[논문]Bispecific Antibody-Bound T Cells as a Novel Anticancer Immunotherapy

Cho, Jaewon; Tae, Nara; Ahn, Jae-Hee; Chang, Sun-Young; Ko, Hyun-Jeong; Kim, Dae Hee

doi:10.4062/biomolther.2022.015

[국내논문] Bispecific Antibody-Bound T Cells as a Novel Anticancer Immunotherapy 원문보기

Biomolecules & therapeutics, v.30 no.5, 2022년, pp.418 - 426

Cho, Jaewon (Department of Pharmacy, Kangwon National University) , Tae, Nara (Kangwon Institute of Inclusive Technology, Kangwon National University) , Ahn, Jae-Hee (Department of Pharmacy, Kangwon National University) , Chang, Sun-Young (College of Pharmacy, Ajou University) , Ko, Hyun-Jeong (Department of Pharmacy, Kangwon National University) , Kim, Dae Hee (Department of Pharmacy, Kangwon National University)

Abstract ▼ AI-Helper

Chimeric antigen receptor T (CAR-T) cell therapy is one of the promising anticancer treatments. It shows a high overall response rate with complete response to blood cancer. However, there is a limitation to solid tumor treatment. Additionally, this currently approved therapy exhibits side effects such as cytokine release syndrome and neurotoxicity. Alternatively, bispecific antibody is an innovative therapeutic tool that simultaneously engages specific immune cells to disease-related target cells. Since programmed death ligand 1 (PD-L1) is an immune checkpoint molecule highly expressed in some cancer cells, in the current study, we generated αCD3xαPD-L1 bispecific antibody (BiTE) which can engage T cells to PD-L1+ cancer cells. We observed that the BiTE-bound OT-1 T cells effectively killed cancer cells in vitro and in vivo. They substantially increased the recruitment of effector memory CD8+ T cells having CD8+CD44+CD62Llow phenotype in tumor. Interestingly, we also observed that BiTE-bound polyclonal T cells showed highly efficacious tumor killing activity in vivo in comparison with the direct intravenous treatment of bispecific antibody, suggesting that PD-L1-directed migration and engagement of activated T cells might increase cancer cell killing. Additionally, BiTE-bound CAR-T cells which targets human Her-2/neu exhibited enhanced killing effect on Her-2-expressing cancer cells in vivo, suggesting that this could be a novel therapeutic regimen. Collectively, our results suggested that engaging activated T cells with cancer cells using αCD3xαPD-L1 BiTE could be an innovative next generation anticancer therapy which exerts simultaneous inhibitory functions on PD-L1 as well as increasing the infiltration of activated T cells having effector memory phenotype in tumor site.

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그림 Fig. 1. Generation of bispecific T-cell engager-bound T cells (BiTE:T cells) as a novel anticancer cell therapy. (A) Bispecific T-cell engager molecules were generated with two single chain fragment variables (scFvs). mCD3xPD-L1 BiTE contains scFvs of anti-CD3ε antibody (145- 2C11) and anti-PD-L1 antibody (KL001-13), which were linked with 'GGGGS' linker. (B) mCD3xPD-L1 BiTE was cultured with CD8+ T cells for 1 h or 48 h in following concentration: 0.01 μg/mL, 0.1 μg/mL, and 1 μg/mL. T cells were stained with 1 μM of CellTraceTM Violet (CTV) and dilution of CTV was measured with FACS to determine T cell proliferation. Non-treated T cells were used as a negative control and indicated it as red line in figure. ***p<0.001, ****p<0.0001 (Student's t-test; n=3/group). (C) Schematic diagram of BiTE-bound T cells (BiTE:T cells) preparation. To prepare BiTE:T cells, CD8+ T cells were isolated from the spleen of mice and incubated for 48 h with 4 μg/mL of αCD3 antibody and 2 μg/mL of αCD28 antibody. Then mCD3xPD-L1 BiTE (0.1 μg/mL) were incubated with the CD8+ T cells for 1 h. BiTE-bound CD8+ T cells (BiTE:T cells) were obtained after removing the unbound BiTEs by washing with PBS. (D) CD8+ T cells from wild type mice isolated using magnetic beads. T cells or BiTE:T cells were prepared following protocol shown in 1C. T cells or BiTE:T cells were incubated with MO5 cells (1×104/well) for 16 h. Represented ratio is Target cell:Effector cell. Cancer killing activity of BiTE:T cells or BiTE:OT-1 cells was measured by CCK8 and CTV assay.
그림 Fig. 2. In vivo antitumor effect of BiTE:OT-1 cells. (A) Scheme of the in vivo experiment for BiTE:OT-1 T cells. Groups of mice were subcutaneously injected with MO5 (2×10⁶/mouse) in their left flank and the tumor size was monitored for 28 days. C57BL/6 mice, which were transplanted with MO5, were injected with OT-1 T cells or BiTE:OT-1 T cells (2×10⁵/mouse) after 10 and 20 days of tumor cell injection. (B) Tumor size was measured in every 2 or 3 days. All groups of mice were sacrificed on day 28. *p<0.05, **p<0.01 (Student's t-test; n=6/ group). (C) CD8+ T cells from wild type mice or OVA-specific OT-1 CD8⁺ T cells (>90% of which were Vα2-positive) isolated from OT-1 mice using magnetic beads. BiTE:T cells or BiTE:OT-1 cells were prepared following protocol shown in 1C. BiTE:T cells or BiTE:OT-1 cells were incubated with MO5 cells for 16 h. Cancer killing activity of BiTE:T cells or BiTE:OT-1 cells were measured by CCK8 and CTV assay. ***p<0.001 (Student's t-test; n=3/group). (D) Tumor-infiltrating lymphocytes (TILs) were analyzed. Effector memory T cells (CD3⁺CD8⁺ CD62L^low) were analyzed by FACS. The population and number of them were shown. **p<0.01, ***p<0.001 (Student';s t-test; n=4/group). These data are representative of three independent experiments.
그림 Fig. 3. Antitumor effect of BiTE:T cells constructed with activated autologous polyclonal CD8⁺ T cells. (A) Scheme of the in vivo experiment for BiTE-bound autologous polyclonal T cells (BiTE:T cells). Groups of mice were subcutaneously injected with MO5 (2×10⁶/mouse) in their left flank and monitored for 30 days. BiTE (2 μg/kg), autologous polyclonal T cells, or BiTE:T cells (2×10⁵/mouse) were intravenously transferred to tumor-bearing mice after 10 days and 20 days of tumor injection. (B) Tumor sizes were monitored for every 2 days and the tumor weights were measured at day 30 after euthanizing the mice. **p<0.01, ***p<0.001 (Student's t-test; n=6/group). (C) CD3⁺CD8⁺CD44⁺CD62L⁺ effector memory T cells and CD3⁺CD8⁺CD44⁺CD62L^- central memory T cells in tumor infiltrating lymphocytes (TILs) of every group of mice were analyzed by FACS. *p<0.05, **p<0.01 (Student's t-test; n=4/group). (D) MO5 cells were grafted in the left flank of C57BL/6 and BiTE (2 μg/kg), and BiTE-bound T cells (BiTE:T) with or without activation through αCD3 and αCD28 were intravenously injected after 10 days and 20 days of tumor challenge. Tumor sizes were monitored for every 2 days or 3 days. Non-activated T cells were also cultured for the same period as the activated T cells. After euthanizing the mice, tumor weight was measured. *p<0.05, **p<0.01 (Student's t-test; n=4/group). All the data shown are representative of three independent experiments.
그림 Fig. 4. Novel antitumor cell therapy of BiTE:ML39 CAR-T. (A) Diagram of the in vivo study for BiTE-bound ML39 CAR-T cells (BiTE:ML39 CAR-T). Groups of mice were grafted with Her-2/CT26 cells (2×10⁶/mouse) into their left flank and monitored for 30 days. ML39 CAR-T cells were generated to express anti-hHer2/neu (ErbB2) for targeting the Her-2/CT26 cells. After 10 and 20 days of tumor injection, mice were injected with control CAR-T cells (CTL CAR-T), ML39 CAR-T cells, or BiTE:ML39 CAR-T cells (2×10⁵/mouse). (B) Confirmation of PD-L1 expression on the surface of Her-2/CT26 cells. (C) Tumor sizes were monitored for every 2 days. *p<0.05, ***p<0.001 (Student's t-test; n=10/ group). (D) After euthanizing every group of mice, the tumor was removed from their body and the tumor mass was measured. *p<0.05, ***p<0.001 (Student's t-test; n=5-8/group). These data are representative of three independent experiments.

참고문헌 (31)

Ahn J. H. Lee B. H. Kim S. E. Kwon B. E. Jeong H. Choi J. R. Kim M. J. Park Y. Kim B. S. Kim D. H. Ko H. J. 2021 A novel anti-PD-L1 antibody exhibits antitumor effects on multiple myeloma in murine models via antibody-dependent cellular cytotoxicity Biomol. Ther. (Seoul) 29 166 174 10.4062/biomolther.2020.131 33139584

원문보기 상세보기
Alegre M. L. Tso J. Y. Sattar H. A. Smith J. Desalle F. Cole M. Bluestone J. A. 1995 An anti-murine CD3 monoclonal antibody with a low affinity for Fc gamma receptors suppresses transplantation responses while minimizing acute toxicity and immunogenicity J. Immunol. 155 1544 1555 7636216

상세보기
Baird J. H. Frank M. J. Craig J. Patel S. Spiegel J. Y. Sahaf B. Oak J. S. Younes S. F. Ozawa M. G. Yang E. Natkunam Y. Tamaresis J. Ehlinger Z. Reynolds W. D. Arai S. Johnston L. Lowsky R. Meyer E. Negrin R. S. Rezvani A. R. Shiraz P. Sidana S. Weng W. K. Davis K. L. Ramakrishna S. Schultz L. Mullins C. Jacob A. Kirsch I. Feldman S. A. Mackall C. L. Miklos D. B. Muffly L. 2021 CD22-directed CAR T-cell therapy induces complete remissions in CD19-directed CAR-refractory large B-cell lymphoma Blood 137 2321 2325 10.1182/blood.2020009432 33512414

상세보기
Choi J. R. Kim M. J. Tae N. Wi T. M. Kim S. H. Lee E. S. Kim D. H. 2020 BLI-based functional assay in phage display benefits the development of a PD-L1-targeting therapeutic antibody Viruses 12 684 10.3390/v12060684 89ef47725f7e408ab4e0d9b3bbb6a168 32630442

상세보기
Chung Y. Kim B. S. Kim Y. J. Ko H. J. Ko S. Y. Kim D. H. Kang C. Y. 2006 CD1d-restricted T cells license B cells to generate long-lasting cytotoxic antitumor immunity in vivo Cancer Res. 66 6843 6850 10.1158/0008-5472.CAN-06-0889 16818662

상세보기
Edeline J. Houot R. Marabelle A. Alcantara M. 2021 CAR-T cells and BiTEs in solid tumors: challenges and perspectives J. Hematol. Oncol. 14 65 10.1186/s13045-021-01067-5 2bbb8d605fec4173a17b12b48f0231fe 33874996

상세보기
Franquiz M. J. Short N. J. 2020 Blinatumomab for the treatment of adult B-cell acute lymphoblastic leukemia: toward a new era of targeted immunotherapy Biol. Targets Ther. 14 23 34 10.2147/BTT.S202746 32103893

상세보기
Geuijen C. Tacken P. Wang L. C. Klooster R. van Loo P. F. Zhou J. Mondal A. Liu Y. B. Kramer A. Condamine T. Volgina A. Hendriks L. J. A. van der Maaden H. Rovers E. Engels S. Fransen F. den Blanken-Smit R. Zondag-van der Zande V. Basmeleh A. Bartelink W. Kulkarni A. Marissen W. Huang C. Y. Hall L. Harvey S. Kim S. Martinez M. O'Brien S. Moon E. Albelda S. Kanellopoulou C. Stewart S. Nastri H. Bakker A. B. H. Scherle P. Logtenberg T. Hollis G. de Kruif J. Huber R. Mayes P. A. Throsby M. 2021 A human CD137×PD-L1 bispecific antibody promotes anti-tumor immunity via context-dependent T cell costimulation and checkpoint blockade Nat. Commun. 12 4445 10.1038/s41467-021-24767-5 4044cbbb9c1d4937ac08f857eba69b30 34290245

상세보기
Horn L. A. Ciavattone N. G. Atkinson R. Woldergerima N. Wolf J. Clements V. K. Sinha P. Poudel M. Ostrand-Rosenberg S. 2017 CD3xPDL1 bi-specific T cell engager (BiTE) simultaneously activates T cells and NKT cells, kills PDL1(+) tumor cells, and extends the survival of tumor-bearing humanized mice Oncotarget 8 57964 57980 10.18632/oncotarget.19865 28938530

상세보기
Jeong S. Park E. Kim H. D. Sung E. Kim H. Jeon J. Kim Y. Jung U. J. Son Y. G. Hong Y. Lee H. Lee S. Lim Y. Won J. Jeon M. Hwang S. Fang L. Jiang W. Wang Z. Shin E. C. Park S. H. Jung J. 2021 Novel anti-4-1BB×PD-L1 bispecific antibody augments anti-tumor immunity through tumor-directed T-cell activation and checkpoint blockade J. Immunother. Cancer 9 e002428 10.1136/jitc-2021-002428 f928715e9bd14a19be6842809c144bf9 34230109

상세보기
Jiang H. Ni H. Zhang P. Guo X. Wu M. Shen H. Wang J. Wu W. Wu Z. Ding J. Tang R. Zhou S. Chen B. Yu M. Jing H. Liu J. 2021 PD-L1/LAG-3 bispecific antibody enhances tumor-specific immunity Oncoimmunology 10 1943180 10.1080/2162402X.2021.1943180 34239776

상세보기
Kershaw M. H. Westwood J. A. Darcy P. K. 2013 Gene-engineered T cells for cancer therapy Nat. Rev. Cancer 13 525 541 10.1038/nrc3565 23880905

상세보기
Kershaw M. H. Westwood J. A. Parker L. L. Wang G. Eshhar Z. Mavroukakis S. A. White D. E. Wunderlich J. R. Canevari S. Rogers-Freezer L. Chen C. C. Yang J. C. Rosenberg S. A. Hwu P. 2006 A phase I study on adoptive immunotherapy using gene-modified T cells for ovarian cancer Clin. Cancer Res. 12 6106 6115 10.1158/1078-0432.CCR-06-1183 17062687

상세보기
Ko H. J. Kim Y. J. Kim Y. S. Chang W. S. Ko S. Y. Chang S. Y. Sakaguchi S. Kang C. Y. 2007 A combination of chemoimmunotherapies can efficiently break self-tolerance and induce antitumor immunity in a tolerogenic murine tumor model Cancer Res. 67 7477 7486 10.1158/0008-5472.CAN-06-4639 17671218

상세보기
Li Y. Kurlander R. J. 2010 Comparison of anti-CD3 and anti-CD28-coated beads with soluble anti-CD3 for expanding human T cells: differing impact on CD8 T cell phenotype and responsiveness to restimulation J. Transl. Med. 8 104 10.1186/1479-5876-8-104 20977748

상세보기
Liu Q. Sun Z. Chen L. 2020 Memory T cells: strategies for optimizing tumor immunotherapy Protein Cell 11 549 564 10.1007/s13238-020-00707-9 f7f324e589c04dc2a6ecb736bbbecb13 32221812

상세보기
Maude S. L. Laetsch T. W. Buechner J. Rives S. Boyer M. Bittencourt H. Bader P. Verneris M. R. Stefanski H. E. Myers G. D. Qayed M. De Moerloose B. Hiramatsu H. Schlis K. Davis K. L. Martin P. L. Nemecek E. R. Yanik G. A. Peters C. Baruchel A. Boissel N. Mechinaud F. Balduzzi A. Krueger J. June C. H. Levine B. L. Wood P. Taran T. Leung M. Mueller K. T. Zhang Y. Sen K. Lebwohl D. Pulsipher M. A. Grupp S. A. 2018 Tisagenlecleucel in children and young adults with B-cell lymphoblastic leukemia N. Engl. J. Med. 378 439 448 10.1056/NEJMoa1709866 29385370

상세보기
Moon E. K. Carpenito C. Sun J. Wang L. C. Kapoor V. Predina J. Powell D. J. Jr. Riley J. L. June C. H. Albelda S. M. 2011 Expression of a functional CCR2 receptor enhances tumor localization and tumor eradication by retargeted human T cells expressing a mesothelin-specific chimeric antibody receptor Clin. Cancer Res. 17 4719 4730 10.1158/1078-0432.CCR-11-0351 21610146

상세보기
Moon E. K. Wang L. C. Dolfi D. V. Wilson C. B. Ranganathan R. Sun J. Kapoor V. Scholler J. Puré E. Milone M. C. June C. H. Riley J. L. Wherry E. J. Albelda S. M. 2014 Multifactorial T-cell hypofunction that is reversible can limit the efficacy of chimeric antigen receptor-transduced human T cells in solid tumors Clin. Cancer Res. 20 4262 4273 10.1158/1078-0432.CCR-13-2627 24919573

상세보기
Neelapu S. S. Locke F. L. Bartlett N. L. Lekakis L. J. Miklos D. B. Jacobson C. A. Braunschweig I. Oluwole O. O. Siddiqi T. Lin Y. Timmerman J. M. Stiff P. J. Friedberg J. W. Flinn I. W. Goy A. Hill B. T. Smith M. R. Deol A. Farooq U. McSweeney P. Munoz J. Avivi I. Castro J. E. Westin J. R. Chavez J. C. Ghobadi A. Komanduri K. V. Levy R. Jacobsen E. D. Witzig T. E. Reagan P. Bot A. Rossi J. Navale L. Jiang Y. Aycock J. Elias M. Chang D. Wiezorek J. Go W. Y. 2017 Axicabtagene ciloleucel CAR T-cell therapy in refractory large B-cell lymphoma N. Engl. J. Med. 377 2531 2544 10.1056/NEJMoa1707447 29226797

상세보기
Newick K. O'Brien S. Moon E. Albelda S. M. 2017 CAR T cell therapy for solid tumors Annu. Rev. Med. 68 139 152 10.1146/annurev-med-062315-120245 27860544

상세보기
Principe N. Kidman J. Goh S. Tilsed C. M. Fisher S. A. Fear V. S. Forbes C. A. Zemek R. M. Chopra A. Watson M. Dick I. M. Boon L. Holt R. A. Lake R. A. Nowak A. K. Lesterhuis W. J. McDonnell A. M. Chee J. 2020 Tumor infiltrating effector memory antigen-specific CD8(+) T cells predict response to immune checkpoint therapy Front. Immunol. 11 584423 10.3389/fimmu.2020.584423 14028b32db2541389131a67ac973ccfb 33262762

상세보기
Qin S. Xu L. Yi M. Yu S. Wu K. Luo S. 2019 Novel immune checkpoint targets: moving beyond PD-1 and CTLA-4 Mol. Cancer 18 155 10.1186/s12943-019-1091-2 7f6ea285e3834011a1ff496a2f1048a8 31690319

상세보기
Teoh P. J. Chng W. J. 2021 CAR T-cell therapy in multiple myeloma: more room for improvement Blood Cancer J. 11 84 10.1038/s41408-021-00469-5 bb326b5565e34c6d95d81ac310641d49 33927192

상세보기
Till B. G. Jensen M. C. Wang J. Qian X. Gopal A. K. Maloney D. G. Lindgren C. G. Lin Y. Pagel J. M. Budde L. E. Raubitschek A. Forman S. J. Greenberg P. D. Riddell S. R. Press O. W. 2012 CD20-specific adoptive immunotherapy for lymphoma using a chimeric antigen receptor with both CD28 and 4-1BB domains: pilot clinical trial results Blood 119 3940 3950 10.1182/blood-2011-10-387969 22308288

상세보기
Turtle C. J. Hay K. A. Hanafi L. A. Li D. Cherian S. Chen X. Wood B. Lozanski A. Byrd J. C. Heimfeld S. Riddell S. R. Maloney D. G. 2017 Durable molecular remissions in chronic lymphocytic leukemia treated with CD19-specific chimeric antigen receptor-modified T cells after failure of ibrutinib J. Clin. Oncol. 35 3010 3020 10.1200/JCO.2017.72.8519 28715249

상세보기
Waldman A. D. Fritz J. M. Lenardo M. J. 2020 A guide to cancer immunotherapy: from T cell basic science to clinical practice Nat. Rev. Immunol. 20 651 668 10.1038/s41577-020-0306-5 32433532

상세보기
You G. Won J. Lee Y. Moon D. Park Y. Lee S. H. Lee S. W. 2021 Bispecific antibodies: a smart arsenal for cancer immunotherapies Vaccines 9 724 10.3390/vaccines9070724 f948b667f9af4ceeb9bfca14fd8e65b8 34358141

상세보기
Yu W. L. Hua Z. C. 2019 Chimeric antigen receptor T-cell (CAR T) therapy for hematologic and solid malignancies: efficacy and safety-a systematic review with meta-analysis Cancers (Basel) 11 47 10.3390/cancers11010047 30621018

상세보기
Zhang C. Liu J. Zhong J. F. Zhang X. 2017 Engineering CAR-T cells Biomarker Res. 5 22 10.1186/s40364-017-0102-y 28652918

상세보기
Zheng P. P. Kros J. M. Li J. 2018 Approved CAR T cell therapies: ice bucket challenges on glaring safety risks and long-term impacts Drug Discov. Today 23 1175 1182 10.1016/j.drudis.2018.02.012 29501911

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