Synthesis and functionalization of protease-activated nanoparticles with tissue plasminogen activator peptides as targeting moiety and diagnostic tool for pancreatic cancer.

BACKGROUND: Functionalized nanoparticles (NPs) are one promising tool for detecting specific molecular targets and combine molecular biology and nanotechnology aiming at modern imaging. We aimed at ligand-directed delivery with a suitable target-biomarker to detect early pancreatic ductal adenocarcinoma (PDAC). Promising targets are galectins (Gal), due to their strong expression in and on PDAC-cells and occurrence at early stages in cancer precursor lesions, but not in adjacent normal tissues. RESULTS: Molecular probes (10-29 AA long peptides) derived from human tissue plasminogen activator (t-PA) were selected as binding partners to galectins. Affinity constants between the synthesized t-PA peptides and Gal were determined by microscale thermophoresis. The 29 AA-long t-PA-peptide-1 with a lactose-functionalized serine revealed the strongest binding properties to Gal-1 which was 25-fold higher in comparison with the native t-PA protein and showed additional strong binding to Gal-3 and Gal-4, both also over-expressed in PDAC. t-PA-peptide-1 was selected as vector moiety and linked covalently onto the surface of biodegradable iron oxide nanoparticles (NPs). In particular, CAN-doped maghemite NPs (CAN-Mag), promising as contrast agent for magnetic resonance imaging (MRI), were selected as magnetic core and coated with different biocompatible polymers, such as chitosan (CAN-Mag-Chitosan NPs) or polylactic co glycolic acid (PLGA) obtaining polymeric nanoparticles (CAN-Mag@PNPs), already approved for drug delivery applications. The binding efficacy of t-PA-vectorized NPs determined by exposure to different pancreatic cell lines was up to 90%, as assessed by flow cytometry. The in vivo targeting and imaging efficacy of the vectorized NPs were evaluated by applying murine pancreatic tumor models and assessed by 1.5 T magnetic resonance imaging (MRI). The t-PA-vectorized NPs as well as the protease-activated NPs with outer shell decoration (CAN-Mag@PNPs-PEG-REGAcp-PEG/tPA-pep1Lac) showed clearly detectable drop of subcutaneous and orthotopic tumor staining-intensity indicating a considerable uptake of the injected NPs. Post mortem NP deposition in tumors and organs was confirmed by Fe staining of histopathology tissue sections. CONCLUSIONS: The targeted NPs indicate a fast and enhanced deposition of NPs in the murine tumor models. The CAN-Mag@PNPs-PEG-REGAcp-PEG/tPA-pep1Lac interlocking steps strategy of NPs delivery and deposition in pancreatic tumor is promising.

Intraductal papillary mucinous neoplasms of the pancreas: radiological predictors of malignant transformation and the introduction of bile duct dilation to current guidelines.

OBJECTIVE: To evaluate the current guidelines as a model to predict malignancy and to determine further radiological predictors of malignancy in intraductal papillary mucinous neoplasms (IPMNs). METHODS: 384 patients who had undergone a pancreatic operation with the pathological diagnosis of IPMN as well as applicable pre-operative imaging (CT/MRI) were included in the study. Images were evaluated retrospectively in consensus by two radiologists, using a standardized checklist. Descriptive statistics, binary logistic regression and receiver operator curve analysis were performed to assess the International Consensus Guidelines and other radiological predictors of clinical malignancy (defined as carcinoma in situ and invasive carcinoma). RESULTS: The best independent predictors of malignancy (n = 191) were solid components [odds ratio (OR) 3.98], parenchymal atrophy with main pancreatic duct dilation 5-9 mm (OR: 5.1) and common bile duct (CBD) dilation (OR: 31.26). >96% of all cases with CBD dilation were malignant IPMNs (positive-predictive value 96.4%; negative-predictive value 63.1%). Analysis of the current guidelines showed a diagnostic improvement with the addition of CBD dilation on determining the malignancy of IPMNs (sensitivity 82.2%/86.9%; specificity 72.7%/74.6%). Subanalysis of branch duct intraductal papillary mucinous neoplasms (BD-IPMNs; n = 168) also resulted in a diagnostic improvement with the addition of CBD dilation (sensitivity 28.6%/45.2%; specificity 92.9%/92.1%). The best independent predictors of malignancy for BD-IPMNs were parenchymal atrophy (OR: 4.00) and CBD dilation (OR: 29.3). Frequency analysis revealed that even small BD-IPMNs had already undergone malignant transformation (≤1 cm: 15%; 1-2 cm: 26%; 2-3 cm: 20%) with about 10% of those having a dilated bile duct. CONCLUSION: CBD dilation was a significant positive predictor of malignancy in IPMNs regardless of their size. ADVANCES IN KNOWLEDGE: Introduction of CBD dilation as a radiological predictor for malignancy might increase the diagnostic accuracy of current imaging-based guidelines.

Cyst Features and Risk of Malignancy in Intraductal Papillary Mucinous Neoplasms of the Pancreas: Imaging and Pathology.

BACKGROUND: Intraductal papillary mucinous neoplasms (IPMNs) display diverse macroscopic, histological, and immunohistochemical characteristics with typical morphological appearance in magnetic resonance imaging. Depending on those, IPMNs may show progression into invasive carcinomas with variable frequency. Overall, IPMN-associated invasive carcinomas are found in about 30% of all IPMNs, revealing phenotpyes comparable with conventional ductal adenocarcinomas or mucinous (colloid) carcinomas of the pancreas. In Sendai-negative side-branch IPMNs, however, the annual risk of the development of invasive cancer is 2%; thus, risk stratification with regard to imaging and preoperative biomarkers and cytology is mandatory. METHODS AND RESULTS: The present study addresses the radiological and interventional preoperative measures including histological features to determine the risk of malignancy and the prognosis of IPMNs. CONCLUSION: While preoperative imaging largely relies on the detection of macroscopic features of IPMNs, which are associated with a divergent risk of malignant behavior, in resected specimens the determination of the grade of dysplasia and the detection of an invasive component are the most important features to estimate the prognosis of IPMNs.