persmedРоссийский журнал персонализированной медициныRussian Journal for Personalized Medicine2782-38062782-3814ФОНД АЛМАЗОВА10.18705/2782-3806-2024-4-5-413-420EENWVApersmed-287Research ArticleОНКОЛОГИЯ, ЛУЧЕВАЯ ТЕРАПИЯONCOLOGY, RADIATION THERAPYСравнительный анализ химерных антигенных рецепторов с двойной специфичностью к CD19 и CD20Comparative Analysis of CAR T Cells with Dual Specificity for CD19 and CD20БеловежецТ. Н.BelovezhetsT. N.

Беловежец Татьяна Николаевна - младший научный сотрудник НИЛ геномного редактирования.

Санкт-Петербург

Belovezhets Tatyana N. - scientist, genome editing group, World Class Research Centre for Personalized Medicine.

Saint Petersburg

ГорчаковА. А.GorchakovA. A.

Горчаков Андрей Александрович - к.б.н., старший научный сотрудник НИЛ геномного редактирования.

Санкт-Петербург

Gorchakov Andrey A. - Candidate of Biological Sciences, senior scientist, genome editing group, World Class Research Centre for Personalized Medicine.

Saint Petersburg

СамочерныхК. А.SamochernykhK. A.

Самочерных Константин Александрович - д.м.н., профессор РАН, директор.

Санкт-Петербург

Samochernykh Konstantin A. - Doctor of Medical Sciences, Professor of the Russian Academy of Sciences, Head.

Saint Petersburg

КулемзинС. В.KulemzinS. V.

Кулемзин Сергей Викторович - к.б.н., старший научный сотрудник НИЛ геномного редактирования НЦМУ «Центр персонализированной медицины».

Ул. Аккуратова, д. 2, Санкт-Петербург, 197341

Kulemzin Sergei V. - Candidate of Biological Sciences, senior scientist, genome editing group, World Class Research Centre for Personalized Medicine.

Akkuratova str., 2, Saint Petersburg, Russia, 197341

skulemzin@mcb.nsc.ruФедеральное государственное бюджетное учреждение «Национальный медицинский исследовательский центр имени В.А. Алмазова» Министерства здравоохранения Российской ФедерацииРоссияAlmazov National Medical Research CentreRussian Federation20241811202445413420Copyright © Беловежец Т.Н., Горчаков А.А., Самочерных К.А., Кулемзин С.В., 20242024Беловежец Т.Н., Горчаков А.А., Самочерных К.А., Кулемзин С.В.Belovezhets T.N., Gorchakov A.A., Samochernykh K.A., Kulemzin S.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://persmed.elpub.ru/jour/article/view/287

CAR T-клеточная терапия онкогематологических заболеваний обладает высокой эффективностью и приемлемым профилем безопасности, однако часть пациентов не отвечает на лечение или быстро возвращается в состояние рецидива. Одной из причин недостаточного ответа на терапию CAR T-клетками может быть возникновение опухолевых эскейперов, не экспрессирующих эпитоп, распознаваемый CAR. Помочь снизить этот риск может использование CAR T-клеток двойной специфичности. В настоящей работе нами были получены CAR T-клетки, специфичные к CD19 и CD20, при этом исследовались четыре варианта организации антиген-распознающих доменов: два в виде biCAR и два в формате dualCAR. Было обнаружено, что наиболее перспективной является экспрессия двух независимых CAR (dualCAR) с одного промотора.

CAR T-cell therapy of patients with B-cell malignancies demonstrates high efficacy and an acceptable safety profile. However, some patients do not respond to treatment or quickly relapse. One of the reasons for an inadequate response to CAR T-cell therapy may be the emergence of cancer cells escape variants that do not express the epitope recognized by the CAR. Using CAR T-cells with dual specificity could help mitigate this issue. In this study, we developed CAR T-cells specific to human CD19 and CD20 by exploring four configurations of antigen-recognition domains: two in the biCAR format and two in the dualCAR format. Expression of two independent CARs (dualCAR) driven by a single promoter was found to be the most promising format.

биспецифические CARклеточная иммунотерапияхимерный антигенный рецепторCD19CD20bispecific CAR TCD19CD20chimeric antigen receptorimmunotherapyРабота выполнена при финансовой поддержке Министерства науки и высшего образования Российской Федерации (соглашение № 075-15-2022-301 от 20.04.2022)The work was carried out with the financial support of the Ministry of Science and Higher Education of the Russian Federation (Agreement No. 075-15-2022301 dated 04/20/2022)ReferencesAwasthi R, Maier HJ, Zhang J, Lim S. Kymriah® (tisagenlecleucel) An overview of the clinical development journey of the first approved CAR-T therapy. Human vaccines & immunotherapeutics. 2023;19(1):2210046. https://doi.org/10.1080/21645515.2023.2210046Awasthi R, Maier HJ, Zhang J, Lim S. Kymriah® (tisagenlecleucel) An overview of the clinical development journey of the first approved CAR-T therapy. Human vaccines & immunotherapeutics. 2023;19(1):2210046. https://doi.org/10.1080/21645515.2023.2210046Ruella M, Korell F, Porazzi P, Maus MV. Mechanisms of resistance to chimeric antigen receptor-T cells in haematological malignancies. Nature reviews. Drug discovery. 2023;22(12):976–995. https://doi.org/10.1038/s41573-023-00807-1Ruella M, Korell F, Porazzi P, Maus MV. Mechanisms of resistance to chimeric antigen receptor-T cells in haematological malignancies. Nature reviews. Drug discovery. 2023;22(12):976–995. https://doi.org/10.1038/s41573-023-00807-1Shah NN, Fry TJ. Mechanisms of resistance to CAR T cell therapy. Nature reviews. Clinical oncology. 2019;16(6): 372–385. https://doi.org/10.1038/s41571019-0184-6Shah NN, Fry TJ. Mechanisms of resistance to CAR T cell therapy. Nature reviews. Clinical oncology. 2019;16(6): 372–385. https://doi.org/10.1038/s41571019-0184-6Cronk RJ, Zurko J, Shah NN. Bispecific Chimeric Antigen Receptor T Cell Therapy for B Cell Malignancies and Multiple Myeloma. Cancers. 2020;12(9):2523. https://doi.org/10.3390/cancers12092523Cronk RJ, Zurko J, Shah NN. Bispecific Chimeric Antigen Receptor T Cell Therapy for B Cell Malignancies and Multiple Myeloma. Cancers. 2020;12(9):2523. https://doi.org/10.3390/cancers12092523Кулемзин С.В., Чикаев Н.А., Волкова О.Ю. и др. Модульная система лентивирусных векторов для работы с химерными антигенными рецепторами. Биоорганическая химия. 2017;43(2):124–132.Kulemzin SV, Chikaev NA, Volkova OYu, et al. A modular system of lentiviral vectors for working with chimeric antigenic receptors. Bioorganicheskaya ximiya=Russian Journal of Bioorganic Chemistry. 2017;43(2):124–132. In RussianBelovezhets T, Kulemzin S, Volkova O, et al. Comparative Pre-Clinical Analysis of CD20-Specific CAR T Cells Encompassing 1F5-, Leu16-, and 2F2-Based Antigen-Recognition Moieties. International journal of molecular sciences. 2023;24(4):3698. https://doi.org/10.3390/ijms24043698Belovezhets T, Kulemzin S, Volkova O, et al. Comparative Pre-Clinical Analysis of CD20-Specific CAR T Cells Encompassing 1F5-, Leu16-, and 2F2-Based Antigen-Recognition Moieties. International journal of molecular sciences. 2023;24(4):3698. https://doi.org/10.3390/ijms24043698Mamonkin M, Rouce RH, Tashiro H, Brenner MK. A T-cell-directed chimeric antigen receptor for the selective treatment of T-cell malignancies. Blood. 2015;126(8):983–992. https://doi.org/10.1182/blood-2015-02-629527Mamonkin M, Rouce RH, Tashiro H, Brenner MK. A T-cell-directed chimeric antigen receptor for the selective treatment of T-cell malignancies. Blood. 2015;126(8):983–992. https://doi.org/10.1182/blood-2015-02-629527Wang L, Fang C, Kang Q, et al. Bispecific CAR-T cells targeting CD19/20 in patients with relapsed or refractory B cell non-Hodgkin lymphoma: a phase I/II trial. Blood cancer journal. 2024;14(1):130. https://doi.org/10.1038/s41408-024-01105-8Wang L, Fang C, Kang Q, et al. Bispecific CAR-T cells targeting CD19/20 in patients with relapsed or refractory B cell non-Hodgkin lymphoma: a phase I/II trial. Blood cancer journal. 2024;14(1):130. https://doi.org/10.1038/s41408-024-01105-8Larson SM, Walthers CM, Ji B, et al. CD19/ CD20 Bispecific Chimeric Antigen Receptor (CAR) in Naive/Memory T Cells for the Treatment of Relapsed or Refractory Non-Hodgkin Lymphoma. Cancer discovery. 2023;13(3):580–597. https://doi.org/10.1158/2159-8290.CD-22-0964Larson SM, Walthers CM, Ji B, et al. CD19/ CD20 Bispecific Chimeric Antigen Receptor (CAR) in Naive/Memory T Cells for the Treatment of Relapsed or Refractory Non-Hodgkin Lymphoma. Cancer discovery. 2023;13(3):580–597. https://doi.org/10.1158/2159-8290.CD-22-0964Shah NN, Johnson BD, Schneider D, et al. Bispecific anti-CD20, anti-CD19 CAR T cells for relapsed B cell malignancies: a phase 1 dose escalation and expansion trial. Nature medicine. 2020;26(10):1569– 1575. https://doi.org/10.1038/s41591-020-1081-3Shah NN, Johnson BD, Schneider D, et al. Bispecific anti-CD20, anti-CD19 CAR T cells for relapsed B cell malignancies: a phase 1 dose escalation and expansion trial. Nature medicine. 2020;26(10):1569– 1575. https://doi.org/10.1038/s41591-020-1081-3Qin H, Ramakrishna S, Nguyen S, et al. Preclinical Development of Bivalent Chimeric Antigen Receptors Targeting Both CD19 and CD22. Molecular therapy oncolytics. 2018;11:127–137. https://doi.org/10.1016/j.omto.2018.10.006Qin H, Ramakrishna S, Nguyen S, et al. Preclinical Development of Bivalent Chimeric Antigen Receptors Targeting Both CD19 and CD22. Molecular therapy oncolytics. 2018;11:127–137. https://doi.org/10.1016/j.omto.2018.10.006Qin H, Nguyen SM, Ramakrishna S, et al. Novel CD19/CD22 Bicistronic Chimeric Antigen Receptors Outperform Single or Bivalent Cars in Eradicating CD19+CD22+, CD19-, and CD22Pre-B Leukemia. Blood. 2017;130:810-810.Qin H, Nguyen SM, Ramakrishna S, et al. Novel CD19/CD22 Bicistronic Chimeric Antigen Receptors Outperform Single or Bivalent Cars in Eradicating CD19+CD22+, CD19-, and CD22Pre-B Leukemia. Blood. 2017;130:810-810.Wang Y, Zhong K, Ke J, et al. Combined 4-1BB and ICOS co-stimulation improves anti-tumor efficacy and persistence of dual anti-CD19/CD20 chimeric antigen receptor T cells. Cytotherapy. 2021;23(8): 715– 723. https://doi.org/10.1016/j.jcyt.2021.02.117Wang Y, Zhong K, Ke J, et al. Combined 4-1BB and ICOS co-stimulation improves anti-tumor efficacy and persistence of dual anti-CD19/CD20 chimeric antigen receptor T cells. Cytotherapy. 2021;23(8): 715– 723. https://doi.org/10.1016/j.jcyt.2021.02.117Larson SM, Walthers CM, Ji B, et al. CD19/ CD20 Bispecific Chimeric Antigen Receptor (CAR) in Naive/Memory T Cells for the Treatment of Relapsed or Refractory Non-Hodgkin Lymphoma. Cancer discovery. 2023;13(3):580–597. https://doi.org/10.1158/2159-8290.CD-22-0964Larson SM, Walthers CM, Ji B, et al. CD19/ CD20 Bispecific Chimeric Antigen Receptor (CAR) in Naive/Memory T Cells for the Treatment of Relapsed or Refractory Non-Hodgkin Lymphoma. Cancer discovery. 2023;13(3):580–597. https://doi.org/10.1158/2159-8290.CD-22-0964Shah NN, Johnson BD, Schneider D, et al. Bispecific anti-CD20, anti-CD19 CAR T cells for relapsed B cell malignancies: a phase 1 dose escalation and expansion trial. Nature medicine. 2020;26(10):1569– 1575. https://doi.org/10.1038/s41591-020-1081-3Shah NN, Johnson BD, Schneider D, et al. Bispecific anti-CD20, anti-CD19 CAR T cells for relapsed B cell malignancies: a phase 1 dose escalation and expansion trial. Nature medicine. 2020;26(10):1569– 1575. https://doi.org/10.1038/s41591-020-1081-3Gomes-Silva D, Mukherjee M, Srinivasan M, et al. Tonic 4-1BB Costimulation in Chimeric Antigen Receptors Impedes T Cell Survival and Is Vector-Dependent. Cell reports. 2017;21(1):17–26. https://doi.org/10.1016/j.celrep.2017.09.015Gomes-Silva D, Mukherjee M, Srinivasan M, et al. Tonic 4-1BB Costimulation in Chimeric Antigen Receptors Impedes T Cell Survival and Is Vector-Dependent. Cell reports. 2017;21(1):17–26. https://doi.org/10.1016/j.celrep.2017.09.015

The authors declare that there are no conflicts of interest present.