Development of a Human Antibody That Exhibits Antagonistic Activity Toward CC Chemokine Receptor 7

Overview

Journal Antib Ther

Publisher Oxford University Press

Date 2022 Aug 15

PMID 35967907

Authors

Moon-Sung Jang

Nurain Syahirah Binti Ismail

Yeon Gyu Yu

Affiliations

Soon will be listed here.

Abstract

Background: CC chemokine receptor 7 (CCR7) is a member of G-protein-coupled receptor family and mediates chemotactic migration of immune cells and different cancer cells induced via chemokine (C-C motif) ligand 19 (CCL19) or chemokine (C-C motif) ligand 21 (CCL21). Hence, the identification of blockade antibodies against CCR7 could lead to the development of therapeutics targeting metastatic cancer.

Methods: CCR7 was purified and stabilized in its active conformation, and antibodies specific to purified CCR7 were screened from the synthetic M13 phage library displaying humanized scFvs. The characterization of selected scFvs identified two scFvs that exhibited CCL19-competitive binding to CCR7. IgG's harboring selected scFv sequences were characterized for binding activity in CCR7 cells, inhibitory activity toward CCR7-dependent cAMP attenuation, and the CCL19 or CCL21-dependent migration of CCR7 cells.

Results: Antibodies specifically binding to purified CCR7 and CCR7 cells were isolated and characterized. Two antibodies, IgG(6RG11) and IgG(72C7), showed ligand-dependent competitive binding to CCR7 with K values of 40 nM and 50 nM, respectively. Particularly, IgG(6RG11) showed antagonistic activity against CCR7, whereas both antibodies significantly blocked the ligand-induced migration and invasion activity of CCR7 cancer cells.

Conclusions: Two antibody clones were successfully identified from a synthetic scFv-displaying phage library using purified recombinant CCR7 as an antigen. Antibodies specifically bound to the surface of CCR7 cells and blocked CCR7 cell migration. Particularly, 6RG11 showed antagonist activity against CCR7-dependent cAMP attenuation.

References

Cuesta-Mateos C, Juarez-Sanchez R, Mateu-Albero T, Loscertales J, Mol W, Terron F . Targeting cancer homing into the lymph node with a novel anti-CCR7 therapeutic antibody: the paradigm of CLL. MAbs. 2021; 13(1):1917484. PMC: 8098074. DOI: 10.1080/19420862.2021.1917484. View

Forster R, Schubel A, Breitfeld D, Kremmer E, Renner-Muller I, Wolf E . CCR7 coordinates the primary immune response by establishing functional microenvironments in secondary lymphoid organs. Cell. 1999; 99(1):23-33. DOI: 10.1016/s0092-8674(00)80059-8. View

Fang L, Kao Y, Chuang Y, Huang H, Tai T . Ets-1 enhances tumor migration through regulation of CCR7 expression. BMB Rep. 2019; 52(9):548-553. PMC: 6774420. View

Forster R, Davalos-Misslitz A, Rot A . CCR7 and its ligands: balancing immunity and tolerance. Nat Rev Immunol. 2008; 8(5):362-71. DOI: 10.1038/nri2297. View

Pan M, Hou M, Chang H, Hung W . Cyclooxygenase-2 up-regulates CCR7 via EP2/EP4 receptor signaling pathways to enhance lymphatic invasion of breast cancer cells. J Biol Chem. 2008; 283(17):11155-63. DOI: 10.1074/jbc.M710038200. View

Goupil E, Laporte S, Hebert T . Functional selectivity in GPCR signaling: understanding the full spectrum of receptor conformations. Mini Rev Med Chem. 2012; 12(9):817-30. DOI: 10.2174/138955712800959143. View

Lacalle R, Blanco R, Carmona-Rodriguez L, Martin-Leal A, Mira E, Manes S . Chemokine Receptor Signaling and the Hallmarks of Cancer. Int Rev Cell Mol Biol. 2017; 331:181-244. DOI: 10.1016/bs.ircmb.2016.09.011. View

Nakata B, Fukunaga S, Noda E, Amano R, Yamada N, Hirakawa K . Chemokine receptor CCR7 expression correlates with lymph node metastasis in pancreatic cancer. Oncology. 2008; 74(1-2):69-75. DOI: 10.1159/000139126. View

Choi H, Song H, Jung Y . The Roles of CCR7 for the Homing of Memory CD8+ T Cells into Their Survival Niches. Immune Netw. 2020; 20(3):e20. PMC: 7327150. DOI: 10.4110/in.2020.20.e20. View

10.

Kang S, Kim N, Yu Y . Identification of novel positive allosteric modulators of GLP1R that stimulate its interaction with ligands and G subunits. Biochem Biophys Res Commun. 2021; 583:162-168. DOI: 10.1016/j.bbrc.2021.10.071. View

11.

Sanchez-Sanchez N, Riol-Blanco L, Rodriguez-Fernandez J . The multiple personalities of the chemokine receptor CCR7 in dendritic cells. J Immunol. 2006; 176(9):5153-9. DOI: 10.4049/jimmunol.176.9.5153. View

12.

Shannon L, Calloway P, Welch T, Vines C . CCR7/CCL21 migration on fibronectin is mediated by phospholipase Cgamma1 and ERK1/2 in primary T lymphocytes. J Biol Chem. 2010; 285(50):38781-7. PMC: 2998076. DOI: 10.1074/jbc.M110.152173. View

13.

Okada T, Cyster J . CC chemokine receptor 7 contributes to Gi-dependent T cell motility in the lymph node. J Immunol. 2007; 178(5):2973-8. DOI: 10.4049/jimmunol.178.5.2973. View

14.

Kim N, Kim K, Shin S, Kim E, Yu Y . Functional expression of human prostaglandin E2 receptor 4 (EP4) in and characterization of the binding property of EP4 with G proteins. Biochem Biophys Rep. 2020; 25:100871. PMC: 7749421. DOI: 10.1016/j.bbrep.2020.100871. View

15.

Webb D, Handel T, Kretz-Rommel A, Stevens R . Opportunities for functional selectivity in GPCR antibodies. Biochem Pharmacol. 2012; 85(2):147-52. PMC: 3729224. DOI: 10.1016/j.bcp.2012.08.021. View

16.

Fowler K, Vasilieva V, Ivanova E, Rimkevich O, Sokolov A, Abbasova S . R707, a fully human antibody directed against CC-chemokine receptor 7, attenuates xenogeneic acute graft-versus-host disease. Am J Transplant. 2019; 19(7):1941-1954. DOI: 10.1111/ajt.15298. View

17.

Heng B, Aubel D, Fussenegger M . An overview of the diverse roles of G-protein coupled receptors (GPCRs) in the pathophysiology of various human diseases. Biotechnol Adv. 2013; 31(8):1676-94. DOI: 10.1016/j.biotechadv.2013.08.017. View

18.

Rizeq B, Malki M . The Role of CCL21/CCR7 Chemokine Axis in Breast Cancer Progression. Cancers (Basel). 2020; 12(4). PMC: 7226115. DOI: 10.3390/cancers12041036. View

19.

Hauser M, Legler D . Common and biased signaling pathways of the chemokine receptor CCR7 elicited by its ligands CCL19 and CCL21 in leukocytes. J Leukoc Biol. 2016; 99(6):869-82. DOI: 10.1189/jlb.2MR0815-380R. View

20.

Ju M, Ahn H, Han S, Ko S, Na J, Jo M . A human antibody against human endothelin receptor type A that exhibits antitumor potency. Exp Mol Med. 2021; 53(9):1437-1448. PMC: 8492878. DOI: 10.1038/s12276-021-00678-9. View