Abstract
A variety of biomarkers are present on cells in peripheral blood of patients with a variety of disorders, including solid tumor malignancies. While rare, characterization of these cells for specific protein levels with the advanced technology proposed, will lead to future validation studies of blood samples as “liquid biopsies” for the evaluation of disease status and therapeutic response. While circulating tumor cells (CTCs) have been isolated in the blood samples of patients with solid tumors, the exact role of CTCs as clinically useful predictive markers is still debated. Current commercial technology has significant bias in that a positive selection technology is used that preassumes specific cell surface markers (such as EpCAM) are present on CTCs. However, CTCs with low EpCAM expression have been experimentally demonstrated to be more likely to be missed by this method. In contrast, this application uses a previously developed, technology that performs a purely negative enrichment methodology on peripheral blood, yielding highly enriched blood samples that contain CTCs as well as other, undefined cell types. The focus of this contribution is the use of multispectral imaging of epifluorescent, microscopic images of these enriched cells in order to help develop clinically relevant liquid biopsies from peripheral blood samples.
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Chambers AF, Groom AC, MacDonald IC (2002) Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer 2:563–572
Fehm T, Mueller V, Marches R, Klein G, Gueckel B, Neubauer H et al (2008) Tumor cell dormancy: implications for the biology and treatment of breast cancer. APMIS 116(7–8):742–753
Pantel K, Alix-Panabieres C, Riethdorf S (2009) Cancer micrometastases. Nat Rev Clin Oncol 6(6):339–351
Pantel K, Brakenhoff RH (2004) Dissecting the metastatic cascade. Nat Rev Cancer 4:448–456
Somlo G, Lau SK, Frankel P, Hsieh HB, Liu X, Yang L et al (2011) Multiple biomarker expression on circulating tumor cells in comparison to tumor tissues from primary and metastatic sites in patients with locally advanced/inflammatory, and stage IV breast cancer, using a novel detection technology. Breast Cancer Res Treat 128(1):155–163
Pestrin M, Bessi S, Galardi F, Truglia M, Biggeri A, Biagioni C et al (2009) Correlation of HER2 status between primary tumors and corresponding circulating tumor cells in advanced breast cancer patients. Breast Cancer Res Treat 118(3):523–530
Hayashi N, Nakamura S, Tokuda Y, Shimoda Y, Yagata H, Yoshida A et al (2012) Prognostic value of HER2-positive circulating tumor cells in patients with metastatic breast cancer. Int J Clin Oncol 17(2):96–104
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
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
Konigsberg R, Obermayr E, Bises G, Pfeiler G, Gneist M, Wrba F et al (2011) Detection of EpCAM positive and negative circulating tumor cells in metastatic breast cancer patients. Acta Oncol 50(5):700–710
Balasubramanian P, Lang JC, Jatana KR, Miller B, Ozer E et al (2012) Multiparameter analysis, including EMT markers, on negatively enriched blood samples from patients with squamous cell carcinoma of the head and neck. PLoS One 7:e42048
Wu Y, Deighan C, Miller B, Balasubramanian P, Lustberg M, Zborowski M et al (2013) Isolation and analysis of rare cells in the blood of cancer patients using a negative depletion methodology. Methods 64(1):169–182
Neher R, Neher E (2004) Optimizing imaging parameters for the separation of multiple labels in a fluorescence image. J Microsc 213(1):46–62
Malik Z, Cabib D, Buckwald RA, Talmi A, Garini Y, Lipson SG (1996) Fourier transform multipixel spectroscopy for quantitative cytology. J Microsc 182(2):133–140
Mansfield JR, Gossage KW, Hoyt CC, Levenson RM (2005) Autofluorescence removal, multiplexing, and automated analysis methods for in-vivo fluorescence imaging. J Biomed Opt 10(4):041207
Schultz RA, Nielsen T, Zavaleta JR, Ruch R, Wyatt R, Garner HR (2001) Hyperspectral imaging: a novel approach for microscopic analysis. Cytometry 43:239–247
Tsurui H, Nishimura H, Hattori S, Hirose S, Okumura K, Shirai T (2000) Seven-color fluorescence imaging of tissue samples based on Fourier spectroscopy and singular value decomposition. J Histochem Cytochem 48(5):653–662
Zimmerman T, Rietdorf J, Pepperkok R (2003) Spectral imaging and it applications in live cell microscopy. FEBS Lett 546:87–92
Zimmerman T (2005) Spectral imaging and linear unmixing in light microscopy. Adv Biochem Eng Biotechnol 95:245–265
Beeckman J, Neyts K, Vanbrabant PJM (2011) Liquid-crystal photonic applications. Opt Eng 50(8):081202
Spring KR, Long JC, Davidson MW (2012) Liquid crystal tunable filters. Microscopy Resource Center, Olympus America, Melville, NY
Miller BL, Chalmers JJ (2012) Cell enrichment from human blood through red cell lysis. In: Michael Conn P (ed) Methods navigator, cell biology protocols module. Elsevier, Amsterdam
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Miller, B., Lustberg, M., Summers, T.A., Chalmers, J.J. (2017). Multispectral Imaging Analysis of Circulating Tumor Cells in Negatively Enriched Peripheral Blood Samples. 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_19
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DOI: https://doi.org/10.1007/978-1-4939-7144-2_19
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Publisher Name: Humana Press, New York, NY
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