<p>Cationic <em>Antheraea pernyi</em> Silk Fibroin-Modified Adenovirus-Mediated ING4 and IL-24 Dual Gene Coexpression Vector Suppresses the Growth of Hepatoma Carcinoma Cells</p>

Abstract

IntroductionCancer gene therapy requires both effective tumor suppressor genes and safe vectors that express target genes efficiently. Inhibitor of growth 4 (ING4) inhibits tumor growth via multiple pathways. Interleukin-24 (IL-24) also has tumor-suppressive activity against a broad spectrum of human cancers. Adenovirus (Ad) vectors exhibit high infection efficiency, but potential toxicity related to high doses of adenovirus has led to careful reconsideration of their use in human clinical trials. Antheraea pernyi silk fibroin (ASF) is a cytocompatible and biodegradable natural polymer, and it possesses Arg–Gly–Asp sequences exhibiting a high binding affinity and selectivity for αvβ3 and αvβ5 integrin receptors, which are overexpressed in tumor vessels and most tumor cells.MethodsIn this study, an Arg-Gly-Asp peptide-modified Ad vector coexpressing ING4 and IL-24 was constructed by homologous recombination of the dual gene coexpression transfer plasmid and RGD-modified pAdEasy-1 adenoviral backbone plasmid. The cationic ASF (CASF) was prepared by modifying ASF with low-molecular-weight PEI. The negatively charged Ad vector was modified with CASF to form a CASF/Ad complex.ResultsHuman hepatoma carcinoma SMMC-7721 cells and normal hepatic L-02 cells were infected with the CASF/Ad complex, which showed significantly higher infection efficiency than the naked Ad. The CASF/Ad complex could effectively mediate the expression of the target gene ING4 in SMMC-7721 cells and the secretion of the target gene IL-24 from SMMC-7721 cells, thus inducing apoptosis of hepatoma carcinoma SMMC-7721 cells. The viability of SMMC-7721 and L-02 cells infected with the CASF/Ad complex was further assessed, and it was found that the growth of SMMC-7721 cells was significantly inhibited but that the growth and proliferation of L-02 cells were not affected.ConclusionThe CASF/Ad complex constructed in this study, showing improved infection efficiency and enhanced suppressive effects on human hepatoma carcinoma SMMC-7721 cells, has the potential to reduce the dose of adenovirus and still maintain high infection efficiency and tumor inhibition.

Antheraea pernyi silk fibroin-coated adenovirus as a VEGF165-Ang-1 dual gene delivery vector

“…And the infection efficiency is also higher than that of gene delivery with PEI-modified Antheraea pernyi silk fibroin coated Ad (92.68%). 36 Particularly, the zeta potential of CASF/Ad complexes prepared by coating Ad with CASF at concentrations of 20 and 50 μg/mL was close to 0 mV. Compared with naked Ad, the significantly reduced surface negative charge of CASF/Ad complexes could reduce the electrostatic repulsion between the complexes and the cell surfaces, facilitating the endocytosis of the complexes by the target cells, and thus facilitating the infection of the target cells by the exogenous genes carried by the complexes.…”

“…The VEGF165 and Ang-1 dual gene coexpression Ad vector containing encoding green fluorescent protein (GFP) gene was obtained according to previously described procedures. 34,36 Briefly, the VEGF165 and Ang-1 cDNA fragments were amplified by polymerase chain reaction (PCR) from pAdTrack-CMV-VEGF165-PolyA-promoter-Ang-1. The fragments of VEGF165 and Ang-1 were subcloned into the pAdTrack-CMV-polyA+promoter transfer plasmid encoding GFP gene at the Bgl II and Sal I sites and the Not I and Xho I sites, respectively.…”

Polyethylenimine-Modified Bombyx mori Silk Fibroin as a Delivery Carrier of the ING4-IL-24 Coexpression Plasmid

“…Importantly, the transfection efficiency of the above proportions of CBSF/pDNA complexes was higher than that of 25kDa PEI/pDNA 11.05% ( Figure 6 E), indicating that CBSF had a higher transfection efficiency to cancer cells than 25kDa PEI. Moreover, the ING4-IL-24 double gene can inhibit the proliferation of cancer cells intracellularly and extracellularly [ 9 ]. The number of round A549 cells in the field of vision increased ( Figure 5 A(b 2 –d 2 )) and the cell viability of the A549 cells significantly decreased ( Figure 7 A) as the mass ratios of CBSF/pDNA increased.…”

“…In addition, IL-24 shows special antitumor activity and can kill nearby tumor cells without toxicity to normal cells. Tumor cells often contain multiple genetic abnormalities, which limit the efficacy of a single gene-mediated cancer therapy [ 9 ]. The coexpression of the ING4 and IL-24 double genes can selectively inhibit tumor angiogenesis and the growth of tumor cells through pathways both inside and outside of the cells.…”

Editorial: Adenoviral Infection and Immunity, and Adenoviral Vectors for Gene Therapy Applications

“…HAdV vector-based gene therapy shows broad application potentials. However, nanomedicine based on HAdV vectors needs to consider the effects of dosage and potential toxicity, which limits their use in clinical trials (Qu et al, 2019 ). Many groups are working on modifying the natural properties of HAdVs to turn them into better tools for gene transfer, oncolytic virotherapy, or vaccines (Yan et al, 2021 ; Zhang et al, 2021a ).…”