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KRAS mutations in pancreatic circulating tumor cells: a pilot study

  • Original Article
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Tumor Biology

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is most often diagnosed in a metastatic stage. Circulating tumor cells (CTC) in the blood are hypothesized as the means of systemic dissemination. We aimed to isolate and characterize CTC to evaluate their significance as prognostic markers in PDAC. Blood obtained from healthy donors and patients with PDAC before therapy was filtered with ScreenCell® filtration devices for size-based CTC isolation. Captured cells were analyzed by immunofluorescence for an epithelial to mesenchymal transition (EMT) marker (zinc finger E-box binding homebox 1 (ZEB1)) and an epithelial antigen (cytokeratin (CK)). Molecular analysis of parallel specimens evaluated the KRAS mutation status of the CTC. The survival of each patient after study was recorded. As demonstrated by either cytology or finding of a KRAS mutation, CTC were detected in 18 of 21 patients (86 %) with proven PDAC: 8 out of 10 patients (80 %) with early stage (UICC IIA/IIB) and 10 out of 11 (91 %) with late stage (UICC III/IV) disease. CTC were not found in any of the 10 control patients (p < 0.001). The presence of CTC did not adversely affect median survival: 16 months in CTC-positive (n = 18) vs. 10 months in CTC-negative (n = 3) patients. Neither ZEB1 nor cytological characteristics correlated with overall survival, although ZEB1 was found almost exclusively in CTC of patients with established metastases. Patients with a CTC KRAS mutation (CTC-KRAS mut) had a substantially better survival, 19.4 vs. 7.4 months than patients with wild type KRAS (p = 0.015). With ScreenCell filtration, CTC are commonly found in PDAC (86 %). Molecular and genetic characterization, including mutations such as KRAS, may prove useful for prognosis.

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Acknowledgments

We thank Silke Hempel for technical assistance in the KRAS-PNA assay.

Author’s contributions

BK designed the study; BK, ASL, JH, and ALW drafted the manuscript; SS performed the experiments and conducted parts of the analysis, PB and MBP performed the histological analysis, TG performed the statistical analysis. All authors critically reviewed the manuscript and approved the final form.

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    Authors and Affiliations

    1. Department of Surgery, University of Freiburg Medical Center, Hugstetter Str. 55, D-79106, Freiburg, Germany

      Birte Kulemann, Sindy Seifert, Torben Glatz & Jens Hoeppner

    2. Department of Surgery & Andrew L. Warshaw, MD Institute for Pancreatic Cancer Research, Massachusetts General Hospital/ Harvard Medical School, Boston, MA, USA

      Andrew S. Liss & Andrew L. Warshaw

    3. Department of Pathology, University of Freiburg Medical Center, Freiburg, Germany

      Peter Bronsert

    4. Comprehensive Cancer Center, University of Freiburg Medical Center, Freiburg, Germany

      Peter Bronsert

    5. Department of Pathology, Massachusetts General Hospital, Boston, MA, USA

      Martha B. Pitman

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    1. Birte Kulemann
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    Correspondence to Birte Kulemann.

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    Martha B. Pitman and Jens Hoeppner contributed equally to this work.

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    Kulemann, B., Liss, A.S., Warshaw, A.L. et al. KRAS mutations in pancreatic circulating tumor cells: a pilot study. Tumor Biol. 37, 7547–7554 (2016). https://doi.org/10.1007/s13277-015-4589-2

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    • DOI: https://doi.org/10.1007/s13277-015-4589-2

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