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

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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.

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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

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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.

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).

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Author notes

  1. Shuguang Zuo, Gongpeng Dai and Liping Wang contributed equally to this work.

Authors and Affiliations

  1. Center for Translational Medicine, Huaihe Hospital of Henan University, Kaifeng, 475001, Henan, China

    Shuguang Zuo, Liping Wang & Xuequn Ren

  2. Institute of Infection and Immunity, Huaihe Hospital of Henan University, Kaifeng, 475001, Henan, China

    Shuguang Zuo

  3. Department of General Surgery, Huaihe Hospital of Henan University, Kaifeng, 475001, Henan, China

    Gongpeng Dai, Yuqing Wen, Wanli Ma & Xuequn Ren

  4. Department of Respiratory Medicine, The First Affiliated Hospital of Henan University, Kaifeng, 475001, Henan, China

    Zhiang Huang

  5. Department of Gastroenterology, The First Affiliated Hospital of Henan University, Kaifeng, 475001, Henan, China

    Wenyi Yang

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  1. Shuguang Zuo
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  2. Gongpeng Dai
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  4. Yuqing Wen
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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.

Corresponding authors

Correspondence to Shuguang Zuo or Xuequn Ren.

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The authors declare that they have no conflicts of interest.

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Handling Editor: Chan-Shing Lin.

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Zuo, S., Dai, G., Wang, L. et al. Suppression of angiogenesis and tumor growth by recombinant T4 phages displaying extracellular domain of vascular endothelial growth factor receptor 2. Arch Virol 164, 69–82 (2019). https://doi.org/10.1007/s00705-018-4026-0

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  • DOI: https://doi.org/10.1007/s00705-018-4026-0

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