Proteolytic enzyme inhibitors as a new strategy in cancer treatment and Danio rerio as a model organism to evaluate the effectiveness and safety of applied therapies.
Proteolytic enzymes in the human body
Proteases play an important role in many human biological processes, such as fertilization, digestion, growth, maturation, aging and death of the body. Based on their mechanism of operation, proteases can be divided into six groups: serine proteases, cysteine proteases, threonine proteases, aspartyl proteases, glutamine proteases and metalloproteases. By controlling the activation, synthesis and degradation of proteins, they regulate numerous physiological processes.
Studies have shown that many human tumors have increased levels of both extracellular and intracellular proteases, and they are believed to be crucial for tumor vascularization and invasion . Proteolytic enzymes play a very important role in the growth of the primary tumor focus as well as in the formation of its metastases; they allow tumor cells to divide and pass into and out of the lumen of blood vessels, allowing them to establish themselves in other organs. The uncontrolled activity of proteolytic enzymes is balanced by endogenous protease inhibitors (e.g., TIMPs, cystatins), but in the microenvironment of cancer, this regulation is disrupted . In this context, lysosomal proteases (including cysteine cathepsins), may represent new pharmacological targets, says Magdalena Rudzińska-Radecka, MD, PhD. When studying human tissues from kidney cancer patients, we noted significantly increased levels of catspsin B in association with one of the natural protease inhibitors, stefin A, which correlated with a worse prognosis for patients and disease progression . In addition, on a molecular basis by overexpressing genes encoding cathepsin B and stefin A in renal cancer cells, we confirmed their reciprocal regulation.
To this day, a number of natural and synthetic products have been investigated that can act as inhibitors of cysteine cathepsins and are promising candidates for anticancer agents. However, most of the research conducted in the field of cathepsin inhibition targets a single protein; in contrast, we have developed inhibitors that are able to inhibit the activity of an entire class of cysteine cathepsins (patent: RU 2752531), and their effect on cancer cell growth inhibition has been confirmed both by bioinformatics simulations and in vitro. These inhibitors result in reduced aggressiveness of kidney cancer cells, which is further confirmed by our observations of ongoing in vivo experiments using a mouse model.
New applications that more specifically modulate proteases and serve their programmed mode of administration are highly valued. In this sense, nanostructures that serve a more controlled drug delivery are attracting our attention. Preliminary studies have shown that the therapeutic capacity of inhibitors is significantly improved when they are formulated in capsules, compared to their free forms [5, 6].
A new way to model the action of inhibitors
In addition, together with Piotr Podlasz, we understand that great opportunities for testing and screening the action of inhibitors can be provided by the model of the zebrafish (Danio rerio). Transparent Danio rerio embryos develop outside the mother’s body and have organ primordia forming as early as 24 hours after fertilization, which will make it an ideal model for toxicological testing. A number of observed morphological and behavioral changes under the influence of various substances are being used to predict the effect of a tested treatment. Recent advances in pharmaceutical screening of zebrafish and its use as a heterotransplant model of human tumors have shown it to be a reliable organism in evaluating the safety of new anti-cancer therapies. It can serve as an ideal alternative to mice in studies of drugs targeting tumor progression. Our studies conducted on the effects of biological anticancer compounds using the zebrafish as a model organism showed comparable results to those obtained using mice.
- A potential impact of A Disintegrin and Metalloproteinase DomainLike Protein Decysin-1 (ADAMDEC1) on clear cell renal cell carcinoma propagation. M. Rudzinska-Radecka. Vol.46, No.8, 2022, pp.1893-1901, doi:10.32604/biocell.2022.019724 · 27 dec 2021.
- The Role of Cysteine Cathepsins in Cancer Progression and Drug Resistance. Magdalena Rudzińska, Alessandro Parodi, Surinder M Soond, Andrey Z Vinarov, Dmitry O Korolev, Andrey O Morozov, Cenk Daglioglu, Yusuf Tutar, Andrey A Zamyatnin Jr. Int J Mol Sci. 2019 Jul 23;20(14):3602. doi: 10.3390/ijms20143602.
- In Silico, In Vitro, and Clinical Investigations of Cathepsin B and Stefin A mRNA Expression and a Correlation Analysis in Kidney Cancer. Magdalena Rudzinska-Radecka, Anastasia S. Frolova, Anastasia V. Balakireva, Neonila V. Gorokhovets, Vadim S. Pokrovsky, Darina V. Sokolova, Dmitry O. Korolev, Natalia V. Potoldykova, Andrey Z. Vinarov, Alessandro Parodi, Andrey A. Zamyatnin, Jr. Cells. 2022 May; 11(9): 1455. Published online 2022 Apr 25. doi: 10.3390/cells11091455.
- Cysteine Cathepsins Inhibition Affects Their Expression and Human Renal Cancer Cell Phenotype. Magdalena Rudzińska, Alessandro Parodi, Valentina D. Maslova, Yuri M. Efremov, Neonila V. Gorokhovets, Vladimir A. Makarov, Vasily A. Popkov, Andrey V. Golovin, Evgeni Y. Zernii, Andrey A. Zamyatnin, Jr., Cancers (Basel). 2020 May 21;12(5):1310. doi: 10.3390/cancers12051310.
- Current Status and Perspectives of Protease Inhibitors and Their Combination with Nanosized Drug Delivery Systems for Targeted Cancer Therapy. Magdalena Rudzińska, Cenk Daglioglu, Lyudmila V Savvateeva, Fatma Necmiye Kaci, Rodolphe Antoine, Andrey A Zamyatnin Jr. Drug Des Devel Ther. 2021; 15: 9–20. Published online 2021 Jan 6. doi: 10.2147/DDDT.S285852.
- Albumin Nanovectors in Cancer Therapy and Imaging.Albumin Nanovectors in Cancer Therapy and Imaging. Alessandro Parodi, Jiaxing Miao, Surinder M. Soond, Magdalena Rudzińska, Andrey A. Zamyatnin, Jr.. Biomolecules. 2019 Jun 5;9(6). pii: E218. doi: 10.3390/biom9060218. · 5 cze 2019.
- The Anti-Tumoral Potential of Phosphonate Analog of Sulforaphane in Zebrafish Xenograft Model. Magdalena Rudzinska-Radecka, Łukasz Janczewski, Anna Gajda, Marlena Godlewska, Malgorzata Chmielewska-Krzesinska, Krzysztof Wasowicz, Piotr Podlasz. Cells 2021, 10(11), 3219; https://doi.org/10.3390/cells10113219.
dr Magdalena Rudzińska-Radecka, dr hab. Piotr Podlasz, Natalia Mikulska