title: Generation of chordoma cell line JHC7 and the identification of Brachyury as a novel molecular target.
abstract: OBJECTIVE
  Chordoma is a malignant bone neoplasm hypothesized to arise from notochordal remnants along the length of the neuraxis. Recent genomic investigation of chordomas has identified T (Brachyury) gene duplication as a major susceptibility mutation in familial chordomas. Brachyury plays a vital role during embryonic development of the notochord and has recently been shown to regulate epithelial-to-mesenchymal transition in epithelial-derived cancers. However, current understanding of the role of this transcription factor in chordoma is limited due to the lack of availability of a fully characterized chordoma cell line expressing Brachyury. Thus, the objective of this study was to establish the first fully characterized primary chordoma cell line expressing gain of the T gene locus that readily recapitulates the original parental tumor phenotype in vitro and in vivo.
  
  
  METHODS
  Using an intraoperatively obtained tumor sample from a 61-year-old woman with primary sacral chordoma, a chordoma cell line (JHC7, or Johns Hopkins Chordoma Line 7) was established. Molecular characterization of the primary tumor and cell line was conducted using standard immunostaining and Western blotting. Chromosomal aberrations and genomic amplification of the T gene in this cell line were determined. Using this cell line, a xenograft model was established and the histopathological analysis of the tumor was performed. Silencing of Brachyury and changes in gene expression were assessed.
  
  
  RESULTS
  The authors report, for the first time, the successful establishment of a chordoma cell line (JHC7) from a patient with pathologically confirmed sacral chordoma. This cell line readily forms tumors in immunodeficient mice that recapitulate the parental tumor phenotype with conserved histological features consistent with the parental tumor. Furthermore, it is demonstrated for the first time that silencing of Brachyury using short hairpin RNA renders the morphology of chordoma cells to a more differentiated-like state and leads to complete growth arrest and senescence with an inability to be passaged serially in vitro.
  
  
  CONCLUSIONS
  This report represents the first xenograft model of a sacral chordoma line described in the literature and the first cell line established with stable Brachyury expression. The authors propose that Brachyury is an attractive therapeutic target in chordoma and that JHC7 will serve as a clinically relevant model for the study of this disease.
authors.affiliation: Department of Neurosurgery and Oncology, Brain Tumor Stem Cell Laboratory, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USA.
authors.forename: Wesley
authors.initials: W
authors.lastname: Hsu
authors.forename: Ahmed
authors.initials: A
authors.lastname: Mohyeldin
authors.forename: Sagar R
authors.initials: SR
authors.lastname: Shah
authors.forename: Colette M
authors.initials: CM
authors.lastname: ap Rhys
authors.forename: Lakesha F
authors.initials: LF
authors.lastname: Johnson
authors.forename: Neda I
authors.initials: NI
authors.lastname: Sedora-Roman
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Kosztowski
authors.forename: Ola A
authors.initials: OA
authors.lastname: Awad
authors.forename: Edward F
authors.initials: EF
authors.lastname: McCarthy
authors.forename: David M
authors.initials: DM
authors.lastname: Loeb
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Wolinsky
authors.forename: Ziya L
authors.initials: ZL
authors.lastname: Gokaslan
authors.forename: Alfredo
authors.initials: A
authors.lastname: Quiñones-Hinojosa
mesh_terms: D000818:Animals; D001859:Bone Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D045744:Cell Line, Tumor / Q000166:cytology* / Q000378:metabolism; D002470:Cell Survival; D002817:Chordoma / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D004195:Disease Models, Animal; D005260:Female; D005326:Fetal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008875:Middle Aged; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism*; D014183:Transplantation, Heterologous; D000098908:Brachyury Protein
chemical_list: D005326:Fetal Proteins; D020825:T-Box Domain Proteins; D000098908:Brachyury Protein