Rapid eradication of colon carcinoma by Clostridium perfringens Enterotoxin suicidal gene therapy

Abstract

BackgroundBacterial toxins have evolved to an effective therapeutic option for cancer therapy. The Clostridium perfringens enterotoxin (CPE) is a pore-forming toxin with selective cytotoxicity. The transmembrane tight junction proteins claudin-3 and -4 are known high affinity CPE receptors. Their expression is highly upregulated in human cancers, including breast, ovarian and colon carcinoma. CPE binding to claudins triggers membrane pore complex formation, which leads to rapid cell death. Previous studies demonstrated the anti-tumoral effect of treatment with recombinant CPE-protein. Our approach aimed at evaluation of a selective and targeted cancer gene therapy of claudin-3- and/or claudin-4- expressing colon carcinoma in vitro and in vivo by using translation optimized CPE expressing vector.MethodsIn this study the recombinant CPE and a translation optimized CPE expressing vector (optCPE) was used for targeted gene therapy of claudin-3 and/or -4 overexpressing colon cancer cell lines. All experiments were performed in the human SW480, SW620, HCT116, CaCo-2 and HT-29 colon cancer and the isogenic Sk-Mel5 and Sk-Mel5 Cldn-3-YFP melanoma cell lines. Claudin expression analysis was done at protein and mRNA level, which was confirmed by immunohistochemistry. The CPE induced cytotoxicity was analyzed by the MTT cytotoxicity assay. In addition patient derived colon carcinoma xenografts (PDX) were characterized and used for the intratumoral in vivo gene transfer of the optCPE expressing vector in PDX bearing nude mice.ResultsClaudin-3 and -4 overexpressing colon carcinoma lines showed high sensitivity towards both recCPE application and optCPE gene transfer. The positive correlation between CPE cytotoxicity and level of claudin expression was demonstrated. Transfection of optCPE led to targeted, rapid cytotoxic effects such as membrane disruption and necrosis in claudin overexpressing cells. The intratumoral optCPE in vivo gene transfer led to tumor growth inhibition in colon carcinoma PDX bearing mice in association with massive necrosis due to the intratumoral optCPE expression.ConclusionsThis novel approach demonstrates that optCPE gene transfer represents a promising and efficient therapeutic option for a targeted suicide gene therapy of claudin-3 and/or claudin-4 overexpressing colon carcinomas, leading to rapid and effective tumor cell killing in vitro and in vivo.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-017-3123-x) contains supplementary material, which is available to authorized users.

Functionalization of gold-nanoparticles by the Clostridium perfringens enterotoxin C-terminus for tumor cell ablation using the gold nanoparticle-mediated laser perforation technique

“…2 ). It was therefore proposed to use CPE for tumor diagnosis and therapy 46 – 50 . Previous experiments demonstrated that CPE by binding to a subset of claudins, e.g.…”

Clostridium perfringens enterotoxin induces claudin-4 to activate YAP in oral squamous cell carcinomas

“…CPE is regarded as a therapeutic tool for epithelial malignant tumors by utilizing the damage of CLDN4 and CLDN3 [11,53,54]. The 50%-inhibitory concentration of CPE for colon cancer cell lines was between 50-100 ng/ml [53].…”

Natural products as reservoirs of novel therapeutic agents

“…Recently, bacterial toxins are gaining interest in targeted cancer therapy (Table 3 (Tab. 3) ; References in Table 3: Foss, 2000[ 46 ]; Karpinski and Szkaradkiewicz, 2013[ 61 ]; Nair at al., 2014[ 80 ]; Pahle et al, 2017[ 153 ]; Patyar et al, 2010[ 155 ]; Yang et al, 2013[ 181 ]; Zahaf and Schmidt, 2017[ 183 ]). They are produced naturally as virulence factors by various strains of pathogenic bacteria.…”