Circulating tumor cells (Ctc) and kras mutant circulating free Dna (cfdna) detection in peripheral blood as biomarkers in patients diagnosed with exocrine pancreatic cancer

Julie Earl, Sandra Garcia-Nieto, Jose Carlos Martinez-Avila, José Montans, Alfonso Sanjuanbenito, Mercedes Rodríguez-Garrote, Eduardo Lisa, Elena Mendía, Eduardo Lobo, Núria Malats, Alfredo Carrato, Carmen Guillen-Ponce, Julie Earl, Sandra Garcia-Nieto, Jose Carlos Martinez-Avila, José Montans, Alfonso Sanjuanbenito, Mercedes Rodríguez-Garrote, Eduardo Lisa, Elena Mendía, Eduardo Lobo, Núria Malats, Alfredo Carrato, Carmen Guillen-Ponce

Abstract

Background: Pancreatic cancer remains one of the most difficult cancers to treat with the poorest prognosis. The key to improving survival rates in this disease is early detection and monitoring of disseminated and residual disease. However, this is hindered due to lack reliable diagnostic and predictive markers which mean that the majority of patients succumb to their condition within a few months.

Methods: We present a pilot study of the detection circulating free DNA (cfDNA) combined with tumor specific mutation detection by digital PCR as a novel minimally invasive biomarker in pancreatic ductal adenocarcinoma (PDAC). This was compared to the detection of CTC by the CellSearch® system and a novel CTC enrichment strategy based on CD45 positive cell depletion. The aim of the study was to assess tumor specific DNA detection in plasma and CTC detection as prognostic markers in PDAC.

Results: We detected KRAS mutant cfDNA in 26% of patients of all stages and this correlated strongly with Overall Survival (OS), 60 days (95% CI: 19-317) for KRAS mutation positive vs 772 days for KRAS mutation negative (95% CI: 416-1127). Although, the presence of CTC detected by the CellSearch® system did correlate significantly with OS, 88 days (95% CI: 27-206) CTC positive vs 393 days CTC negative (95% CI: 284-501), CTC were detected in only 20% of patients, the majority of which had metastatic disease, whereas KRAS mutant cfDNA was detected in patients with both resectable and advanced disease.

Conclusions: Tumor specific cfDNA detection and CTC detection are promising markers for the management of patients with PDAC, although there is a need to validate these results in a larger patient cohort and optimize the detection of CTC in PDAC by applying the appropriate markers for their detection.

Figures

Fig. 1
Fig. 1
Correlation of total cfDNA concentration in plasma with PDAC disease stage. *DNA concentration was estimated by the number of copies of the RNaseP gene in 20 μl of cfDNA in plasma
Fig. 2
Fig. 2
KRAS mutation detection in plasma cfDNA in PDAC cases. a. G12D KRAS mutation detection in plasma and genomic DNA by QX200™ Droplet Digital™ PCR. b. Frequency of mutant KRAS detection in plasma in PDAC. c. Correlation of cfDNA concentration and mutant KRAS detection. *DNA concentration was estimated by the number of copies of the RNaseP gene in 20 μl of cfDNA in plasma. d. Kaplan Meier survival analysis of KRAS mutation status in plasma cfDNA
Fig. 3
Fig. 3
CTC detection whole blood in PDAC cases. a. Frequency of CTC in peripheral blood in PDAC. b. AsPc-1 and PaTu8988S detection in spiked peripheral blood (100 cells/ml) using the CellSearch® system. c. Kaplan Meier survival analysis of CTC status in peripheral blood
Fig. 4
Fig. 4
KRAS mutation detection in CD45 depleted blood. The KRAS G12D mutation was detected in 2 patients that tested negative for CTC by the CellSearch® system

References

    1. Ghaneh P, Costello E, Neoptolemos JP. Biology and management of pancreatic cancer. Postgrad Med J. 2008;84(995):478–497. doi: 10.1136/gut.2006.103333.
    1. Katz MH, Wang H, Fleming JB, Sun CC, Hwang RF, Wolff RA, Varadhachary G, Abbruzzese JL, Crane CH, Krishnan S, et al. Long-term survival after multidisciplinary management of resected pancreatic adenocarcinoma. Ann Surg Oncol. 2009;16(4):836–847. doi: 10.1245/s10434-008-0295-2.
    1. Hidalgo M. Pancreatic cancer. N Engl J Med. 2010;362(17):1605–1617. doi: 10.1056/NEJMra0901557.
    1. Buxbaum JL, Eloubeidi MA. Molecular and clinical markers of pancreas cancer. JOP. 2010;11(6):536–544.
    1. Goonetilleke KS, Siriwardena AK. Systematic review of carbohydrate antigen (CA 19–9) as a biochemical marker in the diagnosis of pancreatic cancer. Eur J Surg Oncol. 2007;33(3):266–270. doi: 10.1016/j.ejso.2006.10.004.
    1. Mayers JR, Wu C, Clish CB, Kraft P, Torrence ME, Fiske BP, Yuan C, Bao Y, Townsend MK, Tworoger SS, et al. Elevation of circulating branched-chain amino acids is an early event in human pancreatic adenocarcinoma development. Nat Med. 2014;20(10):1193–1198. doi: 10.1038/nm.3686.
    1. Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, Miller MC, Reuben JM, Doyle GV, Allard WJ, Terstappen LW, et al. Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med. 2004;351(8):781–791. doi: 10.1056/NEJMoa040766.
    1. de Bono JS, Scher HI, Montgomery RB, Parker C, Miller MC, Tissing H, Doyle GV, Terstappen LW, Pienta KJ, Raghavan D. Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer. Clin Cancer Res. 2008;14(19):6302–6309. doi: 10.1158/1078-0432.CCR-08-0872.
    1. De Giorgi U, Valero V, Rohren E, Dawood S, Ueno NT, Miller MC, Doyle GV, Jackson S, Andreopoulou E, Handy BC, et al. Circulating tumor cells and [18 F]fluorodeoxyglucose positron emission tomography/computed tomography for outcome prediction in metastatic breast cancer. J Clin Oncol. 2009;27(20):3303–3311. doi: 10.1200/JCO.2008.19.4423.
    1. Krebs MG, Sloane R, Priest L, Lancashire L, Hou JM, Greystoke A, Ward TH, Ferraldeschi R, Hughes A, Clack G, et al. Evaluation and prognostic significance of circulating tumor cells in patients with non-small-cell lung cancer. J Clin Oncol. 2011;29(12):1556–1563. doi: 10.1200/JCO.2010.28.7045.
    1. Kurihara T, Itoi T, Sofuni A, Itokawa F, Tsuchiya T, Tsuji S, Ishii K, Ikeuchi N, Tsuchida A, Kasuya K, et al. Detection of circulating tumor cells in patients with pancreatic cancer: a preliminary result. J Hepatobiliary Pancreat Surg. 2008;15(2):189–195. doi: 10.1007/s00534-007-1250-5.
    1. Bidard FC, Huguet F, Louvet C, Mineur L, Bouche O, Chibaudel B, Artru P, Desseigne F, Bachet JB, Mathiot C, et al. Circulating tumor cells in locally advanced pancreatic adenocarcinoma: the ancillary CirCe 07 study to the LAP 07 trial. Annals Oncol. 2013;24(8):2057–2061. doi: 10.1093/annonc/mdt176.
    1. Khoja L, Backen A, Sloane R, Menasce L, Ryder D, Krebs M, Board R, Clack G, Hughes A, Blackhall F, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker. Br J Cancer. 2012;106(3):508–516. doi: 10.1038/bjc.2011.545.
    1. Schwarzenbach H, Hoon DS, Pantel K. Cell-free nucleic acids as biomarkers in cancer patients. Nat Rev Cancer. 2011;11(6):426–437. doi: 10.1038/nrc3066.
    1. Lee YJ, Yoon KA, Han JY, Kim HT, Yun T, Lee GK, Kim HY, Lee JS. Circulating cell-free DNA in plasma of never smokers with advanced lung adenocarcinoma receiving gefitinib or standard chemotherapy as first-line therapy. Clin Cancer Res. 2011;17(15):5179–5187. doi: 10.1158/1078-0432.CCR-11-0400.
    1. Spindler KL, Pallisgaard N, Vogelius I, Jakobsen A. Quantitative cell-free DNA, KRAS, and BRAF mutations in plasma from patients with metastatic colorectal cancer during treatment with cetuximab and irinotecan. Clin Cancer Res. 2012;18(4):1177–1185. doi: 10.1158/1078-0432.CCR-11-0564.
    1. Forbes SA, Bindal N, Bamford S, Cole C, Kok CY, Beare D, Jia M, Shepherd R, Leung K, Menzies A, et al. COSMIC: mining complete cancer genomes in the Catalogue of Somatic Mutations in Cancer. Nucleic Acids Res. 2011;39(Database issue):D945–950. doi: 10.1093/nar/gkq929.
    1. Team RDC. R: A language and environment for statistical computing. 2008.
    1. Inc S: SPSS Statistics for Windows, Version 17.0. Chicago. Released 2008.
    1. Sastre J, Maestro ML, Puente J, Veganzones S, Alfonso R, Rafael S, Garcia-Saenz JA, Vidaurreta M, Martin M, Arroyo M, et al. Circulating tumor cells in colorectal cancer: correlation with clinical and pathological variables. Annals Oncol. 2008;19(5):935–938. doi: 10.1093/annonc/mdm583.
    1. Tol J, Koopman M, Miller MC, Tibbe A, Cats A, Creemers GJ, Vos AH, Nagtegaal ID, Terstappen LW, Punt CJ. Circulating tumour cells early predict progression-free and overall survival in advanced colorectal cancer patients treated with chemotherapy and targeted agents. Annals Oncol. 2010;21(5):1006–1012. doi: 10.1093/annonc/mdp463.
    1. Budd GT, Cristofanilli M, Ellis MJ, Stopeck A, Borden E, Miller MC, Matera J, Repollet M, Doyle GV, Terstappen LW, et al. Circulating tumor cells versus imaging--predicting overall survival in metastatic breast cancer. Clin Cancer Res. 2006;12(21):6403–6409. doi: 10.1158/1078-0432.CCR-05-1769.
    1. Hess KR, Varadhachary GR, Taylor SH, Wei W, Raber MN, Lenzi R, Abbruzzese JL. Metastatic patterns in adenocarcinoma. Cancer. 2006;106(7):1624–1633. doi: 10.1002/cncr.21778.
    1. Rhim AD, Mirek ET, Aiello NM, Maitra A, Bailey JM, McAllister F, Reichert M, Beatty GL, Rustgi AK, Vonderheide RH, et al. EMT and Dissemination Precede Pancreatic Tumor Formation. Cell. 2012;148(1–2):349–361. doi: 10.1016/j.cell.2011.11.025.
    1. Lecharpentier A, Vielh P, Perez-Moreno P, Planchard D, Soria JC, Farace F. Detection of circulating tumour cells with a hybrid (epithelial/mesenchymal) phenotype in patients with metastatic non-small cell lung cancer. Br J Cancer. 2011;105(9):1338–1341. doi: 10.1038/bjc.2011.405.
    1. Bettegowda C, Sausen M, Leary RJ, Kinde I, Wang Y, Agrawal N, Bartlett BR, Wang H, Luber B, Alani RM, et al. Detection of circulating tumor DNA in early- and late-stage human malignancies. Sci TranslMed. 2014;6(224):224ra224.

Source: PubMed

Szponzorok és közreműködők

Egészségi állapot

Kábítószer-beavatkozások

3
Iratkozz fel