Establishment and characterization of a primary human chordoma xenograft model

Laboratory investigation

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Object

Chordomas are rare tumors arising from remnants of the notochord. Because of the challenges in achieving a complete resection, the radioresistant nature of these tumors, and the lack of effective chemotherapeutics, the median survival for patients with chordomas is approximately 6 years. Reproducible preclinical model systems that closely mimic the original patient's tumor are essential for the development and evaluation of effective therapeutics. Currently, there are only a few established chordoma cell lines and no primary xenograft model. In this study, the authors aimed to develop a primary chordoma xenograft model.

Methods

The authors implanted independent tumor samples from 2 patients into athymic nude mice. The resulting xenograft line was characterized by histopathological analysis and immunohistochemical staining. The patient's tumor and serial passages of the xenograft were genomically analyzed using a 660,000 single-nucleotide polymorphism array.

Results

A serially transplantable xenograft was established from one of the 2 patient samples. Histopathological analysis and immunohistochemical staining for S100 protein, epithelial membrane antigen, and cytokeratin AE1/AE3 of the primary patient sample and the xenografts confirmed that the xenografts were identical to the original chordoma obtained from the patient. Immunohistochemical staining and western blot analysis confirmed the presence of brachyury, a recently described marker of chordomas, in the tumor from the patient and each of the xenografts. Genome-wide variation was assessed between the patient's tumor and the xenografts and was found to be more than 99.9% concordant.

Conclusions

To the best of their knowledge, the authors have established the first primary chordoma xenograft that will provide a useful preclinical model for this disease and a platform for therapeutic development.

Abbreviations used in this paperEMA = epithelial membrane antigen; SNP = single-nucleotide polymorphism.

Article Information

Address correspondence to: Gary L. Gallia, M.D., Ph.D., Department of Neurosurgery, Johns Hopkins University School of Medicine, 600 North Wolfe Street, Phipps Building, Room 118, Baltimore, Maryland 21287. email: ggallia1@jhmi.edu.

Please include this information when citing this paper: published online January 27, 2012; DOI: 10.3171/2011.12.JNS111123.

© AANS, except where prohibited by US copyright law.

Headings

Figures

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    Preoperative MR images. Preoperative sagittal T1-weighted image obtained after administration of Gd (A) and axial T2-weighted image (B) demonstrating a large clival chordoma with significant intracranial extension and mass effect on the optic apparatus and the brainstem.

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    Photographs of the primary chordoma xenograft. A: Photograph of a nude mouse with bilateral chordoma xenograft tumors. B: Gross appearance of the bilateral xenograft tumors harvested from the animal shown in panel A demonstrating a lobulated appearance.

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    Histochemical and immunohistochemical evaluation of chordoma xenografts. Paraffin-embedded and formalin-fixed sections from the patient's primary tumor and chordoma xenografts were stained with H & E (A–D), and immunoperoxidase stains specific for cytokeratin (AE1/AE3) (E–H) and S100 protein (I–L). Original magnification × 100.

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    Brachyury expression. Expression of brachyury in the patient tumor (P0), xenograft passage 1, mouse A (P1-A), xenograft passage 1, mouse B (P1-B), and xenograft passage 2 (P2) was determined by immunohistochemical (A) and western blot (B) analyses. A negative control of normal brain (B) was also run in the western blot analysis. Original magnification × 160.

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    Short-term cell line cultures. A: Cells with the typical physaliphorous features characteristic of chordoma. B: Nonvacuolated cells present in the chordoma cultures. C: Growth curves of JHH-2009–011 cultures. D: Western blot analysis for brachyury expression: patient tumor (P0), U87MG glioma cell line, and JHH-2009–011 culture.

References

1

Almefty KPravdenkova SColli BOAl-Mefty OGokden M: Chordoma and chondrosarcoma: similar, but quite different, skull base tumors. Cancer 110:245724672007

2

Butler MGDahir GAHedges LKJuliao SFSciadini MFSchwartz HS: Cytogenetic, telomere, and telomerase studies in five surgically managed lumbosacral chordomas. Cancer Genet Cytogenet 85:51571995

3

Coindre JMRivel JTrojani MDe Mascarel IDe Mascarel A: Immunohistological study in chordomas. J Pathol 150:61631986

4

Cree IAGlaysher SHarvey AL: Efficacy of anti-cancer agents in cell lines versus human primary tumour tissue. Curr Opin Pharmacol 10:3753792010

5

Daniel VCMarchionni LHierman JSRhodes JTDevereux WLRudin CM: A primary xenograft model of small-cell lung cancer reveals irreversible changes in gene expression imposed by culture in vitro. Cancer Res 69:336433732009

6

DeComas AMPenfornis PHarris MRMeyer MSPochampally RR: Derivation and characterization of an extra-axial chordoma cell line (EACH-1) from a scapular tumor. J Bone Joint Surg Am 92:123112402010

7

Hallor KHStaaf JJönsson GHeidenblad MVult von Steyern FBauer HC: Frequent deletion of the CDKN2A locus in chordoma: analysis of chromosomal imbalances using array comparative genomic hybridisation. Br J Cancer 98:4344422008

8

Henderson SRGuiliano DPresneau NMcLean SFrow RVujovic S: A molecular map of mesenchymal tumors. Genome Biol 6:R762005

9

Johannsson OTStaff SVallon-Christersson JKytöla SGudjonsson TRennstam K: Characterization of a novel breast carcinoma xenograft and cell line derived from a BRCA1 germline mutation carrier. Lab Invest 83:3873962003

10

Johnson DR: Hairpin-tail: a case of post-reductional gene action in the mouse egg. Genetics 76:7958051974

11

Kispert AHermann BG: The Brachyury gene encodes a novel DNA binding protein. EMBO J 12:489848991993

12

McMaster MLGoldstein AMBromley CMIshibe NParry DM: Chordoma: incidence and survival patterns in the United States, 1973–1995. Cancer Causes Control 12:1112001

13

Ostroumov EHunter CJ: Identifying mechanisms for therapeutic intervention in chordoma: c-Met oncoprotein. Spine (Phila Pa 1976) 33:277427802008

14

Ostroumov EHunter CJ: The role of extracellular factors in human metastatic chordoma cell growth in vitro. Spine (Phila Pa 1976) 32:295729642007

15

Pandita AAldape KDZadeh GGuha AJames CD: Contrasting in vivo and in vitro fates of glioblastoma cell subpopulations with amplified EGFR. Genes Chromosomes Cancer 39:29362004

16

Presneau NShalaby AYe HPillay NHalai DIdowu B: Role of the transcription factor T (brachyury) in the pathogenesis of sporadic chordoma: a genetic and functional-based study. J Pathol 223:3273352011

17

Ricci-Vitiani LPierconti FFalchetti MLPetrucci GMaira GDe Maria R: Establishing tumor cell lines from aggressive telomerase-positive chordomas of the skull base. Technical note. J Neurosurg 105:4824842006

18

Scheil SBrüderlein SLiehr TStarke HHerms JSchulte M: Genome-wide analysis of sixteen chordomas by comparative genomic hybridization and cytogenetics of the first human chordoma cell line, U-CH1. Genes Chromosomes Cancer 32:2032112001

19

Schwab JHBoland PJAgaram NPSocci NDGuo TO'Toole GC: Chordoma and chondrosarcoma gene profile: implications for immunotherapy. Cancer Immunol Immunother 58:3393492009

20

Vujovic SHenderson SPresneau NOdell EJacques TSTirabosco R: Brachyury, a crucial regulator of notochordal development, is a novel biomarker for chordomas. J Pathol 209:1571652006

21

Yang CHornicek FJWood KBSchwab JHChoy EIafrate J: Characterization and analysis of human chordoma cell lines. Spine (Phila Pa 1976) 35:125712642010

22

Zhou HChen CBLan JLiu CLiu XGJiang L: Differential proteomic profiling of chordomas and analysis of prognostic factors. J Surg Oncol 102:7207272010

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