Abstract 1499
Background
Tumor growth depends on pro-angiogenic vascular endothelial growth factors (VEGF-A/-B) via binding to VEGF receptors (VEGFR1 and VEGFR2) that are biomarkers of tumor angiogenesis. Importantly, interfering with this interaction by small molecules impairs tumor growth and angiogenesis.
Methods
A novel peptide (known as VGB3) was synthesized using a solid-phase peptide synthesis (SPPS) procedure on 2-chlorotrityl chloride (2-CTC) resin. A disulfide bridge was formed in the peptide and it was purified using RP-HPLC. Recombinant His-tagged VEGFR1D2 obtained from inclusion bodies and purified on a Ni2+-NTA agarose resin. The equilibrium constant for the binding of VGB3 to VEGFR1-D2 molecule was determined by microscale thermophoresis (MST). The accumulation of the peptide in 4T1 mammary carcinoma tumors (MCTs) was assessed using 2D optical imaging. Anti-angiogenic activity of VGB3 were investigated using MTT, scratch assay, and two- as well as three-dimensional angiogenesis assays. Downstream signaling pathways were assessed by quantitative estimation of protein expression using western blotting and immunohistochemistry. (IHC).
Results
We have indicated that VGB3 is able to block VEGFR1-D2 in human umbilical vein endothelial cells (HUVECs) and 4T1 mammary carcinoma tumor (MCT) cells. This resulted in inhibition in proliferation of HUVEC, 4T1 MCT, and U87 glioblastoma cells and MST results reveals that VGB3 is a high-affinity (Kd=1.96± 0.69 µM, n = 3, P < 0.0001) binder to VEGFR1-D2. Moreover, fluorescence imaging in Balb/c mice demonstrated that fluorescein isothiocyanate (FITC)-VGB3 accumulated in tumors. VEGFA-stimulated proliferation, migration, and 2 and 3D tube formation in HUVECs were strongly inhibited by VGB3 (n = 3, P < 0.0001). In a murine 4T1 MCT model, VGB3 strongly inhibited AKT and ERK1/2 phosphorylation and tumor cell downstream signaling pathways associated with proliferation, migration, and angiogenesis, and led to the increased apoptosis index and suppression of systematic spreading of the tumor compared to PBS-treated controls.
Conclusions
These results indicate that VGB3 may be applicable for antiangiogenic and antitumor therapy.
Clinical trial identification
Editorial acknowledgement
S. Mohsen Asghari and Prof. Matthew Groves
Legal entity responsible for the study
S. Mohsen Asghari and Matthew Groves.
Funding
University of Guilan.
Disclosure
All authors have declared no conflicts of interest.
Resources from the same session
3909 - Spectrum of pathogenic germline mutations in Chinese lung cancer patients through next-generation sequencing
Presenter: Ying Huang
Session: Poster Display session 1
Resources:
Abstract
3061 - Poor prognostic impact of NTRK2 gene variation in Esophageal Squamous Cell Carcinoma
Presenter: Ye Chen
Session: Poster Display session 1
Resources:
Abstract
4735 - Mutation profile of Tibetan lung cancer revealed by Whole Exome Sequencing
Presenter: Xin Wang
Session: Poster Display session 1
Resources:
Abstract
5236 - Synergistic activity between niraparib and chemotherapy in colorectal cancer: molecular determinants from a preclinical model
Presenter: Pietro Paolo Vitiello
Session: Poster Display session 1
Resources:
Abstract
4051 - cRGDfK (cRGD) conjugated Pyropheophor¬bide-a (Pyro), a new tumor photodynamic agent, is highly accumulated and specific in tumor cell killing
Presenter: Fengwei Wang
Session: Poster Display session 1
Resources:
Abstract
859 - The expression of MMR, CD133 and the presence of p53 wt predict the response to Cabazitaxel in malignant neural tumors cell lines.
Presenter: Kevin Doello
Session: Poster Display session 1
Resources:
Abstract
2497 - IKS01, a next generation antibody drug conjugate (ADC) designed to be efficacious in tumors with low and moderate levels of folate receptor expression
Presenter: Jenny Thirlway
Session: Poster Display session 1
Resources:
Abstract
1636 - Novel Non-Camptothecin Compounds with Antiproliferative Activities against Breast Cancer Cells
Presenter: Wen-shan Li
Session: Poster Display session 1
Resources:
Abstract
3443 - Sensitization of estrogen receptor-positive breast cancer cells to tamoxifen by novel epi-oligomycin A
Presenter: Margarita Yastrebova
Session: Poster Display session 1
Resources:
Abstract
840 - Autophagy inhibition enhances leflunomide-induced cytotoxicity in human bladder cancer cells
Presenter: Li Cheng
Session: Poster Display session 1
Resources:
Abstract