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A Noninvasive and Real-Time Method for Circulating Tumor Cell Detection by In Vivo Flow Cytometry

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Circulating Tumor Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1634))

Abstract

The quantification of circulating tumor cells (CTCs) has been considered a potentially powerful tool in cancer diagnosis and prognosis, as CTCs have been shown to appear very early in cancer development. Great efforts have been made to develop methods that were less invasive and more sensitive to detect CTCs earlier. There is growing evidence that CTC clusters have greater metastatic potential than single CTCs. Therefore, the detection of CTC clusters is also important. This chapter is aimed to introduce a noninvasive technique for CTCs detection named in vivo flow cytometry (IVFC), which has been demonstrated to be capable of monitoring CTCs dynamics continuously. Furthermore, IVFC could be helpful for CTC cluster enumeration.

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References

  1. Crnic I, Christofori G (2004) Novel technologies and recent advances in metastasis research. Int J Dev Biol 48(5–6):573–581

    Article  CAS  PubMed  Google Scholar 

  2. Massagué J, Obenauf AC (2016) Metastatic colonization by circulating tumour cells. Nature 529(7586):298–306

    Article  PubMed  PubMed Central  Google Scholar 

  3. Luzzi KJ, MacDonald IC, Schmidt EE, Kerkvliet N, Morris VL, Chambers AF et al (1998) Multistep nature of metastatic inefficiency: dormancy of solitary cells after successful extravasation and limited survival of early Micrometastases. Am J Pathol 153(3):865–873

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Ashworth T (1869) A case of cancer in which cells similar to those in the tumours were seen in the blood after death. Aust Med J 14(3):146–149

    Google Scholar 

  5. Aceto N, Bardia A, Miyamoto David T, Donaldson Maria C, Wittner Ben S, Spencer Joel A et al (2014) Circulating tumor cell clusters are oligoclonal precursors of breast cancer metastasis. Cell 158(5):1110–1122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Yu M, Bardia A, Wittner BS, Stott SL, Smas ME, Ting DT et al (2013) Circulating breast tumor cells exhibit dynamic changes in epithelial and mesenchymal composition. Science 339(6119):580–584

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Krebs MG, Hou J-M, Sloane R, Lancashire L, Priest L, Nonaka D et al (2012) Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and -independent approaches. J Thorac Oncol 7(2):306–315

    Article  PubMed  Google Scholar 

  8. Ting David T, Wittner Ben S, Ligorio M, Vincent Jordan N, Shah Ajay M, Miyamoto David T et al (2014) Single-cell RNA sequencing identifies extracellular matrix gene expression by pancreatic circulating tumor cells. Cell Rep 8(6):1905–1918

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Clare SE, Sener SF, Wilkens W, Goldschmidt R, Merkel D, Winchester DJ (1997) Prognostic significance of occult lymph node metastases in node-negative breast cancer. Ann Surg Oncol 4(6):447–451

    Article  CAS  PubMed  Google Scholar 

  10. Braun S, Pantel K, Müller P, Janni W, Hepp F, Kentenich CRM et al (2000) Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. N Engl J Med 342(8):525–533

    Article  CAS  PubMed  Google Scholar 

  11. Dirix L, Van Dam P, Vermeulen P (2005) Genomics and circulating tumor cells: promising tools for choosing and monitoring adjuvant therapy in patients with early breast cancer? Curr Opin Oncol 17(6):551–558

    Article  CAS  PubMed  Google Scholar 

  12. Hayes DF, Smerage J (2008) Is there a role for circulating tumor cells in the management of breast cancer? Clin Cancer Res 14(12):3646–3650

    Article  CAS  PubMed  Google Scholar 

  13. Gupta GP, Massagué J (2006) Cancer metastasis: building a framework. Cell 127(4):679–695

    Article  CAS  PubMed  Google Scholar 

  14. Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, Miller MC et al (2004) Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med 351(8):781–791

    Article  CAS  PubMed  Google Scholar 

  15. Vona G, Sabile A, Louha M, Sitruk V, Romana S, Schütze K et al (2000) Isolation by size of epithelial tumor cells: a new method for the Immunomorphological and molecular characterization of circulating tumor cells. Am J Pathol 156(1):57–63

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Vona G, Estepa L, Béroud C, Damotte D, Capron F, Nalpas B et al (2004) Impact of cytomorphological detection of circulating tumor cells in patients with liver cancer. Hepatology 39(3):792–797

    Article  PubMed  Google Scholar 

  17. Riethdorf S, Fritsche H, Muller V, Rau T, Schindlbeck C, Rack B et al (2007) Detection of circulating tumor cells in peripheral blood of patients with metastatic breast cancer: a validation study of the CellSearch system. Clin Cancer Res 13(3):920–928

    Article  CAS  PubMed  Google Scholar 

  18. Allard WJ, Matera J, Miller MC, Repollet M, Connelly MC, Rao C et al (2004) Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res 10(20):6897–6904

    Article  PubMed  Google Scholar 

  19. Nagrath S, Sequist LV, Maheswaran S, Bell DW, Irimia D, Ulkus L et al (2007) Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature 450(7173):1235–1239

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Sieuwerts AM, Kraan J, Bolt J, van der Spoel P, Elstrodt F, Schutte M et al (2009) Anti-epithelial cell adhesion molecule antibodies and the detection of circulating normal-like breast tumor cells. J Natl Cancer Inst 101(1):61–66

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Ozkumur E, Shah AM, Ciciliano JC, Emmink BL, Miyamoto DT, Brachtel E et al (2013) Inertial focusing for tumor antigen–dependent and –independent sorting of rare circulating tumor cells. Sci Transl Med 5(179):179ra147

    Article  Google Scholar 

  22. Krivacic RT, Ladanyi A, Curry DN, Hsieh HB, Kuhn P, Bergsrud DE et al (2004) A rare-cell detector for cancer. Proc Natl Acad Sci U S A 101(29):10501–10504

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Pachmann K, Clement JH, Schneider C-P, Willen B, Camara O, Pachmann U et al (2005) Standardized quantification of circulating peripheral tumor cells from lung and breast cancer. Clin Chem Lab Med 43(6):617–627

    Article  CAS  PubMed  Google Scholar 

  24. Cruz I, Ciudad J, Cruz JJ, Ramos M, Gómez-Alonso A, Adansa JC et al (2005) Evaluation of Multiparameter flow Cytometry for the detection of breast cancer tumor cells in blood samples. Am J Clin Pathol 123(1):66–74

    Article  PubMed  Google Scholar 

  25. Tinhofer I, Hristozova T, Stromberger C, KeilhoIz U, Budach V (2012) Monitoring of circulating tumor cells and their expression of EGFR/Phospho-EGFR during combined radiotherapy regimens in locally advanced squamous cell carcinoma of the head and neck. Int J Radiat Oncol Biol Phys 83(5):e685–e690

    Article  CAS  PubMed  Google Scholar 

  26. Zhang L, Ridgway LD, Wetzel MD, Ngo J, Yin W, Kumar D et al (2013) The identification and characterization of breast cancer CTCs competent for brain metastasis. Sci Transl Med 5(180):180ra148. * Copyright © 2013 American Association for the Advancement of Science

    Article  Google Scholar 

  27. Givan A (2011) Flow Cytometry: an introduction. In: Hawley TS, Hawley RG (eds) Flow cytometry protocols methods in molecular biology. Humana Press, New York, pp 1–29

    Google Scholar 

  28. Robinson JP (2001) Comparative overview of flow and image Cytometry. In: Current protocols in cytometry. John Wiley & Sons Inc., New York

    Chapter  Google Scholar 

  29. Pantel K, Brakenhoff RH, Brandt B (2008) Detection, clinical relevance and specific biological properties of disseminating tumour cells. Nat Rev Cancer 8(5):329–340

    Article  CAS  PubMed  Google Scholar 

  30. Lianidou ES, Markou A (2011) Circulating tumor cells in breast cancer: detection systems, molecular characterization, and future challenges. Clin Chem 57(9):1242–1255

    Article  CAS  PubMed  Google Scholar 

  31. Smith BM, Slade MJ, English J, Graham H, Lüchtenborg M, Sinnett HD et al (2000) Response of circulating tumor cells to systemic therapy in patients with metastatic breast cancer: comparison of quantitative polymerase chain reaction and immunocytochemical techniques. J Clin Oncol Off J Am Soc Clin Oncol 18(7):1432–1439

    Article  CAS  Google Scholar 

  32. Yang M-J, Chiu H-H, Wang H-M, Yen L-C, Tsao D-A, Hsiao C-P et al (2010) Enhancing detection of circulating tumor cells with activating KRAS oncogene in patients with colorectal cancer by weighted chemiluminescent membrane array method. Ann Surg Oncol 17(2):624–633

    Article  PubMed  Google Scholar 

  33. O’Flaherty JD, Gray S, Richard D, Fennell D, O’Leary JJ, Blackhall FH et al (2012) Circulating tumour cells, their role in metastasis and their clinical utility in lung cancer. Lung Cancer 76(1):19–25

    Article  PubMed  Google Scholar 

  34. Miyamoto DT, Zheng Y, Wittner BS, Lee RJ, Zhu H, Broderick KT et al (2015) RNA-Seq of single prostate CTCs implicates noncanonical Wnt signaling in antiandrogen resistance. Science 349(6254):1351–1356

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Novak J, Georgakoudi I, Wei X, Prossin A, Lin CP (2004) In vivo flow cytometer for real-time detection and quantification of circulating cells. Opt Lett 29(1):77

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Georgakoudi I, Solban N, Novak J, Rice WL, Wei X, Hasan T et al (2004) In vivo flow cytometry. Cancer Res 64(15):5044–5047. %* ©2004 American Association for Cancer Research

    Article  CAS  PubMed  Google Scholar 

  37. Galanzha EI, Shashkov EV, Spring PM, Suen JY, Zharov VP (2009) In vivo, noninvasive, label-free detection and eradication of circulating metastatic melanoma cells using two-color photoacoustic flow Cytometry with a diode laser. Cancer Res 69(20):7926–7934

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Galanzha EI, Kim J-W, Zharov VP (2009) Nanotechnology-based molecular photoacoustic and photothermal flow cytometry platform for in-vivodetection and killing of circulating cancer stem cells. J Biophotonics 2(12):725–735

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Xu M, Wang LV (2006) Photoacoustic imaging in biomedicine. Rev Sci Instrum 77(4):041101

    Article  Google Scholar 

  40. Zeidman I, Buss JM (1952) Transpulmonary passage of tumor cell emboli. Cancer Res 12(10):731–733

    CAS  PubMed  Google Scholar 

  41. Cho EH, Wendel M, Luttgen M, Yoshioka C, Marrinucci D, Lazar D et al (2012) Characterization of circulating tumor cell aggregates identified in patients with epithelial tumors. Phys Biol 9(1):016001

    Article  PubMed  PubMed Central  Google Scholar 

  42. Fidler IJ (1973) The relationship of embolic homogeneity, number, size and viability to the incidence of experimental metastasis. Eur J Cancer 9(3):223–227

    Article  CAS  PubMed  Google Scholar 

  43. Molnar B, Ladanyi A, Tanko L, Sréter L, Tulassay Z (2001) Circulating tumor cell clusters in the peripheral blood of colorectal cancer patients. Clin Cancer Res 7(12):4080–4085

    CAS  PubMed  Google Scholar 

  44. Stott SL, Hsu C-H, Tsukrov DI, Yu M, Miyamoto DT, Waltman BA et al (2010) Isolation of circulating tumor cells using a microvortex-generating herringbone-chip. Proc Natl Acad Sci 107(43):18392–18397

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Hou JM, Krebs MG, Lancashire L, Sloane R, Backen A, Swain RK et al (2012) Clinical significance and molecular characteristics of circulating tumor cells and circulating tumor microemboli in patients with small-cell lung cancer. J Clin Oncol 30(5):525–532

    Article  PubMed  Google Scholar 

  46. Sarioglu AF, Aceto N, Kojic N, Donaldson MC, Zeinali M, Hamza B et al (2015) A microfluidic device for label-free, physical capture of circulating tumor cell clusters. Nat Methods 12(7):685–691

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Li Y, Tang Z, Ye S, Liu B, Liu Y, Chen J et al Establishment of a hepatocellular carcinoma cell line with unique metastatic characteristics through in vivo selection and screening for metastasis-related genes through cDNA microarray. J Cancer Res Clin Oncol 129(1):43–51

    Google Scholar 

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Acknowledgments

This work was supported by the grants of the National Major Scientific Research Program of China (Grant Nos. 2011CB910404 and 2012CB966801), the National Nature Science Foundation of China (Grant Nos. 61227017 and 81272818), and the National Science Fund for Distinguished Young Scholars (Grant No. 61425006).

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Authors and Affiliations

  1. Med-X Research Institute and School of Biomedical Engineering, Shanghai Jiao Tong University, 1954 Huashan Rd., Xuhui District, Shanghai, 200030, China

    Xunbin Wei, Xi Zhu, Ping Yang & Qiyan Wang

  2. Liver Cancer Institute, Zhongshan Hospital, Fudan University, 180 Fenglin Rd., Xuhui District, Shanghai, 200032, China

    Jian Zhou & Xinrong Yang

Authors
  1. Xunbin Wei
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  2. Jian Zhou
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  3. Xi Zhu
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  4. Xinrong Yang
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  5. Ping Yang
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  6. Qiyan Wang
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Corresponding authors

Correspondence to Xunbin Wei or Jian Zhou .

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Editors and Affiliations

  1. Division of Hematology Oncology, Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, California, USA

    Mark Jesus M. Magbanua

  2. Division of Hematology Oncology, Helen Diller Family Comprehensive Cancer Center, University of California-San Francisco, San Francisco, California, USA

    John W. Park

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Wei, X., Zhou, J., Zhu, X., Yang, X., Yang, P., Wang, Q. (2017). A Noninvasive and Real-Time Method for Circulating Tumor Cell Detection by In Vivo Flow Cytometry. In: M. Magbanua, M., W. Park, J. (eds) Circulating Tumor Cells. Methods in Molecular Biology, vol 1634. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7144-2_21

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  • DOI: https://doi.org/10.1007/978-1-4939-7144-2_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7143-5

  • Online ISBN: 978-1-4939-7144-2

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