Abstract
The multi-modular glycoprotein thrombospondin-1 (TSP-1) is considered as a key actor within the tumor microenvironment. Besides, TSP-1 binding to CD47 is widely reported to regulate cardiovascular function as it promotes vasoconstriction and angiogenesis limitation. Therefore, many studies focused on targeting TSP-1:CD47 interaction, aiming for up-regulation of physiological angiogenesis to enhance post-ischemia recovery or to facilitate engraftment. Thus, we sought to identify an innovative selective antagonist for TSP-1:CD47 interaction. Protein-protein docking and molecular dynamics simulations were conducted to design a novel CD47-derived peptide, called TAX2. TAX2 binds TSP-1 to prevent TSP-1:CD47 interaction, as revealed by ELISA and co-immunoprecipitation experiments. Unexpectedly, TAX2 inhibits in vitro and ex vivo angiogenesis features in a TSP-1-dependent manner. Consistently, our data highlighted that TAX2 promotes TSP-1 binding to CD36-containing complexes, leading to disruption of VEGFR2 activation and downstream NO signaling. Such unpredicted results prompted us to investigate TAX2 potential in tumor pathology. A multimodal imaging approach was conducted combining histopathological staining, MVD, MRI analysis and μCT monitoring for tumor angiography longitudinal follow-up and 3D quantification. TAX2 in vivo administrations highly disturb syngeneic melanoma tumor vascularization inducing extensive tumor necrosis and strongly inhibit growth rate and vascularization of human pancreatic carcinoma xenografts in nude mice.
Keywords:
CD36; CD47; TSP-1; angiogenesis; cancer.
Publication types
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Research Support, Non-U.S. Gov't
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Video-Audio Media
MeSH terms
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Angiogenesis Inhibitors / chemistry
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Angiogenesis Inhibitors / metabolism
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Angiogenesis Inhibitors / pharmacology*
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Animals
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CD36 Antigens / metabolism
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Carcinoma / blood supply
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Carcinoma / drug therapy*
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Carcinoma / metabolism
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Carcinoma / pathology
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Cell Line, Tumor
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Cell Movement / drug effects
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Cell Proliferation / drug effects
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Computer-Aided Design
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Drug Design
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Human Umbilical Vein Endothelial Cells / drug effects
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Human Umbilical Vein Endothelial Cells / metabolism
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Humans
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Magnetic Resonance Imaging
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Melanoma, Experimental / blood supply
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Melanoma, Experimental / drug therapy*
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Melanoma, Experimental / metabolism
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Melanoma, Experimental / pathology
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Mice, Inbred BALB C
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Mice, Inbred C57BL
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Mice, Nude
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Molecular Docking Simulation
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Molecular Dynamics Simulation
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Molecular Targeted Therapy
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Necrosis
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Neovascularization, Pathologic
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Nitric Oxide / metabolism
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Pancreatic Neoplasms / blood supply
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Pancreatic Neoplasms / drug therapy*
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Pancreatic Neoplasms / metabolism
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Pancreatic Neoplasms / pathology
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Peptides / chemistry
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Peptides / metabolism
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Peptides / pharmacology*
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Peptides, Cyclic / chemistry
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Peptides, Cyclic / metabolism
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Peptides, Cyclic / pharmacology*
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Protein Binding
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Signal Transduction / drug effects
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Thrombospondin 1 / metabolism
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Time Factors
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Transfection
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Tumor Burden / drug effects
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Vascular Endothelial Growth Factor Receptor-2 / metabolism
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X-Ray Microtomography
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Xenograft Model Antitumor Assays
Substances
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Angiogenesis Inhibitors
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CD36 Antigens
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Peptides
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Peptides, Cyclic
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Thrombospondin 1
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cyclic TAX2 peptide
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Nitric Oxide
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KDR protein, human
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Vascular Endothelial Growth Factor Receptor-2