Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 54375 МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MEDICAL BIOPHYSICS AND BIOPHYSICAL CHEMISTRY МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ SYNTHESIS OF LUMINESCENT HER2/neu-SPECIFIC COMPLEXES BASED ON UPCONVERSION NANOPARTICLES AND TARGET PROTEINS СОЗДАНИЕ ЛЮМИНЕСЦЕНТНЫХ HER2/neu-СПЕЦИФИЧНЫХ КОМПЛЕКСОВ НА ОСНОВЕ АНТИСТОКСОВЫХ НАНОФОСФОРОВ И НАПРАВЛЯЮЩИХ БЕЛКОВ Смышляева А С Smyshlyaeva A S smysh.anita@gmail.com Гурьев Е. Л. Guryev E. L. Костюк А Б Kostyuk A B Воденеев В. А. Vodeneev V. A. Деев С М Deev S M Звягин А В Zvyagin A V Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского ru Lobachevsky State University of Nizhny Novgorod ru Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского Нижний Новгород Россия Lobachevsky State University of Nizhny Novgorod Nizhny Novgorod Russian Federation Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского ru Lobachevsky State University of Nizhny Novgorod ru Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского Нижний Новгород Россия Lobachevsky State University of Nizhny Novgorod Nizhny Novgorod Russian Federation Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского; Институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова РАН ru Lobachevsky State University of Nizhny Novgorod; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences ru Национальный исследовательский Нижегородский государственный университет им. Н.И. Лобачевского; Университет Маккуори ru Lobachevsky State University of Nizhny Novgorod; Macquarie University ru 25 12 2018 25 12 2018 3 4 840 846 20 12 2018 20 12 2018 https://rusjbpc.ru/en/nauka/article/54375/view

В настоящее время активно разрабатываются новые стратегии и подходы для биомедицинской диагностики на основе биогибридных фотолюминесцентных наноматериалов. Одним из перспективных направлений является создание тераностических комплексов с целью их направленной доставки и высокочувствительной детекции опухолевых клеток. В настоящей работе были получены двухкомпонентные комплексы на основе антистоксовых нанофосфóров (НАФ), представляющих собой высокоэффективные контрастирующие агенты с уникальными фотолюминесцентными свойствами. В качестве направляющего модуля использован высокоафинный белок неиммуноглобулиновой природы DARPin9-29, специфичный к онкомаркеру HER2/neu. Показано, что комплексы способны специфично связываться с клетками аденокарциномы молочной железы человека SK-BR-3, гиперэкспрессирующих рецептор HER2/neu. Полученные данные позволяют считать предложенную схему биоконъюгации перспективной платформой для создания тераностических агентов.

Modern strategies and approaches for biomedical diagnostics based on biohybrid photoluminescent nanomaterials are being actively developed. One of the promising areas is the synthesis of theranostic complexes for their targeted delivery and highly sensitive detection of tumor cells. In the present work we have created two-component nanocomplexes based on upconversion nanoparticles, highly effective contrast agents with unique photoluminescence properties. A non-immunoglobulin targeting protein DARPin9-29, which can selectively recognize the oncomarker HER2/neu, was used as a targeting module. A specifically binding of created complexes to the human breast adenocarcinoma SK-BR-3 cells overexpressing the HER2/neu-receptor has been shown. The obtained results allow to consider the proposed bioconjugation scheme as a promising platform for creating theranostic agents.

 тераностика антистоксовые нанофосфоры HER2/neu DARPin9-29 theranostics upconversion nanoparticles HER2/neu DARPin9-29

Гребеник Е.А., Генералова А.Н., Нечаев А.В., Хайдуков Е.В., Миронова К.Е., Стремовский О.А., Лебеденко Е.Н., Звягин А.В., Деев С.М. Специфическая визуализация опухолевых клеток с помощью антистоксовых нанофосфóров. Acta naturae, 2014, т. 6, № 4 (23), с. 51-57. [Grebenik E.A., Generalova A.N., Nechaev A.V., Khaydukov E.V., Mironova K.E., Stremovskiy O.A., Lebedenko E.N., Zvyagin A.V., Deyev S.M. Acta naturae, 2014, vol. 6, no. 4 (23), pp. 51-57. (In Russ.)] Grebenik E.A., Generalova A.N., Nechaev A.V., Haydukov E.V., Mironova K.E., Stremovskiy O.A., Lebedenko E.N., Zvyagin A.V., Deev S.M. Specificheskaya vizualizaciya opuholevyh kletok s pomosch'yu antistoksovyh nanofosfórov. Acta naturae, 2014, t. 6, № 4 (23), s. 51-57. [Grebenik E.A., Generalova A.N., Nechaev A.V., Khaydukov E.V., Mironova K.E., Stremovskiy O.A., Lebedenko E.N., Zvyagin A.V., Deyev S.M. Acta naturae, 2014, vol. 6, no. 4 (23), pp. 51-57. (In Russ.)] Friedman A.D., Claypool S.E., Liu R. The Smart Targeting of Nanoparticles. Current Pharmaceutical Design, 2013, 19, pp. 6315-6329. Friedman A.D., Claypool S.E., Liu R. The Smart Targeting of Nanoparticles. Current Pharmaceutical Design, 2013, 19, pp. 6315-6329. Sreenivasan V.K. Luminescent nanoparticles and their applications in the life sciences. Journal of physics. Condensed matter: An Institute of Physics journal, 2013, vol. 25, iss. 19, pp. 1-23. Sreenivasan V.K. Luminescent nanoparticles and their applications in the life sciences. Journal of physics. Condensed matter: An Institute of Physics journal, 2013, vol. 25, iss. 19, pp. 1-23. Grebenik E.A., Kostyuk A.B., Deyev S.M. Upconversion nanoparticles and their hybrid assemblies for biomedical applications. Russ. Chem. Rev., 2016, vol. 85, no. 12, pp. 1277-1296. Grebenik E.A., Kostyuk A.B., Deyev S.M. Upconversion nanoparticles and their hybrid assemblies for biomedical applications. Russ. Chem. Rev., 2016, vol. 85, no. 12, pp. 1277-1296. Kim J. Multifunctional nanostructured materials for multimodal imaging, and simultaneous imaging and therapy. Chem. Soc. Rev., 2009, vol. 38, no. 2, pp. 372-390. Kim J. Multifunctional nanostructured materials for multimodal imaging, and simultaneous imaging and therapy. Chem. Soc. Rev., 2009, vol. 38, no. 2, pp. 372-390. Xie J., Lee S., Chen X. Nanoparticle-based theranostic agents. Advanced Drug Delivery Reviews, 2010, vol. 62, pp. 1064-1079. Xie J., Lee S., Chen X. Nanoparticle-based theranostic agents. Advanced Drug Delivery Reviews, 2010, vol. 62, pp. 1064-1079. Звягин А.В., Панченко В.Я., Нечаев А.В., Шехтер А.Б., Деев С.М., Ахманов А.С., Гулер А.Е., Ивукина Е.И., Генералова А.Н., Семчишен В.А., Хайдуков Е.В. Антистоксовы нанолюминофоры: Перспективы применения в биологии и медицине. Сборник материалов V Троицкой конференции «Медицинская физика и инновации в медицине», 2012, т. 2, c. 8-9. [Zvyagin A.V., Panchenko V.Ya., Nechaev A.V., Shekhter A.B., Deyev S.M., Akhmatova A.S., Guler A.E., Ivukina E.I., Generalova A.N., Semchishen V.A., Khaydukov E.V. Антистоксовы нанолюминофоры: Перспективы применения в биологии и медицине. Sbornik materialov V Troickoj konferencii «Medicinskaya fizika i innovacii v medicine», 2012, vol. 2, pp. 8-9. (In Russ.)] Zvyagin A.V., Panchenko V.Ya., Nechaev A.V., Shehter A.B., Deev S.M., Ahmanov A.S., Guler A.E., Ivukina E.I., Generalova A.N., Semchishen V.A., Haydukov E.V. Antistoksovy nanolyuminofory: Perspektivy primeneniya v biologii i medicine. Sbornik materialov V Troickoy konferencii «Medicinskaya fizika i innovacii v medicine», 2012, t. 2, c. 8-9. [Zvyagin A.V., Panchenko V.Ya., Nechaev A.V., Shekhter A.B., Deyev S.M., Akhmatova A.S., Guler A.E., Ivukina E.I., Generalova A.N., Semchishen V.A., Khaydukov E.V. Antistoksovy nanolyuminofory: Perspektivy primeneniya v biologii i medicine. Sbornik materialov V Troickoj konferencii «Medicinskaya fizika i innovacii v medicine», 2012, vol. 2, pp. 8-9. (In Russ.)] Chen J., Zhao J.X. Upconversion Nanomaterials: Synthesis, Mechanism, and Applications in Sensing. Sensors, 2012, vol. 12, pp. 2414-2435. Chen J., Zhao J.X. Upconversion Nanomaterials: Synthesis, Mechanism, and Applications in Sensing. Sensors, 2012, vol. 12, pp. 2414-2435. Генералова А.Н., Зубов В.П., Хайдуков Е.В. Нанокристаллы с антистоксовой флуоресценцией на пути в медицину. Природа, 2016, т. 11, № 1215, с. 24-32. [Generalova A.N., Zubov V.P., Khaydukov E.V. Nanocrystals with Anti-Stokes Fluorescence on the Way to Medicine. Nature, 2016, vol. 11, no. 1215, pp. 24-32. (In Russ.)] Generalova A.N., Zubov V.P., Haydukov E.V. Nanokristally s antistoksovoy fluorescenciey na puti v medicinu. Priroda, 2016, t. 11, № 1215, s. 24-32. [Generalova A.N., Zubov V.P., Khaydukov E.V. Nanocrystals with Anti-Stokes Fluorescence on the Way to Medicine. Nature, 2016, vol. 11, no. 1215, pp. 24-32. (In Russ.)] Kostyuk A.B., Guryev E.L., Vorotnov A.D., Sencha L.M., Peskova N.N., Sokolova E.A., Liang L., Vodeneev V.A., Balalaeva I.V., Zvyagin A.V. Real-time tracking of Yb3+, Tm3+ doped NaYF4 nanoparticles in living cancer cells. Sovremennye tehnologii v medicine, 2018, vol. 10, no. 1, pp. 57-63. Kostyuk A.B., Guryev E.L., Vorotnov A.D., Sencha L.M., Peskova N.N., Sokolova E.A., Liang L., Vodeneev V.A., Balalaeva I.V., Zvyagin A.V. Real-time tracking of Yb3+, Tm3+ doped NaYF4 nanoparticles in living cancer cells. Sovremennye tehnologii v medicine, 2018, vol. 10, no. 1, pp. 57-63. Хайдуков Е.В, Рочева В.В., Семчишен В.А., Семиногов В.Н., Соколов В.И., Звягин А.В., Ахманов А.С., Панченко В.Я., Нечаев А.В., Генералова А.Н., Шехтер А.Б. Оптическая визуализация опухолевых тканей с применением антистоксовых наночастиц. Вестник российского фонда фундаментальных исследований, 2014, т. 4, № 84, с. 7-17. [Khaydukov E.V., Rocheva V.V., Semchishen V.A., Seminogov V.N., Sokolov V.I., Zvyagin A.V., Akhmanov A.S., Panchenko V. Ya., Nechaev A.V., Generalova A.N., Shekhter A.B. Applications of Upconversion Nanoparticles in Optical Bioimaging of the Tumor Tissue. Vestnik Rossiyskogo fonda fundamental'nykh issledovaniy [RFBR Journal], 2014, vol. 4, no. 84, pp. 7-17. (In Russ.)] Haydukov E.V, Rocheva V.V., Semchishen V.A., Seminogov V.N., Sokolov V.I., Zvyagin A.V., Ahmanov A.S., Panchenko V.Ya., Nechaev A.V., Generalova A.N., Shehter A.B. Opticheskaya vizualizaciya opuholevyh tkaney s primeneniem antistoksovyh nanochastic. Vestnik rossiyskogo fonda fundamental'nyh issledovaniy, 2014, t. 4, № 84, s. 7-17. [Khaydukov E.V., Rocheva V.V., Semchishen V.A., Seminogov V.N., Sokolov V.I., Zvyagin A.V., Akhmanov A.S., Panchenko V. Ya., Nechaev A.V., Generalova A.N., Shekhter A.B. Applications of Upconversion Nanoparticles in Optical Bioimaging of the Tumor Tissue. Vestnik Rossiyskogo fonda fundamental'nykh issledovaniy [RFBR Journal], 2014, vol. 4, no. 84, pp. 7-17. (In Russ.)] Naccache R., Vetrone F., Mahalingam V., Cuccia L.A., Capobianco J.A. Controlled Synthesis and Water Dispersibility of Hexagonal Phase NaGdF4:Ho3+/Yb3+ Nanoparticles. Chemistry of Materials, 2009, vol. 21, no. 4, pp. 717-723. Naccache R., Vetrone F., Mahalingam V., Cuccia L.A., Capobianco J.A. Controlled Synthesis and Water Dispersibility of Hexagonal Phase NaGdF4:Ho3+/Yb3+ Nanoparticles. Chemistry of Materials, 2009, vol. 21, no. 4, pp. 717-723. Wang L.Y., Zhang Y., Zhu Y.Y. One-Pot Synthesis and Strong Near-Infrared Upconversion Luminescence of Poly(acrylic acid)-Functionalized YF3:Yb3+/Er3+ Nanocrystals. Nano Research, 2010, vol. 3, no. 5, pp. 317-325. Wang L.Y., Zhang Y., Zhu Y.Y. One-Pot Synthesis and Strong Near-Infrared Upconversion Luminescence of Poly(acrylic acid)-Functionalized YF3:Yb3+/Er3+ Nanocrystals. Nano Research, 2010, vol. 3, no. 5, pp. 317-325. Deyev S.M., Lebedenko E.N. Targeted Bifunctional Proteins and Hybrid Nanoconstructs for Cancer Diagnostics and Therapies. Molecular Biology, 2017, vol. 51, no. 6, pp. 788-803. Deyev S.M., Lebedenko E.N. Targeted Bifunctional Proteins and Hybrid Nanoconstructs for Cancer Diagnostics and Therapies. Molecular Biology, 2017, vol. 51, no. 6, pp. 788-803. Slamon D.J., Clark G.M., Wong S.G., Levin W.J., Ullrich A., McGuire W.L. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science, 1987, vol. 235, no. 4785, pp. 177-182. Slamon D.J., Clark G.M., Wong S.G., Levin W.J., Ullrich A., McGuire W.L. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science, 1987, vol. 235, no. 4785, pp. 177-182. Ross J.S., Slodkowska E.A., Symmans W.F., Pusztai L., Ravdin P.M., Hortobagyi G.N. The HER-2 receptor and breast cancer: ten years of targeted anti-HER-2 therapy and personalized medicine. Oncologist, 2009, vol. 14, no. 4, pp. 320-368. Ross J.S., Slodkowska E.A., Symmans W.F., Pusztai L., Ravdin P.M., Hortobagyi G.N. The HER-2 receptor and breast cancer: ten years of targeted anti-HER-2 therapy and personalized medicine. Oncologist, 2009, vol. 14, no. 4, pp. 320-368. Mai H.-X., Zhang Y.-W., Sun L.-D., Yan C.-H. Size- and Phase-Controlled Synthesis of Monodisperse NaYF4:Yb,Er Nanocrystals from a Unique Delayed Nucleation Pathway Monitored with Upconversion Spectroscopy. The Journal of Physical Chemistry, 2007, vol. 111, no. 37, pp. 13730-13739. Mai H.-X., Zhang Y.-W., Sun L.-D., Yan C.-H. Size- and Phase-Controlled Synthesis of Monodisperse NaYF4:Yb,Er Nanocrystals from a Unique Delayed Nucleation Pathway Monitored with Upconversion Spectroscopy. The Journal of Physical Chemistry, 2007, vol. 111, no. 37, pp. 13730-13739. Dong A., Ye X., Chen J., Kang Y., Gordon T., Kikkawa J. M., Murray C.B. A Generalized Ligand-Exchange Strategy Enabling Sequential Surface Functionalization of Colloidal Nanocrystals. Journal of the American Chemical Society, 2011, vol. 133, no. 4, pp. 998-1006. Dong A., Ye X., Chen J., Kang Y., Gordon T., Kikkawa J. M., Murray C.B. A Generalized Ligand-Exchange Strategy Enabling Sequential Surface Functionalization of Colloidal Nanocrystals. Journal of the American Chemical Society, 2011, vol. 133, no. 4, pp. 998-1006. Hu M., Zhao J., Ai X., Budanovic M., Mu J., Webster R.D., Cao Q., Mao Z., Xing B. Near infrared light-mediated photoactivation of cytotoxic Re(I) complexes by using lanthanide-doped upconversion nanoparticles. Dalton Trans., 2016, vol. 45, pp. 14101-14108. Hu M., Zhao J., Ai X., Budanovic M., Mu J., Webster R.D., Cao Q., Mao Z., Xing B. Near infrared light-mediated photoactivation of cytotoxic Re(I) complexes by using lanthanide-doped upconversion nanoparticles. Dalton Trans., 2016, vol. 45, pp. 14101-14108. Steiner D., Forrer P., Plückthun A. Efficient selection of DARPins with sub-nanomolar affinities using SRP phage display. J. Mol. Biol., 2008, vol. 382, no. 5, pp. 1211-1227. Steiner D., Forrer P., Plückthun A. Efficient selection of DARPins with sub-nanomolar affinities using SRP phage display. J. Mol. Biol., 2008, vol. 382, no. 5, pp. 1211-1227. Binz H.K., Amstutz P., Kohl A., Stumpp M.T., Briand C., Forrer P., Grutter M.G., Pluckthun A. High-affinity binders selected from designed ankyrin repeat protein libraries. Nature Biotechnology, 2004, vol. 22, no. 5, pp. 575-582. Binz H.K., Amstutz P., Kohl A., Stumpp M.T., Briand C., Forrer P., Grutter M.G., Pluckthun A. High-affinity binders selected from designed ankyrin repeat protein libraries. Nature Biotechnology, 2004, vol. 22, no. 5, pp. 575-582. Sokolova E., Proshkina G., Kutova O., Shilova O., Ryabova A., Schulga A., Stremovskiy O., Zdobnova T., Balalaeva I., Deyev S. Recombinant targeted toxin based on HER2-specific DARPin possesses a strong selective cytotoxic effect in vitro and a potent antitumor activity in vivo. Journal of controlled release: official journal of the Controlled Release Society, 2016, vol. 233, pp. 48-56. Sokolova E., Proshkina G., Kutova O., Shilova O., Ryabova A., Schulga A., Stremovskiy O., Zdobnova T., Balalaeva I., Deyev S. Recombinant targeted toxin based on HER2-specific DARPin possesses a strong selective cytotoxic effect in vitro and a potent antitumor activity in vivo. Journal of controlled release: official journal of the Controlled Release Society, 2016, vol. 233, pp. 48-56. Deyev S.M., Lebedenko E.N., Petrovskaya L.E., Dolgikh D.A., Gabibov A.G., Kirpichnikov M.P. Man-made antibodies and immunoconjugates with desired properties: function optimization using structural engineering. Russian Chemical Reviews, 2015, vol. 84, no. 1, pp. 1-26. Deyev S.M., Lebedenko E.N., Petrovskaya L.E., Dolgikh D.A., Gabibov A.G., Kirpichnikov M.P. Man-made antibodies and immunoconjugates with desired properties: function optimization using structural engineering. Russian Chemical Reviews, 2015, vol. 84, no. 1, pp. 1-26. Interlandi G., Wetzel S.K., Settanni G., PluckthunA., Caflisch A. Characterization and further stabilization of designed ankyrin repeat proteins by combining molecular dynamics simulations and experiments. Journal of Molecular Biology, 2008, vol. 375, no. 3, pp. 837-854. Interlandi G., Wetzel S.K., Settanni G., PluckthunA., Caflisch A. Characterization and further stabilization of designed ankyrin repeat proteins by combining molecular dynamics simulations and experiments. Journal of Molecular Biology, 2008, vol. 375, no. 3, pp. 837-854. Zahnd C., Kawe M., Stumpp M.T., de Pasquale C., Tamaskovic R., Nagy-Davidescu G., Dreier B., Schibli R., BinzH.K., Waibel R., Plückthun A.A. Efficient tumor targeting with high-affinity designed ankyrin repeat proteins: effects of affinity and molecular size. Cancer Research, 2010, vol. 70, no. 4, pp. 1595-1605. Zahnd C., Kawe M., Stumpp M.T., de Pasquale C., Tamaskovic R., Nagy-Davidescu G., Dreier B., Schibli R., BinzH.K., Waibel R., Plückthun A.A. Efficient tumor targeting with high-affinity designed ankyrin repeat proteins: effects of affinity and molecular size. Cancer Research, 2010, vol. 70, no. 4, pp. 1595-1605. Hermanson G. Bioconjugate Techniques. Academic Press, 2008, 1200 р. Hermanson G. Bioconjugate Techniques. Academic Press, 2008, 1200 r. Sokolova E., Guryev E., Yudintsev A., Vodeneev V., Deyev S., Balalaeva I. HER2-specific recombinant immunotoxin 4D5scFv-PE40 passes through retrograde trafficking route and forces cells to enter apoptosis. Oncotarget, 2017, vol. 8, no. 13, pp. 22048-22058. Sokolova E., Guryev E., Yudintsev A., Vodeneev V., Deyev S., Balalaeva I. HER2-specific recombinant immunotoxin 4D5scFv-PE40 passes through retrograde trafficking route and forces cells to enter apoptosis. Oncotarget, 2017, vol. 8, no. 13, pp. 22048-22058.