Download PDF
CC BY 4.0 · Pharmaceutical Fronts 2020; 02(02): e100-e108
DOI: 10.1055/s-0040-1714138
Original Article
Georg Thieme Verlag KG Stuttgart · New York

Generating a Novel Bispecific Nanobody to Enhance Antitumor Activity

Qiuhan Ge
1   College of Pharmacy and Chemistry, Dali University, Yunnan, People's Republic of China
,
Tianyuan Sun
1   College of Pharmacy and Chemistry, Dali University, Yunnan, People's Republic of China
,
Yanlin Bian
2   Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, People's Republic of China
,
Xiaodong Xiao
3   Jecho Laboratories, Inc., Maryland, United States
4   Jecho Biopharmaceuticals Co., Ltd., Tianjin, People's Republic of China
,
Jianwei Zhu
2   Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai, People's Republic of China
3   Jecho Laboratories, Inc., Maryland, United States
4   Jecho Biopharmaceuticals Co., Ltd., Tianjin, People's Republic of China
› Author Affiliations
Further Information

Publication History

Publication Date:
28 July 2020 (online)

Also available at
   Permissions and Reprints

Abstract

Tumor cells express high levels of human epidermal growth factor receptor 2 (HER2) and vascular endothelial growth factor receptor 2 (VEGFR2), which are closely related to their proliferation and survival. Cancer treatments that target a single signaling pathway may result in immune pathway escape or drug resistance. Based on the correlation between the HER2 and VEGFR2 signaling pathways, we speculated that targeting the two pathways simultaneously may produce a synergistic effect and avoid occurrence of drug resistance, resulting in improved efficacy. Anti-VEGFR2 nanobody 3VGR19–3 and anti-HER2 nanobody 2D3 were combined to construct a bispecific nanobody (Bi-Nb). They can recognize both HER2 and VEGFR2 (both highly expressed in HT-29 cells) to simultaneously block the two signaling pathways. We verified the affinity of the Bi-Nb to its targets using the surface plasmon resonance technology, and test its effects to inhibit tumor cell growth and promote cell apoptosis in vitro by the Cell Counting Kit-8 assay and apoptosis assay. In summary, we have successfully constructed a Bi-Nb, and verified its tumor-suppressing effects in vitro. Compared with a single monospecific nanobody, our Bi-Nb showed superior antitumor effect, which provides a new perspective for treatment of tumors with high HER2 and VEGFR2 expression.

Keywords

bispecific nanobody - HER2 - VEGFR2 - colorectal cancer
 

  • References

  • 1 Gupta N, Kupfer SS, Davis AM. Colorectal cancer screening. JAMA 2019; 321 (20) 2022-2023
    CrossrefPubMedSearch in Google Scholar
  • 2 Agus DB, Bunn Jr PA, Franklin W, Garcia M, Ozols RF. HER-2/neu as a therapeutic target in non-small cell lung cancer, prostate cancer, and ovarian cancer. Semin Oncol 2000; 27 (06) (Suppl. 11) 53-63 , discussion 92–100
    PubMedSearch in Google Scholar
  • 3 Smith NR, Baker D, James NH. , et al. Vascular endothelial growth factor receptors VEGFR-2 and VEGFR-3 are localized primarily to the vasculature in human primary solid cancers. Clin Cancer Res 2010; 16 (14) 3548-3561
    CrossrefPubMedSearch in Google Scholar
  • 4 Larsen AK, Ouaret D, El Ouadrani K, Petitprez A. Targeting EGFR and VEGF(R) pathway cross-talk in tumor survival and angiogenesis. Pharmacol Ther 2011; 131 (01) 80-90
    CrossrefPubMedSearch in Google Scholar
  • 5 Mitran B, Güler R, Roche FP. , et al. Radionuclide imaging of VEGFR2 in glioma vasculature using biparatopic affibody conjugate: proof-of-principle in a murine model. Theranostics 2018; 8 (16) 4462-4476
    CrossrefPubMedSearch in Google Scholar
  • 6 Nussenbaum F, Herman IM. Tumor angiogenesis: insights and innovations. J Oncol 2010; 2010: 132641
    PubMedSearch in Google Scholar
  • 7 Zhou Y, Zhen M, Guan M. , et al. Amino acid modified [70] fullerene derivatives with high radical scavenging activity as promising bodyguards for chemotherapy protection. Sci Rep 2018; 8 (01) 16573
    CrossrefPubMedSearch in Google Scholar
  • 8 Zhu J. Mammalian cell protein expression for biopharmaceutical production. Biotechnol Adv 2012; 30 (05) 1158-1170
    CrossrefPubMedSearch in Google Scholar
  • 9 Movahedi MM, Mehdizadeh A, Koosha F. , et al. Investigating the photo-thermo-radiosensitization effects of folate-conjugated gold nanorods on KB nasopharyngeal carcinoma cells. Photodiagn Photodyn Ther 2018; 24: 324-331
    CrossrefPubMedSearch in Google Scholar
  • 10 Roskoski Jr R. Small molecule inhibitors targeting the EGFR/ErbB family of protein-tyrosine kinases in human cancers. Pharmacol Res 2019; 139: 395-411
    CrossrefPubMedSearch in Google Scholar
  • 11 Wang X, Ouyang X, Chen J, Hu Y, Sun X, Yu Z. Nanoparticulate photosensitizer decorated with hyaluronic acid for photodynamic/photothermal cancer targeting therapy. Nanomedicine (Lond) 2019; 14 (02) 151-167
    Search in Google Scholar
  • 12 Warda W, Larosa F, Neto Da Rocha M. , et al. CML hematopoietic stem cells expressing IL1RAP can be targeted by chimeric antigen receptor-engineered T cells. Cancer Res 2019; 79 (03) 663-675
    CrossrefPubMedSearch in Google Scholar
  • 13 Khalil DN, Smith EL, Brentjens RJ, Wolchok JD. The future of cancer treatment: immunomodulation, CARs and combination immunotherapy. Nat Rev Clin Oncol 2016; 13 (05) 273-290
    CrossrefPubMedSearch in Google Scholar
  • 14 Tabas I, Glass CK. Anti-inflammatory therapy in chronic disease: challenges and opportunities. Science 2013; 339 (6116): 166-172
    CrossrefPubMedSearch in Google Scholar
  • 15 Graham BS, Ambrosino DM. History of passive antibody administration for prevention and treatment of infectious diseases. Curr Opin HIV AIDS 2015; 10 (03) 129-134
    CrossrefPubMedSearch in Google Scholar
  • 16 Zhou Y, Zong H, Han L. , et al. A novel bispecific antibody targeting CD3 and prolactin receptor (PRLR) against PRLR-expression breast cancer. J Exp Clin Cancer Res 2020; 39 (01) 87
    CrossrefPubMedSearch in Google Scholar
  • 17 Han L, Chen J, Ding K. , et al. Efficient generation of bispecific IgG antibodies by split intein mediated protein trans-splicing system. Sci Rep 2017; 7 (01) 8360
    CrossrefPubMedSearch in Google Scholar
  • 18 Han L, Zong H, Zhou Y. , et al. Naturally split intein Npu DnaE mediated rapid generation of bispecific IgG antibodies. Methods 2019; 154: 32-37
    CrossrefPubMedSearch in Google Scholar
  • 19 De Groof TWM, Mashayekhi V, Fan TS. , et al. Nanobody-targeted photodynamic therapy selectively kills viral GPCR-expressing glioblastoma cells. Mol Pharm 2019; 16 (07) 3145-3156
    CrossrefPubMedSearch in Google Scholar
  • 20 Kijanka M, Dorresteijn B, Oliveira S, van Bergen en Henegouwen PM. Nanobody-based cancer therapy of solid tumors. Nanomedicine (Lond) 2015; 10 (01) 161-174
    Search in Google Scholar
  • 21 Unciti-Broceta JD, Del Castillo T, Soriano M, Magez S, Garcia-Salcedo JA. Novel therapy based on camelid nanobodies. Ther Deliv 2013; 4 (10) 1321-1336
    CrossrefPubMedSearch in Google Scholar
  • 22 Ayyar BV, Arora S, O'Kennedy R. Coming-of-age of antibodies in cancer therapeutics. Trends Pharmacol Sci 2016; 37 (12) 1009-1028
    CrossrefPubMedSearch in Google Scholar
  • 23 Liu XL, Sun TY, Ge QH. , et al. Construction of novel bispecific single-domain antibodies (BiSdAbs) with potent antiangiogenic activities. Pharml Fronts 2020; 2 (01) e64-e76
    Search in Google Scholar
  • 24 Gao Z, Song C, Li G. , et al. Pyrotinib treatment on HER2-positive gastric cancer cells promotes the released exosomes to enhance endothelial cell progression, which can be counteracted by apatinib. OncoTargets Ther 2019; 12: 2777-2787
    Search in Google Scholar
  • 25 Vaidyanathan G, McDougald D, Choi J. , et al. Preclinical evaluation of 18F-labeled anti-HER2 nanobody conjugates for imaging HER2 receptor expression by immuno-PET. J Nucl Med 2016; 57 (06) 967-973
    CrossrefPubMedSearch in Google Scholar
  • 26 Brenner H, Kloor M, Pox CP. Colorectal cancer. Lancet 2014; 383 (9927): 1490-1502
    CrossrefPubMedSearch in Google Scholar
  • 27 Houlston RS, Cheadle J, Dobbins SE. , et al; COGENT Consortium; CORGI Consortium; COIN Collaborative Group; COINB Collaborative Group. Meta-analysis of three genome-wide association studies identifies susceptibility loci for colorectal cancer at 1q41, 3q26.2, 12q13.13 and 20q13.33. Nat Genet 2010; 42 (11) 973-977
    CrossrefPubMedSearch in Google Scholar
  • 28 Tomlinson IP, Carvajal-Carmona LG, Dobbins SE. , et al; COGENT Consortium; CORGI Collaborators; EPICOLON Consortium. Multiple common susceptibility variants near BMP pathway loci GREM1, BMP4, and BMP2 explain part of the missing heritability of colorectal cancer. PLoS Genet 2011; 7 (06) e1002105
    CrossrefPubMedSearch in Google Scholar
  • 29 Al-Tassan NA, Whiffin N, Hosking FJ. , et al. A new GWAS and meta-analysis with 1000Genomes imputation identifies novel risk variants for colorectal cancer. Sci Rep 2015; 5: 10442
    CrossrefPubMedSearch in Google Scholar
  • 30 Schumacher FR, Schmit SL, Jiao S. , et al. Genome-wide association study of colorectal cancer identifies six new susceptibility loci. Nat Commun 2015; 6: 7138
    CrossrefPubMedSearch in Google Scholar
  • 31 Alameddine RS, Otrock ZK, Awada A, Shamseddine A. Crosstalk between HER2 signaling and angiogenesis in breast cancer: molecular basis, clinical applications and challenges. Curr Opin Oncol 2013; 25 (03) 313-324
    CrossrefPubMedSearch in Google Scholar
  • 32 Yang W, Klos K, Yang Y, Smith TL, Shi D, Yu D. ErbB2 overexpression correlates with increased expression of vascular endothelial growth factors A, C, and D in human breast carcinoma. Cancer 2002; 94 (11) 2855-2861
    CrossrefPubMedSearch in Google Scholar
  • 33 Ishiguro T, Sano Y, Komatsu SI. , et al. An anti-glypican 3/CD3 bispecific T cell-redirecting antibody for treatment of solid tumors. Sci Transl Med 2017; 9 (410) 4291
    Search in Google Scholar
  • 34 Topp MS, Gökbuget N, Zugmaier G. , et al. Phase II trial of the anti-CD19 bispecific T cell-engager blinatumomab shows hematologic and molecular remissions in patients with relapsed or refractory B-precursor acute lymphoblastic leukemia. J Clin Oncol 2014; 32 (36) 4134-4140
    CrossrefPubMedSearch in Google Scholar
  • 35 Yu S, Li A, Liu Q. , et al. Recent advances of bispecific antibodies in solid tumors. J Hematol Oncol 2017; 10 (01) 155
    CrossrefPubMedSearch in Google Scholar
  • 36 Zhu JW. , ed. Update on Production of Recombinant Therapeutic Protein: Transient Gene Expression. Akron, OH: Smithers Rapra Technology; 2013
    Search in Google Scholar
  • 37 Ding K, Han L, Zong H, Chen J, Zhang B, Zhu J. Production process reproducibility and product quality consistency of transient gene expression in HEK293 cells with anti-PD1 antibody as the model protein. Appl Microbiol Biotechnol 2017; 101 (05) 1889-1898
    CrossrefPubMedSearch in Google Scholar
  • 38 Zhang X, Han L, Zong H. , et al. Enhanced production of anti-PD1 antibody in CHO cells through transient co-transfection with anti-apoptotic genes Bcl-x L and Mcl-1. Bioprocess Biosyst Eng 2018; 41 (05) 633-640
    CrossrefPubMedSearch in Google Scholar
  • 39 Luo M, Zhao M, Cagliero C. , et al. A general platform for efficient extracellular expression and purification of Fab from Escherichia coli. Appl Microbiol Biotechnol 2019; 103 (08) 3341-3353
    CrossrefPubMedSearch in Google Scholar
  • 40 Frankel SR, Baeuerle PA. Targeting T cells to tumor cells using bispecific antibodies. Curr Opin Chem Biol 2013; 17 (03) 385-392
    CrossrefPubMedSearch in Google Scholar