TY - JOUR

T1 - Ablation of Red Stable Transfected Claudin Expressing Canine Prostate Adenocarcinoma and Transitional Cell Carcinoma Cell Lines by C-CPE Gold-Nanoparticle-Mediated Laser Intervention

AU - Alnajjar, Suhayla

AU - Nolte, Ingo

AU - Becker, Annegret

AU - Schille, Jan Torben

AU - Trakooljul, Nares

AU - Frank, Marcus

AU - Ngezahayo, Anaclet

AU - Murua Escobar, Hugo

N1 - Funding Information: Funding: The authors wish to thank Aleppo University (Aleppo, Syria) and HGFK (Hannoversche Gesellschaft zur Förderung der Kleintiermeidizin e.V.) (Hannover, Germany) for supporting SA with a scholarship. Acknowledgments: The authors would like to thank Alexander Heisterkamp, Institute of Quantum Optics, Leibniz University, Hannover, for cell killing data interpretation and Karoline Schulz and Armin Springer, Medical Biology and Electron Microscopy Centre, University of Rostock, for technical support during electron microscopy analysis. The publication of this article was funded by the Open Access Fund of the Leibniz Universität Hannover.

PY - 2021/11/13

Y1 - 2021/11/13

N2 - 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.

AB - 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.

KW - Adenocarcinoma/metabolism

KW - Animals

KW - Carcinoma, Transitional Cell/metabolism

KW - Cell Line, Tumor

KW - Clostridium perfringens/chemistry

KW - Dog Diseases/metabolism

KW - Dogs

KW - Enterotoxins/chemistry

KW - Gold/chemistry

KW - Laser Therapy

KW - Male

KW - Metal Nanoparticles/chemistry

KW - Prostatic Neoplasms/metabolism

KW - C-CPE

KW - GNOME-LP

KW - Cell lines

KW - Prostate cancer

KW - Gold nanoparticle

UR - http://www.scopus.com/inward/record.url?scp=85118927674&partnerID=8YFLogxK

U2 - 10.3390/ijms222212289

DO - 10.3390/ijms222212289

M3 - Article

C2 - 34830170

VL - 22

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1422-0067

IS - 22

M1 - 12289

ER -