Gene-viro-therapy targeting liver cancer by a dual-regulated oncolytic adenoviral vector harboring IL-24 and TRAIL

Abstract

Cancer-targeting gene-viro-therapy is a promising cancer therapeutic strategy that strengthens the antitumor effect of oncolytic viruses by expressing an inserted foreign antitumor gene. To achieve liver cancer targeting and to improve the safety of the ZD55 vector (a widely-used E1B55KD gene-deleted oncolytic adenoviral vector (OV), we previously constructed), we designed a novel OV named Ad Á AFP Á D55 that selectively replicates in hepatocellular carcinoma (HCC) cells by replacing the E1A promoter with the liver-cancer specific a-Fetoprotein (AFP) promoter based on the ZD55 vector. We found that the oncolytic adenoviruses Ad Á AFP Á D55-IL-24 and Ad Á AFP Á D55-TRAIL express tumor-suppressor gene interleukin-24 (IL-24) and tumor necrosis factorrelated apoptosis-inducing ligand (TRAIL), respectively, significantly suppressed the HCC cell growth in vitro by inducing apoptosis by the caspase-8 and mitochondria-dependent caspase-9 signaling pathways. Furthermore, the combined treatment of Ad Á AFP Á D55-IL-24 and Ad Á AFP Á D55-TRAIL showed strong antitumor effects in vivo by significantly inhibiting the tumor growth in HCC HuH-7 cell xenograft mice, and markedly increasing animal survival rate. Therefore, this novel HCC cell-targeting OV carrying tumor-suppressor genes may provide a promising approach for liver cancer gene therapy.

Inhibition of kinesin family member 20B sensitizes hepatocellular carcinoma cell to microtubule‐targeting agents by blocking cytokinesis

“…Recombinant adenoviruses were generated by respective homologous recombination between the shuttle plasmids and the packaging plasmid pBHGE3 in HEK293 cells. Generation, identification, production, purification, and titration of the recombinant adenovirus were carried out as previously described …”

“…Generation, identification, production, purification, and titration of the recombinant adenovirus were carried out as previously described. 18,19…”

Liver cell-targeted delivery of therapeutic molecules

“…HCC-targeted oncolytic viruses contain HCC-specific promoters (e.g. a-fetoprotein promoter) and/or mutated genes (Altomonte et al, 2010;Liu et al, 2012b;Zhang et al, 2007b) or recognize an HCC-specific membrane marker (e.g. CD46) (Blechacz et al, 2006).…”

MPHOSPH1: A Potential Therapeutic Target for Hepatocellular Carcinoma

“…The sections were washed with PBS, treated with 3% H 2 O 2 , and blocked with a blocking solution. These steps were followed by overnight incubation with primary antibody at the proper dilution for IHC or hematoxylin and eosin (H&E) staining assays as previously described (10). To avoid experimental bias, the clinical human samples were scored from 0 to 3, based on the IHC staining of MPHOSPH1, by two people using a double-blind procedure.…”