. 2016 Jun 14;114(12):1367-75.

doi: 10.1038/bjc.2016.121.

Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer

J S Ankeny^{1

2}, C M Court^{1

2}, S Hou^{1

3}, Q Li³, M Song³, D Wu³, J F Chen³, T Lee⁴, M Lin³, S Sho^{1

2}, M M Rochefort¹, M D Girgis¹, J Yao³, Z A Wainberg^{5

6}, V R Muthusamy^{5

7}, R R Watson^{5

7}, T R Donahue^{1

5}, O J Hines^{1

5}, H A Reber^{1

5}, T G Graeber³, H R Tseng³, J S Tomlinson^{1

2

5}

Affiliations

¹ Department of Surgery, University of California Los Angeles, 575 Westwood Plaza, Los Angeles, CA 90095, USA.
² Veteran's Health Administration, Greater Los Angeles, Department of Surgery, 11301 Wilshire Boulevard, Los Angeles, CA 90073, USA.
³ Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, California NanoSystems Institute, University of California, Los Angeles, 570 Westwood Plaza, Los Angeles, CA 90095-1770, USA.
⁴ California NanoSystems Institute, University of California, 570 Westwood Plaza, Los Angeles, CA 90095, USA.
⁵ UCLA Center for Pancreatic Diseases, 575 Westwood Plaza, Los Angeles, CA 90095, USA.
⁶ Department of Hematology/Oncology, University of California Los Angeles, 575 Westwood Plaza, Los Angeles, CA 90095, USA.
⁷ Department of Gastroenterology, University of California Los Angeles, 575 Westwood Plaza, Los Angeles, CA 90095, USA.

PMID: 27300108
PMCID: PMC4984454
DOI: 10.1038/bjc.2016.121

Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer

J S Ankeny et al. Br J Cancer. 2016.

. 2016 Jun 14;114(12):1367-75.

doi: 10.1038/bjc.2016.121.

Authors

Affiliations

¹ Department of Surgery, University of California Los Angeles, 575 Westwood Plaza, Los Angeles, CA 90095, USA.
² Veteran's Health Administration, Greater Los Angeles, Department of Surgery, 11301 Wilshire Boulevard, Los Angeles, CA 90073, USA.
³ Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, California NanoSystems Institute, University of California, Los Angeles, 570 Westwood Plaza, Los Angeles, CA 90095-1770, USA.
⁴ California NanoSystems Institute, University of California, 570 Westwood Plaza, Los Angeles, CA 90095, USA.
⁵ UCLA Center for Pancreatic Diseases, 575 Westwood Plaza, Los Angeles, CA 90095, USA.
⁶ Department of Hematology/Oncology, University of California Los Angeles, 575 Westwood Plaza, Los Angeles, CA 90095, USA.
⁷ Department of Gastroenterology, University of California Los Angeles, 575 Westwood Plaza, Los Angeles, CA 90095, USA.

PMID: 27300108
PMCID: PMC4984454
DOI: 10.1038/bjc.2016.121

Abstract

Background: Current diagnosis and staging of pancreatic ductal adenocarcinoma (PDAC) has important limitations and better biomarkers are needed to guide initial therapy. We investigated the performance of circulating tumour cells (CTCs) as an adjunctive biomarker at the time of disease presentation.

Methods: Venous blood (VB) was collected prospectively from 100 consecutive, pre-treatment patients with PDAC. Utilising the microfluidic NanoVelcro CTC chip, samples were evaluated for the presence and number of CTCs. KRAS mutation analysis was used to compare the CTCs with primary tumour tissue. CTC enumeration data was then evaluated as a diagnostic and staging biomarker in the setting of PDAC.

Results: We found 100% concordance for KRAS mutation subtype between primary tumour and CTCs in all five patients tested. Evaluation of CTCs as a diagnostic revealed the presence of CTCs in 54/72 patients with confirmed PDAC (sensitivity=75.0%, specificity=96.4%, area under the curve (AUROC)=0.867, 95% CI=0.798-0.935, and P<0.001). Furthermore, a cut-off of ⩾3 CTCs in 4 ml VB was able to discriminate between local/regional and metastatic disease (AUROC=0.885; 95% CI=0.800-0.969; and P<0.001).

Conclusion: CTCs appear to function well as a biomarker for diagnosis and staging in PDAC.

PubMed Disclaimer

Conflict of interest statement

Hsian-Rong Tseng, Co-Founder, Cytolumina Technologies Corp. Inc. The remaining authors declare no conflict of interest.

Figures

**Figure 1**
**Study design and CTC detection principles.** (A) Flow diagram depicting VB draws from our study cohort followed by CTC enumeration on NanoVelcro Chips and subsequent correlation with diagnostic and staging information. (B) Schematic depicting CTC identification via a 4-colour ICC approach in conjunction with high-resolution fluorescent microscopy. Representative images of 2 common CTC and WBC staining patterns are shown at × 400 magnification.

**Figure 2**
**Study cohort characteristics.** Diagnostic and staging flowchart of enrolled patients in the study.

**Figure 3**
**Validation of ICC and evaluation of tumour origin for CTCs by *KRAS* mutational analysis.** (A) Flow diagram depicting the confirmation of tumour origin by isolating and sequencing CTCs and patient matched primary tumour tissue. (B) Sanger sequencing results for *KRAS* codon-12 mutations. CTCs, WBCs, and primary tumour tissue for two of the patients are depicted. Patient A has a G12V mutation and patient B has a G12D mutation. Both patients' WBCs were found to have wild-type *KRAS* sequences.

**Figure 4**
**CTCs as a diagnostic biomarker.** (A) Comparison of CTC enumeration in PDAC and non-adenocarcinoma diseases. (B) ROC curve for illustration of CTC performance in the discrimination of PDAC from non-adenocarcinoma diseases. CTC AUROC=0.867 (95% CI 0.798–0.935, P<0.001).

**Figure 5**
**CTCs as a staging biomarker.** (A) CTC enumeration showing correlation with PDAC stage. (B) CTC enumeration in local/regional (stages I–III) and metastatic (stage IV) disease. (C) Comparison of the performance of CTCs and CA19-9, in discriminating local/regional from metastatic disease. The CTC AUROC was 0.885 (95% CI=0.800–0.969 and P<0.001), the CA19-9 AUROC was 0.690 (95% CI=0.551–0.829 and P=0.014).

See this image and copyright information in PMC

References

1. Alix-Panabieres C, Pantel K (2014) Challenges in circulating tumour cell research. Nat Rev Cancer 14(9): 623–631. - PubMed
1. Bidard FC, Huguet F, Louvet C, Mineur L, Bouche O, Chibaudel B, Artru P, Desseigne F, Bachet JB, Mathiot C, Pierga JY, Hammel P (2013) Circulating tumor cells in locally advanced pancreatic adenocarcinoma: the ancillary CirCe 07 study to the LAP 07 trial. Ann Oncol 24(8): 2057–2061. - PubMed
1. Bryant KL, Mancias JD, Kimmelman AC, Der CJ (2014) KRAS: feeding pancreatic cancer proliferation. Trends Biochem Sci 39(2): 91–100. - PMC - PubMed
1. Cameron JL, He J (2015) Two thousand consecutive pancreaticoduodenectomies. J Am Coll Surg 220(4): 530–536. - PubMed
1. Chen JF, Ho H, Lichterman J, Lu YT, Zhang Y, Garcia MA, Chen SF, Liang AJ, Hodara E, Zhau HE, Hou S, Ahmed RS, Luthringer DJ, Huang J, Li KC, Chung LW, Ke Z, Tseng HR, Posadas EM (2015) Subclassification of prostate cancer circulating tumor cells by nuclear size reveals very small nuclear circulating tumor cells in patients with visceral metastases. Cancer 121(18): 3240–3251. - PMC - PubMed

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Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer

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Circulating tumour cells as a biomarker for diagnosis and staging in pancreatic cancer

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