Laurenzana Anna, Cencetti Francesca, Serratì Simona, Bruno Gennaro, Japtok Lukasz, Bianchini Francesca, Torre Eugenio, Fibbi Gabriella, Del Rosso Mario, Bruni Paola, Donati Chiara
Dipartimento di Scienze Biomediche Sperimentali e Cliniche "Mario Serio", Università di Firenze, Viale G.B. Morgagni 50, 50134, Florence, Italy.
Department of Experimental Oncology, Hematology Unit, Advanced Cellular Therapy Centre, Bari, Italy.
J Mol Med (Berl). 2015 Oct;93(10):1145-57. doi: 10.1007/s00109-015-1292-0. Epub 2015 May 9.
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 role on 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.
• 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.
内皮细胞与周细胞之间的相互作用对于血管生成过程中新形成血管的稳定至关重要。了解调节周细胞募集的机制可能为血管相关疾病开辟治疗前景。鞘氨醇-1-磷酸（S1P）是一种生物活性鞘脂，源自鞘磷脂分解代谢，可调节细胞存活、增殖和分化中的生物学功能。在本研究中，我们旨在确定S1P轴在人骨髓间充质祖细胞中胚层血管母细胞（MAB）与内皮细胞（人微血管内皮细胞（H-MVEC））之间的细胞间通讯在毛细血管样结构形成中的作用。我们证明，由鞘氨醇激酶（SK）SK1和SK2以及H-MVEC中的spinster同源物2（SPNS2）转运体引发的S1P生物合成途径对于通过Boyden小室测量的MAB迁移以及在3D基质胶培养中毛细血管样结构的形成和稳定至关重要。此外，从SK1、SK2和SPNS2被下调的H-MVEC中收集的条件培养基（CM）对MAB毛细血管形态发生和迁移的影响显著减弱。值得注意的是，我们证明S1P1和S1P3受体积极参与CM诱导的毛细血管样形成和迁移，而S1P2对CM诱导的MAB迁移作用发挥负性作用。最后，SK抑制以及MAB中S1P1和S1P3的下调显著损害了H-MVEC-MAB的相互作用，通过基质胶栓塞试验评估，体内血管生成明显减少。这些发现为MAB在血管相关病理状况中的应用确定了新的靶点。
• 与MAB共培养时，H-MVEC中SK1/2的下调损害血管形成。
• H-MVEC的SPNS2对于H-MVEC CM诱导的MAB形态发生和迁移至关重要。
• 来自SK1-和SK2-siRNA H-MVEC的CM损害MAB的形态发生和迁移。
• S1P1/3参与CM诱导的MAB形态发生和迁移。
• 基质胶栓塞试验显示S1P轴在MAB-内皮细胞相互作用中的作用。
