Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4
Abstract Background: Antibodies and derivative drugs targeting immune checkpoints have been approved for the treatment of several malignancies, but there are fewer responses in patients with pancreatic cancer. Here, we designed a nanobody molecule with bi-targeting on PD-L1 and CXCR4, as both target...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-10-01
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| Series: | BMC Cancer |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12885-022-10165-7 |
| _version_ | 1797991613522247680 |
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| author | Shuai Hao Shuyi Xu Liangzhu Li Yaxian Li Meiqi Zhao Junsheng Chen Shunying Zhu Yueqing Xie Hua Jiang Jianwei Zhu Mingyuan Wu |
| author_facet | Shuai Hao Shuyi Xu Liangzhu Li Yaxian Li Meiqi Zhao Junsheng Chen Shunying Zhu Yueqing Xie Hua Jiang Jianwei Zhu Mingyuan Wu |
| author_sort | Shuai Hao |
| collection | DOAJ |
| description | Abstract Background: Antibodies and derivative drugs targeting immune checkpoints have been approved for the treatment of several malignancies, but there are fewer responses in patients with pancreatic cancer. Here, we designed a nanobody molecule with bi-targeting on PD-L1 and CXCR4, as both targets are overexpressed in many cancer cells and play important roles in tumorigenesis. We characterized the biochemical and anti-tumour activities of the bispecific nanobodies in vitro and in vivo. Methods: A nanobody molecule was designed and constructed. The nanobody sequences targeting PD-L1 and CXCR4 were linked by the (G4S)3 flexible peptide to construct the anti-PD-L1/CXCR4 bispecific nanobody. The bispecific nanobody was expressed in E. coli cells and purified by affinity chromatography. The purified nanobody was biochemically characterized by mass spectrometry, Western blotting and flow cytometry to confirm the molecule and its association with both PD-L1 and CXCR4. The biological function of the nanobody and its anti-tumour effects were examined by an in vitro tumour cell-killing assay and in vivo tumour inhibition in mouse xenograft models. Results: A novel anti-PD-L1/CXCR4 bispecific nanobody was designed, constructed and characterized. The molecule specifically bound to two targets on the surface of human cancer cells and inhibited CXCL12-induced Jurkat cell migration. The bispecific nanobody increased the level of IFN-γ secreted by T-cell activation. The cytotoxicity of human peripheral blood mononuclear cells (hPBMCs) against pancreatic cancer cells was enhanced by the molecule in combination with IL-2. In a human pancreatic cancer xenograft model, the anti-PD-L1/CXCR4 nanobody markedly inhibited tumour growth and was superior to the combo-treatment by anti-PD-L1 nanobody and anti-CXCR4 nanobody or treatment with atezolizumab as a positive control. Immunofluorescence and immunohistochemical staining of xenograft tumours showed that the anti-tumour effects were associated with the inhibition of angiogenesis and the infiltration of immune cells. Conclusion: These results clearly revealed that the anti-PD-L1/CXCR4 bispecific nanobody exerted anti-tumour efficacy in vitro and inhibited tumour growth in vivo. This agent can be further developed as a therapeutic reagent to treat human pancreatic cancer by simultaneously blocking two critical targets. |
| first_indexed | 2024-04-11T08:55:00Z |
| format | Article |
| id | doaj.art-3e00bfdc6861413c8cde585309b0cf39 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-04-11T08:55:00Z |
| publishDate | 2022-10-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-3e00bfdc6861413c8cde585309b0cf392022-12-22T04:33:19ZengBMCBMC Cancer1471-24072022-10-0122111210.1186/s12885-022-10165-7Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4Shuai Hao0Shuyi Xu1Liangzhu Li2Yaxian Li3Meiqi Zhao4Junsheng Chen5Shunying Zhu6Yueqing Xie7Hua Jiang8Jianwei Zhu9Mingyuan Wu10Engineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityInstitute of Translational Medicine, Shanghai Jiao Tong UniversityJecho Laboratories, IncJecho Laboratories, IncEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityEngineering Research Center of Cell and Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong UniversityAbstract Background: Antibodies and derivative drugs targeting immune checkpoints have been approved for the treatment of several malignancies, but there are fewer responses in patients with pancreatic cancer. Here, we designed a nanobody molecule with bi-targeting on PD-L1 and CXCR4, as both targets are overexpressed in many cancer cells and play important roles in tumorigenesis. We characterized the biochemical and anti-tumour activities of the bispecific nanobodies in vitro and in vivo. Methods: A nanobody molecule was designed and constructed. The nanobody sequences targeting PD-L1 and CXCR4 were linked by the (G4S)3 flexible peptide to construct the anti-PD-L1/CXCR4 bispecific nanobody. The bispecific nanobody was expressed in E. coli cells and purified by affinity chromatography. The purified nanobody was biochemically characterized by mass spectrometry, Western blotting and flow cytometry to confirm the molecule and its association with both PD-L1 and CXCR4. The biological function of the nanobody and its anti-tumour effects were examined by an in vitro tumour cell-killing assay and in vivo tumour inhibition in mouse xenograft models. Results: A novel anti-PD-L1/CXCR4 bispecific nanobody was designed, constructed and characterized. The molecule specifically bound to two targets on the surface of human cancer cells and inhibited CXCL12-induced Jurkat cell migration. The bispecific nanobody increased the level of IFN-γ secreted by T-cell activation. The cytotoxicity of human peripheral blood mononuclear cells (hPBMCs) against pancreatic cancer cells was enhanced by the molecule in combination with IL-2. In a human pancreatic cancer xenograft model, the anti-PD-L1/CXCR4 nanobody markedly inhibited tumour growth and was superior to the combo-treatment by anti-PD-L1 nanobody and anti-CXCR4 nanobody or treatment with atezolizumab as a positive control. Immunofluorescence and immunohistochemical staining of xenograft tumours showed that the anti-tumour effects were associated with the inhibition of angiogenesis and the infiltration of immune cells. Conclusion: These results clearly revealed that the anti-PD-L1/CXCR4 bispecific nanobody exerted anti-tumour efficacy in vitro and inhibited tumour growth in vivo. This agent can be further developed as a therapeutic reagent to treat human pancreatic cancer by simultaneously blocking two critical targets.https://doi.org/10.1186/s12885-022-10165-7PD-L1CXCR4AntibodyBispecific nanobodyImmunotherapyPancreatic cancer |
| spellingShingle | Shuai Hao Shuyi Xu Liangzhu Li Yaxian Li Meiqi Zhao Junsheng Chen Shunying Zhu Yueqing Xie Hua Jiang Jianwei Zhu Mingyuan Wu Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4 BMC Cancer PD-L1 CXCR4 Antibody Bispecific nanobody Immunotherapy Pancreatic cancer |
| title | Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4 |
| title_full | Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4 |
| title_fullStr | Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4 |
| title_full_unstemmed | Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4 |
| title_short | Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4 |
| title_sort | tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting pd l1 and cxcr4 |
| topic | PD-L1 CXCR4 Antibody Bispecific nanobody Immunotherapy Pancreatic cancer |
| url | https://doi.org/10.1186/s12885-022-10165-7 |
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