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

Ali Mansour; Walaa Darwiche; Linda Yaker; Sophie Da Nascimento; Cathy Gomila; Claire Rossi; Vincent Jung; Pascal Sonnet; Saïd Kamel; Ida Chiara Guerrera; Agnès Boullier; Jérôme Ausseil

doi:10.3389/fcell.2020.589761

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

Front Cell Dev Biol. 2020 Nov 30:8:589761. doi: 10.3389/fcell.2020.589761. eCollection 2020.

Authors

Ali Mansour¹, Walaa Darwiche¹, Linda Yaker¹, Sophie Da Nascimento², Cathy Gomila¹, Claire Rossi³, Vincent Jung⁴, Pascal Sonnet², Saïd Kamel^{1

5}, Ida Chiara Guerrera⁴, Agnès Boullier^{1

5}, Jérôme Ausseil^{1

6

7}

Affiliations

¹ MP3CV-UR7517, CURS-Université de Picardie Jules Verne, Amiens, France.
² AGIR, UR4294, UFR de Pharmacie, Université de Picardie Jules Verne, Amiens, France.
³ Alliance Sorbonne Université, Université de Technologie de Compiègne, UMR7025 CNRS Enzyme and Cell Engineering Laboratory, Compiègne, France.
⁴ Plateforme protéomique Necker, Faculté de Médecine Paris Descartes, Université de Paris - Structure Fédérative de Recherche Necker, INSERM US24/CNRS UMS3633, Paris, France.
⁵ Laboratoire de Biochimie, Centre Hospitalier Universitaire d' Amiens, Amiens, France.
⁶ Centre de Physiopathologie Toulouse Purpan, INSERM UMR1043 - CNRS UMR5282 - Université Toulouse III, Toulouse, France.
⁷ CHU Toulouse - Institut Fédératif de Biologie, Laboratoire de Biochimie, Toulouse, France.

Abstract

Objective: Vascular 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.

Approach: We 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.

Results: Calcification 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.

Conclusion: GFOGER peptide reduces vascular calcification by modifying the protein content of the subsequently released EVs, in particular by decreasing osteogenicswitching in VSMCs.

Keywords: GFOGER sequence; extracellular vesicle; oligogalacturonic acid; osteogenic switch; type I collagen; vascular calcification.