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Design and synthesis of multi-targeted nanoparticles for gene delivery to breast cancer tissues

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Abstract

Biocompatibility of nanoparticles is the most essential factor in their use in clinical applications. In this study, hyperbranched spermine (HS), hyperbranched spermine-polyethylene glycol-folic acid (HSPF), and hyperbranched spermine-polyethylene glycol-glucose (HSPG) were synthesized for DNA protection and gene delivery to breast cancer cells. The synthesis of HSPG and HSPF was confirmed using proton nuclear magnetic resonance (H-NMR), Fourier-transform infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA) spectroscopy. The HS/DNA, HSPF/DNA, HSPG/DNA, and hyperbranched spermine-polyethylene glycol-folic acid/glucose/DNA (HSPFG/DNA) nanoparticles were prepared by combining different concentrations of HS, HSPF, and HSPG with the same amount of DNA. The ability of HS, HSPF, and HSPG to interact with DNA and protect it against plasm digestion was evaluated using agarose gel. Moreover, in vivo and in vitro biocompatibility of HSPF/DNA, HSPG/DNA, and HSPFG/DNA was investigated using MTT assay and calculating weight change and survival ratio of BALB/c mice, respectively. The results of agarose gel electrophoresis showed that HS, HSPF, and HSPG have the high ability to neutralize the negative charge of DNA and protect it against plasma degradation. The results of in vivo cytotoxicity assay revealed that the HSPF/DNA, HSPG/DNA, and HSPFG/DNA nanoparticles have good biocompatibility on female BALB/c mice. In vitro and in vivo transfection assays revealed that functionalization of the surface of HS using polyethylene glycol-folic acid (HSPF) and polyethylene glycol-glucose (HSPG) significantly increases gene delivery efficiency in vitro and in vivo. These results also showed that gene transfer using both HSPF and HSPG copolymers increases gene transfer efficiency compared to when only one of them is used. The HSPFG/DNA nanoparticles have a high potential for use in therapeutic applications because of their excellent biocompatibility and high gene transfer efficiency to breast cancer tissue.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The author would like to thank Prof. Danièle Werck-Reichhart; Institute of Plant Molecular Biology, CNRS, University of Strasbourg, Strasbourg, France; Dr. Maris A Cinelli of the Northern Michigan University; Dr. Zohreh Amoozgar of the MGH, Harvard Medical School; and Dr. Farnaz Ahmadi-Nouraldinvand of the University of Mohaghegh Ardabili for the helpful advice on various technical issues examined in this paper.

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Authors and Affiliations

  1. Department of Plant Production and Genetics, Faculty of Agriculture and Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran

    Mehdi Afrouz, Amin Amani & Mohammad Taghi Alebrahim

  2. Department of Pharmaceutics, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran

    Amin Amani & Zainab Ahmadian

  3. Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, 5166414766, Iran

    Amin Amani

  4. UMR 8516, Laboratoire de Spectroscopie Pour Les Interactions, la Reactivite et l’Environment (LASIRE), CNRS, Université Lille, 59000, Lille, France

    Ali Eftekhari

  5. IMRCP, CNRS UMR5623, UPS, Université de Toulouse, 118 route de Narbonne, 31062, Toulouse, France

    Christophe Coudret

  6. Department of Crop and Soil Science, Seed Science and Technology, Oregon State University, Corvallis, OR, USA

    Sabry G. Elias

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  1. Mehdi Afrouz
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by M. A., S. E., A. A., and C. C. M. A. wrote the first draft of the manuscript and all authors commented on previous versions of the manuscript. All authors read and approved the manuscript and all data were generated in-house and that no paper mill was used. The authors declare that all data were generated in-house and that no paper mill was used.

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Correspondence to Mohammad Taghi Alebrahim.

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Ethics approval

In this study in vivo cytotoxicity assay was performed on female BALB/c mice in accordance with the U.K. Animals (Scientific Procedures) Act 1986, consistent with the Guide for animal experiments (EU Directive 2010/63/EU) and approved by the Ethics Committee of Tabriz University of Medical Science (Approval ID: IR.TBZMED.VCR.REC.1397.054).

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Highlights

• Functionalization of the surface of nanoparticles with two different ligands increases gene delivery efficiency.

• HSPFG/DNA nanoparticles are novel and biocompatible nanoparticles with excellent potential to deliver DNA to breast cancer tissues.

• HSPFG/DNA nanoparticles protect the nucleic acid against restriction enzymes and increase its transfer efficiency in plasma.

• Functionalization of HS using PEG-folic acid and PEG-glucose increases gene transfer efficiency in vivo.

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Afrouz, M., Amani, A., Eftekhari, A. et al. Design and synthesis of multi-targeted nanoparticles for gene delivery to breast cancer tissues. Naunyn-Schmiedeberg's Arch Pharmacol 396, 121–137 (2023). https://doi.org/10.1007/s00210-022-02303-6

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  • DOI: https://doi.org/10.1007/s00210-022-02303-6

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