Prognostic value of circulating tumour DNA in patients undergoing curative resection for pancreatic cancer

Naoto Hadano, Yoshiaki Murakami, Kenichiro Uemura, Yasusi Hashimoto, Naru Kondo, Naoya Nakagawa, Taijiro Sueda, Eiso Hiyama, Naoto Hadano, Yoshiaki Murakami, Kenichiro Uemura, Yasusi Hashimoto, Naru Kondo, Naoya Nakagawa, Taijiro Sueda, Eiso Hiyama

Abstract

Background: Pancreatic ductal adenocarcinoma (PDAC) is frequently diagnosed at an advanced stage, leading to a poor prognosis. Therefore, interest in the development of non-invasive biomarkers for prognostic prediction has grown rapidly. Here, we assessed the clinical implications of v-Ki-ras2 kirsten rat sarcoma viral oncogene homolog (KRAS)-mutated circulating tumour DNA (ctDNA) as a useful surrogate biomarker in patients with resectable PDAC.

Methods: We used droplet digital polymerase chain reaction to detect rare mutant tumour-derived KRAS genes in plasma cell-free DNA (cfDNA) as ctDNA. Samples were collected from 105 patients who underwent pancreatoduodenectomy for PDAC at a single institution. Overall survival (OS) was analysed according to the presence of ctDNA.

Results: Among the 105 cases, ctDNA was detected in 33 (31%) plasma samples. The median OS durations were 13.6 months for patients with ctDNA (ctDNA+) and 27.6 months for patients without ctDNA. Patients who were ctDNA+ had a significantly poorer prognosis with respect to OS (P<0.0001).

Conclusions: Our findings suggested that the presence of ctDNA in plasma samples could be an important and powerful predictor of poor survival in patients with PDAC. Accordingly, ctDNA detection might be a promising approach with respect to PDAC treatment.

Figures

Figure 1
Figure 1
Overview of droplet digital PCR assay.(A) Schematic representation of the droplet digital PCR (ddPCR) assay, which is based on nanolitre-sized water-in-oil emulsion droplet technology. In this assay, target DNA molecules are uniformly distributed across thousands of emulsified droplets, after which PCR amplification is performed in each partitioned droplet. After amplification, reactions containing one or more target DNA molecules represent the positive end-point, whereas those without target DNA molecules represent the negative end-point. The number of target DNA molecules present can be calculated from the fraction of positive end-point reactions using Poisson statistics. (B) Two-dimensional histogram of ddPCR assay for KRAS amplification. FAM (blue) and HEX (green) fluorescence levels were plotted for each droplet. Clusters in the upper and right halves of the plot (dashed circle and solid circle) represent the positive mutant and wild-type KRAS end-point results, respectively.
Figure 2
Figure 2
The results of the ctDNA detection.(A) Frequency of KRAS mutations in all primary tumour specimens and plasma samples. (B) Scattergram of ctDNA concentrations in all patients subdivided according to the Union for International Cancer Control (UICC) classification.
Figure 3
Figure 3
Overall survival curves according to the (A) presence of ctDNA and (B) KRAS mutation subtypes of ctDNA.Overall survival did not differ significantly according to the KRAS mutation subtypes of ctDNA. In contrast, significant differences in OS were observed according the categorisation of patients into ctDNA+ and ctDNA− groups.

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