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Mechanism assay of interaction between blood vessels-near infrared probe and cell surface marker proteins of endothelial cells.

ABSTRACT: In vivo blood vessels imaging is crucial to study blood vessels related diseases in real-time. For this purpose, fluorescent based imaging is one of the utmost techniques for imaging a living system. The discovery of a new near-infrared probe (CyA-B2) by screening chemical probe library in our previous report which showed the most specific binding on the blood capillaries of the 3D-tissue models give us interest to study more about the binding site of this probe to the surface of endothelial cells main component cell of blood capillaries. By studying the competition assays of CyA-B2 using several potential surface markers of endothelial cells found through the chemical database (ChEMBL) and manually selected, CD133 gave the lowest IC₅₀ (half maximal inhibitory concentration) value. Hence, CD133 protein which is expressed on the endothelial cell membrane was postulated to be the binding site due to the suppression of CyA-B2 on the blood capillaries by the competition assays. Since, CD133 is also expressed on many types of cancer cells, it would be useful to use CyA-B2 as a bioprobe to monitor or diagnostic tumor growth.

SUBMITTER: Abdul Sisak MA

PROVIDER: S-EPMC9251809 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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Mechanism assay of interaction between blood vessels-near infrared probe and cell surface marker proteins of endothelial cells.

Abdul Sisak Muhammad Asri MA Louis Fiona F Miyao Tomoyuki T Lee Sun Hyeok SH Chang Young-Tae YT Matsusaki Michiya M

Materials today. Bio 20220623

<i>In vivo</i> blood vessels imaging is crucial to study blood vessels related diseases in real-time. For this purpose, fluorescent based imaging is one of the utmost techniques for imaging a living system. The discovery of a new near-infrared probe (CyA-B2) by screening chemical probe library in our previous report which showed the most specific binding on the blood capillaries of the 3D-tissue models give us interest to study more about the binding site of this probe to the surface of endothel ...[more]

PMID: 35795137

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Project description:Tumor Endothelial Marker 8/Anthrax Toxin Receptor 1 (TEM8/ANTXR1) expression is induced in the vascular compartment of multiple tumors and therefore, is a candidate molecule to target tumor therapies. This cell surface molecule mediates anthrax toxin internalization, however, its physiological function in blood vessels remains largely unknown. We identified the chicken chorioallantoic membrane (CAM) as a model system to study the endogenous function of TEM8 in blood vessels as we found that TEM8 expression was induced transiently between day 10 and 12 of embryonic development, when the vascular tree is undergoing final development and growth. We used the cell-binding component of anthrax toxin, Protective Antigen (PA), to engage endogenous TEM8 receptors and evaluate the effects of PA-TEM8 complexes on vascular development. PA applied at the time of highest TEM8 expression reduced vascular density and disrupted hierarchical branching as revealed by quantitative morphometric analysis of the vascular tree after 48h. PA-dependent reduced branching phenotype was partially mimicked by Wnt3a application and ameliorated by the Wnt antagonist, Dikkopf-1. These results implicate TEM8 expression in endothelial cells in regulating the canonical Wnt signaling pathway at this day of CAM development. Consistent with this model, PA increased beta catenin levels acutely in CAM blood vessels in vivo and in TEM8 transfected primary human endothelial cells in vitro. TEM8 expression in Hek293 cells, which neither express endogenous PA-binding receptors nor Wnt ligands, stabilized beta catenin levels and amplified beta catenin-dependent transcriptional activity induced by Wnt3a. This agonistic function is supported by findings in the CAM, where the increase in TEM8 expression from day 10 to day 12 and PA application correlated with Axin 2 induction, a universal reporter gene for canonical Wnt signaling. We postulate that the developmentally controlled expression of TEM8 modulates endothelial cell response to canonical Wnt signaling to regulate vessel patterning and density.

Dataset Information

Mechanism assay of interaction between blood vessels-near infrared probe and cell surface marker proteins of endothelial cells.

Publications

Mechanism assay of interaction between blood vessels-near infrared probe and cell surface marker proteins of endothelial cells.

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