title: Combined therapy with oncolytic adenoviruses encoding TRAIL and IL-12 genes markedly suppressed human hepatocellular carcinoma both in vitro and in an orthotopic transplanted mouse model.
abstract: BACKGROUND
  Gene-based virotherapy mediated by oncolytic viruses is currently experiencing a renaissance in cancer therapy. However, relatively little attention has been given to the potentiality of dual gene virotherapy strategy as a novel therapeutic approach to mediate triplex anticancer combination effects, particularly if the two suitable genes are well chosen. Both tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and interleukin-12 (IL-12) have been emerged as promising pharmacological candidates in cancer therapy; however, the combined efficacy of TRAIL and IL-12 genes for treatment of human hepatocellular carcinoma (HCC) remains to be determined.
  
  
  METHODS
  Herein, we investigated the therapeutic efficacy of concurrent therapy with two armed oncolytic adenoviruses encoding human TRAIL gene (Ad-ΔB/TRAIL) and IL-12 gene (Ad-ΔB/IL-12), respectively, on preclinical models of human HCC, and also elucidated the possible underlying mechanisms. The effects of Ad-ΔB/TRAIL+Ad-ΔB/IL-12 combination therapy were assessed both in vitro on Hep3B and HuH7 human HCC cell lines and in vivo on HCC-orthotopic model established in the livers of athymic nude mice by intrahepatic implantation of human Hep3B cells.
  
  
  RESULTS
  Compared to therapy with non-armed control Ad-ΔB, combined therapy with Ad-ΔB/TRAIL+Ad-ΔB/IL-12 elicited profound anti-HCC killing effects on Hep3B and HuH7 cells and on the transplanted Hep3B-orthotopic model. Efficient viral replication and TRAIL and IL-12 expression were also confirmed in HCC cells and the harvested tumor tissues treated with this combination therapy. Mechanistically, co-therapy with Ad-ΔB/TRAIL+Ad-ΔB/IL-12 exhibited an enhanced effect on apoptosis promotion, activation of caspase-3 and-8, generation of anti-tumor immune response evidenced by upregulation of interferon gamma (IFN-γ) production and infiltration of natural killer-and antigen presenting cells, and remarkable repression of intratumor vascular endothelial growth factor (VEGF) and cluster of differentiation 31 (CD31) expression and tumor microvessel density.
  
  
  CONCLUSIONS
  Overall, our data showed a favorable therapeutic effect of Ad-ΔB/TRAIL+Ad-ΔB/IL-12 combination therapy against human HCC, and may therefore constitute a promising and effective therapeutic strategy for treating human HCC. However, further studies are warranted for its reliable clinical translation.
authors.affiliation: Department of Laboratory Medicine, Faculty of Applied Medical Sciences, Umm Al-Qura University, PO Box 7607, Holy Makkah, Saudi Arabia. dr_adel_elshemy2006@yahoo.com.|Department of Pharmacology, Faculty of Medicine, Assiut University, Assiut, Egypt. dr_adel_elshemy2006@yahoo.com.
authors.forename: Adel Galal
authors.initials: AG
authors.lastname: El-Shemi
authors.affiliation: Department of Laboratory Medicine, Faculty of Applied Medical Sciences, Umm Al-Qura University, PO Box 7607, Holy Makkah, Saudi Arabia.
authors.forename: Ahmad Mohammed
authors.initials: AM
authors.lastname: Ashshi
authors.affiliation: Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsinmi-ro, Seongdong-gu, Seoul, Korea.
authors.forename: Youjin
authors.initials: Y
authors.lastname: Na
authors.affiliation: Graduate Program for Nanomedical Science, Yonsei University, Seoul, Korea.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Laboratory Medicine, Faculty of Applied Medical Sciences, Umm Al-Qura University, PO Box 7607, Holy Makkah, Saudi Arabia.|Department of Pathology, Faculty of Medicine, Umm Al-Qura University, Holy Makkah, Saudi Arabia.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Basalamah
authors.affiliation: Science and Technology Unit & Department of Medical Genetics, Faculty of Medicine, Umm Al-Qura University, Holy Makkah, Saudi Arabia.
authors.forename: Faisal Ahmad
authors.initials: FA
authors.lastname: Al-Allaf
authors.affiliation: Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsinmi-ro, Seongdong-gu, Seoul, Korea.
authors.forename: Eonju
authors.initials: E
authors.lastname: Oh
authors.affiliation: Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsinmi-ro, Seongdong-gu, Seoul, Korea.
authors.forename: Bo-Kyeong
authors.initials: BK
authors.lastname: Jung
authors.affiliation: Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsinmi-ro, Seongdong-gu, Seoul, Korea. chaeok@hanyang.ac.kr.
authors.forename: Chae-Ok
authors.initials: CO
authors.lastname: Yun
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000628:therapy*; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000235:genetics*; D008113:Liver Neoplasms / Q000378:metabolism / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D050130:Oncolytic Virotherapy / Q000379:methods*; D050504:Oncolytic Viruses / Q000235:genetics; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
chemical_list: C508594:IL2 protein, human; D007376:Interleukin-2; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human
keywords: Dual gene virotherapy; Hepatocellular carcinoma; Interleukin-12; Oncolytic adenoviruses; Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)