GFOGER Peptide Modifies the Protein Content of Extracellular Vesicles and Inhibits Vascular Calcification

Abstract

ObjectiveVascular calcification (VC) is an active process during which vascular smooth muscle cells (VSMCs) undergo an osteogenic switch and release extracellular vesicles (EVs). In turn, the EVs serve as calcification foci via interaction with type 1 collagen (COL1). We recently showed that a specific, six-amino-acid repeat (GFOGER) in the sequence of COL1 was involved in the latter’s interaction with integrins expressed on EVs. Our main objective was to test the GFOGER ability to inhibit VC.ApproachWe synthesized the GFOGER peptide and tested its ability to inhibit the inorganic phosphate (Pi)-induced calcification of VSMCs and aortic rings. Using mass spectrometry, we studied GFOGER’s effect on the protein composition of EVs released from Pi-treated VSMCs.ResultsCalcification of mouse VSMCs (MOVAS-1 cells), primary human VSMCs, and rat aortic rings was lower in the presence of GFOGER than with Pi alone (with relative decreases of 66, 58, and 91%, respectively; p < 0.001 for all) (no effect was observed with the scramble peptide GOERFG). A comparative proteomic analysis of EVs released from MOVAS-1 cells in the presence or absence of Pi highlighted significant differences in EVs’ protein content. Interestingly, the expression of some of the EVs’ proteins involved in the calcification process (such as osteogenic markers, TANK-binding kinase 1, and casein kinase II) was diminished in the presence of GFOGER peptide (data are available via ProteomeXchange with identifier PXD018169∗). The decrease of osteogenic marker expression observed in the presence of GFOGER was confirmed by q-RT-PCR analysis.ConclusionGFOGER peptide reduces vascular calcification by modifying the protein content of the subsequently released EVs, in particular by decreasing osteogenicswitching in VSMCs.

Extracellular Vesicles From LPS-Treated Macrophages Aggravate Smooth Muscle Cell Calcification by Propagating Inflammation and Oxidative Stress

“…Furthermore, an increase in reactive oxygen species (ROS) production is involved in VSMC osteochondrogenic trans-differentiation during the VC process ( Tóth et al, 2020 ; Hu et al, 2021 ). Recent studies have highlighted the role of extracellular vesicles (EVs) in VC ( Hodroge et al, 2017 ; Mansour et al, 2020 ; Yaker et al, 2020 ; Qin et al, 2021 ). These membrane-bound vesicles, secreted by prokaryotic and eukaryotic cells ( Woith et al, 2019 ), can be of various origins, depending on their mode of biogenesis.…”

Effects of Chronic Kidney Disease and Uremic Toxins on Extracellular Vesicle Biology

“…These findings were confirmed by Mansour et al, who showed that a synthetic GFOGER peptide could inhibit VC in VSMCs and aortic rings [129]. This effect was accompanied by a change in the EVs' protein content [129].…”

“…Moreover, an oligogalacturonic acid (DP8) was able to block the GFOGER interaction between type I collagen and EVs and thus to reduce VC and VSMC osteogenic switching [128]. These findings were confirmed by Mansour et al, who showed that a synthetic GFOGER peptide could inhibit VC in VSMCs and aortic rings [129]. This effect was accompanied by a change in the EVs' protein content [129].…”

Cellular Crosstalk in the Vascular Wall Microenvironment: The Role of Exosomes in Vascular Calcification

“…Clinically, circulating as well as cell-derived exosomes with a low fetuin-A level play a detrimental role in VC ( 58 , 79 ). While the GFOGER peptide, a six amino-acid repeat present in type 1 collagen, alleviated VC by reducing the content of osteogenic switching–related proteins, such as TANK-binding kinase 1 and casein kinase II, in VSMC-derived EVs ( 80 ). These pieces of evidence remind us that the cargo loaded by exosomes secreted from VSMCs changes dynamically with the progression of VC, which provides potential diagnostic biomarkers and targets for treatment.…”