Modulating Sphingosine-1-Phosphate receptors to improve chemotherapy delivery to Ewing sarcoma

ABSTRACTTumor vasculature is innately dysfunctional. Poorly functional tumor vessels inefficiently deliver chemotherapy to tumor cells; vessel hyper-permeability promotes chemotherapy delivery primarily to a tumor’s periphery. Here we identify a method for enhancing chemotherapy delivery and efficacy in Ewing sarcoma (ES) in mice by modulating tumor vessel permeability. Vessel permeability is partially controlled by the G protein-coupled Sphinosine-1-phosphate receptors 1 and 2 (S1PR1 and S1PR2) on endothelial cells. S1PR1 promotes endothelial cell junction integrity while S1PR2 destabilizes it. We hypothesize that an imbalance of S1PR1:S1PR2 is partially responsible for the dysfunctional vascular phenotype characteristic of ES and that by altering the balance in favor of S1PR1, ES vessel hyper-permeability can be reversed. In this study, we demonstrate that pharmacologic activation of S1PR1 by SEW2871 or inhibition of S1PR2 by JTE-013 caused more organized, mature, and functional tumor vessels. Importantly, S1PR1 activation or S1PR2 inhibition improved chemotherapy delivery to the tumor and anti-tumor efficacy. Our data suggests that pharmacologic targeting of S1PR1 and S1PR2 may be a useful adjuvant to standard chemotherapy for ES patients.NOVELTY AND IMPACTThis study demonstrates that Sphingosine-1-Phosphate (S1P) receptors are potential novel targets for tumor vasculature remodeling and adjuvant therapy for the treatment of Ewing Sarcoma. Unlike receptor tyrosine kinases that have already been extensively evaluated for use as vascular normalizing agents in oncology, S1P receptors are G protein-coupled receptors, which have not been well studied in tumor endothelium. Pharmacologic activators and inhibitors of S1P receptors are currently in clinical trials for treatment of auto-immune and cardiovascular diseases, indicating potential for clinical translation of this work.

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Sphingosine 1-Phosphate (S1P) Induces Migration and ERK/MAP-Kinase-Dependent Proliferation in Chronic Lymphocytic Leukemia (B-CLL) Due to Expression of the G Protein-Coupled Receptors S1P1/4.

Blood ◽

10.1182/blood.v106.11.4996.4996 ◽

2005 ◽

Vol 106 (11) ◽

pp. 4996-4996

Author(s):

Gabriele Seitz ◽

Sedat Yildirim ◽

Andreas M. Boehmler ◽

Lothar Kanz ◽

Robert Möhle

Keyword(s):

Chronic Lymphocytic Leukemia ◽

G Protein ◽

Cell Lines ◽

Peripheral Blood ◽

G Protein Coupled Receptors ◽

Lymphocytic Leukemia ◽

S1p Receptors ◽

Sphingosine 1 Phosphate ◽

G Protein Coupled

Abstract Egress of lymphocytes from lymphoid organs into the circulation has been shown to depend on the presence of the lipid mediator sphingosine 1-phosphate (S1P) in the peripheral blood, and expression of corresponding S1P receptors (i.e., S1P1), that belong to the family of 7-transmembrane G protein-coupled receptors (GPCR). As circulating lymphocytic lymphoma cells are a hallmark of chronic lymphocytic leukemia, we analyzed expression of different S1P receptors and the effects of S1P on B-CLL cells. By qualitative and quantitative (TaqMan) RT-PCR, significant mRNA expression of S1P1 and S1P4 was found in CLL cell lines (EHEB, MEC-1) and in most samples (S1P1 in 88%, S1P4 in 100%) of primary CD19+ cells isolated from the peripheral blood of untreated B-CLL patients. mRNA of other S1P receptors (S1P2, S1P3, S1P5) was less consistently detected. Normal, nonmalignant B cells were strongly positive for S1P1, while other S1P receptors were weakly expressed or negative. S1P induced typical effects of chemotactic GPCR, such as actin polymerization (analyzed by flow cytometry) and chemotaxis (measured in a modified Boyden chamber assay) in CLL cell lines and primary B-CLL cells. After serum deprivation in vitro, S1P induced phosphorylation of ERK/MAP-kinase as analyzed by Western blot, demonstrating that S1P receptors expressed in CLL were able to activate signaling pathways of GPCR not only related to cell migration and chemotaxis, but also to cell proliferation. Of note, the S1P1 ligand FTY720, which induces receptor internalization after prolonged exposure and acts as an antagonist, resulted in apoptosis in CLL cell lines and primary CLL cells in vitro, as measured by MTT-test and staining with Annexin-FITC, respectively. We conclude that sphingosine 1-phosphate, which is present in the peripheral blood in considerable amounts, contributes to the trafficking of B-CLL cells expressing the GPCRs S1P1/4, and to their prolonged survival.

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Sphingosine 1-phosphate signalling in mammalian cells

Biochemical Journal ◽

10.1042/bj3490385 ◽

2000 ◽

Vol 349 (2) ◽

pp. 385-402 ◽

Cited By ~ 260

Author(s):

Susan PYNE ◽

Nigel J. PYNE

Keyword(s):

G Protein ◽

Tyrosine Kinases ◽

Mammalian Cells ◽

Receptor Tyrosine Kinases ◽

Cytoskeletal Proteins ◽

Mitogen Activated Protein Kinase ◽

G Protein Coupled Receptor ◽

Biological Responses ◽

Sphingosine 1 Phosphate ◽

G Protein Coupled

Sphingosine 1-phosphate is formed in cells in response to diverse stimuli, including growth factors, cytokines, G-protein-coupled receptor agonists, antigen, etc. Its production is catalysed by sphingosine kinase, while degradation is either via cleavage to produce palmitaldehyde and phosphoethanolamine or by dephosphorylation. In this review we discuss the most recent advances in our understanding of the role of the enzymes involved in metabolism of this lysolipid. Sphingosine 1-phoshate can also bind to members of the endothelial differentiation gene (EDG) G-protein-coupled receptor family [namely EDG1, EDG3, EDG5 (also known as H218 or AGR16), EDG6 and EDG8] to elicit biological responses. These receptors are coupled differentially via Gi, Gq, G12/13 and Rho to multiple effector systems, including adenylate cyclase, phospholipases C and D, extracellular-signal-regulated kinase, c-Jun N-terminal kinase, p38 mitogen-activated protein kinase and non-receptor tyrosine kinases. These signalling pathways are linked to transcription factor activation, cytoskeletal proteins, adhesion molecule expression, caspase activities, etc. Therefore sphingosine 1-phosphate can affect diverse biological responses, including mitogenesis, differentiation, migration and apoptosis, via receptor-dependent mechanisms. Additionally, sphingosine 1-phosphate has been proposed to play an intracellular role, for example in Ca2+ mobilization, activation of non-receptor tyrosine kinases, inhibition of caspases, etc. We review the evidence for both intracellular and extracellular actions, and extensively discuss future approaches that will ultimately resolve the question of dual action. Certainly, sphingosine 1-phosphate will prove to be unique if it elicits both extra- and intra-cellular actions. Finally, we review the evidence that implicates sphingosine 1-phosphate in pathophysiological disease states, such as cancer, angiogenesis and inflammation. Thus there is a need for the development of new therapeutic compounds, such as receptor antagonists. However, identification of the most suitable targets for drug intervention requires a full understanding of the signalling and action profile of this lysosphingolipid. This article describes where the research field is in relation to achieving this aim.

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G-protein–coupled formyl peptide receptors play a dual role in neutrophil chemotaxis and bacterial phagocytosis

Molecular Biology of the Cell ◽

10.1091/mbc.e18-06-0358 ◽

2019 ◽

Vol 30 (3) ◽

pp. 346-356 ◽

Cited By ~ 7

Author(s):

Xi Wen ◽

Xuehua Xu ◽

Wenxiang Sun ◽

Keqiang Chen ◽

Miao Pan ◽

...

Keyword(s):

G Protein ◽

Receptor Tyrosine Kinase ◽

Tyrosine Kinases ◽

Actin Polymerization ◽

G Protein Coupled Receptors ◽

Peptide Receptors ◽

Tyrosine Kinase Signaling ◽

Formyl Peptide Receptors ◽

Formyl Peptide ◽

G Protein Coupled

A dogma of innate immunity is that neutrophils use G-protein–coupled receptors (GPCRs) for chemoattractant to chase bacteria through chemotaxis and then use phagocytic receptors coupled with tyrosine kinases to destroy opsonized bacteria via phagocytosis. Our current work showed that G-protein–coupled formyl peptide receptors (FPRs) directly mediate neutrophil phagocytosis. Mouse neutrophils lacking formyl peptide receptors (Fpr1/2–/–) are defective in the phagocytosis of Escherichia coli and the chemoattractant N-formyl-Met-Leu-Phe (fMLP)-coated beads. fMLP immobilized onto the surface of a bead interacts with FPRs, which trigger a Ca2+response and induce actin polymerization to form a phagocytic cup for engulfment of the bead. This chemoattractant GPCR/Gi signaling works independently of phagocytic receptor/tyrosine kinase signaling to promote phagocytosis. Thus, in addition to phagocytic receptor-mediated phagocytosis, neutrophils also utilize the chemoattractant GPCR/Gi signaling to mediate phagocytosis to fight against invading bacteria.

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Signaling of sphingosine-1-phosphate via the S1P/EDG-family of G-protein-coupled receptors

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids ◽

10.1016/s1388-1981(02)00139-7 ◽

2002 ◽

Vol 1582 (1-3) ◽

pp. 72-80 ◽

Cited By ~ 227

Author(s):

M Kluk

Keyword(s):

G Protein ◽

G Protein Coupled Receptors ◽

Sphingosine 1 Phosphate ◽

G Protein Coupled

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Ligand bias in receptor tyrosine kinase signaling

Journal of Biological Chemistry ◽

10.1074/jbc.rev120.015190 ◽

2020 ◽

Vol 295 (52) ◽

pp. 18494-18507

Author(s):

Kelly Karl ◽

Michael D. Paul ◽

Elena B. Pasquale ◽

Kalina Hristova

Keyword(s):

G Protein ◽

Receptor Tyrosine Kinase ◽

Tyrosine Kinases ◽

Receptor Tyrosine Kinases ◽

G Protein Coupled Receptors ◽

Operational Model ◽

Tyrosine Kinase Signaling ◽

Rtk Signaling ◽

Ligand Bias ◽

G Protein Coupled

Ligand bias is the ability of ligands to differentially activate certain receptor signaling responses compared with others. It reflects differences in the responses of a receptor to specific ligands and has implications for the development of highly specific therapeutics. Whereas ligand bias has been studied primarily for G protein–coupled receptors (GPCRs), there are also reports of ligand bias for receptor tyrosine kinases (RTKs). However, the understanding of RTK ligand bias is lagging behind the knowledge of GPCR ligand bias. In this review, we highlight how protocols that were developed to study GPCR signaling can be used to identify and quantify RTK ligand bias. We also introduce an operational model that can provide insights into the biophysical basis of RTK activation and ligand bias. Finally, we discuss possible mechanisms underpinning RTK ligand bias. Thus, this review serves as a primer for researchers interested in investigating ligand bias in RTK signaling.

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Sphingosine-1-Phosphate as a Ligand for the G Protein-Coupled Receptor EDG-1

Science ◽

10.1126/science.279.5356.1552 ◽

1998 ◽

Vol 279 (5356) ◽

pp. 1552-1555 ◽

Cited By ~ 701

Author(s):

M. Lee

Keyword(s):

G Protein ◽

G Protein Coupled Receptor ◽

Sphingosine 1 Phosphate ◽

G Protein Coupled

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Sphingosine 1-phosphate signalling through the G-protein-coupled receptor Edg-1

Biochemical Journal ◽

10.1042/bj3300605 ◽

1998 ◽

Vol 330 (2) ◽

pp. 605-609 ◽

Cited By ~ 181

Author(s):

C. M. Gerben ZONDAG ◽

R. Friso POSTMA ◽

Ingrid VAN ETTEN ◽

Ingrid VERLAAN ◽

H. Wouter MOOLENAAR

Keyword(s):

Adenylate Cyclase ◽

Map Kinase ◽

G Protein ◽

G Protein Coupled Receptors ◽

Lpa Receptor ◽

Kinase Activation ◽

Protein Map ◽

Mitogen Activated Protein ◽

Sphingosine 1 Phosphate ◽

G Protein Coupled

Sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) are structurally related lipid mediators that act on distinct G-protein-coupled receptors to evoke similar responses, including Ca2+ mobilization, adenylate cyclase inhibition, and mitogen-activated protein (MAP) kinase activation. However, little is still known about the respective receptors. A recently cloned putative LPA receptor (Vzg-1/Edg-2) is similar to an orphan Gi-coupled receptor termed Edg-1. Here we show that expression of Edg-1 in Sf9 and COS-7 cells results in inhibition of adenylate cyclase and activation of MAP kinase (Gi-mediated), but not Ca2+ mobilization, in response to S1P. These responses are specific in that (i) S1P action is not mimicked by LPA, and (ii) Vzg-1/Edg-2 cannot substitute for Edg-1. Thus the Edg-1 receptor is capable of mediating a subset of the cellular responses to S1P.

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G-Protein-Coupled Receptor Cell Signaling Pathways Mediating Embryonic Chick Retinal Growth Cone Collapse Induced by Lysophosphatidic Acid and Sphingosine-1-Phosphate

Developmental Neuroscience ◽

10.1159/000364858 ◽

2014 ◽

Vol 36 (6) ◽

pp. 443-453 ◽

Cited By ~ 18

Author(s):

Jarod Fincher ◽

Canaan Whiteneck ◽

Eric Birgbauer

Keyword(s):

G Protein ◽

Lysophosphatidic Acid ◽

Growth Cone ◽

Receptor Cell ◽

Embryonic Chick ◽

Growth Cone Collapse ◽

Retinal Growth ◽

Sphingosine 1 Phosphate ◽

G Protein Coupled ◽

Cell Signaling Pathways

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Lysophospholipid Growth Factors and Their G Protein-Coupled Receptors in Immunity, Coronary Artery Disease, and Cancer

The Scientific World JOURNAL ◽

10.1100/tsw.2002.124 ◽

2002 ◽

Vol 2 ◽

pp. 324-338 ◽

Cited By ~ 21

Author(s):

Edward J. Goetzl ◽

Markus Graeler ◽

Mei-Chuan Huang ◽

Geetha Shankar

Keyword(s):

Growth Factors ◽

Tumor Cells ◽

G Protein ◽

Neural Development ◽

Cellular Proliferation ◽

Biological Fluids ◽

Functional Responses ◽

Cancer Data ◽

S1p Receptors ◽

G Protein Coupled

The physiological lysophospholipids (LPLs), exemplified by lysophosphatidic acid (LPA) and sphingosine 1-phosphate (S1P), are omnific mediators of normal cellular proliferation, survival, and functions. Although both LPA and S1P attain micromolar concentrations in many biological fluids, numerous aspects of their biosynthesis, transport, and metabolic degradation are unknown. Eight members of a new subfamily of G protein-coupled LPA/S1P receptors, originally termed Edg Rs, bind either LPA or S1P with high affinity and transduce a series of growth-related and/or cytoskeleton-based functional responses. The most critical areas of LPL biology and pathobiology are neural development and neurodegeneration, immunity, atherosclerosis and myocardial injury, and cancer. Data from analyses of T cells established two basic points: (1) the plasticity and adaptability of expression of LPA/S1P Rs by some cells as a function of activation, and (2) the role of opposing signals from two different receptors for the same ligand as a mechanism for fine control of effects of LPLs. In the heart, LPLs may promote coronary atherosclerosis, but are effectively cytoprotective for hypoxic cardiac myocytes and those exposed to oxygen free radicals. The findings of production of LPA by some types of tumor cells, overexpression of selected sets of LPA receptors by the same tumor cells, and augmentation of the effects of protein growth factors by LPA have suggested pathogenetic roles for the LPLs in cancer. The breadth of physiologic and pathologic activities of LPLs emphasizes the importance of developing bioavailable nonlipid agonists and antagonists of the LPA/S1P receptors for diverse therapeutic applications.

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