wu J, Sawa T et al. Tumor vascular permeability and the EPR effect in macromolecular therapeutics: a review. J Control Release 2000; 65 (1–2): 271–284.

9. Fang J, Nakamura H, Maeda H. The EPR effect: Unique features of tumor blood vessels for drug delivery, factors involved, and limitations and augmentation of the effect. Adv Drug Deliv Rev 2011; 63 (3): 136–151. doi: 10.1016/j.addr.2010.04.009.

10. Krenn L, Paper DH. Inhibition of angiogenesis and inflammation by an extract of red clover (Trifolium pratense L.). Phytomedicine 2009; 16 (12): 1083–1088. doi: 10.1016/j.phymed.2009.05.017.

11. AshaRani PV, Low Kah Mun G, Hande MP et al. Cyto-toxicity and genotoxicity of silver nanoparticles in human cells. ACS nano 2009; 3 (2): 279–290. doi: 10.1021/nn800596w.

12. Li M, Wu D, Chen Y et al. Apoferritin nanocages with Au nanoshell coating as drug carrier for multistimuli-responsive drug release. Mater Sci Eng C Mater Biol Appl 2019; 95: 11–18. doi: 10.1016/j.msec.2018.10.060.

13. Sukirtha R, Priyanka KM, Antony JJ et al. Cytotoxic effect of Green synthesized silver nanoparticles using Melia azedarach against in vitro HeLa cell lines and lymphoma mice model. Process Biochem 2012; 47 (2): 273–279.

14. Piao MJ, Kang KA, Lee IK et al. Silver nanoparticles induce oxidative cell damage in human liver cells through inhibition of reduced glutathione and induction of mitochondria-involved apoptosis. Toxicol Lett 2011; 201 (1): 92–100. doi: 10.1016/j.toxlet.2010.12.010.

15. Švihovec J, Bultas J, Anzenbacher P et al. Farmakologie. 1. vyd. Praha: Grada Publishing 2018: 636–637.

16. Ruozi B, Veratti P, Vandelli MA et al. Apoferritin nanocage as streptomycin drug reservoir: Technological optimization of a new drug delivery system. Int J Pharm 2017; 518 (1–2): 281–288. doi: 10.1016/j.ijpharm.2016.12.038.