Analysis of in vitro cytotoxicity of human NK cell line co-expressing a PSMA-specific CAR and an antitumor agent lactaptin

Abstract

Development of efficient antitumor modalities remains a priority of modern translation medicine. Adoptive transfer of autologous T cells expressing chimeric antigen receptors (CARs) specific for tumor-associated surface targets of B cells has demonstrated encouraging results in recent clinical trials. It is tempting therefore to adapt this approach to solid cancers and move it to universal/allogeneic formats. For this reason, it appears very attractive to use human NK cell lines as CAR carriers. Using lentiviral delivery, human NK cell line YT was modified to express a PSMA-specific CAR and to produce an anticancer peptide lactaptin. "Armored” CAR-YT-Lact cell line obtained shows cytotoxicity against PSMA-positive prostate adenocarcinoma cells. Accordingly, YT-Lact cells display a moderate and non-specific activity against PSMA-negative target cells, which may translate into a tighter control of tumor cell escape via antigen loss.

Keywords

YT / lactaptin / NK cell lines / YT / CAR

References

[1] June C.H., Sadelain M. Chimeric Antigen Receptor Therapy. N. Engl. J. Med. 2018; 379(1): 64-73.

[2] Кулемзин С.В., Кузнецова В.В., Мамонкин М. и др. CAR T-клеточная терапия: баланс эффективности и безопасности. Молекулярная биология 2017; 51(2): 274-87.

[3] D'Aloia M.M., Zizzari I.G., Sacchetti B. et al. CAR-T cells: the long and winding road to solid tumors. Cell Death Dis. 2018; 9(3): 282.

[4] Al-Zoughbi W., Huang J., Paramasivan G.S. et al. Tumor macroenvironment and metabolism. Semin. Oncol. 2014; 41(2): 281-95.

[5] Denko N.C. Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat. Rev. Cancer 2008; 8(9): 705-13.

[6] Brown J.M., Giaccia A.J. The unique physiology of solid tumors: opportunities (and problems) for cancer therapy. Cancer Res. 1998; 58(7): 1408-16.

[7] O'Rourke D.M., Nasrallah M.P., Desai A. et al. A single dose of peripherally infused EGFRvIII-directed CAR T cells mediates antigen loss and induces adaptive resistance in patients with recurrent glioblastoma. Sci. Transl. Med. 2017; 9(399): pii: eaaa0984.

[8] Yeku O.O., Purdon T.J., Koneru M. et al. Armored CAR T cells enhance antitumor efficacy and overcome the tumor microenvironment. Sci. Rep. 2017; 7(1): 10541.

[9] Li S., Siriwon N., Zhang X. et al. Enhanced Cancer Immunotherapy by Chimeric Antigen Receptor-Modified T Cells Engineered to Secrete Check-point Inhibitors. Clin. Cancer Res. 2017; 23(22): 6982-92.

[10] Pizzo E., Di Maro A. A new age for biomedical applications of Ribosome Inactivating Proteins (RIPs): from bioconjugate to nanoconstructs. J. Biomed. Sci. 2016; 23(1): 54.

[11] Ohba M., Mizuki E., Uemori A. Parasporin, a new anticancer protein group from Bacillus thuringiensis. Anticancer Res. 2009; 29(1): 427-33.

[12] Ni Y., Schwaneberg U., Sun Z.H. Arginine deiminase, a potential anti-tumor drug. Cancer Lett. 2008; 261(1): 1-11.

[13] Pritsa A.A., Papazisis K.T., Kortsaris A.H. et al. Antitumor activity of L-asparaginase from Thermus thermophilus. Anticancer Drugs 2001; 12(2): 137-42.

[14] Koval O.A., Fomin A.S., Kaledin V.I. et al. A novel pro-apoptotic effector lactaptin inhibits tumor growth in mice models. Biochimie 2012, 94(12): 2467-74.

[15] Koval O.A., Tkachenko A.V., Fomin A.S. et al. Lactaptin induces p53-independent cell death associated with features of apoptosis and autophagy and delays growth of breast cancer cells in mouse xenografts. PLoS One 2014; 9(4): e93921.

[16] Yodoi J., Teshigawara K., Nikaido T. et al. TCGF (IL 2)-receptor inducing factor(s). I. Regulation of IL 2 receptor on a natural killer-like cell line (YT cells). J. Immunol. 1985; 134(3): 1623-30.

[17] Коваль О.А., Волкова О.Ю., Горчаков А.А. и др. Сравнительный анализ активности лактаптина, полученного в про- и эукариотических системах экспрессии. Вавиловский журнал генетики и селекции 2017; 21(7): 764-9.

[18] Кулемзин 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): 124-32.

[19] Kingston R.E., Chen C.A., Rose J.K. Calcium phosphate transfection. Curr. Protoc. Mol. Biol. 2003; 9: 9.1.

[20] O'Doherty U., Swiggard W.J., Malim M.H. Human immunodeficiency virus type 1 spinoculation enhances infection through virus binding. J. Virol. 2000; 74(21): 10074-80.

[21] Zheng Z., Chinnasamy N., Morgan R.A. Protein L: a novel reagent for the detection of chimeric antigen receptor (CAR) expression by flow cytometry. J. Transl. Med. 2012; 10: 29.

[22] Golubovskaya V., Berahovich R., Zhou H. et al. CD47-CAR-T Cells Effectively Kill Target Cancer Cells and Block Pancreatic Tumor Growth. Cancers 2017; 9(10): pii: E139.

[23] Koval O.A., Sakaeva G.R., Fomin A.S. et al. Sensitivity of endometrial cancer cells from primary human tumor samples to new potential anticancer peptide lactaptin. J. Cancer Res. Ther. 2015; 11(2): 345-51.

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