Cancer Therapeutic Trispecific Antibodies Recruiting Both T and Natural Killer Cells to Cancer Cells

Overview

Journal Oncol Rep

Specialty Oncology

Date 2023 Oct 20

PMID 37859608

Authors

Kouki Kimura

Atsushi Kuwahara

Saori Suzuki

Takeshi Nakanishi

Izumi Kumagai

Ryutaro Asano

Affiliations

Soon will be listed here.

Abstract

T cells and natural killer (NK) cells are major effector cells recruited by cancer therapeutic bispecific antibodies; however, differences in the populations of these cells in individual tumors limit the general use of these antibodies. In the present study, trispecific antibodies were created, namely T cell and NK cell engagers (TaKEs), that recruit both T cells and NK cells. Notably, three Fc‑fused TaKEs were designed, TaKE1‑Fc, TaKE2‑Fc and TaKE3‑Fc, using variable fragments targeting the epidermal growth factor receptor on tumor cells, CD3 on T cells, and CD16 on NK cells. Among them, TaKE1‑Fc was predicted to form a circular tetrabody‑like configuration and exhibited the highest production and greatest cancer growth inhibitory effects. TaKE1 was prepared from TaKE1‑Fc by digesting the Fc region for further functional evaluation. The resulting TaKE1 exhibited trispecificity via its ability to bind cancer cells, T cells and NK cells, as well as comparable or greater cancer growth inhibitory effects to those of two bispecific antibodies that recruit T cells and NK cells, respectively. A functional trispecific antibody with the potential to exert strong therapeutic effects independent of T cell and NK cell populations was developed.

References

Jaiswal D, Verma S, Nair D, Salunke D . Antibody multispecificity: A necessary evil?. Mol Immunol. 2022; 152:153-161. DOI: 10.1016/j.molimm.2022.10.012. View

Schlereth B, Fichtner I, Lorenczewski G, Kleindienst P, Brischwein K, da Silva A . Eradication of tumors from a human colon cancer cell line and from ovarian cancer metastases in immunodeficient mice by a single-chain Ep-CAM-/CD3-bispecific antibody construct. Cancer Res. 2005; 65(7):2882-9. DOI: 10.1158/0008-5472.CAN-04-2637. View

Li S, Schmitz K, Jeffrey P, Wiltzius J, Kussie P, Ferguson K . Structural basis for inhibition of the epidermal growth factor receptor by cetuximab. Cancer Cell. 2005; 7(4):301-11. DOI: 10.1016/j.ccr.2005.03.003. View

Wu L, Seung E, Xu L, Rao E, Lord D, Wei R . Trispecific antibodies enhance the therapeutic efficacy of tumor-directed T cells through T cell receptor co-stimulation. Nat Cancer. 2022; 1(1):86-98. DOI: 10.1038/s43018-019-0004-z. View

Gall F, Reusch U, Moldenhauer G, Little M, Kipriyanov S . Immunosuppressive properties of anti-CD3 single-chain Fv and diabody. J Immunol Methods. 2004; 285(1):111-27. DOI: 10.1016/j.jim.2003.11.007. View

Pei X, Holliger P, Murzin A, Williams R . The 2.0-A resolution crystal structure of a trimeric antibody fragment with noncognate VH-VL domain pairs shows a rearrangement of VH CDR3. Proc Natl Acad Sci U S A. 1997; 94(18):9637-42. PMC: 23241. DOI: 10.1073/pnas.94.18.9637. View

Zhong X, DAntona A . Recent Advances in the Molecular Design and Applications of Multispecific Biotherapeutics. Antibodies (Basel). 2021; 10(2). PMC: 8103270. DOI: 10.3390/antib10020013. View

Asano R, Sone Y, Makabe K, Tsumoto K, Hayashi H, Katayose Y . Humanization of the bispecific epidermal growth factor receptor x CD3 diabody and its efficacy as a potential clinical reagent. Clin Cancer Res. 2006; 12(13):4036-42. DOI: 10.1158/1078-0432.CCR-06-0059. View

Beha N, Harder M, Ring S, Kontermann R, Muller D . IL15-Based Trifunctional Antibody-Fusion Proteins with Costimulatory TNF-Superfamily Ligands in the Single-Chain Format for Cancer Immunotherapy. Mol Cancer Ther. 2019; 18(7):1278-1288. DOI: 10.1158/1535-7163.MCT-18-1204. View

10.

Ecker D, Jones S, Levine H . The therapeutic monoclonal antibody market. MAbs. 2014; 7(1):9-14. PMC: 4622599. DOI: 10.4161/19420862.2015.989042. View

11.

Meermeier E, Welsh S, Sharik M, Du M, Garbitt V, Riggs D . Tumor burden limits bispecific antibody efficacy through T cell exhaustion averted by concurrent cytotoxic therapy. Blood Cancer Discov. 2021; 2(4):354-369. PMC: 8266040. DOI: 10.1158/2643-3230.BCD-21-0038. View

12.

Arlotta K, Owen S . Antibody and antibody derivatives as cancer therapeutics. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2019; 11(5):e1556. DOI: 10.1002/wnan.1556. View

13.

Kuwahara A, Nagai K, Nakanishi T, Kumagai I, Asano R . Functional Domain Order of an Anti-EGFR × Anti-CD16 Bispecific Diabody Involving NK Cell Activation. Int J Mol Sci. 2020; 21(23). PMC: 7727810. DOI: 10.3390/ijms21238914. View

14.

Asano R, Shimomura I, Konno S, Ito A, Masakari Y, Orimo R . Rearranging the domain order of a diabody-based IgG-like bispecific antibody enhances its antitumor activity and improves its degradation resistance and pharmacokinetics. MAbs. 2014; 6(5):1243-54. PMC: 4623410. DOI: 10.4161/mabs.29445. View

15.

Asano R, Ikoma K, Sone Y, Kawaguchi H, Taki S, Hayashi H . Highly enhanced cytotoxicity of a dimeric bispecific diabody, the hEx3 tetrabody. J Biol Chem. 2010; 285(27):20844-9. PMC: 2898345. DOI: 10.1074/jbc.M110.120444. View

16.

Asano R, Kumagai T, Nagai K, Taki S, Shimomura I, Arai K . Domain order of a bispecific diabody dramatically enhances its antitumor activity beyond structural format conversion: the case of the hEx3 diabody. Protein Eng Des Sel. 2013; 26(5):359-67. DOI: 10.1093/protein/gzt009. View

17.

Wang J, Kang G, Yuan H, Cao X, Huang H, de Marco A . Research Progress and Applications of Multivalent, Multispecific and Modified Nanobodies for Disease Treatment. Front Immunol. 2022; 12:838082. PMC: 8804282. DOI: 10.3389/fimmu.2021.838082. View

18.

Asano R, Kuroki Y, Honma S, Akabane M, Watanabe S, Mayuzumi S . Comprehensive study of domain rearrangements of single-chain bispecific antibodies to determine the best combination of configurations and microbial host cells. MAbs. 2018; 10(6):854-863. PMC: 6152445. DOI: 10.1080/19420862.2018.1476815. View

19.

Maejima A, Suzuki S, Makabe K, Kumagai I, Asano R . Incorporation of a repeated polypeptide sequence in therapeutic antibodies as a universal masking procedure: A case study of T cell-engaging bispecific antibodies. N Biotechnol. 2023; 77:80-89. DOI: 10.1016/j.nbt.2023.07.004. View

20.

Hudson P, Kortt A . High avidity scFv multimers; diabodies and triabodies. J Immunol Methods. 2000; 231(1-2):177-89. DOI: 10.1016/s0022-1759(99)00157-x. View