Comparison of in vitro and in vivo approaches to studying brain colonization by breast cancer cells

Abstract

Brain metastases occur in 20 to 40% of patients with metastatic breast cancer. The process is complex and depends on successful cancer cell evasion from the primary tumor, distribution and survival within the blood stream and cerebral microvasculature, penetration of the blood brain barrier and proliferation within the brain microenvironment. The initial steps of brain colonization are difficult to study in vivo. Therefore, in vitro assays have been developed to mimic this process. Most commonly, in vitro studies of brain colonization focus on tumor cell adhesion to brain endothelial cells and transendothelial migration. We previously investigated breast cancer brain colonization from the blood stream in vivo and defined the time and process of brain entry for five different cancer cell lines in a mouse model. We now investigated if in vitro approaches can reliably emulate the initial steps that determine successful brain colonization in vivo. To this end, we optimized an in vitro model of the vascular blood brain barrier and compared the brain invasion properties of the in vivo characterized cell models with their ability to interact with and penetrate the blood brain barrier model in vitro. Our results show that the in vitro findings correlate only poorly with the vivo results. The limitations of the in vitro approaches are discussed in light of the in vivo processes. We conclude that investigation of mechanisms supporting the earliest steps of breast cancer brain metastasis from the blood stream will depend on in vivo analyses.

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References

  1. 1.

    Al-Shamy G, Sawaya R (2009) Management of brain metastases: the indispensable role of surgery. J Neurooncol 92(3):275–282. doi:10.1007/s11060-009-9839-y

    PubMed  Article  Google Scholar 

  2. 2.

    Santarelli JG, Sarkissian V, Hou LC, Veeravagu A, Tse V (2007) Molecular events of brain metastasis. Neurosurg Focus 22(3):E1

    PubMed  Article  Google Scholar 

  3. 3.

    Grewal J, Kesari S (2008) Breast cancer surface receptors predict risk for developing brain metastasis and subsequent prognosis. Breast Cancer Res 10(2):104

    PubMed  Article  Google Scholar 

  4. 4.

    Grewal J, Zhou H, Factor R, Kesari S (2010) Isolated loss of hormonal receptors in leptomeningeal metastasis from estrogen receptor- and progesterone receptor-positive lobular breast cancer. J Clin Oncol 28(13):200–202. doi:10.1200/JCO.2009.25.3518

    Article  Google Scholar 

  5. 5.

    DeAngelis LM (2008) Treatment of brain metastasis. J Support Oncol 6(2):87–88

    Google Scholar 

  6. 6.

    Gerrard GE, Franks KN (2004) Overview of the diagnosis and management of brain, spine, and meningeal metastases. J Neurol Neurosurg Psychiatry 75(Suppl2):ii37–ii42

    PubMed  Article  Google Scholar 

  7. 7.

    Norden AD, Wen PY, Kesari S (2005) Brain metastases. Curr Opin Neurol 18(6):654–661

    PubMed  Google Scholar 

  8. 8.

    Ballinger WE Jr, Schimpff RD (1979) An experimental model for cerebral metastasis: preliminary light and ultrastructural studies. J Neuropathol Exp Neurol 38(1):19–34

    PubMed  Article  Google Scholar 

  9. 9.

    Kawaguchi T, Tobai S, Nakamura K (1982) Extravascular migration of tumor cells in the brain: an electron microscopic study. Invasion Metastasis 2(1):40–50

    PubMed  CAS  Google Scholar 

  10. 10.

    Paku S, Dome B, Toth R, Timar J (2000) Organ-specificity of the extravasation process: an ultrastructural study. Clin Exp Metastasis 18(6):481–492

    PubMed  Article  CAS  Google Scholar 

  11. 11.

    Lorger M, Felding-Habermann B (2010) Capturing changes in the brain microenvironment during initial steps of breast cancer brain metastasis. Am J Pathol 176(6):2958–2971. doi:10.2353/ajpath.2010.090838

    PubMed  Article  Google Scholar 

  12. 12.

    Kieda C (2003) How endothelial cell organo-specificity mediates circulating cell homing. Arch Immunol Ther Exp (Warsz) 51(2):81–89

    CAS  Google Scholar 

  13. 13.

    Nicolson GL, Menter DG, Herrmann J, Cavanaugh P, Jia L, Hamada J, Yun Z, Nakajima M, Marchetti D (1994) Tumor metastasis to brain: role of endothelial cells, neurotrophins, and paracrine growth factors. Crit Rev Oncog 5(5):451–471

    PubMed  CAS  Google Scholar 

  14. 14.

    Pauli BU, Lee CL (1988) Organ preference of metastasis. The role of organ-specifically modulated endothelial cells. Lab Invest 58(4):379–387

    PubMed  CAS  Google Scholar 

  15. 15.

    Palmieri D, Chambers AF, Felding-Habermann B, Huang S, Steeg PS (2007) The biology of metastasis to a sanctuary site. Clin Cancer Res 13(6):1656–1662. doi:10.1158/1078-0432.CCR-06-2659

    PubMed  Article  CAS  Google Scholar 

  16. 16.

    Palmieri D, Smith QR, Lockman PR, Bronder J, Gril B, Chambers AF, Weil RJ, Steeg PS (2006) Brain metastases of breast cancer. Breast Dis 26:139–147

    PubMed  CAS  Google Scholar 

  17. 17.

    Bechmann I, Galea I, Perry VH (2007) What is the blood-brain barrier (not)? Trends Immunol 28(1):5–11. doi:10.1016/j.it.2006.11.007

    PubMed  Article  CAS  Google Scholar 

  18. 18.

    Lee SW, Kim WJ, Park JA, Choi YK, Kwon YW, Kim KW (2006) Blood-brain barrier interfaces and brain tumors. Arch Pharm Res 29(4):265–275

    PubMed  Article  CAS  Google Scholar 

  19. 19.

    Reichel A, Begley DJ, Abbott NJ (2003) An overview of in vitro techniques for blood-brain barrier studies. Methods Mol Med 89:307–324. doi:10.1385/1-59259-419-0:307

    PubMed  CAS  Google Scholar 

  20. 20.

    Haseloff RF, Blasig IE, Bauer HC, Bauer H (2005) In search of the astrocytic factor(s) modulating blood-brain barrier functions in brain capillary endothelial cells in vitro. Cell Mol Neurobiol 25(1):25–39

    PubMed  Article  CAS  Google Scholar 

  21. 21.

    Ribeiro MM, Castanho MA, Serrano I (2010) In vitro blood-brain barrier models–latest advances and therapeutic applications in a chronological perspective. Mini Rev Med Chem 10(3):262–270

    PubMed  Article  CAS  Google Scholar 

  22. 22.

    Deli MA, Abraham CS, Kataoka Y, Niwa M (2005) Permeability studies on in vitro blood-brain barrier models: physiology, pathology, and pharmacology. Cell Mol Neurobiol 25(1):59–127

    PubMed  Article  Google Scholar 

  23. 23.

    Yoneda T, Williams PJ, Hiraga T, Niewolna M, Nishimura R (2001) A bone-seeking clone exhibits different biological properties from the MDA-MB-231 parental human breast cancer cells and a brain-seeking clone in vivo and in vitro. J Bone Miner Res 16(8):1486–1495

    PubMed  Article  CAS  Google Scholar 

  24. 24.

    Fulcher CA, Zimmerman TS (1982) Characterization of the human factor VIII procoagulant protein with a heterologous precipitating antibody. Proc Natl Acad Sci USA 79(5):1648–1652

    PubMed  Article  CAS  Google Scholar 

  25. 25.

    Zimmerman TS, Ruggeri ZM, Fulcher CA (1983) Factor VIII/von Willebrand factor. Prog Hematol 13:279–309

    PubMed  CAS  Google Scholar 

  26. 26.

    Bellail AC, Hunter SB, Brat DJ, Tan C, Van Meir EG (2004) Microregional extracellular matrix heterogeneity in brain modulates glioma cell invasion. Int J Biochem Cell Biol 36(6):1046–1069. doi:10.1016/j.biocel.2004.01.013

    PubMed  Article  CAS  Google Scholar 

  27. 27.

    Giese A, Westphal M (1996) Glioma invasion in the central nervous system. Neurosurgery 39(2):235–250 discussion 250–232

    PubMed  Article  CAS  Google Scholar 

  28. 28.

    Gaillard PJ, Voorwinden LH, Nielsen JL, Ivanov A, Atsumi R, Engman H, Ringbom C, de Boer AG, Breimer DD (2001) Establishment and functional characterization of an in vitro model of the blood-brain barrier, comprising a co-culture of brain capillary endothelial cells and astrocytes. Eur J Pharm Sci 12(3):215–222

    PubMed  Article  CAS  Google Scholar 

  29. 29.

    Rubin LL, Hall DE, Porter S, Barbu K, Cannon C, Horner HC, Janatpour M, Liaw CW, Manning K, Morales J et al (1991) A cell culture model of the blood-brain barrier. J Cell Biol 115(6):1725–1735

    PubMed  Article  CAS  Google Scholar 

  30. 30.

    Kis B, Deli MA, Kobayashi H, Abraham CS, Yanagita T, Kaiya H, Isse T, Nishi R, Gotoh S, Kangawa K, Wada A, Greenwood J, Niwa M, Yamashita H, Ueta Y (2001) Adrenomedullin regulates blood-brain barrier functions in vitro. Neuroreport 12(18):4139–4142

    PubMed  Article  CAS  Google Scholar 

  31. 31.

    Schulze C, Smales C, Rubin LL, Staddon JM (1997) Lysophosphatidic acid increases tight junction permeability in cultured brain endothelial cells. J Neurochem 68(3):991–1000

    PubMed  Article  CAS  Google Scholar 

  32. 32.

    Carbonell WS, Ansorge O, Sibson N, Muschel R (2009) The vascular basement membrane as “soil” in brain metastasis. PLoS One 4(6):e5857. doi:10.1371/journal.pone.0005857

    PubMed  Article  Google Scholar 

  33. 33.

    Bergers G, Brekken R, McMahon G, TH Vu, Itoh T, Tamaki K, Tanzawa K, Thorpe P, Itohara S, Werb Z, Hanahan D (2000) Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol 2(10):737–744. doi:10.1038/35036374

    PubMed  Article  CAS  Google Scholar 

  34. 34.

    Bos PD, Zhang XH, Nadal C, Shu W, Gomis RR, Nguyen DX, Minn AJ, van de Vijver MJ, Gerald WL, Foekens JA, Massague J (2009) Genes that mediate breast cancer metastasis to the brain. Nature 459(7249):1005–1009. doi:10.1038/nature08021

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

Supported by grants from the National Institute of Health (CA112287 to BFH), Department of Defense (W81XWH-08-1-0468 to BFH), California Breast Cancer Research Program (15IB-0050, 16IB-0052 to BFH and 16IB-0030 to ML), and a Susan G. Komen Breast Cancer Foundation fellowship (PDF0707935 to ML).

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Correspondence to B. Felding-Habermann.

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Lorger, M., Lee, H., Forsyth, J.S. et al. Comparison of in vitro and in vivo approaches to studying brain colonization by breast cancer cells. J Neurooncol 104, 689–696 (2011). https://doi.org/10.1007/s11060-011-0550-4

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Keywords

  • Brain metastasis
  • Breast cancer
  • In vitro
  • In vivo
  • Blood brain barrier
  • Transendothelial migration
  • Extravasation
  • Brain metastasis model