Angiogenesis Inhibition by a Short 13 Amino Acid Peptide Sequence of Tetrastatin, the α4(IV) NC1 Domain of Collagen IV.

Alexia Vautrin-Glabik,Camille Bour,Stéphanie Baud,Aurélie Dupont-Deshorgue,Christèle Sellier,Jérôme Devy,Laurence Choulier,Jean Claude Monboisse,Jean-Baptiste Oudart,Sylvie Brassart-Pasco,Anthony Hoarau,Bertrand Brassart,Laurent Ramont

doi:10.3389/fcell.2020.00775

Abstract

Angiogenesis is defined as the formation of new capillaries by sprouting from the pre-existing microvasculature. It occurs in physiological and pathological processes particularly in tumor growth and metastasis. α1, α2, α3, and α6 NC1 domains from type IV collagen were reported to inhibit tumor angiogenesis. We previously demonstrated that the α4 NC1 domain from type IV collagen, named Tetrastatin, inhibited tumor growth in a mouse melanoma model. The inhibitory activity was located in a 13 amino acid sequence named QS-13. In the present paper, we demonstrate that QS-13 decreases VEGF-induced-angiogenesis in vivo using the Matrigel plug model. Fluorescence molecular tomography allows the measurement of a 65% decrease in Matrigel plug angiogenesis following QS-13 administration. The results are confirmed by CD31 microvessel density analysis on Matrigel plug slices. QS-13 peptide decreases Human Umbilical Vein Endothelial Cells (HUVEC) migration and pseudotube formation in vitro. Relevant QS-13 conformations were obtained from molecular dynamics simulations and docking. A putative interaction of QS-13 with α5β1 integrin was investigated. The interaction was confirmed by affinity chromatography, solid phase assay, and surface plasmon resonance. QS-13 binding site on α5β1 integrin is located in close vicinity to the RGD binding site, as demonstrated by competition assays. Collectively, our results suggest that QS-13 exhibits a mighty anti-angiogenic activity that could be used in cancer treatment and other pathologies with excessive angiogenesis such as hemangioma, psoriasis or diabetes.

Highlights

Angiogenesis is the formation of new blood vessels and capillaries from sprouting of pre-existing blood vessels
As proliferation of endothelial cells plays an essential role in angiogenesis, Human Umbilical Vein Endothelial Cells (HUVEC) proliferation under the influence of QS13 was assessed at 24, 48, 72, and 96 h using the WST-1 method
Angiogenesis plays critical roles in human physiological processes. This is a complex phenomenon regulated in a spatial and temporal manner that depends on the cooperation between angiogenic factors, extracellular matrix (ECM) components, and endothelial cells

Summary

Introduction

Angiogenesis is the formation of new blood vessels and capillaries from sprouting of pre-existing blood vessels. It normally occurs during wound healing and embryonic development, but it is required for tumor growth and metastasis. This process is regulated by growth factors, such as vascular endothelial growth factors (VEGFs), which bind to their receptors on the normal. QS-13 Peptide Inhibits Angiogenesis endothelial cell surface, induce transduction pathways and promote proliferation and migration of endothelial cells and vascular tube formation (Teleanu et al, 2019). Several BM components were reported to largely participate in the regulation of tumor angiogenesis (Kalluri, 2003)

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