Cell-free DNA promoter hypermethylation in plasma as a diagnostic marker for pancreatic adenocarcinoma

Abstract

BackgroundPancreatic cancer has a 5-year survival rate of only 5–7%. Difficulties in detecting pancreatic cancer at early stages results in the high mortality and substantiates the need for additional diagnostic tools. Surgery is the only curative treatment and unfortunately only possible in localized tumours. A diagnostic biomarker for pancreatic cancer will have a major impact on patient survival by facilitating early detection and the possibility for curative treatment. DNA promoter hypermethylation is a mechanism of early carcinogenesis, which can cause inactivation of tumour suppressor genes. The aim of this study was to examine promoter hypermethylation in a panel of selected genes from cell-free DNA, as a diagnostic marker for pancreatic adenocarcinoma.MethodsPatients with suspected or biopsy-verified pancreatic cancer were included prospectively and consecutively. Patients with chronic/acute pancreatitis were included as additional benign control groups. Based on an optimized accelerated bisulfite treatment protocol, methylation-specific PCR of a 28 gene panel was performed on plasma samples. A diagnostic prediction model was developed by multivariable logistic regression analysis using backward stepwise elimination.ResultsPatients with pancreatic adenocarcinoma (n = 95), chronic pancreatitis (n = 97) and acute pancreatitis (n = 59) and patients screened, but negative for pancreatic adenocarcinoma (n = 27), were included. The difference in mean number of methylated genes in the cancer group (8.41 (95% CI 7.62–9.20)) vs the total control group (4.74 (95% CI 4.40–5.08)) was highly significant (p < 0.001). A diagnostic prediction model (age >65, BMP3, RASSF1A, BNC1, MESTv2, TFPI2, APC, SFRP1 and SFRP2) had an area under the curve of 0.86 (sensitivity 76%, specificity 83%). The model performance was independent of cancer stage.ConclusionsCell-free DNA promoter hypermethylation has the potential to be a diagnostic marker for pancreatic adenocarcinoma and differentiate between malignant and benign pancreatic disease. This study brings us closer to a clinical useful diagnostic marker for pancreatic cancer, which is urgently needed. External validation is, however, required before the test can be applied in the clinic.Trial registrationClinicalTrials.gov, NCT02079363 Electronic supplementary materialThe online version of this article (doi:10.1186/s13148-016-0286-2) contains supplementary material, which is available to authorized users.

Promoter hypermethylation in plasma‐derived cell‐free DNA as a prognostic marker for pancreatic adenocarcinoma staging

“…American Society of Anaesthesiologists (ASA) score and WHO performance status (PS) were registered at time of inclusion. Data from the same patients were used in a previous study …”

“…We previously performed a literature review to identify genes aberrantly methylated and detectable in cell‐free DNA from patients with pancreatic cancer . Additionally, we examined the hypermethylation of plasma‐derived cell‐free DNA as a diagnostic marker for pancreatic adenocarcinoma and found a panel of eight genes that could discriminate patients with pancreatic adenocarcinoma from patients with benign pancreatic disease or symptoms mimicking upper gastrointestinal cancer . The aim of this present study was to develop prognostic prediction models for pancreatic adenocarcinoma according to stage, based on a selected panel of hypermethylated promoter regions in plasma‐derived cell‐free DNA.…”

Epigenetics and Precision Oncology

“…There is potential for DNA methylation analysis to aid in early diagnosis and decrease cancer-specific mortality in this disease. A clinical cohort study published in 2016 demonstrated that interrogation of circulating blood DNA for methylation can identify patients with pancreatic adenocarcinoma (34). By examining methylation status at BMP3 , RASSF1A , BNC1 , MESTv2 , TFPI2 , APC , SFRP1 , and SFRP2, overt cancer was distinguished from healthy controls, as well as patients with chronic or acute pancreatitis with a sensitivity of 76% and specificity of 83% (34).…”

“…A clinical cohort study published in 2016 demonstrated that interrogation of circulating blood DNA for methylation can identify patients with pancreatic adenocarcinoma (34). By examining methylation status at BMP3 , RASSF1A , BNC1 , MESTv2 , TFPI2 , APC , SFRP1 , and SFRP2, overt cancer was distinguished from healthy controls, as well as patients with chronic or acute pancreatitis with a sensitivity of 76% and specificity of 83% (34). Diagnostic predictive accuracy was independent of tumor stage, suggesting that circulating DNA methylation may aid in early diagnosis of pancreatic cancer.…”

Alterations of Epigenetic Regulators in Pancreatic Cancer and Their Clinical Implications

“…The aberrant methylation of some of the genes, such as BNC1 and ADAMTS1 , was shown to be detected in the serum of patients with PDA, which could be served as potential early diagnostic biomarkers for PDAs [23]. Cell-free DNA promoter hypermethylation in patient plasma was also shown to be significantly different between pancreatic cancer patients and control group [24]. Another study has examined DNA methylation differences in leukocyte DNA between PDA patients and controls [25].…”