Identifying intact and fibrotic parenchyma in pancreatic ductal adenocarcinomas using compression optical coherence elastography.

Intraoperative assessment of pancreatic quality, followed by sampling for the potential isolation of Langerhans islets for subsequent autotransplantation, is currently a key component of post-total pancreatectomy diabetes mellitus treatment. The aim of this study was to quantitatively evaluate pancreatic parenchymal stiffness using optical coherence elastography (OCE) imaging, and to investigate the utility of the OCE method as a potential indicator of islet yield after pancreatectomy. A total of 41 freshly excised human pancreatic specimens, containing pancreatic ductal adenocarcinoma (PDAC) and surrounding non-tumorous tissues post-pancreatectomy, were studied. In this research, the stiffness (Young’s modulus, kPa) and its color-coded 2D distribution were calculated for various pancreatic samples using compression OCE. Stiffness values were compared between intact pancreatic parenchyma (islet-poor and islet-rich) and pancreatic lesion groups (parenchymal fibrosis and/or PDAC invasion). The data were confirmed by histological analysis. In addition, the measured stiffness values for various morphological groups of the pancreatic samples were compared with the number of isolated islets obtained from pancreatic samples after collagenase treatment. The study demonstrated that OCE can effectively distinguish areas of pancreatic lesions and identify intact pancreatic parenchyma containing Langerhans islets. A highly significant increase in mean stiffness (p < 0.0001) was observed in postoperative pancreatic samples exhibiting signs of parenchymal fibrosis or PDAC invasion compared to unaffected, intact pancreatic parenchyma. For the first time, a relationship between stiffness values and the number of isolated pancreatic islets was demonstrated; in particular, the number of isolated islets significantly decreased (≤110 pcs/g) in samples exhibiting stiffness values above 150 kPa and below 75 kPa. The optimal stiffness range for the efficient isolation of islets (≥120 pcs/g) from pancreatic tissue was identified as 75–150 kPa. The study introduces a novel approach for rapid and objective intraoperative assessment of pancreatic tissue quality using real-time OCE data. This technique facilitates the identification of regions affected by pancreatic lesions and supports the selection of intact pancreatic parenchyma, potentially enhancing the accuracy of Langerhans islet yield predictions during surgical resection.

Decision letter: Neutrophil-mediated fibroblast-tumor cell il-6/stat-3 signaling underlies the association between neutrophil-to-lymphocyte ratio dynamics and chemotherapy response in localized pancreatic cancer: A hybrid clinical-preclinical study

Objective To explore the quantitative characteristics of pancreatic ductal adenocarcinoma (PDAC) in single-source dual energy spectral CT imaging. Methods From January 2013 to December 2014, 113 patients underwent dual phase contrast-enhanced gemstone spectral imaging (GSI) on Discovery CT 750 HD. All diagnoses were pathologically confirmed by surgery or biopsy. The spectral HU curves of PDAC were observed, the monochromatic CT values, the effective atomic number (Zeff), the iodine concentration (IC), water concentration (WC), and the corresponding normalized values (normalized monochromatic CT values, normalized Zeff, normalized IC, normalized WC) of the lesion and the pancreatic parenchyma in late arterial phase (AP) and portal venous phase (PP) were recorded. The measurements were performed three times repeatedly. Paired t test (normal distribution) or Wilcoxon test (non-normal distribution) were used for analyzing the differences between the two phases and between PDAC and pancreatic parenchyma. Results The monochromatic CT values of PDAC in AP were lower than in PP at each energy level and the difference was more marked at lower energy. The normalized monochromatic CT values increased with the increase of energy level in both AP and PP and the difference was more distinct at lower energy. The Zeff, IC and normalized IC of PDAC all had significant differences (P<0.05), while the WC, normalized Zeff, and normalized WC had no difference between AP and PP. The Zeff and IC of pancreatic parenchyma had significant differences (P<0.05), while the WC had no difference between AP and PP. The differences of Zeff, IC, and WC between PDAC and pancreatic parenchyma were significant in both two phases (P<0.05). Conclusions Dual phase CT spectral imaging showed characteristic quantitative parameters of pancreatic ductal adenocarcinoma. The monochromatic CT values, Zeff, and iodine concentration of PDAC were lower than those of pancreatic parenchyma in both AP and PP. The monochromatic CT values, Zeff, and iodine concentration of PDAC in late arterial phase were lower than those in portal venous phase. The differences were all more distinct at lower energy. Key words: Pancreatic neoplasms; Tomography, X-ray computed; Comparative study

Background: Pancreatic cancer is associated with 6.9% and 4% of all cancer-related deaths in the United States and Brazil, respectively. Pancreatic ductal carcinoma comprises 90% of cases, the majority being of adenocarcinoma subtype. Approximately 12% of periampullary tumors are adenocarcinomas of Vater papilla (ampullary adenocarcinomas); ampullary tumors are often associated with a better prognosis than ductal adenocarcinomas. Although genetic alterations were previously identified in pancreatic carcinomas, there is still a lack of effective treatment strategies. Therefore, the identification of new biomarkers, such as alterations in non-coding RNAs, is urgently needed for the development of novel molecularly targeted therapies for these cancers. microRNAs (miRNAs) are frequently deregulated and contribute to cancer development and progression and have potential prognostic and predictive value. Global miRNA expression profiling analysis in pancreatic cancer, followed by the identification of miRNA target genes may lead to the identification of clinically applicable biomarkers. The novel aspect of our work is the investigation of pancreatic tumors from Brazilian patients, with the inclusion of ampullary adenocarcinomas, a rare subtype. Objectives: To identify global miRNA expression profiles and miRNA target genes in pancreatic ductal and ampullary adenocarcinomas compared to paired histologically normal pancreatic tissue. Patients and Methods: 30 formalin fixed, paraffin embedded (FFPE) pancreatic carcinoma samples were used, including 24 pancreatic ductal adenocarcinomas (PDAC) and 6 ampullary adenocarcinomas (AMP). Paired histologically normal pancreatic tissues were used as controls. All tumor and normal tissues were needle microdissected (Leica EZ4 stereomicroscope). Global miRNA expression profiles were determined using the TaqMan Array Human MicroRNA Cards (TLDA) (card A, v3.0) (Life Technologies) platform. Data analysis was performed using the ExpressionSuite Software v1.0.3. Statistical analysis was performed to correlate miRNA expression with relevant clinical data, using SAS 9.3 software. Computational bioinformatics analysis was performed to identify miRNA target genes, as well as to construct protein-protein interaction and miRNA-gene targets networks. Results and Discussion: We identified 63 significantly deregulated (FC≥2 and p&amp;lt;0.05) miRNAs in PDAC (33 over- and 30 under-expressed) compared to paired histologically normal pancreatic tissue. In AMP, a group of 7 miRNAs was significantly deregulated (4 over- and 3 under-expressed) compared to normal pancreas. Our results showed differentially expressed miRNAs and a complexity of miRNA changes potentially associated to PDAC and AMP tumorigenesis. 3/7 miRNAs (miR-222, 148a and 375) were commonly deregulated in PDAC and AMP tumors. Furthermore, miRNA-gene targets networks were distinct in these different histological subtypes of pancreatic carcinomas. Global miRNA expression profiles showed that PDAC have a significantly higher number of altered miRNAs and a higher number of predicted miRNA target genes than AMP tumors, which could be potentially associated to disease progression and tumor aggressiveness in PDAC compared to AMP. Although these tumors have biological differences, commonly deregulated miRNAs in PDAC and AMP suggest that PDAC and AMP tumorigenesis may share commonly deregulated pathways. Conclusion: miRNAs identified herein may be associated to the biology of PDAC and AMP. Among the miRNAs exclusively deregulated in PDAC, we identified known and not previously reported (novel) miRNAs. In addition, we identified several miRNA target genes associated with tumor invasion, metastasis and poor patient prognosis. Functional in vitro and in vivo validation studies may elucidate the role of identified miRNAs as modulators of oncogenesis mechanisms in PDAC and AMP. T. Felix was funded through São Paulo Research Foundation (FAPESP), MSc. fellowship (2014/00367-4) Citation Format: Tainara F. Felix, Tomas Tokar, Maria A. M. Rodrigues, Rogerio A. Oliveira, Claudia N. Hasimoto, Juan C. Llanos, Robson F. Carvalho, Silvia R. Rogatto, Wan Lam, Igor Jurisica, Sandra A. Drigo, Patricia P. Reis.{Authors}. Differentially expressed microRNA profiles in pancreatic ductal and ampullary adenocarcinomas. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A28.

Objective To investigate the expression of KL-6 mucin in pancreatic ducatal tissue,and to explore its clinical significance.Methods All specimens form 38 patients who were diagnosed as pancreatic ductal adenocarcinoma were subjected to immunohistochemical analysis using KL-6 monoclonal antibody.Results 100％ specimens of pancreatic adenocarcinoma were found positive staining,and marked positive staining was 78.9％,remarkable positive staining was not found in the surrounding noncancer regions of the pancreatic tissues (72.3％).KL-6 mucin expression was statistical significance with the tumor development and differentiation.Conclusion Most of all pancreatic adenocarcinoma cases showed remarkable positive staining for KL-6 mucin,while the majority of surrounding non-cancer regions did not.The aberrant expression of KL-6 mucin is significantly related to aggressive tumor behaviors of pancreatic adenocarcinoma.KL-6 mucin may be a specific target to detect the pancreatic adenocarcinoma in molecular imaging study. Key words: Pancreatic ductal adenocarcinoma; KL-6 mucin; Immunohistochemical

Background: Zinc content and its flux from the exocrine pancreas are both novel candidates for molecular imaging of pancreatic ductal adenocarcinoma (PDAC) due to the markedly dysregulated homeostasis in PDAC lesions. Here, we introduce the use of a Gd-based zinc probe (GdL) to monitor stimulated exocrine function in the healthy and PDAC bearing mouse pancreas by MRI. Methods: Male adult mice were implanted with syngeneic PDAC cells into the pancreas and allowed to form PDAC for 1 and 2 weeks. Healthy mice were used as controls. Mice were fasted overnight and imaged in a 4.7 T scanner. Two 2D T1-weighted gradient echo scans were obtained as baseline (TE/TR = 2.9/125 ms, Avg = 4). Mice then received either 1) 0.07mmol/kg GdL (i.v.) plus 10 ug/kg caerulein (i.p.), 2) 0.07mmol/kg GdL (i.v.) plus Saline (i.p.), or 3) 0.01 mmol/kg Gd-DOTA (i.v.). Immediately following injections, serial 2D T1-weighted gradient echo scans were obtained for 25 minutes. PDAC mice receiving saline were considered as negative controls. Change in contrast to-noise ratio (CNR) of tumor and pancreas was quantified and reported as a function of time. Tumor and pancreas tissue were then fixed, and histological analysis was done to characterize the expression of zinc transporters in healthy pancreas and PDAC tissues. Results: Secretagogue-stimulated zinc secretion (SSZS) MRI of the exocrine pancreas relies on evoking zinc release from intracellular stores from acinar and ductal cells. After i.p. injection of the secretagogue caerulein in fasted healthy animals, we observed only a small enhancement in the pancreas. However, both groups of animals carrying orthotopically implanted PDAC cells (1-wk and 2-wks) responded in hyper-secretory fashion. Negative controls (saline group) did not show significant pancreatic or tumor enhancement. Importantly, the effect of caerulein in PDAC animals and the increase in overall pancreatic enhancement were only found in PDAC tumor bearing animals. Dysregulation of zinc trafficking was confirmed by immunohistochemistry staining for the zinc import transporters ZIP3 and ZIP4. These transporters were upregulated only in PDAC-bearing pancreas and the positive staining was concentrated mostly in the tumor parenchyma and gradually dissipated in the surrounding pancreatic tissue. Conclusions: Zinc flux is markedly upregulated in PDAC. By stimulating the natural zinc secretion from exocrine pancreatic cells using an exogenous secretagogue, caerulein, we can non-invasively monitor early malignant transformations in pancreatic parenchyma and tumor cells by MRI with a Gd-based zinc sensitive probe. Citation Format: Mozhdeh Sojoodi, Ian Ramsay, Eric Gale, Stephen C. Barrett, Motaz Qadan, Peter Caravan, Kenneth K. Tanabe, Veronica Clavijo Jordan. Imaging pancreatic ductal adenocarcinoma using a zinc-sensitive MRI contrast agent: A novel method to detect early-stage PDAC lesions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2454.

Increased Serotonin Signaling Contributes to the Warburg Effect in Pancreatic Tumor Cells Under Metabolic Stress and Promotes Growth of Pancreatic Tumors in Mice

Bombesin Receptors in Distinct Tissue Compartments of Human Pancreatic Diseases

S100P-Binding Protein, S100PBP, Mediates Adhesion through Regulation of Cathepsin Z in Pancreatic Cancer Cells

The purpose of this study was to evaluate the diagnostic performance of magnetization transfer (MT) imaging and multigradient echo magnetic resonance imaging (MRI) to quantify pancreatic fibrosis and lipomatosis in patients before pancreatoduodenectomy for postoperative risk stratification with histopathology as the reference standard. Twenty-four patients (age, 68 ± 8 years, 16 males) prospectively underwent quantitative MT imaging using a 2-dimensional gradient echo sequence with and without MT prepulse and multigradient echo imaging on a 3 T MRI 1 day before pancreatoduodenectomy due to adenocarcinoma of the pancreatic head region (n = 20), neuroendocrine tumor (n = 3), or intraductal papillary mucinous neoplasm (n = 1). Magnetization transfer ratio (MTR) and proton density fat fraction (PDFF) were measured in pancreatic tail (PT) and at the resection margin (RM). Histopathologically, pancreatic fibrosis was graded as mild, moderate, or severe (F1-F3), lipomatosis was graded as 0% to 10%, 11% to 30%, and greater than 30% fat deposition (L1-L3). In addition, MTR and histopathologic fibrosis was assessed in pancreatic adenocarcinoma. Mann-Whitney U test and Spearman correlation were used. Patients with advanced pancreatic fibrosis (F3) showed a significantly higher MTR compared with the F1 group at the RM and PT (38 ± 4 vs 32.3 ± 1.6, P = 0.018 and 39.7 ± 5.5 vs 31.2 ± 1.7, P = 0.001). Spearman correlation coefficient of MTR and fibrosis grade was r = 0.532 (P = 0.011) and 0.554 (P = 0.008), respectively. Pancreatic parenchyma with advanced fat deposition (L2-L3) showed significantly higher PDFF compared with lipomatosis grade L1 (RM: P = 0.002 and PT: P = 0.001). Proton density fat fraction of pancreatic parenchyma exhibited a moderate and significant correlation with histopathologic lipomatosis grade (RM: r = 0.668 and PT: r = 0.707, P < 0.001). Magnetization transfer ratio was significantly higher in pancreatic adenocarcinoma compared with pancreatic parenchyma (44 ± 5.5 vs 36.0 ± 4.4 and 37.4 ± 5.4, P = 0.004). Multiparametric MRI of the pancreas including MTR and PDFF maps may provide quantitative and noninvasive information on pancreatic fibrosis and lipomatosis before surgery.

A positive microscopic margin (R1) following resection of pancreatic ductal adenocarcinoma (PDAC) can occur in up to 80% of patients and is associated with reduced survival and increased recurrence. Our aim was to characterize the impact of neoadjuvant therapy (NAT) on survival and recurrence in patients with PDAC following an R1 resection. A retrospective analysis of patients with PDAC who underwent pancreatectomy from 2008 to 2017 was performed. Patients were staged according to the American Joint Committee on Cancer 8th edition and stratified based on resection margin (R0 vs. R1) and treatment sequence (NAT vs. surgery first [SF]). Conditional survival analysis was performed using Cox regression and inverse probability weighted estimates. Among 580 patients, 59% received NAT and 41% underwent SF. On final pathology, the NAT cohort had smaller tumors and less lymph node (LN) positivity (p<0.05). NAT was not associated with an R1 resection (50%, p=0.653). Compared with the R1 cohort, the R0 cohort had a higher median overall survival (OS; 39.6 vs. 22.8months; hazard ratio [HR] 1.6, p<0.001) and disease-free survival (DFS; 19 vs. 13months; HR 1.35, p=0.004). After risk adjustment, NAT was not associated with OS, regardless of margin status (R0, 95% confidence interval [CI] (-)7.31-27.07, p=0.26; or R1, 95% CI (-)36.99-15.25, p=0.42). However, NAT was associated with improved DFS in the R1 cohort (95% CI 1.79-11.91, p=0.008) but not in the R0 cohort (95% CI (-)11.22-10.54, p=0.95). An R0 resection remains an important determinant of overall and disease-free survival, even when NAT is administered. For patients with an R1 resection, receipt of NAT may prolong DFS.

Adhesion molecules are cell surface glycoproteins that are important in cell-to-cell and cell-to-extracellular matrix interactions. In the present study, we analyzed the adhesion molecules ICAM-1 (intercellular adhesion molecule-1), VCAM-1 (vascular cell adhesion molecule-1), and ELAM-1 (endothelial leukocyte adhesion molecule-1) in human pancreatic cancer. ICAM-1, VCAM-1, and ELAM-1 were analyzed in 20 pancreatic cancer specimens and 20 normal pancreatic tissues. mRNA expression encoding ICAM-, VCAM-1, and ELAM-1 was assessed with Northern blot analysis. The distribution and localization of ICAM-1, VCAM-1, and ELAM-1 was determined in the pancreatic specimens by immunohistochemistry. Northern blot analysis revealed a 5.4-fold increase of ICAM-1 (P<0.01) and a 3.7-fold increase in VCAM-1 (P<0.01) mRNA expression in cancer samples in comparison with normal controls. In contrast, ELAM-1 mRNA levels did not show significant differences between the cancer and the normal tissues. Immunohistochemical analysis of cancer tissues showed strong immunostaining for ICAM-1 and VCAM-1, and faint immunostaining for ELAM-1 in the pancreatic cancer cells. Fibrotic or noncancerous pancreatic tissue adjacent to the cancer mass was devoid of any immunoreactivity for ICAM-1, ELAM-1, and VCAM-1. In contrast, the normal pancreas exhibited no immunoreactivity of ICAM-1, ELAM-1, and VCAM-1. Enhanced expression of ICAM-1 and VCAM-1 in human pancreatic cancers suggests a role in tumor pathogenesis. The increase of these adhesion molecules might influence the detachment of cancer cells in the primary tumor, might contribute to cancer cell migration and the spread of cancer cells to distant organs, or both.

395 Background: Neoadjuvant therapy (NAT) for pancreatic ductal adenocarcinoma (PDAC) is associated with improved overall survival (OS), and this has led to its rising use. The aim of this study was to evaluate the correlation between use of NAT and OS among patients with PDAC. Methods: This population-level study evaluated the Spearman correlation between the annual proportion of patients receiving NAT and the annual 1-year and 5-year OS, respectively, using the 2004-2015 National Cancer Database. Annual 1-year and 5-year OS was calculated from year of diagnosis using Kaplan-Meier survival analysis. All patients with a confirmed diagnosis of PDAC (histology code 8500), without any metastasis, and who underwent an R0 or R1 resection were included. Results: A total of 18,852 patients (median age 67 (IQR 60–74); 49.4% female) with PDAC underwent an R0/R1 resection from 2004 to 2015. Among these patients, there was a significantly positive correlation between the proportion of patients who received NAT (12.1%; n = 2,133) and 1-year OS (Spearman’s rho = 0.9091; P = 0.0001) and 5-year OS (Spearman’s rho = 0.7833; P = 0.01), respectively. Patients who underwent R0 resection (n = 14,547; median age 67 (IQR 60-74); 49.9% female) had a significantly positive correlation between those who received NAT (13.1%; n = 1,773) and 1-year OS (Spearman’s rho = 0.8818; P = 0.0003) and 5-year OS (Spearman’s rho = 0.7333; P = 0.02), respectively. Among 9,142 patients who had upfront resectable disease with R0 resection margin status (median age 68 (IQR 60–75); 49.8% female), there was a significantly positive correlation between proportion of patients who received NAT (9.1%; n = 781) and 1-year OS (Spearman’s rho = 0.7273; P = 0.01) and 5-year OS (Spearman’s rho = 0.8000; P = 0.0096), respectively. Conclusions: Between 2004 and 2015 there has been an increase in the use of NAT for patients with PDAC. Concurrently, the OS has also increased during this time period. This study demonstrates that there is a statistically significant and positive correlation between the proportion of patients with R0/R1 resected PDAC who received NAT and 1-year OS and 5-year OS, respectively.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease associated with a poor prognosis and high morbidity. Early diagnosis and complete tumor removal are still the principal factors extending life expectancy in PDAC patients. Here, the relationship of neurexophillin 1 (NXPH1) and NXPH2 expressions with clinicopathological parameters of PDAC was evaluated. Immunohistochemical analysis of NXPH1 and NXPH2 was performed in one tissue microarray of 96 surgical specimens, including normal pancreatic duct tissue (n = 5), PDACs at various stages of differentiation (n = 77), and pancreatic neuroendocrine tumors (n = 14). The intensity of both NXPH1 and NXPH2 staining was weak in only a small proportion of benign pancreatic ductal epithelial cells and was significantly higher in most PDAC specimens than in non-neoplastic pancreatic tissue specimens. However, no correlation of the expression of these PDAC biomarkers with tumor grades, T, N, and American Joint Committee on Cancer (AJCC) pathologic stages was established. Contrary to our expectation, the immunohistochemical results of NXPH1 and NXPH2 were inversely correlated with N stage in PDAC. NXPH1 and NXPH2 as PDAC biomarkers may be used to identify patients with this tumor, help delineate appropriate surgical margins, and identify lymph node metastasis in imaging studies.

Invasive intraductal papillary mucinous neoplasms (I-IPMNs) are considered more indolent than conventional pancreatic ductal adenocarcinoma (PDAC). Although neoadjuvant therapy (NAT) is widely adopted in PDAC, its role in I-IPMN remains unclear. This study aimed to evaluate the impact of NAT on survival in I-IPMN compared with PDAC. The study enrolled I-IPMN and PDAC patients undergoing resection at the University of Colorado Hospital between 2013 and 2023. Prognostic factors for overall survival (OS) were identified using Cox models. OS was compared between histology/NAT subgroups using the Kaplan-Meier method and the log-rank test, stratified by resectability. Of 500 patients (413 PDAC, 87 I-IPMN), 289 PDAC and 34 I-IPMN patients received NAT, and the I-IPMN patients showed slightly longer median OS than the PDAC patients (30.2 vs 28.1 months; p = 0.04). In the entire cohort, worse OS was associated with borderline resectable (BR)/locally advanced (LA) disease, elevated cancer antigen 19-9 (CA19-9), node-positive disease, lymphovascular invasion, perineural invasion, and absence of adjuvant therapy. In the resectable cohort (n = 311), NAT was associated with longer OS in PDAC (61.9 vs 34.2 months; p < 0.001), but not in I-IPMN (not reached vs 47.9 months; p = 0.74). Survival did not differ between NAT-treated resectable I-IPMN and PDAC (p = 0.95). In the BR/LA cohort treated with NAT (n = 183), OS was similar between I-IPMN and PDAC (17.1 vs 22.0 months; p = 0.69). In resectable I-IPMN, NAT was not associated with improved survival. Comparable survival between NAT-treated BR/LA I-IPMN and PDAC suggests a need for further research on treatment outcomes.