title: NYESO-1/LAGE-1s and PRAME are targets for antigen specific T cells in chondrosarcoma following treatment with 5-Aza-2-deoxycitabine.
abstract: BACKGROUND
  Chondrosarcoma has no proven systemic option in the metastatic setting. The development of a non-cross-resistant strategy, such as cellular immunotherapy using antigen-specific T cells would be highly desirable. NY-ESO-1 and PRAME are members of the Cancer Testis Antigen (CTA) family that have been identified as promising targets for T cell therapy. LAGE-1 is a cancer testis antigen 90% homologous to NY-ESO-1, sharing the 157-165 A*0201 NY-ESO-1 epitope with its transcript variant, LAGE-1s. A number of CTA's have been induced using 5-Aza-2-Deoxycitabine (5-Aza-dC) in other cancers. We sought to evaluate the feasibility of targeting chondrosarcoma tumors using NY-ESO-1/LAGE-1s and PRAME specific T cells using 5-Aza-dC to induce antigen expression.
  
  
  METHODS
  We used 11 flash frozen tumors from the University of Washington tumor bank to test for the expression of NY-ESO-1, PRAME, LAGE-1s and LAGE-1L in chondrosarcoma tumors. Using four chondrosarcoma cell lines we tested the expression of these CTA's with and without 5-Aza-dC treatments. Finally, using NY-ESO-1/LAGE-1s and PRAME specific effectors that we generated from sarcoma patients, we evaluated the ability of these T cells to lyse A*0201 expressing chondrosarcoma cell lines in vitro both with and without 5-Aza-dC treatment.
  
  
  RESULTS
  A minority (36%) of chondrosarcoma tumors expressed either NY-ESO-1 or LAGE-1s at >10% of our reference value and none expressed PRAME at that level. However, in all four of the chondrosarcoma cell lines tested, NY-ESO-1 and PRAME expression could be induced following treatment with 5-Aza-dC including in cell lines where expression was absent or barely detectable. Furthermore, NY-ESO-1/LAGE-1s and PRAME specific CD8+ effector T cells were able to specifically recognize and lyse A*0201 expressing chondrosarcoma cell lines following 5-Aza-dC treatment.
  
  
  CONCLUSIONS
  These data suggest that adoptive immunotherapy in combination with 5-Aza-dC may be a potential strategy to treat unresectable or metastatic chondrosarcoma patients where no proven systemic therapies exist.
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America. spollac@fhcrc.org
authors.forename: Seth M
authors.initials: SM
authors.lastname: Pollack
authors.forename: Yonqing
authors.initials: Y
authors.lastname: Li
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Blaisdell
authors.forename: Erik A
authors.initials: EA
authors.lastname: Farrar
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Chou
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Hoch
authors.forename: Elizabeth T
authors.initials: ET
authors.lastname: Loggers
authors.forename: Eve
authors.initials: E
authors.lastname: Rodler
authors.forename: Janet F
authors.initials: JF
authors.lastname: Eary
authors.forename: Ernest U
authors.initials: EU
authors.lastname: Conrad
authors.forename: Robin L
authors.initials: RL
authors.lastname: Jones
authors.forename: Cassian
authors.initials: C
authors.lastname: Yee
mesh_terms: D000941:Antigens / Q000737:chemistry; D000951:Antigens, Neoplasm / Q000096:biosynthesis / Q000737:chemistry*; D000954:Antigens, Surface / Q000737:chemistry*; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D001374:Azacitidine / Q000494:pharmacology*; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism; D002813:Chondrosarcoma / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D000939:Epitopes / Q000737:chemistry; D006680:HLA Antigens; D006801:Humans; D016130:Immunophenotyping; D007167:Immunotherapy / Q000379:methods; D053645:Interleukin-2 Receptor alpha Subunit / Q000096:biosynthesis; D008297:Male; D008565:Membrane Proteins / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D060888:Real-Time Polymerase Chain Reaction; D013737:Testis / Q000378:metabolism
chemical_list: D000941:Antigens; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D000964:Antimetabolites, Antineoplastic; C104948:CTAG1B protein, human; C510162:CTAG2 protein, human; D017931:DNA Primers; D000939:Epitopes; D006680:HLA Antigens; D053645:Interleukin-2 Receptor alpha Subunit; D008565:Membrane Proteins; C404944:PRAME protein, human; D010455:Peptides; D001374:Azacitidine