caa a22 4500 605543526 CHVBK 20210128100927.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00109-015-1292-0 doi (NATIONALLICENCE)springer-10.1007/s00109-015-1292-0 Endothelial sphingosine kinase/SPNS2 axis is critical for vessel-like formation by human mesoangioblasts [Elektronische Daten] [Anna Laurenzana, Francesca Cencetti, Simona Serratì, Gennaro Bruno, Lukasz Japtok, Francesca Bianchini, Eugenio Torre, Gabriella Fibbi, Mario Del Rosso, Paola Bruni, Chiara Donati] The interaction between endothelial cells and pericytes is crucial for the stabilization of newly formed vessels in angiogenesis. The comprehension of the mechanisms regulating pericyte recruitment might open therapeutical perspectives on vascular-related pathologies. Sphingosine 1-phosphate (S1P) is a bioactive sphingolipid that derives from sphingomyelin catabolism and regulates biological functions in cell survival, proliferation, and differentiation. In this study, we aimed to identify the role of S1P axis in the intercellular communication between human mesenchymal progenitor mesoangioblasts (MAB) and endothelial cells (human microvascular endothelial cells (H-MVEC)) in the formation of capillary-like structures. We demonstrated that the S1P biosynthetic pathway brought about by sphingosine kinases (SK) SK1 and SK2 as well as spinster homolog 2 (SPNS2) transporter in H-MVEC is crucial for MAB migration measured by Boyden chambers and for the formation and stabilization of capillary-like structures in a 3D Matrigel culture. Moreover, the conditioned medium (CM) harvested from H-MVEC, where SK1, SK2, and SPNS2 were down-regulated, exerted a significantly diminished effect on MAB capillary morphogenesis and migration. Notably, we demonstrated that S1P1 and S1P3 receptors were positively involved in CM-induced capillary-like formation and migration, while S1P2 exerted a negative roleon CM-induced migratory action of MAB. Finally, SK inhibition as well as MAB S1P1 and S1P3 down-regulation impaired H-MVEC-MAB cross-talk significantly reducing in vivo angiogenesis evaluated by Matrigel plug assay. These findings individuate novel targets for the employment of MAB in vascular-related pathologic conditions. Key message: • Down-regulation of SK1/2 in H-MVEC impaired vessel formation when cultured with MAB. • H-MVEC SPNS2 is critical for morphogenesis and migration induced by H-MVEC CM of MAB. • CM from SK1- and SK2-siRNA H-MVEC impaired morphogenesis and migration of MAB. • S1P1/3 were involved on CM-induced morphogenesis and migration of MAB. • Matrigel plug assay showed the role of S1P axis in MAB-endothelial cell interaction. Springer-Verlag Berlin Heidelberg, 2015 Mesoangioblasts nationallicence Endothelial cells nationallicence Sphingosine kinase nationallicence Migration nationallicence Morphogenesis nationallicence SK : Sphingosine kinase nationallicence S1P : Sphingosine 1-phosphate nationallicence FCS : Fetal calf serum nationallicence siRNA : Short-interfering RNA nationallicence SCR : Scrambled nationallicence CM : Conditioned medium nationallicence PBS : Phosphate-buffered saline nationallicence DMEM : Dulbecco's modified Eagle's medium nationallicence BSA : Bovine serum albumin nationallicence TGFβ : Transforming growth factor beta nationallicence SPNS2 : Spinster homolog 2 nationallicence bFGF : Basic fibroblast growth factor nationallicence VEGF : Vascular endothelial growth factor nationallicence MAPK : Mitogen-activated protein kinase nationallicence PI3K : Phosphatidylinositol 3-kinase nationallicence SCID : Severe combined immunodeficiency nationallicence Laurenzana Anna Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Cencetti Francesca Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Serratì Simona Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Bruno Gennaro Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Japtok Lukasz Faculty of Mathematics and Natural Science, Institute of Nutritional Science, Department of Toxicology, University of Potsdam, Arthur-Scheunert Allee 114-116, 14558, Potsdam, Nuthetal, Germany aut Bianchini Francesca Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Torre Eugenio Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Fibbi Gabriella Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Del Rosso Mario Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Bruni Paola Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Donati Chiara Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut Journal of Molecular Medicine Springer Berlin Heidelberg 93/10(2015-10-01), 1145-1157 0946-2716 93:10<1145 2015 93 109 https://doi.org/10.1007/s00109-015-1292-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00109-015-1292-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Laurenzana Anna Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Cencetti Francesca Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Serratì Simona Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Bruno Gennaro Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Japtok Lukasz Faculty of Mathematics and Natural Science, Institute of Nutritional Science, Department of Toxicology, University of Potsdam, Arthur-Scheunert Allee 114-116, 14558, Potsdam, Nuthetal, Germany aut NATIONALLICENCE 700 1- Bianchini Francesca Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Torre Eugenio Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Fibbi Gabriella Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Del Rosso Mario Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Bruni Paola Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 700 1- Donati Chiara Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio”, Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy aut NATIONALLICENCE 773 0- Journal of Molecular Medicine Springer Berlin Heidelberg 93/10(2015-10-01), 1145-1157 0946-2716 93:10<1145 2015 93 109