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Non-phosphorylated FTY720 Induces Apoptosis of Human Microglia by Activating SREBP2

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Abstract

A synthetic analog of sphingosine named FTY720 (Fingolimod), phosphorylated by sphingosine kinase-2, interacts with sphingosine-1-phosphate (S1P) receptors expressed on various cells. FTY720 suppresses the disease activity of multiple sclerosis (MS) chiefly by inhibiting S1P-dependent egress of autoreactive T lymphocytes from secondary lymphoid organs, and possibly by exerting anti-inflammatory and neuroprotective effects directly on brain cells. However, at present, biological effects of FTY720 on human microglia are largely unknown. We studied FTY720-mediated apoptosis of a human microglia cell line HMO6. The exposure of HMO6 cells to non-phosphorylated FTY720 (FTY720-non-P) induced apoptosis in a dose-dependent manner with IC50 of 10.6 ± 2.0 μM, accompanied by the cleavage of caspase-7 and caspase-3 but not of caspase-9. The apoptosis was inhibited by Z-DQMD-FMK, a caspase-3 inhibitor, but not by Pertussis toxin, a Gi protein inhibitor, suramin, a S1P3/S1P5 inhibitor, or W123, a S1P1 competitive antagonist, although HMO6 expressed S1P1, S1P2, and S1P3. Furthermore, both phosphorylated FTY720 (FTY720-P) and SEW2871, S1P1 selective agonists, did not induce apoptosis of HMO6. Genome-wide gene expression profiling and molecular network analysis indicated activation of transcriptional regulation by sterol regulatory element-binding protein (SREBP) in FTY720-non-P-treated HMO6 cells. Western blot verified activation of SREBP2 in these cells, and apoptosis was enhanced by pretreatment with simvastatin, an activator of SREBP2, and by overexpression of the N-terminal fragment of SREBP2. These observations suggest that FTY720-non-P-induced apoptosis of HMO6 human microglia is independent of S1P receptor binding, and positively regulated by the SREBP2-dependent proapoptotic signaling pathway.

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Abbreviations

CNS:

Central nervous system

DAVID:

Database for Annotation, Visualization, and Integrated Discovery

EDG:

Endothelial differentiation gene

FTY720-non-P:

Non-phosphorylated form of FTY720

FTY720-P:

Phosphorylated form of FTY720

G3PDH:

Glyceraldehyde-3-phosphate dehydrogenase

GPCR:

G-protein-coupled receptor

INSIG1:

Insulin-induced gene 1

LDLR:

Low density lipoprotein receptor

MS:

Multiple sclerosis

OPC:

Oligodendroglial progenitor cell

S1P:

Sphingosine-1-phosphate

SPHK2:

Sphingosine kinase-2

SREBP:

Sterol regulatory element-binding protein

PARP:

Poly-ADP-ribose-polymerase

PTX:

Pertussis toxin

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Acknowledgments

This work was supported by grants from the Research on Intractable Diseases, the Ministry of Health, Labour and Welfare, Japan (H22-Nanchi-Ippan-136; H21-Nanchi-Ippan-201; H21-Nanchi Ippan-217; H21-Kokoro-Ippan-018) and the High-Tech Research Center Project (S0801043) and the Grant-in-Aid (C22500322), the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. The microarray data are available from Gene Expression Omnibus (GEO) under the accession number GSE28642.

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Authors and Affiliations

  1. Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan

    Takashi Yoshino, Hiroko Tabunoki & Jun-ichi Satoh

  2. Department of Chemistry of Functional Molecules, Meiji Pharmaceutical University, 2-522-1 Noshio, Kiyose, Tokyo, 204-8588, Japan

    Shigeo Sugiyama & Keitaro Ishii

  3. Division of Neurology, Department of Medicine, University of British Columbia Hospital, University of British Columbia, Vancouver, BC, Canada

    Seung U. Kim

  4. Medical Research Institute, Chung-Ang University College of Medicine, Seoul, Korea

    Seung U. Kim

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  1. Takashi Yoshino
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  2. Hiroko Tabunoki
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  3. Shigeo Sugiyama
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  4. Keitaro Ishii
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  6. Jun-ichi Satoh
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Correspondence to Jun-ichi Satoh.

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Yoshino, T., Tabunoki, H., Sugiyama, S. et al. Non-phosphorylated FTY720 Induces Apoptosis of Human Microglia by Activating SREBP2. Cell Mol Neurobiol 31, 1009–1020 (2011). https://doi.org/10.1007/s10571-011-9698-x

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