Transcript Abstract 392

Gene-Expression Pattern and Differentiation Potential of
Synovial Sarcoma Suggest its Cellular Origin as
Neural Crest-Derived Cells
Tatsuya Ishibe, Tomitaka Nakayama, Satoshi Nagayama,
Takeshi Okamoto, Tomoki Aoyama, Koichi Nishijo,
Takashi Nakamura, and Junya Toguchida
Department of Orthopaedic Surgery,
Institute for Frontier Medical Sciences,
Kyoto University, Japan
12th CTOS meeting at Boca Raton, FL.
Why “synovial” sarcoma ?
1865 First report
Simon G., Arch Klin Chir
・ arises in para-articular region
・ microscopic resemblance to synovium
1934 “Synovialsarcome”
Sabrazes J. Gaz. Hebd. Sc. Med. Bordeaux
・ rarely arises in joint cavities.
・ differs from normal synovium by
immunohistochemical and ultrastructural study
1984 “Synovial sarcoma” is a misnomer.
Billings SD, 2004 Mod Pathol
Billings SD, 2004 Mod Pathol
Miettinen M. Am J Pathol
The cell-of-origin of synovial sarcoma is still unknown.
Backgrounds
We previously performed gene expression profiling of 47
soft tissue sarcomas by cDNA microarray (23,040 genes) , and
reported that synovial sarcoma was clustered into the same
group with malignant peripheral nerve sheath tumor which
is one of the neural crest-derived tumor.
Nagayama et al. Cancer Res. 2002
SS＋
MPNST
Same cluster
Hypothesis: Synovial sarcoma is neural crest-derived tumor
Neural crest (stem) cells: NCSC
- unique cell population of the vertebrate embryo -
arise from the neural ectoderm
and recognizable after neural
tube closure
Head and neck
Cartilage
& Bone
Neuron
Neural
tube
Trunk
Glia
Melanocyte
Joseph NM
Development, 2004
NCSCs have multiple differentiation potential
Does synovial sarcoma have similar potential as NCSC?
Objective
To estimate the origin of synovial sarcoma (SS) by
investigating its gene expression and differentiation potential.
Materials and Methods
Fourteen cell lines including 5 SS cells
Twenty-nine tumor tissues including 18 SS tumors
1) Gene expression of neural crest-related genes by RT-PCR
2) Chondrogenic, osteogenic and adipogenic inductions
: by the same methods as mesenchymal stem cells (MSCs)
3) Neurogenic induction by FGF2, ATRA, or BMP4
Differentiation markers in tumor tissues
Synovial sarcoma
neuronal
osteo chondro
Biphasic
SYT-SSX1(+)
Monophasic
SYT-SSX1(+)
Monophasic
SYT-SSX2(+)
SOX9
COL2A1
COL9A1
OSF2
ALP
MASH1
NGFR
NFH
adipo
glial
melano
CRABP1
EDN3
EDNRB
HES1
S100b
PPARg
Genes related to neural crest-derived cell lineages are expressed in SS
b actin
Chondrogenesis: Pellet and matrix formation assay
5x105 cells
3D-culture
for 7 days
Synovial sarcoma cells
HS-SY-II
SYO-1
Fuji
1273/99
MSC
HE stain
YaFuSS
Blue
Alcian
loose
Pellet formation with positive Alcian blue was observed in four SS cells
Chondrogenesis: Gene expression before and after induction
Induction
YaFuSS
HS-SY-II
SYO-1
-
-
-
+
+
+
Fuji
-
1273/99
+
-
+
MSC
-
+
SOX9
COMP
COL2A1
COL9A1
COL11A1
COL11A2
AGC
b actin
COMP: cartilage oligomeric matrix protein
Chondro-related genes were significantly upregulated.
Osteogenesis: Gene expression with/without induction
- Induction
+ Induction
for a week
+ AA + Dex + b-GP
or
Induction
YaFuSS
HS-SY-II
SYO-1
-
-
-
+
+
+
Fuji
-
1273/99
+
-
+
OSF2
ALP
COL1A1
OC
b actin
Osteo-related genes were upregulated after induction.
+
Osteogenesis: ALPase activity and Alizarin Red staining
SYO-1
Fuji
1273/99
MSC
control
Osteo
induction
control
HS-SY-II
Osteo
induction
Alizarin red stain
ALPase stain
YaFuSS
At least one cell line shows strong osteogenic differentiation.
Adipogenesis
Induction
YaFuSS
HS-SY-II
SYO-1
-
-
-
+
+
+
Fuji
-
1273/99
+
-
+
MSC
-
+
PPARg
AP2
ADP
b actin
SYO-1
Fuji
1273/99
MSC
control
HS-SY-II
Adipo
induction
Oil-red-O stain
YaFuSS
SS cells have poor or no potential for adipogenic induction
Neurogenesis: neuron-related genes
Without induction
NFH
NFM
NFL
MAP2
Nestin
b actin
ATRA
YaFuSS
HS-SY-II
SYO-1
-
-
-
+
+
+
Fuji
-
1273/99
+
-
+
ATRA, 7days
MASH1
b actin
BMP4
MASH1
b actin
YaFuSS
HS-SY-II
SYO-1
-
-
-
+
+
+
Fuji
-
1273/99
+
-
+
BMP4, 48ｈｒｓ
Immunostain of Neuron-specific beta III tubulin (Tuj1)
HS-SY-II
SYO-1
Fuji
1273/99
PC12h
(positive ctrl)
BMP4
FGF2
ATRA
vehicle
YaFuSS
Tuj1 / DAPI
Synovial sarcoma cells
Neural
Crest Cell
YaFuSS
HS-SY-II
SYO-1
Fuji
Gene exp. (growing phase)
Gene exp. (after induc.)
Pellet formation
Alcian blue stain
++
++
+
+
++
++
+
+
++
++
+
+
++
++
++
++
Gene exp. (growing phase)
Gene exp. (after induc.)
ALP stain
Alizarin red stain
-
-
+
±
±
±
+
+
-
-
+
-
±
±
Gene exp. (growing phase)
Tuj-1 stain (after FGF2)
Gene exp. (after ATRA)
Tuj-1 (after ATRA)
Gene exp. (after BMP4)
Tuj-1 (after BMP4)
++
++
+
+
-
++
+
+
-
++
+
+
-
++
+
+
++
+
++
-
Glial
Gene exp. (growing phase)
S100b (after 5-aza-dC)
+
+
+
+
+
-
Glia
Melano
Gene exp. (after 5-aza-dC)
Stress fiber reorgan.(FSK)
Melanogenesis.(FSK)
±
+
-
+
-
+
+
-+
-
Melanocyte
Gene exp. (growing phase)
Gene exp. (after induc.)
Oil-red-O stain
±
±
-
-
±
-
-
±
±
±
±
±
Chondro
Osteo
Neuro
Adipo
-
±
1273/99
±
++
Cartilage
±
+
+
++
++
Bone
Similar
potential
Neuron
-
These spectra of differentiation are similar to that of neural crest
Discussions from pathological point of view
May form ossification or calcification
・About 30% of synovial sarcomas have calcified focus.
(Enzinger FM, Cancer, 1982)
May arise from cartilage in head and neck region, but
not arise from articular cartilage which is not neural
crest origin.
・Arytenoid-epiglotic rim (Szuhai K, Cancer Genet Cytogenet, 2004)
・Cricoid cartilage, Monophasic SS (Kikuchi I, Acta Cytol, 2003)
May arise from neural tissue.
・C7 spinal root (Chu PG, Mod Pathol, 2004);
・Common digital nerve (O’Connell JX, Mod Pathol, 1996)
・Radial nerve (Cugola L, J Hand Surg, 1985)
These data also support our hypothesis that “the cell-of-origin of
synovial sarcoma is neural crest-derived cells.”
Conclusion
Synovial sarcoma expressed the genes related to
chondrocyte, osteocyte, neuron, glia, and melanocyte
which are known to originate from neural crest stem
cells.
Synovial sarcoma cells had potential to differentiate
into chondrocyte, osteocyte, and neuron to various
extent, but not into adipocyte. This spectrum of
differentiation is similar to that of neural crest stem
cells, supporting our hypothesis that “the cell-of-origin
of synovial sarcoma is neural crest-derived cells”.