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Development of Rous sarcoma Virus-like Particles Displaying hCC49 scFv for Specific Targeted Drug Delivery to Human Colon Carcinoma Cells

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Abstract

Purpose

Virus-like particles (VLPs) have been used as drug carriers for drug delivery systems. In this study, hCC49 single chain fragment variable (scFv)-displaying Rous sarcoma virus-like particles (RSV VLPs) were produced in silkworm larvae to be a specific carrier of an anti-cancer drug.

Method

RSV VLPs displaying hCC49 scFv were created by the fusion of the transmembrane and cytoplasmic domains of hemagglutinin from influenza A (H1N1) virus and produced in silkworm larvae. The display of hCC49 scFv on the surface of RSV VLPs was confirmed by enzyme-linked immunosorbent assay using tumor-associated glycoprotein-72 (TAG-72), fluorescent microscopy, and immunoelectron microscopy. Fluorescein isothiocyanate (FITC) or doxorubicin (DOX) was incorporated into hCC49 scFv-displaying RSV VLPs by electroporation and specific targeting of these VLPs was investigated by fluorescent microscopy and cytotoxicity assay using LS174T cells.

Results

FITC was delivered to LS174T human colon adenocarcinoma cells by hCC49 scFv-displaying RSV VLPs, but not by RSV VLPs. This indicated that hCC49 scFv allowed FITC-loaded RSV VLPs to be delivered to LS174T cells. DOX, which is an anti-cancer drug with intrinsic red fluorescence, was also loaded into hCC49 scFv-displaying RSV VLPs by electroporation; the DOX-loaded hCC49 scFv-displaying RSV VLPs killed LS174T cells via the specific delivery of DOX that was mediated by hCC49 scFv. HEK293 cells were alive even though in the presence of DOX-loaded hCC49 scFv-displaying RSV VLPs.

Conclusion

These results showed that hCC49 scFv-displaying RSV VLPs from silkworm larvae offered specific drug delivery to colon carcinoma cells in vitro. This scFv-displaying enveloped VLP system could be applied to drug and gene delivery to other target cells.

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Abbreviations

CLSM:

Confocal laser scanning microscope

BmNPV:

Bombyx mori nucleopolyhedrovirus

BmNPV/RSV-gag-577:

BmNPV bacmid encoding the RSV gag protein gene

BSA:

Bovine serum albumin

DOX:

Doxorubicin

DLS:

Dynamic light scattering

ELISA:

Enzyme-linked immunosorbent assay

FITC:

Fluorescein isothiocyanate

gag:

Group antigen protein

GPI:

Glycosylphosphatidylinositol

HA:

Hemagglutinin

hCC49:

Humanized CC49 antibody

HRP:

Horseradish peroxidase

MTT:

3-(4,5-di-methylthiazol-2-2yl)-2,5-diphenyltetrazolium bromide

PBS:

Phosphate-buffered saline

RSV:

Rous sarcoma virus

RSV VLPs:

Rous sarcoma virus-like particles

scFv:

Single-chain variable fragment

TAG-72:

Tumor associated glycoparticle-72

VLPs:

Virus-like particles

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ACKNOWLEDGMENTS AND DISCLOSURES

We thank Professor Hiroshi Ueda (Tokyo Institute of Technology, Japan) for the contribution of the plasmid carrying scFv cDNA. This work was supported by Grant-in-Aid for Scientific Research (A) Grant No.22248009 and by Promotion of Nanobio-Technology Research to Support Aging and Welfare Society from the Ministry of Education, Culture, Sports, Science and Technology, Japan. No additional external funding was received for this study.

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Authors and Affiliations

  1. Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka, 422-8529, Japan

    Tatsuya Kato, Megumi Yui & Enoch Y. Park

  2. Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Shizuoka, 422-8529, Japan

    Tatsuya Kato, Vipin Kumar Deo & Enoch Y. Park

  3. Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan

    Tatsuya Kato & Enoch Y. Park

Authors
  1. Tatsuya Kato
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  2. Megumi Yui
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  4. Enoch Y. Park
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Corresponding author

Correspondence to Enoch Y. Park.

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Kato, T., Yui, M., Deo, V.K. et al. Development of Rous sarcoma Virus-like Particles Displaying hCC49 scFv for Specific Targeted Drug Delivery to Human Colon Carcinoma Cells. Pharm Res 32, 3699–3707 (2015). https://doi.org/10.1007/s11095-015-1730-2

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  • DOI: https://doi.org/10.1007/s11095-015-1730-2

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