Predicting the Grade of Dysplasia of Pancreatic Cystic Neoplasms Using Cyst Fluid DNA Methylation Markers

Abstract

Purpose Pancreatic cysts are common and pose diagnostic and management challenges. Pancreatic cyst fluid markers have the potential to aid in the management of cysts with concerning imaging findings. Our aim was to evaluate cyst fluid methylated DNA markers for their accuracy for predicting the histologic grade of neoplastic pancreatic cysts. Experimental Design Pancreatic cyst fluid samples from 183 patients (29 discovery, 154 validation) aspirated after surgical resection were analyzed for methylated DNA at selected genes (SOX17, BNIP3, FOXE1, PTCHD2, SLIT2, EYA4 and SFRP1) using methylation-specific droplet-digital PCR (dd-QMSP). Methylated DNA levels were evaluated for their accuracy at predicting the grade of dysplasia of the pancreatic cyst. Results All six markers evaluated in the validation set could accurately distinguish high-risk cystic neoplasms (with high-grade dysplasia and/or associated invasive cancer) from low-risk cysts (lower grades of dysplasia) with accuracies from 79.8 to 83.6%. Methylated SOX17 had the highest overall accuracy as a single marker (sensitivity, 78.4%; specificity, 85.6%; accuracy 83.6%, cut-off; 25 methylated DNA molecules/μL cyst fluid). The best four-gene combination had 84.3% sensitivity, 89.4% specificity and 88.0% accuracy at distinguishing cysts with high-grade dysplasia and/or invasive cancer from those without. All six markers were independent predictors of having invasive cancer/high-grade dysplasia after adjusting for clinical/imaging factors known to be associated with grade of dysplasia. The combination of methylated SOX17 with cytology better predicted neoplastic grade than cytology alone. Conclusion A panel of methylated gene markers quantified by dd-QMSP can be used to predict the grade of dysplasia of pancreatic cysts.

Early Detection of Pancreatic Cancer: Opportunities and Challenges

“…98 Promising biomarker results with PCF have also been reported for differentially methylated DNA, telomerase activity, protease expression, and the overexpression of Das-1. [99][100][101][102] However, the majority of these biomarkers have not been rigorous validated in a diverse cohort of pancreatic cysts. Hence, this is the goal of the Pancreatic Cyst Biomarker Validation Study, an ongoing double-blinded PCF biomarker study that is sponsored by the National Cancer Institute Early Detection Research Network.…”

Cyst Fluid Biomarkers - Diagnosis and Prediction of Malignancy for Cystic Lesions of the Pancreas

“…Alterations in the phosphatidylinositol-3-kinase (PI3K) pathway, including somatic mutations in the oncogenes PIK3CA and AKT1 as well as in the tumor suppressor gene PTEN , have been reported but are rare in IPMNs [8,14]. Additional markers which have shown promise for identifying high-grade dysplasia in IPMNs include hypermethylation of SOX17 , which in one study was associated with an accuracy of 84% for identifying IPMNs with high-grade dysplasia [15]. Elevated cyst fluid telomerase levels have also been associated with an increased risk of high-grade dysplasia [16].…”

EYA4 inhibits hepatocellular carcinoma by repressing MYCBP by dephosphorylating β‐catenin at Ser552

“…5 Recent studies have shown that aberrant methylation of EYA4 is a potential biomarker in colorectal cancer, urothelial bladder cancer, esophageal squamous cell carcinoma and pancreatic cystic neoplasms. [6][7][8][9][10] In our previous studies, we demonstrated that EYA4 was highly methylated in HCC and related to the prognosis of HCC patients. 11 We also clarified that EYA4 functioned as a tumor suppressor gene in HCC.…”