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Archives of Virology

, Volume 164, Issue 1, pp 69–82 | Cite as

Suppression of angiogenesis and tumor growth by recombinant T4 phages displaying extracellular domain of vascular endothelial growth factor receptor 2

  • Shuguang ZuoEmail author
  • Gongpeng Dai
  • Liping Wang
  • Yuqing Wen
  • Zhiang Huang
  • Wenyi Yang
  • Wanli Ma
  • Xuequn RenEmail author
Original Article

Abstract

Tumor growth, invasion and metastasis are dependent on angiogenesis. The Vascular endothelial growth factor (VEGF)/VEGF receptor 2 (VEGFR2) signaling pathway plays a pivotal role in tumor angiogenesis and therefore represents a reasonable target for anti-angiogenesis/anti-tumor therapy. In the present study, we generated T4 recombinant phages expressing the extracellular domain of VEGFR2 (T4-VEGFR2) and investigated their anti-angiogenic activity. The T4-VEGFR2 phages were able to bind to VEGF specifically and inhibit VEGF-mediated phosphorylation of VEGFR2 and its downstream kinases such as extracellular signal-regulated kinase (ERK) and p38 mitogen activated protein kinase (MAPK). The in vitro experiments showed that the T4-VEGFR2 phages could inhibit VEGF-stimulated cell proliferation and migration of endothelial cells. Finally, administration of T4-VEGFR2 phages was able to suppress tumor growth and decrease microvascular density in murine models of Lewis lung carcinoma and colon carcinoma, and prolong the survival of tumor bearing mice. In conclusion, this study reveals that the recombinant T4-VEGFR2 phages generated using T4-based phage display system can inhibit VEGF-mediated tumor angiogenesis and the T4 phage display technology can therefore be used for the development of novel anti-cancer strategies.

Abbreviations

ANOVA

Analysis of variance

ATCC

American Type Culture Collection

bp

Base pair

BSA

Bovine serum albumin

CC

Colon carcinoma

CSFV

Swine fever virus

DMSO

Dimethyl sulfoxide

DNA

Deoxyribonucleic acid

ECGS

Endothelial cell growth supplement

ELISA

Enzyme-linked immunosorbent assay

ERK

Extracellular-signal-regulated kinase

FBS

Fetal bovine serum

FLK1

Fetal liver kinase 1

FLT1

fms-like tyrosine kinase-1

FLT4

fms-like tyrosine kinase-4

FMDV

Foot and mouth disease virus

HIV

Human immunodeficiency virus

HOC

Highly antigenic outer protein capsid

HRP

Horseradish peroxidase

HUVEC

Human umbilical vein endothelial cell

Ig

Immunoglobulin

IHC

Immunohistochemistry

KDR

Kinase insert domain receptor

LLC

Lewis lung carcinoma

MAPK

Mitogen-activated protein kinase

MFI

Mean fluorescence intensity

MOI

Multiplicity of infection

MTT

Methylthiazolyldiphenyl-tetrazolium bromide

MVD

Microvascular density

OD

Optical density

PBS

Phosphate-buffered saline

PCR

Polymerase chain reaction

pERK

Phospho-extracellular-signal-regulated kinase

PI3K

Phosphoinositide-3-kinase

PKB

Protein kinase B

PlGF

Placental growth factor

pVEGFR2

Phospho-vascular endothelial growth factor receptor 2

SD

Standard deviation

SOC

Small outer capsid

SPSS

Statistical Package for Social Sciences

T4-W

Wild-type T4

VEGF

Vascular endothelial growth factor

VEGFR

Vascular endothelial growth factor receptor

Notes

Acknowledgements

We thank Dr. Zhaojun Ren (VersatileBio LLC, MD, USA) for providing the pJKS plasmid and the T4-e-phage. We also thank MogoEdit (http://www.mogoedit.com) for its linguistic assistance during the preparation of this manuscript.

Author contributions

SZ, GD and LW contributed equally to this work. SZ and XR designed the study. SZ analyzed the data, and wrote the manuscript. GD, LW and YW performed the experiments. ZH and WY provided critical review and comments. WM supervised the study.

Funding

This work was supported by Science and Technology Development Program of Henan Province (No. 132300410274), National Natural Science Foundation of China (No. 81301963), Natural Science Foundation of Henan Province (No. 162300410040) and Outstanding Youth Science Foundation of Henan University (No. yqpy20140036).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Center for Translational MedicineHuaihe Hospital of Henan UniversityKaifengChina
  2. 2.Institute of Infection and ImmunityHuaihe Hospital of Henan UniversityKaifengChina
  3. 3.Department of General SurgeryHuaihe Hospital of Henan UniversityKaifengChina
  4. 4.Department of Respiratory MedicineThe First Affiliated Hospital of Henan UniversityKaifengChina
  5. 5.Department of GastroenterologyThe First Affiliated Hospital of Henan UniversityKaifengChina

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