Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
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”.