ABSTRACT
Central nervous system (CNS) inflammation is primarily driven by microglial cells which secrete proinflammatory cytokines and undergo proliferation upon activation, as it occurs in neurodegenerative diseases. Uncontrolled or prolonged CNS inflammation is potentially harmful and can result in cellular damage. Recently, many studies have focused on human adipose tissue as an attractive source of cytokines with immunosuppressive properties that potentially modulate inflammation. Our study aimed to evaluate if different methods of human tissue collection could affect adipose mesenchymal stem cell (ADSC)-derived cytokine secretion and investigate the effects of ADSC secretome in modulating microglia activation and the possible implication of sphingosine-1-phosphate (S1P) in these effects. Our results demonstrate that the conditioned medium (CM) of ADSCs isolated by two different processing methods (lipoaspirate and Lipogems) significantly inhibited the lipopolysaccharide (LPS)-induced effects on microglia activation, including microglial expression of CD68, cytokine secretion, proliferation, and migration. Pulse studies with radiolabeled sphingosine demonstrated that LPS treatment of resting microglia induced a significant increase of both cellular and extracellular S1P. Moreover, and of relevance, FTY720, a functional antagonist of S1P receptor, inhibited the multiple LPS-induced proinflammatory effects on microglia, and S1P suppressed the anti-inflammatory effect of ADSC-CM. This suggests that LPS-mediated microglial activation is countered by ADSC-CM through the modulation of sphingosine kinase/S1P signalling.
Subject(s)
Adipose Tissue/cytology , Inflammation/pathology , Lysophospholipids/metabolism , Mesenchymal Stem Cells/metabolism , Microglia/pathology , Proteome/metabolism , Signal Transduction , Sphingosine/analogs & derivatives , Adult , Cell Proliferation/drug effects , Cell Separation , Chemotaxis/drug effects , Culture Media, Conditioned/pharmacology , Cytokines/metabolism , Down-Regulation/drug effects , Female , Fingolimod Hydrochloride/pharmacology , Humans , Inflammation/metabolism , Lipopolysaccharides , Male , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Microglia/metabolism , Middle Aged , Phenotype , Signal Transduction/drug effects , Sphingosine/metabolismABSTRACT
Beyond key functions in hemostasis and thrombosis, platelets are recognized as key players of inflammation, an underlying feature of a variety of diseases. In this regard, platelets act as a circulating source of several pro- and anti-inflammatory molecules, which are secreted from their intracellular stores upon activation. Among them, mounting evidence highlights a crucial role of sphingosine-1-phosphate (S1P), a multifunctional sphingoid mediator. S1P-induced pleiotropic effects include those crucial in inflammatory processes, such as the maintenance of the endothelial barrier integrity, and leukocyte activation and recruitment at the injured site. This review outlines the peculiar features and molecular mechanisms that allow platelets for acting as a unique factory that produces and stores S1P in large quantities. A particular emphasis is placed on the autocrine and paracrine roles of S1P derived from the "inflamed" platelets, highlighting the role of its cross-talk with endothelial and blood cells involved in inflammation, and the mechanisms of its contribution to the development and progression of inflammatory diseases. Finally, potential clinical implications of platelet-derived S1P as diagnostic tool of inflammatory severity, and as therapeutic target in inflammation are discussed.
Subject(s)
Blood Platelets/metabolism , Inflammation/metabolism , Lysophospholipids/metabolism , Sphingosine/analogs & derivatives , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Autocrine Communication , Biological Transport , Blood Platelets/drug effects , Endothelium, Vascular/metabolism , Endothelium, Vascular/pathology , Humans , Inflammation/blood , Inflammation/diagnosis , Inflammation/drug therapy , Leukocytes/metabolism , Lysophospholipids/antagonists & inhibitors , Lysophospholipids/blood , Molecular Targeted Therapy , Platelet Activation , Signal Transduction , Sphingosine/antagonists & inhibitors , Sphingosine/blood , Sphingosine/metabolism , Thrombosis/blood , Thrombosis/metabolismABSTRACT
Accumulating reports suggest that human glioblastoma contains glioma stem-like cells (GSCs) which act as key determinants driving tumor growth, angiogenesis, and contributing to therapeutic resistance. The proliferative signals involved in GSC proliferation and progression remain unclear. Using GSC lines derived from human glioblastoma specimens with different proliferative index and stemness marker expression, we assessed the hypothesis that sphingosine-1-phosphate (S1P) affects the proliferative and stemness properties of GSCs. The results of metabolic studies demonstrated that GSCs rapidly consume newly synthesized ceramide, and export S1P in the extracellular environment, both processes being enhanced in the cells exhibiting high proliferative index and stemness markers. Extracellular S1P levels reached nM concentrations in response to increased extracellular sphingosine. In addition, the presence of EGF and bFGF potentiated the constitutive capacity of GSCs to rapidly secrete newly synthesized S1P, suggesting that cooperation between S1P and these growth factors is of central importance in the maintenance and proliferation of GSCs. We also report for the first time that S1P is able to act as a proliferative and pro-stemness autocrine factor for GSCs, promoting both their cell cycle progression and stemness phenotypic profile. These results suggest for the first time that the GSC population is critically modulated by microenvironmental S1P, this bioactive lipid acting as an autocrine signal to maintain a pro-stemness environment and favoring GSC proliferation, survival and stem properties.
