The Pyrazolyl-Urea Gege3 Inhibits the Activity of ANXA1 in the Angiogenesis Induced by the Pancreatic Cancer Derived EVs.

Raffaella Belvedere,Chiara Brullo,Olga Bruno,Antonello Petrella,Silvana Morello,Elva Morretta,Nunzia Novizio

doi:10.3390/biom11121758

Raffaella Belvedere, Chiara Brullo + Show 5 more

Open Access

https://doi.org/10.3390/biom11121758

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Journal: Biomolecules	Publication Date: Nov 24, 2021
Citations: 7	License type: CC BY 4.0

Affiliation: University of Salerno, University of Genoa

Abstract

The pyrazolyl-urea Gege3 molecule has shown interesting antiangiogenic effects in the tumor contest. Here, we have studied the role of this compound as interfering with endothelial cells activation in response to the paracrine effects of annexin A1 (ANXA1), known to be involved in promoting tumor progression. ANXA1 has been analyzed in the extracellular environment once secreted through microvesicles (EVs) by pancreatic cancer (PC) cells. Particularly, Gege3 has been able to notably prevent the effects of Ac2-26, the ANXA1 mimetic peptide, and of PC-derived EVs on endothelial cells motility, angiogenesis, and calcium release. Furthermore, this compound also inhibited the translocation of ANXA1 to the plasma membrane, otherwise induced by the same ANXA1-dependent extracellular stimuli. Moreover, these effects have been mediated by the indirect inhibition of protein kinase Cα (PKCα), which generally promotes the phosphorylation of ANXA1 on serine 27. Indeed, by the subtraction of intracellular calcium levels, the pathway triggered by PKCα underwent a strong inhibition leading to the following impediment to the ANXA1 localization at the plasma membrane, as revealed by confocal and cytofluorimetry analysis. Thus, Gege3 appeared an attractive molecule able to prevent the paracrine effects of PC cells deriving ANXA1 in the tumor microenvironment.

Highlights

It is known that the initiation of tumor angiogenesis is required for tumor progression, arousing a considerable interest from the research community stimulating extensive efforts based on an anti-angiogenic therapy to treat cancer [1]
The Annexin A1 (ANXA1) fluorescence intensity at the different levels in endothelial cells has been analyzed as described in the Materials and Methods sections and the results reported in Supplementary Figure S2A, further highlighted the increase of ANXA1 signal at the plasma membrane in presence of Ac2-26 and the following rescue when the peptide has been administered together with Gege3
The promising antiangiogenic activity of the compound Gege3 has been studied in in vitro and in vivo systems highlighting the ability of this molecule to interfere with the activation of several kinases as ERK, Akt, and dystrophia myotonica protein kinase (DMPK)1, known to be some of the intermediate elements of VEGF signaling in endothelial cells

Summary

Introduction

It is known that the initiation of tumor angiogenesis is required for tumor progression, arousing a considerable interest from the research community stimulating extensive efforts based on an anti-angiogenic therapy to treat cancer [1] During this pathological process, pro-angiogenic factors are initially secreted into the extracellular fluid to activate endothelial cells, which form a functional vascular network [2]. Pro-angiogenic factors are initially secreted into the extracellular fluid to activate endothelial cells, which form a functional vascular network [2] Among these factors, the tumor microenvironment consists of VEGF (Vascular Endothelial Growth Factor), bFGF (basic Fibroblast Growth Factor), and PDGF (Platelet-Derived Growth Factor), cytokines, small non-coding RNAs which strongly acts to promote angiogenesis mainly in a hypoxic environment [3].

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