Fluid biopsy in patients with metastatic prostate, pancreatic and breast cancers - PubMed (original) (raw)

doi: 10.1088/1478-3975/9/1/016003. Epub 2012 Feb 3.

Kelly Bethel, Anand Kolatkar, Madelyn S Luttgen, Michael Malchiodi, Franziska Baehring, Katharina Voigt, Daniel Lazar, Jorge Nieva, Lyudmila Bazhenova, Andrew H Ko, W Michael Korn, Ethan Schram, Michael Coward, Xing Yang, Thomas Metzner, Rachelle Lamy, Meghana Honnatti, Craig Yoshioka, Joshua Kunken, Yelena Petrova, Devin Sok, David Nelson, Peter Kuhn

Affiliations

Fluid biopsy in patients with metastatic prostate, pancreatic and breast cancers

Dena Marrinucci et al. Phys Biol. 2012 Feb.

Abstract

Hematologic spread of carcinoma results in incurable metastasis; yet, the basic characteristics and travel mechanisms of cancer cells in the bloodstream are unknown. We have established a fluid phase biopsy approach that identifies circulating tumor cells (CTCs) without using surface protein-based enrichment and presents them in sufficiently high definition (HD) to satisfy diagnostic pathology image quality requirements. This 'HD-CTC' assay finds >5 HD-CTCs mL(-1) of blood in 80% of patients with metastatic prostate cancer (n = 20), in 70% of patients with metastatic breast cancer (n = 30), in 50% of patients with metastatic pancreatic cancer (n = 18), and in 0% of normal controls (n = 15). Additionally, it finds HD-CTC clusters ranging from 2 HD-CTCs to greater than 30 HD-CTCs in the majority of these cancer patients. This initial validation of an enrichment-free assay demonstrates our ability to identify significant numbers of HD-CTCs in a majority of patients with prostate, breast and pancreatic cancers.

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Figures

Figure 1

Figure 1

Gallery of representative HD-CTCs found in cancer patients. Each HD-CTC is cytokeratin positive (red), CD45 negative (green), contains a DAPI nucleus (blue), and is morphologically distinct from surrounding white blood cells.

Figure 2

Figure 2

Mean observed SKBR3s plotted against expected SKBR3s. Four aliquots of normal control blood was spiked with varying numbers of SKBR2 cells to produce 4 slides with approximately 10, 30, 100, and 300 cancer cells per slide. The mean of each quadruplicate is displayed as well as error bars noting standard deviation.

Figure 3

Figure 3

Comparison of CTC counts between two separate processors on 9 different cancer patient samples. CTC/mL counts ranged from 0 to 203.

Figure 4

Figure 4

Gallery of representative clusters that are found in most patients with cancer. Clusters range from 2 to over 30 HD-CTCs. Each HD-CTC is cytokeratin positive (red), CD45 negative (green), contains a DAPI nucleus (blue), and is morphologically distinct from surrounding nucleated cells.

Figure 5

Figure 5

Gallery of candidate HD-CTCs that were excluded because they lacked various morphologic or morphometric inclusion criteria. A) Cytokeratin intensity too dim. B) Nuclear size too small. C) Cytokeratin insufficiently circumferential (surrounds less than 2/3 of nucleus) D) Cytokeratin too dim, although appears to be a cluster of two very large cells. E) Nucleus shows apoptotic disintegration changes. F) Nucleus too small and cytoplasm insufficiently circumferential; appears to be a cell in late apoptosis. G) Nucleus too small (same size as surrounding WBC nuclei). H. Cytokeratin present, but not circumferential. I) Cytoplasm insufficiently circumferential, nucleus too small.

Figure 6

Figure 6

Representative images of types of suspected CTCs found in a single prostate cancer patient. Top Row: Suspected CTC that is negative for Cytokeratin and CD45, but has a nucleus that is large and looks like other HD-CTCs found in this patient. Middle Row: Typical HD-CTC that is cytokeratin positive, CD45 negative, with a DAPI nucleus. Bottom Row: HD-CTC cluster of 4 cells.

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