1 / 22

Synovial Sarcoma (SS)

SS18-SSX IS A CELL-CONTEXT-DEPEDENT EPIGENETIC REGULATOR: IMPLICATION FOR CELL-OF-ORIGIN OF SYNOVIAL SARCOMAS. Sakura Tamaki 1,2 , Makoto Fukuta 1,2,3 , Kazuo Hayakawa 1,2,3 , Tomohisa Kato Jr. 1 , and Junya Toguchida 1,2,4.

eagan-holden
Download Presentation

Synovial Sarcoma (SS)

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. SS18-SSX IS A CELL-CONTEXT-DEPEDENT EPIGENETIC REGULATOR: IMPLICATION FOR CELL-OF-ORIGIN OF SYNOVIAL SARCOMAS Sakura Tamaki1,2, Makoto Fukuta1,2,3, Kazuo Hayakawa1,2,3, Tomohisa Kato Jr.1, and Junya Toguchida1,2,4 1Dept. Tissue Regeneration, Inst. Frontier Med. Sci., Kyoto Univ. 2Dept. Growth and Differentiation, CiRA, Kyoto Univ. 3Dept. Orthop. Surg., Grad. School Med. Sci, Nagoya City Univ. 4Dept. Orthop, Surg., Grad. School Med., Kyoto Univ.

  2. Synovial Sarcoma (SS) SSX SS18 (SSX1, SSX2, SSX4) Xp11 18q11 18q11-qter Xp11-qter SS18-SSX

  3. Function of SS18-SSX protein Transcriptional activator Transcriptional repressor BRM PcG p300 SS18-SSX DD ◆No DNA binding motif NH2 SNH QPGY RD COOH Chromatin modulator Up- or down-regulate target genes

  4. Search for genes up-regulated by SS18-SSX ◆FZD10 is expressed only in placenta among adult normal tissues. (Sited from LSBM) ◆FZD10 is specifically up-regulated in SS, but not in other types of soft tissue sarcomas(STS). MSC LS SS SS cell lines MFH LMS MPNST 6 1 2 3 4 5 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 FZD10 b2MG The FZD10 gene is a candidate as gene up-regulated by SS18-SSX [Nagayama et.al., Cancer Res., 2002]

  5. Objective Understand how SS18-SSX regulates the downstream genes using the FZD10 gene as a representative

  6. SS18-SSX binds core promoter region of FZD10 Luc Luc Luc Luc Luc Luc Luc Luc FZD10 (-3029) FZD10 (-2715) FZD10 (-1332) FZD10 (-780) FZD10 (-360) FZD10 (-118) SYO-1 FZD10 (-65) ChIP on FZD10 core promoter Relative Luc activity U2OS Relative to Input (%) FZD10 SS18-SSX2 ** Relative expression to SYO-1 HA rIgG IP: U2OS/EV SS18- SSX2 SS18- SSX2 U2OS/HA-SS18-SSX2 ( ** p<0.01) EV EV

  7. SS18-SSX directly regulates the FZD10 gene SS cell lines 1273/99 SYO-1 SS18-SSX2 FZD10 ( * P<0.05, ** p<0.01)

  8. SS18-SSX failed to induce FZD10 in hDF hDF49 SS-18SSX1 YaFu-SS SS18-SSX2 SYO-1 EV SS18-SSX SS18-SSX1 SS18-SSX2 FZD10 Human dermalfibroblast ACTB (FZD10-negative) → Are hDFs not appropriate cells?→ If so, what kind of cells are appropriate?

  9. SS shares the gene expression profile with MPNST ↓ Create “Cluster Trees” 1204 genes 577 558 391 606 712 446 298 623 357 697 406 679 734 508 330 407 551 181 327 543 341 416 580 602 325 456 370 267 473 403 350 188 646 487 582 334 190 737 259 248 213 438 397 316 53 97 44 SS SS+MPNST MPNST MFH LMS PLS or DLS Expressed in >90% of 47 STS cases Those expression ratio varied by SD>1 ↓ 1,204 Genes (Nagayama et al., Cancer Res., 2002)

  10. SS shares the protein expression profile with CCS (Suehara et al., Proteomics 6: 4402-9, 2006) Clear cell sarcoma (CCS) Synovial sarcoma MPNST

  11. SS is also a neural crest-derive tumor? ? ? Neural crest SSprecursor Schwann cell Melanocyte SS MPNST CCS

  12. Application of iPS cells for sarcoma research : investigation for the cell-of-origin of synovial sarcoma Makoto Fukuta1,2,3, Kazuo Hayakawa1,2,3, Sakura Tamaki1,2, Kazuya Sekiguchi1,2, Shingo Sato1,Makoto Ikeya2, Takanobu Otsuka3, Knut Woltjen4,Junya Toguchida1,2 SS18-SSX2 SS18-SSX2 SS18-SSX2 NCC Skin fibroblast + Dox + Dox + Dox Bone Fat Muscle NC-derived Mesenchymalstem cell Pluripotent stem cell (PSC) Neural crest cell Cartilage NC-MSC

  13. Property of NCC CD271(p75) Relative expression PAX6/Ap2α/DAPI Relative expression

  14. Property of NC-MSC Osteogenic differentiation Chondrogenic differentiation Alucian Blue staining Alizarin Red staining

  15. Induction of the FZD10 in ESC and NCC Stuffer SS18-SSX2 (** p<0.01) KhES1 SS18-SSX2 FZD10 ** ** ** Relative expression to SYO-1 DOX (ug/ml) 0 0.03 0.1 1.0 0 0.03 0.1 1.0 0.3 0.3 NCC-KhES1 SS18-SSX2 FZD10 ** ** Relative expression to SYO-1 0 0.1 0.3 1.0 0 0.1 0.3 1.0

  16. Induction of the FZD10 in NC-MSC NC-MSC-KhES1 Stuffer SS18-SSX2 SS18-SSX2 FZD10 Relative expression to SYO-1 DOX (ug/ml) 0.1 0.3 1.0 5.0 0 0.1 0.3 1.0 5.0 0 SYO-1 Relative expression to SYO-1 DOX (ug/ml) 0 0 1.0 1.0 0 0 0 1.0 1.0 1.0 0 1.0 SYO-1 (hr) 48 72 96 48 72 96

  17. Change in H3K27me3 at the FZD10 locus by SS18-SSX2 ATG -2kb -1kb +457 ORF FZD10 5’UTR 1 2 3 4 H3K27me3 Stuffer SS18-SSX2 NCC-KhES1 KhES1 NC-MSC-KhES1 FZD10; Induced Induced Not induced Relative to Input (%) 1 2 3 4 1 2 3 4 Region 1 2 3 4

  18. Change in H3K4me3 at the FZD10 locus by SS18-SSX2 ATG -2kb -1kb +457 ORF FZD10 5’UTR 1 2 3 4 Stuffer SS18-SSX2 H3K4me3 NCC-KhES1 KhES1 NC-MSC-KhES1 FZD10; Induced Induced Not induced Relative to Input (%) 1 2 3 4 1 2 3 4 Region 1 2 3 4

  19. Change in H3Ac at the FZD10 locus by SS18-SSX2 ATG -2kb -1kb +457 ORF FZD10 5’UTR 1 2 3 4 H3Ac Stuffer SS18-SSX2 NCC-KhES1 KhES1 NC-MSC-KhES1 FZD10; Induced Induced Not induced Relative to Input (%) 1 2 3 4 1 2 3 4 Region 1 2 3 4

  20. NCC Importance of H3Ac for the FZD10 induction TSA (HDAC inhibitor) Occupancy of H3Ac at FZD10 locus Relative expression (to SYO-1) Relative to Input (%) TSA (uM); 0 0.01 0.03 0.1 0.3 1.0 Region 1 2 3 4 SS18-SSX2 FZD10 DMSO TSA

  21. Conclusion SS18-SSX2 SS18-SSX2 SS18-SSX2 NC-MSC ESC NCC (-) (-) (-) FZD10 (+) (+) (-) FZD10 H3K4me3 H3K27me H3Ac

  22. Toguchida’s Lab. in IFMS & CiRA, Kyoto Univ.

More Related