Chick chorioallantoic membrane (CAM) assay for the evaluation of the antitumor and antimetastatic activity of platinum-based drugs in association with the impact on the amino acid metabolism

Katerina Mitrevska; Miguel Angel Merlos Rodrigo; Natalia Cernei; Hana Michalkova; Zbynek Splichal; David Hynek; Ondrej Zitka; Zbynek Heger; Pavel Kopel; Vojtech Adam; Vedran Milosavljevic

doi:10.1016/j.mtbio.2023.100570

Chick chorioallantoic membrane (CAM) assay for the evaluation of the antitumor and antimetastatic activity of platinum-based drugs in association with the impact on the amino acid metabolism

Mater Today Bio. 2023 Jan 31:19:100570. doi: 10.1016/j.mtbio.2023.100570. eCollection 2023 Apr.

Authors

Katerina Mitrevska¹, Miguel Angel Merlos Rodrigo¹, Natalia Cernei¹, Hana Michalkova^{1

2}, Zbynek Splichal^{1

2}, David Hynek^{1

2}, Ondrej Zitka^{1

2}, Zbynek Heger^{1

2}, Pavel Kopel³, Vojtech Adam^{1

2}, Vedran Milosavljevic¹

Affiliations

¹ Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00, Brno, Czech Republic.
² Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00, Brno, Czech Republic.
³ Department of Inorganic Chemistry, Faculty of Science, Palacky University, 17. Listopadu 12, CZ-779 00, Olomouc, Czech Republic.

Abstract

The combination of in ovo and ex ovo chorioallantoic membrane (CAM) assay provides an excellent platform which extends its relevance in studying carcinogenesis to the field of screening of anticancer activity of platinum nanoparticles (PtNPs) and further study of the amino acids' fluctuations in liver and brain. PtNPs are promising candidates for replacing cisplatin (CDDP); however, insufficient data of their antitumor efficiency and activity on the cancer-related amino acid metabolism are available, and the assessment of the in vivo performance has barely scratched the surface. Herein, we used CAM assay as in vivo model for screening of novel therapeutic modalities, and we conducted a comparative study of the effects of CDDP and polyvinylpyrrolidone coated PtNPs on MDA-MB-231 breast cancer xenograft. PtNPs showed a higher efficiency to inhibit the tumor growth and metastasis compared to CDDP. The amino acids profiling in the MDA-MB-231 cells revealed that the PtNPs had an overall depleting effect on the amino acids content. Noteworthy, more side effects to amino acid metabolism were deduced from the depletion of the amino acids in tumor, brain, and liver upon CDDP treatment. Different sets of enzymes of the tricarboxylic acid (TCA) cycle were targeted by PtNPs and CDDP, and while mRNA encoding multiple enzymes was downregulated by PtNPs, the treatment with CDDP affected only two TCA enzymes, indicating a different mechanism of action. Taken together, CAM assay represents and invaluable model, demonstrating the PtNPs capability of repressing angiogenesis, decrease amino acid contents and disrupt the TCA cycle.

Keywords: Amino acids metabolism; Breast cancer; CAM assay; Cisplatin; Platinum nanoparticles; TCA cycle.