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Fas Ligand: switching signals from
tumor immune erosion to metastasis
Bei-Chang Yang, Ph.D. (楊倍昌)
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Professor, Department of Microbiology and Immunology,
College of Medicine, National Cheng Kung University
(NCKU), Tainan, Taiwan.
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Director, Research Center for Society, Technology and
Medicine (STM center); NCKU Tainan, Taiwan.
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Fas+
On Tumor microenvironment and infiltrating T cells
FasL+
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Novel FasL signaling mechanism!
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Why are T cells trapped in peripheral area of tumor?
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(Tenascin C) Huang JY, et al (2010) J Immunol. 185:1450-1459.
(collagen) Lin YP, et al. (2009) Mol Immunol. Mol Immunol. 46:3328-3335.
Are those infiltrating T cells alive ?
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(PI3K) Su CC, et al (2007) J Immunol. 179:4589-4597.
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(FasL) Hor WS, et al. (2003) J Leukocyte Biol. 73: 363-368.
Do infiltrating T cells affect tumor growth or metastasis?
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(FasL) Lin HC, et al (2013) J Biol Chem. 287:20664–20673.
(depletion study) Chen YL, et al (2002) Br J Cancer 87:359-365.
(Fas signaling) Chen YL, et al. (2003) J Immunol. 171:1183-1192.
Can they modulate consequent local immune reaction?
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(IL-10 & TGF-b) Yang BC, et al. (2003) J Immunol. 171:3947-3954.
(Fas and Th17) Su CC, et al (2013) Cell Immunol. 281:101–110
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apoptosis
Fas/Fas-ligand : the major signaling
system for cell death.
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Fas (1991): Nagata, S.;
Krammer, P.
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FasL (1993): Nagata, S. &
Goldstein, P.
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CD95/CD95-L
長田 重一
P Krammer
Fas:
Itoh N, et al. (1991) Cell 66: 233–243 (Nagata S)
Oehm A , et al. (1992) J. Biol. Chem. 267: 10709–10715 (Krammer P)
FasL:
Suda T, et al. (1993) Cell 75: 1169-1178 (Nagata S; Goldstein P)
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Nature 407, 789 – 795(2000)
Apaf-1
Caspase 9
Caspase 3
Death
substrates
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FasL reverse signaling
(FasL by itself acts as a receptor)
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Suzuki I. and Fink PJ, Maximal proliferation of cytotoxic T
lymphocytes requires reverse signaling through Fas ligand. J.
Exp. Med., 187:123, 1998.
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Sun M, Ames KT, Suzuki I,. Fink PJ, The Cytoplasmic
Domain of Fas Ligand Costimulates TCR Signals, J.
Immunol., 177, 1481, 2006.
Pamela J. Fink, h.D.
Department of Immunology
The cytoplasmic domain of FasL is
sufficient to mediate costimulation
(FasL cross-linking is required).
University of Washington
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BC Yang
Conventional view of receptor-ligand interaction
FasL
FasL
Fas
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Take home massage: Fas ligand hijack the Met receptor on lipid raft.
Protein-protein interaction in lipid raft is an
alternative way for cell signaling.
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The structure of FasL
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Due to unstable protein region
(A). Full-length and truncated FasL. Adapted from Orlinick JR, et al. J Biol Chem
1997, 272:32221-29. Jodo S, et al. J Immunol 2005, 174:4470-4474. FasL-△70: deleting
the N-terminal 2-70 aa; FasL-△33: deleting the N-terminal 2-33 aa.
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FasL enhances cell migration and invasion.
A
B
(A) In the single cell motility assay, cell migration distance in 6 h was recorded every
5 min by time lapse video under a microscope. Total distance of cell migration was
taken to represent cell mobility. (B) In invasion assay, cells were seeded on matrigel
(1 mg/ml) coated transwell plates in 16 h. Invading cells were calculated for
quantification. Values was shown the mean ± SEM. (* p<0.05, ** p<0.001 compared
with N1, # p<0.05 compared with FasL)
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Truncated FasL promote cell transformation (enhances
lung metastasis, anchorage-indecent growth)
Lung metastasis in eight-week-old
BALB/cAnN.Cg-Foxn1nu/CrlNarl
nude mice
anchorage-independent growth.
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Q: Why FasL without intracellular domain still promotes tumor metastasis?
The Met tyrosine kinase receptor is a high-affinity receptor for hepatocyte
growth factor/scatter factor (HGF/SF). HGF/SF-Met system is necessary
for embryonic development, and aberrant MET signaling favors
tumorigenesis and metastasis.
cyclinD1
Metastasis
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Association of FasL and Met in lipid raft
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Location of Met is not changed
Lipid raft formation is required
MbCD:methyl-beta-cyclodextrin, a specific cholesterol-binding agent
extracting cholesterol from plasma membranes
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Confocal imaging reveals colocalization of FasL and Met
labeled cholera toxin B
MF/M: Met/FasL complex/total Met
FL/L:FasL in lipid raft/lipid raft
ML/L:Met in lipid raft/lipid raft
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A FasL domain between 70-130 is important for
complex with Met.
NIH3T3/ aFasL (C-terminal, BD)
MRC5 cells/aFasL (N-20, Senta Cruz)
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FasL105-130
Note: full length FasL triggered profound apoptosis in MRC5
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The Met tyrosine kinase receptor is a high-affinity receptor for hepatocyte
growth factor/scatter factor (HGF/SF). HGF/SF-Met system is necessary
for embryonic development, and aberrant MET signaling favors
tumorigenesis and metastasis.
cyclinD1
Metastasis
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•
Met signal is initiated and mediates transformed phenotype.
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Stat3 is essential for the enhanced cell migration.
Met inhibitor: PHA665752
Stst3 inhibitor: AG490
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FasL/Met complex in human tumor cells
B
A
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The complex of FasL and Met is detected in various tumor cell
lines.
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Suppression of FasL by specific ribozyme reduces the
phosphorylation of Met.
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Fas/FasL signaling revised:
There are three different pathways initiated by FasL.
FasL-associated metastasis
Autoimmunity?
lipid raft
Fas counter-attack mechanism/
Tissue destruction?
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Mother nature works in a way that is
not always as been told.
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