Cancer Res Treat.  2020 Oct;52(4):1219-1228. 10.4143/crt.2019.688.

Evaluation of Circulating Tumor DNA in Patients with Ovarian Cancer Harboring Somatic PIK3CA or KRAS Mutations

• Ogasawara A
• Hihara T
• Shintani D
• Yabuno A
• Ikeda Y
• Tai K
• Fujiwara K
• Watanabe K
• Hasegawa K

• KMID: 2507947
• DOI: http://doi.org/10.4143/crt.2019.688

Abstract

Purpose
Circulating tumor DNA (ctDNA) is an attractive source for liquid biopsy to understand molecular phenotypes of a tumor non-invasively, which is also expected to be both a diagnostic and prognostic marker. PIK3CA and KRAS are among the most frequently mutated genes in epithelial ovarian cancer (EOC). In addition, their hotspot mutations have already been identified and are ready for a highly sensitive analysis. Our aim is to clarify the significance of PIK3CA and KRAS mutations in the plasma of EOC patients as tumor-informed ctDNA.
Methods
We screened 306 patients with ovarian tumors for somatic PIK3CA or KRAS mutations. A total of 85 EOC patients had somatic PIK3CA and/or KRAS mutations, and the corresponding mutations were subsequently analyzed using a droplet digital polymerase chain reaction in their plasma.
Results
The detection rates for ctDNA were 27% in EOC patients. Advanced stage and positive peritoneal cytology were associated with higher frequency of ctDNA detection. Preoperative ctDNA detection was found to be an indicator of outcomes, and multivariate analysis revealed that ctDNA remained an independent risk factor for recurrence (p=0.010). Moreover, we assessed the mutation frequency in matched plasma before surgery and at recurrence from 17 patients, and found six patients had higher mutation rates in cell-free DNA at recurrence compared to that at primary diagnosis.
Conclusion
The presence of ctDNA at diagnosis was an indicator for recurrence, which suggests potential tumor spread even when tumors were localized at the time of diagnosis.

Keyword

Ovarian neoplasms; ctDNA; Biomarker

Figure

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  Fig. 1. Flow diagram. Numbers of patients who were included in the analyses.

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  Fig. 2. Survival curves according to circulating tumor DNA (ctDNA) status, positive or negative. Progression-free survival (A) and overall survival (B) in all epithelial ovarian cancer (EOC) patients as to ctDNA status for PIK3CA and/or KRAS mutations (p=0.0001 and p=0.017, respectively). Progression-free survival (C) and overall survival (D) in all EOC patients as to ctDNA status for PIK3CA mutations (p=0.008 and p=0.118, respectively). Progression-free survival (E) and overall survival (F) in all EOC patients as to ctDNA status for KRAS mutations (p=0.004 and p=0.072, respectively). Recurrence-free survival (G) and overall survival (H) in stage I/II EOC patients as to ctDNA status for PIK3CA and/or KRAS mutations (p=0.010 and p=0.888, respectively). Recurrence-free survival (I) and overall survival (J) in stage I/II EOC patients as to ctDNA status for PIK3CA mutations (p=0.071 and p=0.725, respectively). Recurrence-free survival (K) and overall survival (L) in stage I/II EOC patients as to ctDNA status for KRAS mutations (p=0.050 and p=0.464, respectively).

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  Fig. 3. Quantitative analysis of circulating tumor DNA (cfDNA) at the time of primary treatment and at recurrence. (A) Total mutation copies in plasma. (B) Mutation frequency in cell-free DNA (cfDNA) ([mutated copies/wild type copies]×100). (C) Levels of serum cancer antigen 125 (CA125). (D) Tumor size.

Reference

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