Rahimizadeh Parastou, Kim Seheon, Yoon Byeong Jun, Jeong Younsik, Lim Seoyoen, Jeon Hyeyoon, Lim Hyung Jin, Park Sang Ho, Park Sang-In, Kong Deok-Hoon, Park Jeong-Ran, Song Yong Bhum
Division of Research Center, Scripps Korea Antibody Institute, Chuncheon 24341, Republic of Korea.
Division of Research Center, Scripps Korea Antibody Institute, Chuncheon 24341, Republic of Korea.
Biomed Pharmacother. 2025 Apr;185:117966. doi: 10.1016/j.biopha.2025.117966. Epub 2025 Mar 9.
G protein-coupled receptors (GPCRs) are crucial in several physiological and pathological processes and are associated with numerous diseases across all therapeutic areas. Antibodies are well-established therapeutics with better selectivity, stability, and half-life than small molecules and peptides. CXC motif chemokine receptor 2 (CXCR2) and its ligands play functional roles in the progression and metastasis of tumors and activation and trafficking of inflammatory mediators. Several chemical-based antagonists that inhibit the CXCLs/CXCR2 signaling axis are under clinical study in cancer or inflammatory disease research; however, therapeutic antibodies have not yet been successfully established. In this study, we used phage display technology to identify single-chain variable fragment proteins that target the N-terminal domain of human CXCR2. Subsequently, we performed an enzyme-linked immunosorbent assay to validate the interaction of these IgG1 candidates and bio-layer interferometry to determine their affinity. The association of these antibodies, IgG18 and IgG56, with stable HEK293 cell lines expressing hCXCR2 and various cancer cell lines, including pancreatic cancer cells, was further analyzed. We found that IgG18 and IgG56 antibodies antagonized CXCL8-induced CXCR2 signaling, suppressed CXCL8-mediated cell proliferation and migration, and induced apoptosis in pancreatic cancer cells. In a xenograft model of pancreatic cancer, CXCR2 antibodies, IgG18 and IgG56, decreased tumor growth and induced apoptosis in vivo. Our results indicate that CXCR2 inhibitory antibodies attenuate tumor progression and may be potential candidates for anti-tumor therapeutics.
G蛋白偶联受体（GPCRs）在多种生理和病理过程中起着关键作用，并且与所有治疗领域的众多疾病相关。抗体是成熟的治疗药物，与小分子和肽相比，具有更好的选择性、稳定性和半衰期。CXC基序趋化因子受体2（CXCR2）及其配体在肿瘤的进展和转移以及炎症介质的激活和运输中发挥功能作用。几种抑制CXCLs/CXCR2信号轴的基于化学的拮抗剂正在癌症或炎症性疾病研究中进行临床研究；然而，治疗性抗体尚未成功建立。在本研究中，我们使用噬菌体展示技术来鉴定靶向人CXCR2 N端结构域的单链可变片段蛋白。随后，我们进行了酶联免疫吸附测定以验证这些IgG1候选物的相互作用，并进行生物层干涉术以确定它们的亲和力。进一步分析了这些抗体IgG18和IgG56与表达hCXCR2的稳定HEK293细胞系以及包括胰腺癌细胞在内的各种癌细胞系的结合情况。我们发现IgG18和IgG56抗体拮抗CXCL8诱导的CXCR2信号传导，抑制CXCL8介导的细胞增殖和迁移，并诱导胰腺癌细胞凋亡。在胰腺癌异种移植模型中，CXCR2抗体IgG18和IgG56在体内降低肿瘤生长并诱导凋亡。我们的结果表明，CXCR2抑制性抗体可减弱肿瘤进展，可能是抗肿瘤治疗的潜在候选药物。
