In vivo immunotherapy of lung cancer using cross-species reactive vascular endothelial growth factor nanobodies

Document Type : Original Article

Authors

1 Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab., Pasteur Institute of Iran, Tehran, Iran

2 National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran

Abstract

Objective(s): Lung cancer is the main leading cause of cancer death worldwide. Angiogenesis is the main step in proliferation and spreading of tumor cells. Targeting vascular endothelial growth factor (VEGF) is an effective approach for inhibition of cancer angiogenesis. Nanobodies (NBs) are a novel class of antibodies derived from the camel. Unique characteristics of Nbs like their small size and good penetration to tumor tissues makes them promising tools in drug development.  Development of NBs targeting both human and mouse VEGF is required for understanding their in vivo functions.  Therefore, development of cross-species reactive anti-VEGF Nbs for immunotherapy of lung cancer was the main aim of the current study.
Materials and Methods: Here we developed NBs from Camelus dromedarius library with high specificity and binding affinity to both human and mouse VEGF. In vitro and In vivo function of developed NB was evaluated on human endothelial cells and lung epithelial tumor cells (TC-1).
Results: A nanobody showed the highest affinity to human and mouse VEGF and potently inhibited VEGF in the ELISA experiment. Anti-VEGF NBs significantly inhibited in vitro human endothelial cell migration through blockade of VEGF (P=0.045). Anti-VEGF NBs also significantly inhibited in vivo TC-1 growth in a dose-dependent manner (P=0.001) and resulted in higher survival rate in the nanobody treated group
Conclusion: These findings demonstrate the potential of anti-VEGF NBsin tumor growth inhibition and are promising as novel cancer therapeutic candidate.

Keywords

References

1. Cheng T-YD, Cramb SM, Baade PD, Youlden DR, Nwogu C, Reid ME. The international epidemiology of lung cancer: latest trends, disparities, and tumor characteristics. J Thorac Oncol 2016; 11:1653-1671
2. Adnan K, Zahra EM, Sharareh S, Shirin K, Habib E, Kian K. Clinicopathological  characteristics of Iranian patients with lung cancer: a single institute experience. Asian Pac J Cancer Prev 2016; 17:3815-3820.
3. Tulotta C, He S, van der Ent W, Chen L, Groenewoud A, Spaink H, et al. Imaging cancer angiogenesis and metastasis in a zebrafish embryo model. Cancer and Zebrafish: Springer; 2016. p. 239-263.
4. O'Donnell RK, Falcon B, Hanson J, Goldstein WE, Perruzzi C, Rafii S, et al. VEGF-A/VEGFR inhibition restores hematopoietic homeostasis in the bone marrow and attenuates tumor growth. Cancer Res 2016; 76:517-524.
5. Shojaei F. Anti-angiogenesis therapy in cancer: Current challenges and future perspectives. Cancer Lett 2012; 320:130-137.
6. Bhattacharya R, Xia L, Fan F, Wang R, Boulbes D, Ye X-C, et al. Inhibition of intracrine VEGF signaling prevents colorectal cancer cell migration and invasion. Cancer Res 2016; 76:3255-.
7. Gille H, Hülsmeyer M, Trentmann S, Matschiner G, Christian HJ, Meyer T, et al. Functional characteri-zation of a VEGF-A-targeting Anticalin, prototype of a novel therapeutic human protein class. Angiogenesis 2016; 19:79-94.
8. Cosmai L, Gallieni M, Liguigli W, Porta C. Renal toxicity of anticancer agents targeting vascular endothelial growth factor (VEGF) and its receptors (VEGFRs). J Nephrol 2016; 30:171-180.
9. Hamilton JL, Nagao M, Levine BR, Chen D, Olsen BR, Im HJ. Targeting VEGF and its receptors for the treatment of osteoarthritis and associated pain. J Bone Miner Res 2016; 31:911-924.
10. Hurwitz H, Fehrenbacher L, Novotny W, Cartwright T, Hainsworth J, et al. Bevacizumab plus irinotecan, fluorouracil, and leucovorin for metastatic colorectal cancer. N Engl J Med 2004; 350:2335-2342.
11. Miller K, Wang M,   Gralow J,  Dickler M,  Cobleigh M,  Perez EA,  Shenkier T,Cella D, Davidson  NE. Paclitaxel plus bevacizumab versus paclitaxel alonefor metastatic breast cancer. N Engld J Med 2007; 357:2666-2676.
12. Ellis LM, HicklinDJ. Pathways mediating resistance to vascular endothelial growth factor-targeted therapy. Clin Cancer Res 2008; 14:6371-6375.
13. Huang T, Wang H, Chen NG, Frentzen A, Minev B, Szalay AA. Expression of anti-VEGF antibody together with anti-EGFR or anti-FAP enhances tumor regression as a result of vaccinia virotherapy. Mol Ther Oncol 2015; 2:15003.
14. Senturk B, Cubuk MO, Ozmen MC, Aydin B, Guler MO, Tekinay AB. Inhibition of VEGF mediated corneal neovascularization by anti-angiogenic peptide nanofibers. Biomaterials 2016 107:124-132.
15. Behdani M, Zeinali S,  Khanahmad H,  Karimipour M, Asadzadeh N,  et al. Generation and characteri-zation of a functional Nanobody against the vascular endothelial growth factor receptor-2; angiogenesis cell receptor. Mol Immunol 2012; 50:35-41.
16. Kolkman JA, Law DA. Nanobodies-from llamas to therapeutic proteins. Drug Discov Today Technol 2010; 7:139-146.
17. Gerber HP, Wu X, Yu L, Wiesmann C, Liang XH, Lee CV, et al. Mice expressing a humanized form of VEGF-A may provide insights into the safety and efficacy of anti-VEGF antibodies. Proc Natl Acad Sci U S A 2007; 104:3478-3483.
18. Heist RS, Duda DG, Sahani DV, Ancukiewicz M, Fidias P, Sequist LV, et al. Improved tumor vascularization after anti-VEGF therapy with carboplatin and nab-paclitaxel associates with survival in lung cancer. Proc Natl Acad Sci 2015; 112:1547-1552.
19. Kazemi-Lomedasht F, Behdani M, Pooshang Bagheri K, Habibi-Anbouhi M, Abolhassani M, Arezumand R, Shahbazzadeh D, Mirzahoseini H. Inhibition of angiogenesis in human endothelial cell using VEGF specific nanobody. Mol Immunol 2015; 65:58-67.
20. Beatty JD, Beatty BG, Vlahos WG. Measurement of monoclonal antibody affinity by non-competitive enzyme immunoassay. J Immunol Methods 1987; 100:173-139.
21. Han M, Wang H, Zhang H-T, Han Z. Expression of Tax-interacting protein 1 (TIP-1) facilitates angiogenesis and tumor formation of human glioblastoma cells in nude mice. Cancer Lett  2013; 328:55-64.
22. Yardehnavi N, Behdani M, Pooshang  Bagheri K, Mahmoodzadeh A, Khanahmad H, Shahbazzadeh D,  et al. A camelid antibody candidate for development of a therapeutic agent against Hemiscorpius lepturus envenomation. FASEB J  2014; 28:4004-4014.
23. Xanthopoulos JM, Romano AE,   MajumdarSK. Response of mouse breast cancer cells to anastrozole, tamoxifen, and the combination. J Biomed Biotechnol 2005; 1:10-19.
24. Jamnani FR, Rahbarizadeh F, Shokrgozar MA, Ahmadvand D, Mahboudi F, Sharifzadeh Z. Targeting high affinity and epitope-distinct oligoclonal nanobodies to HER2 over-expressing tumor cells. Exp Cell Res 2012; 318:1112-1124.
25. Muyldermans S. Single domain camel antibodies: current status. J Biotechnol 2001; 74:277-302.
26. Zhou R, Curry J, Roy L, Grover P, Haider J, Moore L, et al. A novel association of neuropilin-1 and MUC1 in pancreatic ductal adenocarcinoma: role in induction of VEGF signaling and angiogenesis. Oncogene 2016; 35:5608-5618.
27. Simons M, Gordon E, Claesson-Welsh L. Mechanisms and regulation of endothelial VEGF receptor signalling. Nat Rev Mol Cell Biol 2016; 17:611-625.
28. García‐Caballero M, Blacher S, Paupert J, Quesada
A, Medina M, Noël A. Novel application assigned            to toluquinol: inhibition of lymphangiogenesis by interfering with VEGF‐C/VEGFR‐3 signalling pathway. Br J Pharmacol 2016; 173:1966-1987.
29. Liang WC, Wu X, Peale FV, Lee CV, Meng YG, Gutierrez J, et al. Cross-species vascular endothelial growth factor(VEGF)-blocking antibodies completely inhibit the growth of human tumor xenografts and measure the contribution of stromal VEGF. J Biol Chem 2006; 281:951-961.
30. McMurphy T, Xiao R, Magee D, Slater A, Zabeau L, Tavernier J, et al.The anti-tumor activity of a neutralizing nanobody targeting leptin receptor in a mouse model of melanoma. PLoS One 2014; 9:e89895.
31. Vosjan MJ, Vercammen J, Kolkman JA, Stigter-van Walsum M, Revets H, van Dongen GA.  Nanobodies targeting the hepatocyte growth factor: potential new drugs for molecular cancer therapy. Mol Cancer Ther 2012; 11:1017-1025.
32. Inoue M, Hager JH, Ferrara N, Gerber HP, Hanahan D. VEGF-A has a critical, nonredundant role in angiogenic switching and pancreatic beta cell carcinogenesis. Cancer Cell 2002; 1:193-202.
33. Ferrara N, Hillan KJ, Gerber HP, Novotny W. Discovery and development of bevacizumab, an anti-VEGF antibody for treating cancer. Nat Rev Drug Discov 2004; 3:391-400.
Iranian Journal of Basic Medical Sciences
Volume 20, Issue 5
May 2017
Pages 489-496

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