Mutant TP53 in Duodenal Samples of Pancreatic Juice From Patients With Pancreatic Cancer or High-Grade Dysplasia

doi:10.1016/j.cgh.2012.11.016

Clinical Gastroenterology and Hepatology

Volume 11, Issue 6, June 2013, Pages 719-730.e5

https://doi.org/10.1016/j.cgh.2012.11.016 Get rights and content

Background & Aims

Imaging tests can identify patients with pancreatic neoplastic cysts but not microscopic dysplasia. We investigated whether mutant TP53 can be detected in duodenal samples of secretin-stimulated pancreatic juice, and whether this assay can be used to screen for high-grade dysplasia and invasive pancreatic cancer.

Methods

We determined the prevalence of mutant TP53 in microdissected pancreatic intraepithelial neoplasias (PanINs), intraductal papillary mucinous neoplasms (IPMNs), and invasive adenocarcinomas. TP53 mutations were quantified by digital high-resolution melt-curve analysis and sequencing of secretin-stimulated pancreatic juice samples, collected from duodena of 180 subjects enrolled in Cancer of the Pancreas Screening trials; patients were enrolled because of familial and/or inherited predisposition to pancreatic cancer, or as controls.

Results

TP53 mutations were identified in 9.1% of intermediate-grade IPMNs (2 of 22), 17.8% of PanIN-2 (8 of 45), 38.1% of high-grade IPMNs (8 of 21), 47.6% of PanIN-3 (10 of 21), and 75% of invasive pancreatic adenocarcinomas (15 of 20); no TP53 mutations were found in PanIN-1 lesions or low-grade IPMNs. TP53 mutations were detected in duodenal samples of pancreatic juice from 29 of 43 patients with pancreatic ductal adenocarcinoma (67.4% sensitivity; 95% confidence interval, 0.52–0.80) and 4 of 8 patients with high-grade lesions (PanIN-3 and high-grade IPMN). No TP53 mutations were identified in samples from 58 controls or 55 screened individuals without evidence of advanced lesions.

Conclusions

We detected mutant TP53 in secretin-stimulated pancreatic juice samples collected from duodena of patients with high-grade dysplasia or invasive pancreatic cancer. Tests for mutant TP53 might be developed to improve the diagnosis of and screening for pancreatic cancer and high-grade dysplasia. Clinical Trial numbers: NCT00438906 and NCT00714701.

Section snippets

Materials and Methods

All elements of this investigation were approved by The Johns Hopkins Medical Institutional Review Board and written informed consent was obtained from all patients. All authors had access to the study data and reviewed and approved the final manuscript.

Prevalence of TP53 Mutations in Pancreatic Intraepithelial Neoplasias, Intraductal Papillary Mucinous Neoplasms, and Ductal Adenocarcinomas

TP53 mutations were detected in 9.1% of intermediate-grade IPMNs, 17.8% of intermediate-grade PanINs (PanIN-2), 38.1% of high-grade IPMNs, 47.6% of PanIN-3, and 75% of 20 invasive ductal adenocarcinomas (one cancer had 2 mutations) (Table 1). No TP53 mutations were detected in low-grade PanINs (PanIN-1) or IPMNs.

Detection of Mutant TP53 in Duodenal Collections of Pancreatic Juice

We first evaluated the limit of detection of our digital HRM assay. Mutant TP53 reliably could detect 0.1% to 10% concentrations of mutant to wild-type DNA (Supplementary Figure 1).

Discussion

Our results indicate that mutant TP53 detected in duodenal collections of secretin-stimulated pancreatic juice may provide evidence that the pancreas contains either microscopic PanIN-3 IPMNs with high-grade dysplasia, or invasive pancreatic ductal adenocarcinoma. Mutant TP53 was detected in the pancreatic juice of only 1 of 102 individuals not known to have high-grade dysplasia, and that was a patient with a 6-cm IPMN with intermediate-grade dysplasia. The prevalence of mutant TP53 detected in

References (31)

C. Shi et al.
Intraductal papillary mucinous neoplasm
Hum Pathol
(2012)
K. de Jong et al.
High prevalence of pancreatic cysts detected by screening magnetic resonance imaging examinations
Clin Gastroenterol Hepatol
(2010)
M.I. Canto et al.
Frequent detection of pancreatic lesions in asymptomatic high-risk individuals
Gastroenterology
(2012)
M.I. Canto et al.
Screening for early pancreatic neoplasia in high-risk individuals: a prospective controlled study
Clin Gastroenterol Hepatol
(2006)
M.I. Canto et al.
Screening for pancreatic neoplasia in high-risk individuals: an EUS-based approach
Clin Gastroenterol Hepatol
(2004)
L. Yan et al.
Molecular analysis to detect pancreatic ductal adenocarcinoma in high-risk groups
Gastroenterology
(2005)
M. Löhr et al.
p53 and K-ras mutations in pancreatic juice samples from patients with chronic pancreatitis
Gastrointest Endosc
(2001)
M. Tanaka et al.
International consensus guidelines 2012 for the management of IPMN and MCN of the pancreas
Pancreatology
(2012)
H. Haeno et al.
Computational modeling of pancreatic cancer reveals kinetics of metastasis suggesting optimum treatment strategies
Cell
(2012)
S. Raimondi et al.
Epidemiology of pancreatic cancer: an overview
Nat Rev Gastroenterol Hepatol
(2009)

M. Goggins

Markers of pancreatic cancer: working toward early detection

Clin Cancer Res

(2011)

E. Ludwig et al.

Feasibility and yield of screening in relatives from familial pancreatic cancer families

Am J Gastroenterol

(2011)

E.C. Verna et al.

Pancreatic cancer screening in a prospective cohort of high-risk patients: a comprehensive strategy of imaging and genetics

Clin Cancer Res

(2010)

P. Langer et al.

Five years of prospective screening of high-risk individuals from families with familial pancreatic cancer

Gut

(2009)

J.W. Poley et al.

The yield of first-time endoscopic ultrasonography in screening individuals at a high risk of developing pancreatic cancer

Am J Gastroenterol

(2009)

Cited by (143)

Biological characteristics of pancreatic ductal adenocarcinoma: Initiation to malignancy, intracellular to extracellular
2023, Cancer Letters
Pancreatic ductal adenocarcinoma (PDAC) is a highly life-threatening tumour with a low early-detection rate, rapid progression and a tendency to develop resistance to chemotherapy. Therefore, understanding the regulatory mechanisms underlying the initiation, development and metastasis of pancreatic cancer is necessary for enhancing therapeutic effectiveness. In this review, we summarised single-gene mutations (including KRAS, CDKN2A, TP53, SMAD4 and some other less prevalent mutations), epigenetic changes (including DNA methylation, histone modifications and RNA interference) and large chromosome alterations (such as copy number variations, chromosome rearrangements and chromothripsis) associated with PDAC. In addition, we discussed variations in signalling pathways that act as intermediate oncogenic factors in PDAC, including PI3K/AKT, MAPK/ERK, Hippo and TGF-β signalling pathways. The focus of this review was to investigate alterations in the microenvironment of PDAC, particularly the role of immunosuppressive cells, cancer-associated fibroblasts, lymphocytes, other para-cancerous cells and tumour extracellular matrix in tumour progression. Peripheral axons innervating the pancreas have been reported to play a crucial role in the development of cancer. In addition, tumour cells can influence the behaviour of neighbouring non-tumour cells by secreting certain factors, both locally and at a distance. In this review, we elucidated the alterations in intracellular molecules and the extracellular environment that occur during the progression of PDAC. Altogether, this review may enhance the understanding of the biological characteristics of PDAC and guide the development of more precise treatment strategies.
Diagnostic accuracy of the AGA, IAP, and European guidelines for detecting advanced neoplasia in intraductal papillary mucinous neoplasm/neoplasia
2023, Pancreatology
Follow-up in patients with intraductal papillary mucinous neoplasm (IPMN) aims to detect advanced neoplasia (high-grade dysplasia/cancer) in an early stage. The 2015 American Gastroenterological Association (AGA), 2017 International Association of Pancreatology (IAP), and the 2018 European Study Group on Cystic tumours of the Pancreas (European) guidelines differ in their recommendations on indications for surgery. However, it remains unclear which guideline is most accurate in predicting advanced neoplasia in IPMN.
Patients who underwent surgery were extracted from a prospective database (January 2006–January 2021). In patients with IPMN, final pathology was compared with the indication for surgery according to the guidelines. ROC-curves were calculated to determine the diagnostic accuracy for each guideline.
Overall, 247 patients underwent surgery for cystic lesions. In 145 patients with IPMN, 52 had advanced neoplasia, of which the AGA guideline would have advised surgery in 14 (27%), the IAP and European guideline in 49 (94%) and 50 (96%). In 93 patients without advanced neoplasia, the AGA, IAP, and European guidelines would incorrectly have advised surgery in 8 (8.6%), 77 (83%) and 71 (76%).
The European and IAP guidelines are clearly superior in detecting advanced neoplasia in IPMN as compared to the AGA, albeit at the cost of a higher rate of unnecessary surgery. To harmonize care and to avoid confusion caused by conflicting statements, a global evidence-based guideline for PCN in collaboration with the various guidelines groups is required once the current guidelines require an update.
Simultaneous and sequential combination of genetic and epigenetic biomarkers for the presence of high-grade dysplasia in patients with pancreatic cyst: Discovery in cyst fluid and test in pancreatic juice
2023, Pancreatology
Screening patients with intraductal papillary mucinous neoplasms (IPMN) has the primary goal of identifying potentially curable noninvasive precursors. We aimed to evaluate the diagnostic impact of genetic and epigenetic biomarkers in the presence of noninvasive precursors.
Mutated KRAS/GNAS and methylated SOX17/TBX15/BMP3/TFPI2 DNA were assessed by droplet digital PCR in a discovery cohort of 70 surgically aspirated cyst fluids, and diagnostic performances for differentiating high-grade dysplasia (HGD) from low-grade dysplasia (LGD) was evaluated. We then tested these markers using an independent test cohort consisting of 156 serially collected pancreatic juice samples from 30 patients with IPMN.
Mutated KRAS and GNAS are specific for IPMNs but are not helpful for the prediction of histological grades. Cyst fluids from IPMN with HGD showed higher methylation levels of SOX17 (median, 0.141 vs. 0.021; P = 0.086) and TBX15 (median, 0.030 vs. 0.003; P = 0.028) than those with LGD. The combination of all tested markers yielded a diagnostic performance with sensitivity of 69.6%, and specificity of 90.0%. Among the 30 pancreatic juice samples exhibiting the highest abundance of KRAS/GNAS mutations in each patient in the test cohort, patients with histologically proven HGD due to pancreatic resection had a significantly higher prevalence (100% vs. 31%, P = 0.018) and abundance (P = 0.037) of methylated TBX15 than those without cytohistological diagnosis undergoing surveillance.
A simultaneous and sequential combination of mutated and methylated DNA markers in pancreatic cyst fluid and juice sample markers can help detect noninvasive pancreatic precursor neoplasms.
Systematic review and meta-analysis: Diagnostic performance of DNA alterations in pancreatic juice for the detection of pancreatic cancer
2022, Pancreatology
Citation Excerpt :
The initial search identified 580 studies, of which 504 were excluded based on title and abstract. After full-text review, 41 studies were included: 32 concerning DNA mutations, 14 DNA methylations and five describing both [12–52] (Fig. 1). For each study, distinct collection methods and DNA analysis methods were reported.
Pancreatic cancer has a dismal prognosis. So far, imaging has been proven incapable of establishing an early enough diagnosis. Thus, biomarkers are urgently needed for early detection and improved survival. Our aim was to evaluate the pooled diagnostic performance of DNA alterations in pancreatic juice.
A systematic literature search was performed in EMBASE, MEDLINE Ovid, Cochrane CENTRAL and Web of Science for studies concerning the diagnostic performance of DNA alterations in pancreatic juice to differentiate patients with high-grade dysplasia or pancreatic cancer from controls. Study quality was assessed using QUADAS-2. The pooled prevalence, sensitivity, specificity and diagnostic odds ratio were calculated.
Studies mostly concerned cell-free DNA mutations (32 studies: 939 cases, 1678 controls) and methylation patterns (14 studies: 579 cases, 467 controls). KRAS, TP53, CDKN2A, GNAS and SMAD4 mutations were evaluated most. Of these, TP53 had the highest diagnostic performance with a pooled sensitivity of 42% (95% CI: 31–54%), specificity of 98% (95%-CI: 92%–100%) and diagnostic odds ratio of 36 (95% CI: 9–133).
Of DNA methylation patterns, hypermethylation of CDKN2A, NPTX2 and ppENK were studied most. Hypermethylation of NPTX2 performed best with a sensitivity of 39–70% and specificity of 94–100% for distinguishing pancreatic cancer from controls.
This meta-analysis shows that, in pancreatic juice, the presence of distinct DNA mutations (TP53, SMAD4 or CDKN2A) and NPTX2 hypermethylation have a high specificity (close to 100%) for the presence of high-grade dysplasia or pancreatic cancer. However, the sensitivity of these DNA alterations is poor to moderate, yet may increase if they are combined in a panel.
Integrating Molecular Analysis into the Pathologic Evaluation of Pancreatic Cysts
2022, Surgical Pathology Clinics
Molecular Pathology of Pancreatic Cystic Lesions with a Focus on Malignant Progression
2024, Cancers

View all citing articles on Scopus

: Conflicts of interest These authors disclose the following: Michael Goggins and Ralph Hruban have a licensing agreement with Myriad Genetics for the discovery of PALB2 as a pancreatic cancer susceptibility gene. The remaining authors disclose no conflicts.

: None of the companies involved had any part in the design of this study, analysis or interpretation of data, or in the writing of this manuscript. The corresponding author had full access to all of the data and takes full responsibility for the veracity of the data and statistical analysis.

: Funding Supported by National Institutes of Health grants (CA62924, R01CA120432, and RC2CA148376), the Lustgarten Foundation for Pancreatic Cancer Research, the Jimmy V Foundation, Susan Wojcicki and Dennis Troper, the Michael Rolfe Foundation, the Alan Graff Foundation, Karp Family H.H. Metals, Inc, Fund for Cancer Research, Michael Hooven and Susan Spies, and Hugh and Rachel Victor. Recombinant secretin was provided for this study by ChiRhoClin, Inc, and Repligen.

View full text

Original articlePancreas, biliary tract, and liverMutant TP53 in Duodenal Samples of Pancreatic Juice From Patients With Pancreatic Cancer or High-Grade Dysplasia

Background & Aims

Methods

Results

Conclusions

Section snippets

Materials and Methods

Prevalence of TP53 Mutations in Pancreatic Intraepithelial Neoplasias, Intraductal Papillary Mucinous Neoplasms, and Ductal Adenocarcinomas

Detection of Mutant TP53 in Duodenal Collections of Pancreatic Juice

Discussion

Hum Pathol

Clin Gastroenterol Hepatol

Gastroenterology

Clin Gastroenterol Hepatol

Clin Gastroenterol Hepatol

Gastroenterology

Gastrointest Endosc

Pancreatology

Cell

Epidemiology of pancreatic cancer: an overview

Nat Rev Gastroenterol Hepatol

Markers of pancreatic cancer: working toward early detection

Clin Cancer Res

Feasibility and yield of screening in relatives from familial pancreatic cancer families

Am J Gastroenterol

Pancreatic cancer screening in a prospective cohort of high-risk patients: a comprehensive strategy of imaging and genetics

Clin Cancer Res

Five years of prospective screening of high-risk individuals from families with familial pancreatic cancer

Gut

The yield of first-time endoscopic ultrasonography in screening individuals at a high risk of developing pancreatic cancer

Am J Gastroenterol

Original article
Pancreas, biliary tract, and liver
Mutant TP53 in Duodenal Samples of Pancreatic Juice From Patients With Pancreatic Cancer or High-Grade Dysplasia