title: Soluble CD146 boosts therapeutic effect of endothelial progenitors through proteolytic processing of short CD146 isoform.
abstract: Endothelial colony-forming cells (ECFC) constitute an endothelial progenitor fraction with a promising interest for the treatment of ischaemic cardiovascular diseases. As soluble CD146 (sCD146) is a new factor promoting angiogenesis, we examined whether sCD146 priming could improve the therapeutic potential of ECFC and defined the involved mechanism.
  
  
  
  We investigated the effects of sCD146 priming on regenerative properties of ECFC in vivo. In a mouse model of hindlimb ischaemia, the homing of radiolabelled cells to ischaemic tissue was assessed by SPECT-CT imaging. Soluble CD146 priming did not modify the number of engrafted ECFC but improved their survival capacity, leading to an enhanced revascularization. The mechanism of action of sCD146 on ECFC was studied in vitro. We showed that sCD146 acts in ECFC through a signalosome, located in lipid rafts, containing angiomotin, the short isoform of CD146 (shCD146), VEGFR1, VEGFR2, and presenilin-1. Soluble CD146 induced a sequential proteolytic cleavage of shCD146, with an extracellular shedding followed by an intramembrane cleavage mediated by matrix metalloprotease (MMP)/ADAM and presenilin-1, respectively. The generated intracellular part of shCD146 was directed towards the nucleus where it associated with the transcription factor CSL and modulated the transcription of genes involved in cell survival (FADD, Bcl-xl) and angiogenesis (eNOS). This effect was dependent on both VEGFR1 and VEGFR2, which were rapidly phosphorylated by sCD146.
  
  
  
  These findings establish that activation of the proteolytic processing of shCD146, in particular by sCD146, constitutes a promising pathway to improve endothelial progenitors' regenerative properties for the treatment of cardiovascular diseases.
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Jimmy
authors.initials: J
authors.lastname: Stalin
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Karim
authors.initials: K
authors.lastname: Harhouri
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Lucas
authors.initials: L
authors.lastname: Hubert
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France CERIMED (European Center of Research in Medical Imaging), Aix-Marseille University, Marseille, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Garrigue
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Nollet
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Amel
authors.initials: A
authors.lastname: Essaadi
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Muller
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Alexandrine
authors.initials: A
authors.lastname: Foucault-Bertaud
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Richard
authors.initials: R
authors.lastname: Bachelier
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France Laboratory of Culture and Cell Therapy, INSERM CIC-BT510, CHU Conception, Marseille, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Sabatier
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Pisano
authors.affiliation: INSERM UMR-S 1062, Aix-Marseille University, Marseille, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Peiretti
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Aurélie S
authors.initials: AS
authors.lastname: Leroyer
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France CERIMED (European Center of Research in Medical Imaging), Aix-Marseille University, Marseille, France.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Guillet
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bardin
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Dignat-George
authors.affiliation: INSERM UMR-S 1076, Aix-Marseille University, UFR Pharmacy, 27 Bd J. Moulin, 13005 Marseille, France marcel.blot-chabaud@laposte.net.
authors.forename: Marcel
authors.initials: M
authors.lastname: Blot-Chabaud
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000818:Animals; D051929:CD146 Antigen / Q000378:metabolism; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D004195:Disease Models, Animal; D066026:Endothelial Progenitor Cells / Q000201:enzymology / Q000637:transplantation*; D053200:Fas-Associated Death Domain Protein / Q000235:genetics / Q000378:metabolism; D006614:Hindlimb; D007511:Ischemia / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology / Q000601:surgery*; D053504:Matrix Metalloproteinases, Membrane-Associated / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism; D051379:Mice; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D018482:Muscle, Skeletal / Q000098:blood supply*; D018919:Neovascularization, Physiologic; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D053764:Presenilin-1 / Q000378:metabolism; D020033:Protein Isoforms; D059748:Proteolysis; D012038:Regeneration; D015398:Signal Transduction; D013997:Time Factors; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D051020:bcl-X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: C494812:Bcl2l1 protein, mouse; D051929:CD146 Antigen; C505534:Fadd protein, mouse; D053200:Fas-Associated Death Domain Protein; C497903:Mcam protein, mouse; D009124:Muscle Proteins; D053764:Presenilin-1; D020033:Protein Isoforms; C499035:Smpx protein, mouse; D051020:bcl-X Protein; C580804:presenilin 1, mouse; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse; C501163:Flt1 protein, mouse; C000592875:Kdr protein, mouse; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2; D051722:ADAM Proteins; D053504:Matrix Metalloproteinases, Membrane-Associated
keywords: Angiogenesis; Apoptosis; Endothelial progenitor; Hindlimb ischaemia; SPECT-CT