Membrane and Cell BiologyMembrane and Cell BiologyБиологические мембраны0233-47553034-5219The Russian Academy of Sciences66738510.31857/S0233475523010073NTUFGRArticlesСТАТЬИProduction of Reactive Oxygen Species by Genetically Encoded Photosensitizers 4D5scFv-miniSOG and DARPin-miniSOG in Living CellsВыработка активных форм кислорода генетически кодируемыми фотосенсибилизаторами 4D5scFv-miniSOG и DARPin-miniSOG внутри живых клетокProshkinaG. M.ПрошкинаГ. М.gmb@ibch.ruShramovaE. I.ШрамоваЕ. И.gmb@ibch.ruDeyevS. M.ДеевС. М.gmb@ibch.ruShemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of SciencesИнститут биоорганической химии им. М.М. Шемякина и Ю.А. Овчинникова РАНSechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation (Sechenov University)Первый Московский государственный медицинский университет им. И.М. Сеченова Минздрава России (Сеченовский Университет)01012023401616526022025Copyright ©; 2023, The Russian Academy of SciencesCopyright ©; 2023, Российская академия наук2023The Russian Academy of SciencesРоссийская академия наукhttps://journals.eco-vector.com/0233-4755/article/view/667385

Currently, experimental oncology is developing approaches based on the development of targeted photosensitizers that can be delivered specifically to a tumor of a certain molecular profile. This work is devoted to the study of the generation of reactive oxygen species by photosensitizers of protein nature 4D5scFv-miniSOG and DARPin-miniSOG, specific to the oncomarker HER2. It was found that despite the presence of the same phototoxic module in their composition, these photosensitizers are characterized by different levels of ROS production, which may be explained by the different rate of receptor-mediated internalization of 4D5scFv-miniSOG and DARPin-miniSOG.

В настоящее время в экспериментальной онкологии развиваются подходы, основанные на разработке адресных фотосенсибилизаторов, которые можно доставлять специфично к опухоли определенного молекулярного профиля. Данная работа посвящена изучению генерации активных форм кислорода фотосенсибилизаторами белковой природы 4D5scFv-miniSOG и DARPin-miniSOG, специфичными к онкомаркеру HER2. Установлено, что, несмотря на наличие одного и того же фототоксического модуля в своем составе, данные фотосенсибилизаторы характеризуются различным уровнем производства АФК, что, возможно, объясняется различной скоростью рецептор-опосредованной интернализации 4D5scFv-miniSOG и DARPin-miniSOG.

reactive oxygen speciesgenetically encoded photosensitizersphotodynamic therapyактивные формы кислородагенетически кодируемые фотосенсибилизаторыфотодинамическая терапияDougherty T.J., Kaufman J.E., Goldfarb A., Weishaupt K.R., Boyle D., Mittleman A. 1978. Photoradiation therapy for the treatment of malignant tumors. Cancer Res. 38 (8), 2628–2635.Castano A.P., Mroz P., Hamblin M.R. 2006. Photodynamic therapy and anti-tumour immunity. Nat Rev Cancer. 6 (7), 535–545.Slamon D.J., Clark G.M., Wong S.G., Levin W.J., Ullrich A., McGuire W.L. 1987. Human breast cancer: Correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science. 235 (4785), 177–182.Gusterson B.A., Gelber R.D., Goldhirsch A., Price K.N., Save-Soderborgh J., Anbazhagan R., Styles J., Rudenstam C.M., Golouh R., Reed R. 1992. Prognostic importance of c-erbB-2 expression in breast cancer. International (Ludwig) Breast Cancer Study Group. J. Clin. Oncol. 10 (7), 1049–1056.Bulina M.E., Chudakov D.M., Britanova O.V., Yanushevich Y.G., Staroverov D.B., Chepurnykh T.V., Merzlyak E.M., Shkrob M.A., Lukyanov S., Lukyanov K.A. 2006. A genetically encoded photosensitizer. Nat. Biotechnol. 24 (1), 95–99.Shu X., Lev-Ram V., Deerinck T.J., Qi Y., Ramko E.B., Davidson M.W., Jin Y., Ellisman M.H., Tsien R.Y. 2011. A genetically encoded tag for correlated light and electron microscopy of intact cells, tissues, and organisms. PLoS Biol. 9 (4), e1001041.Serebrovskaya E.O., Edelweiss E.F., Stremovskiy O.A., Lukyanov K.A., Chudakov D.M., Deyev S.M. 2009. Targeting cancer cells by using an antireceptor antibody-photosensitizer fusion protein. Proc. Natl. Acad. Sci. USA. 106 (23), 9221–9225.Shipunova V.O., Deyev S.M. 2022. Artificial scaffold polypeptides as an efficient tool for the targeted delivery of nanostructures in vitro and in vivo. Acta Naturae. 14 (1), 54–72.Tolmachev V.M., Chernov V.I., Deyev S.M. 2022. Targeted nuclear mediine. Seek and destroy. Russ. Chem. Rev. 91, RCR5034. https://doi.org/10.1070/RCR5034Shramova E.I., Chumakov S.P., Shipunova V.O., Ryabova A.V., Telegin G.B., Kabashin A.V., Deyev S.M., Proshkina G.M. 2022. Genetically encoded BRET-activated photodynamic therapy for the treatment of deep-seated tumors. Light Sci. Appl. 11 (1), 38.Zhai Y., Huang X., Zhang K., Huang Y., Jiang Y., Cui J., Zhang Z., Chiu C.K.C., Zhong W., Li G. 2022. Spatiotemporal-resolved protein networks profiling with photoactivation dependent proximity labeling. Nat. Commun. 13 (1), 4906.Mironova K.E., Proshkina G.M., Ryabova A.V., Stremovskiy O.A., Lukyanov S.A., Petrov R.V., Deyev S.M. 2013. Genetically encoded immunophotosensitizer 4D5scFv-miniSOG is a highly selective agent for targeted photokilling of tumor cells in vitro. Theranostics. 3 (11), 831–840.Proshkina G.M., Shilova O.N., Ryabova A.V., Stremovskiy O.A., Deyev S.M. 2015. A new anticancer toxin based on HER2/neu-specific DARPin and photoactive flavoprotein miniSOG. Biochimie. 118, 116–122.Worn A., Plückthun A. 2001. Stability engineering of antibody single-chain Fv fragments. J. Mol. Biol. 305 (5), 989–1010.Binz H.K., Amstutz P., Kohl A., Stumpp M.T., Briand C., Forrer P., Grutter M.G., Plückthun A. 2004. High-affinity binders selected from designed ankyrin repeat protein libraries. Nat. Biotechnol. 22 (5), 575–582.Studier F.W. 2014. Stable expression clones and auto-induction for protein production in E. coli. Methods Mol. Biol. 1091, 17–32.Subik K., Lee J.F., Baxter L., Strzepek T., Costello D., Crowley P., Xing L., Hung M.C., Bonfiglio T., Hicks D.G., Tang P. 2010. The expression patterns of ER, PR, HER2, CK5/6, EGFR, Ki-67 and AR by immunohistochemical analysis in breast cancer cell lines. Breast Cancer (Auckl). 4, 35–41.Кузичкина Е.О., Шилова О.Н., Деев С.М. 2018. Механизм тушения флуоресценции белковых фотосенсибилизаторов на основе miniSOG в процессе интернализации рецептора HER2. Acta Naturae. 10 (4), 87–94.Шилова О.Н., Прошкина Г.М., Лебеденко Е.Н., Деев С.М. 2015. Интернализация и рециркуляция рецептора HER2 при взаимодействии адресного фототоксичного белка DARPin-miniSOG с клетками аденокарциномы молочной железы человека. Acta Naturae. 7 (3), 141–148.Girotti A.W. 2001. Photosensitized oxidation of membrane lipids: Reaction pathways, cytotoxic effects, and cytoprotective mechanisms. J. Photochem. Photobiol. B. 63 (1–3), 103–113.Finkel T. 2011. Signal transduction by reactive oxygen species. J. Cell Biol. 194 (1), 7–15.