KillerRed and miniSOG as genetically encoded photosensitizers for photodynamic therapy of cancer

Marina V. Shirmanova; Ekaterina O. Serebrovskaya; Ludmila B. Snopova; Maria M. Kuznetsova; Alina P. Ryumina; Ilya V. Turchin; Ekaterina A. Sergeeva; Nadezhda I. Ignatova; Natalia V. Klementieva; Konstantin A. Lukyanov; Sergey A. Lukyanov; Elena V. Zagaynova

doi:10.1117/12.2032552

24 June 2013 KillerRed and miniSOG as genetically encoded photosensitizers for photodynamic therapy of cancer

Marina V. Shirmanova, Ekaterina O. Serebrovskaya, Ludmila B. Snopova, Maria M. Kuznetsova, Alina P. Ryumina, Ilya V. Turchin, Ekaterina A. Sergeeva, Nadezhda I. Ignatova, Natalia V. Klementieva, Konstantin A. Lukyanov, Sergey A. Lukyanov, Elena V. Zagaynova

Author Affiliations +

Proceedings Volume 8803, Medical Laser Applications and Laser-Tissue Interactions VI; 88030L (2013) https://doi.org/10.1117/12.2032552
Event: European Conferences on Biomedical Optics, 2013, Munich, Germany

Abstract

Despite of the success of photodynamic therapy (PDT) in cancer treatment, the problems of low selective accumulation of a photosensitizer in a tumor and skin phototoxicity have not resolved yet. The idea of encoding of a photosensitizer in genome of cancer cells is attractive, particularly because it can provide highly selective light induced cell killing. This work is aimed at the development of new approach to PDT of cancer, namely to using genetically encoded photosensitizers. A phototoxicity of red fluorescent GFP-like protein KillerRed and FMN-binding protein miniSOG was investigated on HeLa tumor xenografts in nude mice. The tumors were generated by subcutaneous injection of HeLa cells stably expressing the phototoxic proteins. The tumors were irradiated with 594 nm or 473 nm laser at 150 mW/cm² for 20 or 30 min, repeatedly. Fluorescence intensity of the tumors was measured in vivo before and after each treatment procedure. Detailed pathomorphological analysis was performed 24 h after the therapy. On the epi-fluorescence images in vivo photobleaching of both proteins was observed indicating photodynamic reaction. Substantial pathomorphological abnormalities were found in the treated KillerRed-expressing tumor tissue, such as vacuolization of cytoplasm, cellular and nuclear membrane destruction, activation of apoptosis. In contrast, miniSOG-expressing tumors displayed no reaction to PDT, presumably due to the lack of FMN cofactor needed for fluorescence recovery of the flavoprotein. The results are of interest for photodynamic therapy as a proof of possibility to induce photodamages in cancer cells in vivo using genetically encoded photosensitizers.

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Marina V. Shirmanova, Ekaterina O. Serebrovskaya, Ludmila B. Snopova, Maria M. Kuznetsova, Alina P. Ryumina, Ilya V. Turchin, Ekaterina A. Sergeeva, Nadezhda I. Ignatova, Natalia V. Klementieva, Konstantin A. Lukyanov, Sergey A. Lukyanov, and Elena V. Zagaynova "KillerRed and miniSOG as genetically encoded photosensitizers for photodynamic therapy of cancer", Proc. SPIE 8803, Medical Laser Applications and Laser-Tissue Interactions VI, 88030L (24 June 2013); https://doi.org/10.1117/12.2032552

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