Effect of Diphtheria Toxin-Based Gene Therapy for Hepatocellular Carcinoma

Abstract

Hepatocellular carcinoma (HCC) is a major global malignancy, responsible for >90% of primary liver cancers. Currently available therapeutic options have poor performances due to the highly heterogeneous nature of the tumor cells; recurrence is highly probable, and some patients develop resistances to the therapies. Accordingly, the development of a novel therapy is essential. We assessed gene therapy for HCC using a diphtheria toxin fragment A (DTA) gene-expressing plasmid, utilizing a non-viral hydrodynamics-based procedure. The antitumor effect of DTA expression in HCC cell lines (and alpha-fetoprotein (AFP) promoter selectivity) is assessed in vitro by examining HCC cell growth. Moreover, the effect and safety of the AFP promoter-selective DTA expression was examined in vivo using an HCC mice model established by the hydrodynamic gene delivery of the yes-associated protein (YAP)-expressing plasmid. The protein synthesis in DTA transfected cells is inhibited by the disappearance of tdTomato and GFP expression co-transfected upon the delivery of the DTA plasmid; the HCC cell growth is inhibited by the expression of DTA in HCC cells in an AFP promoter-selective manner. A significant inhibition of HCC occurrence and the suppression of the tumor marker of AFP and des-gamma-carboxy prothrombin can be seen in mice groups treated with hydrodynamic gene delivery of DTA, both 0 and 2 months after the YAP gene delivery. These results suggest that DTA gene therapy is effective for HCC.

In vivo gene delivery mediated by non-viral vectors for cancer therapy

“…In a study conducted by Kamimura and his colleagues, a diphtheria toxin fragment A (DTA) gene-expressing plasmid was transferred using the hydrodynamics-based procedure. The results demonstrated a substantial inhibition of hepatocellular carcinoma occurrence in mice treated with hydrodynamic-based gene therapy 0 and 2 months after gene delivery [128]. Since ultrasound microbubble sonoporation have shown great potential for gene delivery, a ternary nanodroplet composed of perfluoropentane/C9F17-PAsp(DET)/miR-122/PGA-g-mPEG (PFP-7TNDs/miR-122) was prepared and evaluated for the transferring of microRNA-122 (miR-122) for hepatocellular carcinoma treatment.…”

Establishment of a pancreatic cancer animal model using the pancreas-targeted hydrodynamic gene delivery method

“…With this aim, we applied the hydrodynamic gene delivery (HGD) method, which has been used successfully to develop mice liver cancer in vivo by delivering oncogenes, including Yap , Ras , and Myc, to the liver 11 , 12 , 13 . To utilize this method, we established pancreas-targeted HGD to wild-type rats.…”

HBx and YAP expression could promote tumor development and progression in HBV-related hepatocellular carcinoma

“…Indeed, we have shown that YAP overexpressed liver in wild-type mice showed the development of huge HCC in the short term and the tumors were heterogeneous, histologically [ 18 ]. Interestingly, HBx transgenic mice increase YAP protein expression postnatally in the hepatocytes, and YAP accumulation in the nucleus resulted in the hepatocytes’ carcinogenesis [ 12 ].…”

“…Interestingly, HBx transgenic mice increase YAP protein expression postnatally in the hepatocytes, and YAP accumulation in the nucleus resulted in the hepatocytes’ carcinogenesis [ 12 ]. Therefore, it is reasonable to hypothesize that the HBx-YAP axis is involved in the HBV-related HCC occurrence and development of huge tumors and poor prognosis in HBV-related HCC [ 18 ]. Since there are no reports assessing the HBx and YAP expression patterns in HCC tumor and peritumor tissue with clinical information, we have assessed the HBx and YAP expression in human HBV-related HCC samples and their association with tumor size, histological findings, serum tumor markers, and prognosis, comparing them with HCC related to other etiologies.…”