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Novel engineered HER2 specific recombinant protein nanocages for targeted drug delivery

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Abstract

Protein nanocages resemble natural biomimetic carriers and can be engineered to act as targeted delivery systems, making them an attractive option for various drug delivery and biomedical applications. Our research investigated the genetic link of a specific anti-HER2 peptide (LTVSPWY) to the exposed N-terminal region of the maize (Zea mays) ferritin 1 (ZmFer1) protein nanocage, employing either a 7-amino acid (for LTVS-ZmFer1) or 16-amino acid (for LTVS-L-ZmFer1) linker. We utilized a heat treatment method to load the chemotherapeutic drug doxorubicin into the protein nanocage. The construct with the longer linker (LTVS-L) produced a greater amount of soluble protein nanocage and was selected for further experiments. The average size, polydispersity index, and zeta potential of the engineered protein nanocage were 19.01 nm, 0.168, and − 2.13 mV, respectively. The LTVS-L-ZmFer1 protein nanocage exhibited excellent thermal stability, withstanding temperatures up to 100 °C with only partial denaturation. Furthermore, we observed that cellular uptake of the LTVS-L-ZmFer1 protein nanocages in HER2-positive breast cancer cells was significantly higher compared to ZmFer1 after labeling with FITC (fluorescein isothiocyanate) (P-value = 0.0001). In addition, we observed a significant decrease in the viability of SKBR3 cells when treated with DOX-loaded LTVS-L-ZmFer1 protein nanocages compared to cells treated with DOX-loaded ZmFer1 protein nanocages. Therefore, this new treatment strategy may prove to be an effective way to reduce both the side effects and toxicity associated with conventional cancer treatments in patients with HER2-positive breast cancer.

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Data availability

No datasets were generated or analysed during the current study.

Abbreviations

BC:

breast cancer

HER2:

human epidermal growth factor receptor-2

PAGE:

polyacrylamide gel electrophoresis

CBB:

coomassie brilliant blue

DLS:

dynamic light scattering

cDNA:

complementary DNA

IPTG:

isopropyl β-D-1-thiogalactopyranoside

DOX:

doxorubicin

FITC:

fluorescein isothiocyanate

HFt:

human ferritin

ZmFer1:

Zea mays ferritin 1.

HCC:

hepatocellular carcinoma

ERP:

enhanced permeability and retention

PfTrx:

Pyrococcus furiosus thioredoxin

FTH1:

ferritin heavy chain 1

RBCs:

red blood cells

BSA:

bovine serum albumin

PBS:

phosphate buffered saline

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Funding

This study was financially supported by the Vice-chancellor for Research and Technology, Hamadan University of Medical Sciences, Hamadan, Iran (Grant No. 140010148491).

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Author notes

  1. Javad Kheshti and Mohammad Ahmadyousefi contributed equally.

Authors and Affiliations

  1. Department of Pharmaceutical Biotechnology, School of Pharmacy, Hamadan University of Medical Sciences, Hamadan, Iran

    Javad Kheshti & Meysam Soleimani

  2. Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical Sciences, Hamadan, Iran

    Mohammad Ahmadyousefi

  3. Research Center for Molecular Medicine, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

    Mohammad Ahmadyousefi

Authors
  1. Javad Kheshti
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  2. Mohammad Ahmadyousefi
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Contributions

Javad Kheshti: Methodology, Data curation, Writing - original draft. Meysam Soleimani: Resources, Validation, Writing - review & editing, Supervision, Project administration, Funding acquisition. Mohammad Ahmadyousefi: Conceptualization, Methodology, Data curation, Investigation, Formal analysis, Visualization, Validation, Writing - original draft.

Corresponding author

Correspondence to Meysam Soleimani.

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Kheshti, J., Ahmadyousefi, M. & Soleimani, M. Novel engineered HER2 specific recombinant protein nanocages for targeted drug delivery. Mol Biol Rep 51, 773 (2024). https://doi.org/10.1007/s11033-024-09636-w

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