Display options
Share it on

J Bone Oncol. 2021 Mar 24;28:100357. doi: 10.1016/j.jbo.2021.100357. eCollection 2021 Jun.

Using a combination of gangliosides and cell surface vimentin as surface biomarkers for isolating osteosarcoma cells in microfluidic devices.

Journal of bone oncology

Henrietta O Fasanya, Pablo J Dopico, Zachary Yeager, Z Hugh Fan, Dietmar W Siemann

Affiliations

  1. Department of Radiation Oncology, University of Florida, Gainesville, FL, USA.
  2. College of Medicine MD-PhD Program, University of Florida, Gainesville, FL, USA.
  3. Interdisciplinary Microsystems Group, Department of Mechanical and Aerospace Engineering, Gainesville, FL, USA.
  4. J. Crayton Pruitt Family Department of Biomedical Engineering, Gainesville, FL, USA.

PMID: 33912384 PMCID: PMC8065304 DOI: 10.1016/j.jbo.2021.100357

Abstract

BACKGROUND: Osteosarcoma (OS) is the most common primary bone tumor and the third leading cause of pediatric cancer deaths. Liquid biopsies are an alternative to current diagnostic imaging modalities that can be used to monitor treatment efficacy and the development of metastases. This study addresses the use of novel biomarkers to detect circulating osteosarcoma cells.

PROCEDURES: Flow cytometry was used to evaluate the relative expression of epithelial cell adhesion molecule (EpCAM), ganglioside 2 and 3 (GD2/3), and cell surface vimentin (CSV) on a panel of OS cell lines. A microfluidic device was used to affirm the efficacy of GD2/3 and CSV to capture CTCs. Once captured, CTCs on the device are enumerated and the capture efficiency for each marker is measured. Patient samples were captured using the LFAM chip.

RESULTS: We report the evaluation of GD2, GD3, and CSV as markers for OS cell capture in cell lines and in patient samples. The results of our capture studies correlate with our flow cytometry data and have shown a low capture efficiency of OS cells using EpCAM antibodies, while showing a moderate capture efficiency of OS cells using the GD2, GD3, and CSV antibodies independently. The combination of biomarkers demonstrate a high capture efficiency of approximately 80%. This is further supported by the detection of 1-1.5 CTCs per mL of blood using GD2 + CSV in OS patient samples.

CONCLUSIONS: The combination of GD2 + CSV significantly increased the capture efficacy of OS cells. The detection of CTCs through routine blood sampling may be used clinically for earlier detection of metastases and monitoring the therapeutic effect of treatments in metastatic osteosarcomas.

© 2021 Published by Elsevier GmbH.

Keywords: CK, Cytokeratin; CSV, Cell Surface Vimentin; CTC, Circulating Tumor Cell; Circulating tumor cells; DAPI, 4′,6-diamidino-2-phenylindole; EpCAM, Epithelial Cell Adhesion Molecule; GD2, Ganglioside 2; GD3, Ganglioside 3; Ganglioside GD2; Ganglioside GD3; IHC, Immunohistochemistry; OS, Osteosarcoma; Osteosarcoma; PET, Positron Emission Tomography; Vimentin; mL, Milliliter

Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References

  1. Cancer Res. 2014 Mar 15;74(6):1645-50 - PubMed
  2. Cancer. 2014 Feb 15;120(4):548-54 - PubMed
  3. Cancer Discov. 2016 May;6(5):479-91 - PubMed
  4. Biochim Biophys Acta. 1994 Sep 15;1214(2):115-23 - PubMed
  5. Mol Imaging Radionucl Ther. 2012 Apr;21(1):23-8 - PubMed
  6. PLoS One. 2014 Oct 31;9(10):e111597 - PubMed
  7. Cytometry A. 2015 Feb;87(2):137-44 - PubMed
  8. N Engl J Med. 2004 Aug 19;351(8):781-91 - PubMed
  9. J Cancer Res Clin Oncol. 1989;115(6):531-6 - PubMed
  10. Oncotarget. 2017 Jul 18;8(29):47454-47473 - PubMed
  11. Cancer Sci. 2012 Sep;103(9):1656-64 - PubMed
  12. Pediatr Blood Cancer. 2016 Oct;63(10):1780-5 - PubMed
  13. J Clin Oncol. 2004 Nov 15;22(22):4463-73 - PubMed
  14. Cancer Manag Res. 2018 Sep 10;10:3333-3339 - PubMed
  15. Cancer Res. 1996 Jun 15;56(12):2844-8 - PubMed
  16. Oncotarget. 2017 May 9;8(19):31592-31600 - PubMed
  17. Cancer. 2012 Feb 1;118(3):740-9 - PubMed
  18. J Hematol Oncol. 2020 Dec 10;13(1):172 - PubMed
  19. Clin Cancer Res. 2004 Oct 15;10(20):6897-904 - PubMed
  20. Prog Clin Biol Res. 1994;385:319-28 - PubMed
  21. Clin Sarcoma Res. 2015 Jan 24;5(1):4 - PubMed
  22. Cell Mol Life Sci. 2011 Sep;68(18):3033-46 - PubMed
  23. Adv Exp Med Biol. 2020;1257:55-66 - PubMed
  24. Orthopade. 2017 Jun;46(6):484-497 - PubMed
  25. Cancer Res. 1987 Oct 15;47(20):5377-81 - PubMed
  26. Eur J Surg Oncol. 2017 Jun;43(6):1050-1055 - PubMed
  27. Vet Surg. 2017 Jan;46(1):75-80 - PubMed
  28. J Clin Oncol. 2012 Sep 10;30(26):3264-70 - PubMed
  29. Clin Cancer Res. 2009 Sep 1;15(17):5323-37 - PubMed
  30. Mol Clin Oncol. 2015 Jan;3(1):31-36 - PubMed
  31. Eur J Haematol. 2003 Jan;70(1):43-52 - PubMed
  32. Curr Cancer Drug Targets. 2010 Mar;10(2):200-9 - PubMed
  33. Oncotarget. 2017 Jun 6;8(23):36936-36949 - PubMed
  34. Melanoma Res. 1992 Dec;2(5-6):355-62 - PubMed
  35. Cancer Res. 1984 Nov;44(11):5262-5 - PubMed
  36. Clin Cancer Res. 2010 May 15;16(10):2769-80 - PubMed
  37. Cancer Biol Ther. 2011 May 1;11(9):812-5 - PubMed
  38. Angew Chem Int Ed Engl. 2019 Jun 3;58(23):7606-7610 - PubMed
  39. Lab Chip. 2014 Jan 7;14(1):89-98 - PubMed
  40. Expert Opin Ther Targets. 2015 Mar;19(3):349-62 - PubMed
  41. Ergeb Physiol. 1966;57:190-222 - PubMed
  42. J Clin Oncol. 2008 Jul 1;26(19):3213-21 - PubMed
  43. Cancer Res. 2001 May 15;61(10):4244-52 - PubMed
  44. Med Oncol. 2015 Jan;32(1):430 - PubMed
  45. Int J Cancer. 1988 Mar 15;41(3):336-43 - PubMed
  46. Blood. 2007 May 15;109(10):4245-8 - PubMed
  47. BioDrugs. 2003;17(3):155-67 - PubMed
  48. Cancer Immunol Immunother. 2012 Dec;61(12):2261-71 - PubMed
  49. Indian J Med Paediatr Oncol. 2017 Jan-Mar;38(1):33-43 - PubMed
  50. Cancer Res. 1992 Aug 15;52(16):4342-7 - PubMed
  51. Chem Phys Lipids. 1986 Dec 15;42(1-3):209-33 - PubMed
  52. N Engl J Med. 2010 Sep 30;363(14):1324-34 - PubMed
  53. Jpn J Cancer Res. 2002 Jul;93(7):816-24 - PubMed
  54. Methods Mol Biol. 2017;1634:33-53 - PubMed
  55. J Biol Chem. 2005 Aug 19;280(33):29828-36 - PubMed
  56. Oncotarget. 2017 Aug 24;8(45):78965-78977 - PubMed
  57. Cancer Treat Res. 2009;152:3-13 - PubMed
  58. Hum Antibodies Hybridomas. 1992 Jan;3(1):19-24 - PubMed
  59. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1972-6 - PubMed
  60. J Clin Invest. 2009 Jul;119(7):1763-71 - PubMed
  61. J Oncol. 2010;2010:617421 - PubMed

Publication Types