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Ablation of Red Stable Transfected Claudin Expressing Canine Prostate Adenocarcinoma and Transitional Cell Carcinoma
Suhayla Alnajjar1,2, Ingo Nolte1, Annegret Becker3
1Small Animal Clinic, University of Veterinary Medicine Hannover, 30559 Hannover, Germany.
Abstract:
Claudin (CLDN) proteins are commonly expressed in cancers and targeted in novel therapeutic approaches. The C-terminal of Clostridium perfringens enterotoxin (C-CPE) efficiently binds several claudins. In this study, recombinant C-CPE conjugated to gold nanoparticles (AuNPs) has been used for prostate adenocarcinoma (PAC) and transitional cell carcinoma (TCC) cell killing in vitro using gold-nanoparticle-mediated laser perforation (GNOME-LP). A PAC and TCC cell lines, as well as red fluorescence variants, allowing deep tissue imaging, were used. CLDN-3, -4, and -7 expression was confirmed by qPCR and immunofluorescences. The binding of C-CPE-AuNPs complexes on the cell surface was examined by scanning electron microscopy (SEM). Further, transcriptome analysis was carried out to evaluate the effect of C-CPE binder on the biological response of treated cells. Directed C-CPE-AuNP binding verified the capability to target CLDN receptors. Transcriptome analysis showed that C-CPE binding may activate immune and inflammatory responses but does not directly affect cell survival. Cancer cells ablation was demonstrated using a combination of GNOME-LP and C-CPE-AuNPs treatment reducing tumor cell viability to less than 10% depending on cell line. The fluorescent cell lines and the verified proof of concept in vitro provide the basis for perspective xenograft studies in an animal model.
Insights
Gold nanoparticles conjugated with Clostridium perfringens enterotoxin (C-CPE) target claudin proteins on cancer cells. This novel approach, combined with laser perforation, effectively destroys prostate and transitional cell carcinoma cells in vitro.
Area of Science:
- Oncology
- Nanotechnology
- Biochemistry
Background:
- Claudin (CLDN) proteins are frequently overexpressed in various cancers.
- Targeting CLDNs presents a promising avenue for novel cancer therapeutics.
- Clostridium perfringens enterotoxin's C-terminal (C-CPE) specifically binds to several CLDNs.
Purpose of the Study:
- To investigate the efficacy of C-CPE conjugated to gold nanoparticles (AuNPs) for cancer cell ablation.
- To evaluate the targeting capability of C-CPE-AuNPs on prostate adenocarcinoma (PAC) and transitional cell carcinoma (TCC) cells.
- To assess the biological response of cancer cells to C-CPE binding.
Main Methods:
- Utilized C-CPE conjugated to gold nanoparticles (AuNPs) for cancer cell treatment.
- Employed gold-nanoparticle-mediated laser perforation (GNOME-LP) for in vitro cell killing.
- Confirmed CLDN expression via qPCR and immunofluorescence; examined C-CPE-AuNP binding using SEM.
- Conducted transcriptome analysis to understand cellular responses to C-CPE binding.
Main Results:
- C-CPE-AuNPs demonstrated directed binding to CLDN receptors on PAC and TCC cells.
- Transcriptome analysis indicated C-CPE binding may trigger immune/inflammatory responses without directly impacting cell survival.
- Combined GNOME-LP and C-CPE-AuNPs treatment reduced cancer cell viability to below 10% in vitro.
- Established fluorescent cell lines for deep tissue imaging and future in vivo studies.
Conclusions:
- C-CPE-AuNPs are effective in targeting CLDN-expressing cancer cells.
- The GNOME-LP technique combined with C-CPE-AuNPs shows significant potential for cancer cell ablation.
- This study provides a strong foundation for future xenograft studies in animal models.
