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Published on: November 5, 2020
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Preoperative next-generation sequencing of pancreatic cyst fluid is highly accurate in cyst classification and
Aatur D Singhi1, Kevin McGrath2, Randall E Brand2
1Department of Pathology, University of Pittsburgh Medical Center Health System, Pittsburgh, Pennsylvania, USA.
Gut
|October 4, 2017
Summary
Next-generation sequencing (NGS) of pancreatic cyst fluid (PCF) DNA accurately detects KRAS/GNAS mutations, improving diagnosis of mucinous pancreatic cysts. Combined with TP53/PIK3CA/PTEN analysis, NGS also identifies advanced neoplasia, outperforming traditional Sanger sequencing.
Area of Science:
- Oncology
- Gastroenterology
- Molecular Diagnostics
Background:
- Pancreatic cysts (PCs) require accurate evaluation to differentiate neoplastic subtypes and assess malignant potential.
- Pancreatic cyst fluid (PCF) DNA testing, particularly for KRAS/GNAS and TP53/PIK3CA/PTEN mutations, offers a minimally invasive diagnostic approach.
- Next-generation sequencing (NGS) provides a comprehensive molecular profiling of PCF DNA.
Purpose of the Study:
- To evaluate the utility of preoperative PCF DNA testing using NGS for diagnosing pancreatic cysts.
- To compare the diagnostic performance of NGS with Sanger sequencing for KRAS/GNAS mutations.
- To assess the combined diagnostic value of KRAS/GNAS and TP53/PIK3CA/PTEN alterations for advanced neoplasia.
Main Methods:
- A prospective study involving 626 PCF specimens from 595 patients analyzed by targeted NGS.
- Correlation of molecular findings with endoscopic ultrasound (EUS) features, ancillary studies, and patient follow-up.
- A separate cohort of 159 PCF specimens was analyzed for KRAS/GNAS mutations using Sanger sequencing for comparison.
Main Results:
- NGS detected KRAS/GNAS mutations in 49% of PCs, with high sensitivity (89%) and specificity (100%) for mucinous PCs, significantly outperforming Sanger sequencing (65% sensitivity).
- Alterations in TP53/PIK3CA/PTEN were found in 6% of cases and, when combined with KRAS/GNAS mutations, demonstrated 89% sensitivity and 100% specificity for advanced neoplasia.
- Traditional EUS findings like ductal dilatation and mural nodules showed lower sensitivity and specificity compared to molecular markers.
Conclusions:
- Preoperative NGS of PCF DNA for KRAS/GNAS mutations is highly sensitive and specific for diagnosing mucinous pancreatic cysts, surpassing Sanger sequencing.
- The combined analysis of KRAS/GNAS and TP53/PIK3CA/PTEN alterations via NGS serves as a valuable preoperative marker for advanced pancreatic neoplasia.
