A novel tetravalent bispecific antibody targeting programmed death 1 and tyrosine-protein kinase Met for treatment of gastric cancer
Background Redirecting T cells to tumor cells using bispecific antibodies (BsAbs) is emerging as a potent cancer therapy. The main concept of this strategy is to cross-link tumor cells and T cells by simultaneously binding to cell surface tumor-associated antigen (TAA) and the CD3ƹ chain. However, immune checkpoint programmed cell death ligand-1 (PD-L1) on tumor cells or other myeloid cells upreglulated remarkablely after the treatment of CD3-binding BsAbs, leads to the generation of suppressed microenvironment for immune evasion and tumor progression. Although this resistance could be partially reversed by anti-PD-L1 treatment, targeting two pathways through one antibody-based molecule may provide a strategic advantage over the combination of BsAbs and immune checkpoint inhibitors. Methods We developed two novel BsAbs PD-1/c-Met DVD-Ig and IgG-scFv both targeting PD-1 to restore the immune effector function of T cells and engaging them to tumor cells via binding to cellular-mesenchymal to epithelial transition factor (c-Met). Binding activities, T cell activation and proliferation were analyzed by flow cytometry. Cell Cytotoxicity and cytokine release were measured using LDH release assay and ELISA, respectively. Anti-tumor response in vivo was evaluated by generate xenograft models in NOD-SCID mice. Results These bispecific antibodies exhibited effective antitumor activity against high- and low- c-Met-expressing gastric cancer cell lines in vitro and mediated strong tumor growth inhibition in human gastric cancer xenograft models. Conclusion The engagement of the PD-1/PD-L1 blockade to c-Met-overexpressing cancer cells is a promising strategy for the treatment of gastric cancer and potentially other malignancies.
KeywordsPD-1 C-MET Bispecific antibody Cancer immunotherapy
This work was supported financially by the National Key Research Project Bio-safety Key Technology Development Program 2016YFC1201501.
Compliance with ethical standards
Conflict of interest
All procedures involving human blood products were in accordance with the 1964 Helsinki declaration and its later amendments. Study involving mice use was carried out according to the protocols approved by Institutional Animal Care and Use Committee of Fudan University.
All individual participants who donated blood samples for analysis signed an informed consent form.
- 5.Lennerz JK, Kwak EL, Ackerman A, Michael M, Fox SB, Bergethon K, Lauwers GY, Christensen JG, Wilner KD, Haber DA, Salgia R, Bang YJ, Clark JW, Solomon BJ, Iafrate AJ (2011) MET amplification identifies a small and aggressive subgroup of esophagogastric adenocarcinoma with evidence of responsiveness to crizotinib. J Clin Oncol Off J Am Soc Clin Oncol 29(36):4803–4810. https://doi.org/10.1200/JCO.2011.35.4928 CrossRefGoogle Scholar
- 6.Erichsen R, Kelsh MA, Oliner KS, Nielsen KB, Froslev T, Laenkholm AV, Vyberg M, Acquavella J, Sorensen HT (2016) Prognostic impact of tumor MET expression among patients with stage IV gastric cancer: a Danish cohort study. Ann Epidemiol 26(7):500–503. https://doi.org/10.1016/j.annepidem.2016.05.002 CrossRefGoogle Scholar
- 9.Dreier T, Lorenczewski G, Brandl C, Hoffmann P, Syring U, Hanakam F, Kufer P, Riethmuller G, Bargou R, Baeuerle PA (2002) Extremely potent, rapid and costimulation-independent cytotoxic T-cell response against lymphoma cells catalyzed by a single-chain bispecific antibody. Int J Cancer 100(6):690–697. https://doi.org/10.1002/ijc.10557 CrossRefGoogle Scholar
- 13.Cartellieri M, Arndt C, Feldmann A, von Bonin M, Ewen EM, Koristka S, Michalk I, Stamova S, Berndt N, Gocht A, Bornhauser M, Ehninger G, Schmitz M, Bachmann M (2013) TCR/CD3 activation and co-stimulation combined in one T cell retargeting system improve anti-tumor immunity. Oncoimmunology 2(12):e26770. https://doi.org/10.4161/onci.26770 CrossRefGoogle Scholar
- 14.Bargou R, Leo E, Zugmaier G, Klinger M, Goebeler M, Knop S, Noppeney R, Viardot A, Hess G, Schuler M, Einsele H, Brandl C, Wolf A, Kirchinger P, Klappers P, Schmidt M, Riethmuller G, Reinhardt C, Baeuerle PA, Kufer P (2008) Tumor regression in cancer patients by very low doses of a T cell-engaging antibody. Science 321(5891):974–977. https://doi.org/10.1126/science.1158545 CrossRefGoogle Scholar
- 15.Osada T, Patel SP, Hammond SA, Osada K, Morse MA, Lyerly HK (2015) CEA/CD3-bispecific T cell-engaging (BiTE) antibody-mediated T lymphocyte cytotoxicity maximized by inhibition of both PD1 and PD-L1. Cancer Immunol Immunother: CII 64(6):677–688. https://doi.org/10.1007/s00262-015-1671-y CrossRefGoogle Scholar
- 17.Taube JM, Anders RA, Young GD, Xu H, Sharma R, McMiller TL, Chen S, Klein AP, Pardoll DM, Topalian SL, Chen L (2012) Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape. Sci Transl Med 4(127):127ra137. https://doi.org/10.1126/scitranslmed.3003689 CrossRefGoogle Scholar
- 19.Junttila TT, Li J, Johnston J, Hristopoulos M, Clark R, Ellerman D, Wang BE, Li Y, Mathieu M, Li G, Young J, Luis E, Lewis Phillips G, Stefanich E, Spiess C, Polson A, Irving B, Scheer JM, Junttila MR, Dennis MS, Kelley R, Totpal K, Ebens A (2014) Antitumor efficacy of a bispecific antibody that targets HER2 and activates T cells. Cancer Res 74(19):5561–5571. https://doi.org/10.1158/0008-5472.CAN-13-3622-T CrossRefGoogle Scholar
- 20.Sun LL, Wang P, Clark R, Hristopoulos M, Ellerman D, Mathieu M, Chu Y-W, Wang H, Totpal K, Ebens AJ, Polson AG, Gould S (2016) Preclinical characterization of combinability and potential synergy of anti-CD20/CD3 T-cell dependent bispecific antibody with chemotherapy and PD-1/PD-L1 blockade. Blood 128(22):4168–4168Google Scholar
- 23.Eto S, Yoshikawa K, Nishi M, Higashijima J, Tokunaga T, Nakao T, Kashihara H, Takasu C, Iwata T, Shimada M (2016) Programmed cell death protein 1 expression is an independent prognostic factor in gastric cancer after curative resection. Gastric Cancer 19(2):466–471. https://doi.org/10.1007/s10120-015-0519-7 CrossRefGoogle Scholar
- 25.Wu C, Ying H, Grinnell C, Bryant S, Miller R, Clabbers A, Bose S, McCarthy D, Zhu RR, Santora L, Davis-Taber R, Kunes Y, Fung E, Schwartz A, Sakorafas P, Gu J, Tarcsa E, Murtaza A, Ghayur T (2007) Simultaneous targeting of multiple disease mediators by a dual-variable-domain immunoglobulin. Nat Biotechnol 25(11):1290–1297. https://doi.org/10.1038/nbt1345 CrossRefGoogle Scholar
- 28.Nagai T, Arao T, Furuta K, Sakai K, Kudo K, Kaneda H, Tamura D, Aomatsu K, Kimura H, Fujita Y, Matsumoto K, Saijo N, Kudo M, Nishio K (2011) Sorafenib inhibits the hepatocyte growth factor-mediated epithelial mesenchymal transition in hepatocellular carcinoma. Mol Cancer Ther 10(1):169–177. https://doi.org/10.1158/1535-7163.MCT-10-0544 CrossRefGoogle Scholar
- 36.Klein C, Sustmann C, Thomas M, Stubenrauch K, Croasdale R, Schanzer J, Brinkmann U, Kettenberger H, Regula JT, Schaefer W (2012) Progress in overcoming the chain association issue in bispecific heterodimeric IgG antibodies. mAbs 4(6):653–663. https://doi.org/10.4161/mabs.21379 CrossRefGoogle Scholar
- 41.Michaud NR, Jani JP, Hillerman S, Tsaparikos KE, Barbacci-Tobin EG, Knauth E, Putz H Jr, Campbell M, Karam GA, Chrunyk B, Gebhard DF, Green LL, Xu JJ, Dunn MC, Coskran TM, Lapointe JM, Cohen BD, Coleman KG, Bedian V, Vincent P, Kajiji S, Steyn SJ, Borzillo GV, Los G (2012) Biochemical and pharmacological characterization of human c-Met neutralizing monoclonal antibody CE-355621. mAbs 4(6):710–723. https://doi.org/10.4161/mabs.22160 CrossRefGoogle Scholar
- 42.Pacchiana G, Chiriaco C, Stella MC, Petronzelli F, De Santis R, Galluzzo M, Carminati P, Comoglio PM, Michieli P, Vigna E (2010) Monovalency unleashes the full therapeutic potential of the DN-30 anti-Met antibody. J Biol Chem 285(46):36149–36157. https://doi.org/10.1074/jbc.M110.134031 CrossRefGoogle Scholar