Introduction

​Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer-related deaths, with a five-year survival rate under 10% [1]. Its lethality stems from its aggressive progression and strong chemotherapy resistance, both closely linked to epithelial-mesenchymal transition (EMT). EMT enables epithelial cells to gain migratory mesenchymal traits and is regulated by transcription factors such as SNAIL, SLUG, ZEB1/2, and TWIST [2]. Among these, SNAIL plays a major role in metastasis [3], while ZEB1 is associated with poor prognosis [4]. EMT is tightly modulated by networks including ELAVL1, which encodes the RNA-binding protein HuR. HuR stabilizes mRNAs by binding AU-rich elements (AREs) in their 3′ untranslated regions (3′UTRs), promoting their expression [5].

Aims & Methods

This study aimed to identify post-transcriptional interactions between ELAVL1 and EMT-related genes and assess their prognostic value in PDAC. mRNA was extracted from cancerous and adjacent normal tissues (n=65), converted to cDNA, and analyzed by RT-PCR. BxPC-3, MiaPaCa-2, and Su.86.86 cell lines were cultured under standard conditions. Immunoprecipitation was performed on 1–2×10⁷ cells using a mouse monoclonal anti-HuR antibody, with IgG as control. qRT-PCR assessed mRNA enrichment. Binding sites were annotated using CISBP-RNA, considering only RNAdirect-confirmed 3′UTR motifs. Target 3′UTR sequences were retrieved from GENCODE V47. Statistical analysis was performed using GraphPad Prism, applying the Wilcoxon signed-rank test, Kruskal-Wallis test with Dunn’s correction, and Spearman correlation. Data were presented as median ± interquartile range, with p<0.05 considered significant.

Results

​Expression levels of EMT-related genes and ELAVL1 varied according to survival outcomes. In short-term survivors (1–12 months), SNAIL and SLUG were significantly upregulated (2.19- and 1.92-fold), while ZEB2 was downregulated (0.65-fold). Medium-term survivors (13–35 months) showed reduced ZEB1 (0.53), SNAIL (0.30), SLUG (0.35), and TWIST (0.68). Long-term survivors (36–125 months) had the lowest levels of ZEB1 (0.24), ZEB2 (0.50), SLUG (0.24), TWIST (0.35), and ELAVL1 (0.72). ELAVL1 showed strong positive correlations with ZEB1, SNAIL, and SLUG (r = 0.74–0.76), whereas overall survival was weakly negatively correlated with the expression of ZEB1, SNAIL, SLUG, and TWIST (r = -0.25 to -0.32). Although ZEB1, SNAIL, and TWIST did not show significant survival differences individually, low SLUG predicted better early survival (36 vs. 15 months, p = 0.0278). ELAVL1 expression was stratified as high and low. High ELAVL1 corresponded to a 71.65-fold increase in ZEB1, whereas high ZEB1 reduced ELAVL1 by 96.1%. High ELAVL1 also led to a 312.35-fold increase in SNAIL, while low ELAVL1 still supported elevated SNAIL expression (8.45-fold). High SNAIL expression reduced ELAVL1 by 90%, though simultaneous high expression of both resulted in a 3.71-fold ELAVL1 increase. High SLUG reduced ELAVL1 by 93%, whereas low SLUG led to a 198.11-fold increase. Binding site analysis revealed ELAVL1 targets within 3′UTRs: 34 in ZEB1, 1 in SNAIL, and 9 in SLUG.

Conclusion

ELAVL1 (HuR) likely plays a central role in regulating EMT in PDAC through stabilization of key transcription factor mRNAs. High levels of ELAVL1, SNAIL, SLUG, and ZEB1 were associated with worse outcomes, particularly in short-term survivors. Conversely, low expression of SLUG was linked to improved survival. The strong correlations and validated binding sites suggest ELAVL1 as a promising prognostic biomarker and potential therapeutic target in PDAC.

References

1. Nakaoka K, Ohno E, Kawabe N, Kuzuya T, Funasaka K, Nakagawa Y, et al. Current status of the diagnosis of early-stage pancreatic ductal adenocarcinoma. Diagnostics (Basel). 2023;13(2).
2. Kaşıkcı E, Aydemir E, Bayrak ÖF, Şahin F. Inhibition of migration, invasion and drug resistance of pancreatic adenocarcinoma cells - role of Snail, Slug and Twist and small molecule inhibitors. Onco Targets Ther. 2020;13:5763-77.
3. Tang X, Sui X, Weng L, Liu Y. SNAIL1: Linking tumor metastasis to immune evasion. Front Immunol. 2021;12:724200.
4. Liu M, Zhang Y, Yang J, Zhan H, Zhou Z, Jiang Y, et al. Zinc-dependent regulation of ZEB1 and YAP1 coactivation promotes epithelial-mesenchymal transition plasticity and metastasis in pancreatic cancer. Gastroenterology. 2021;160(5):1771-1783.e1.
5. Majumder M, Chakraborty P, Mohan S, Mehrotra S, Palanisamy V. HuR as a molecular target for cancer therapeutics and immune-related disorders. Adv Drug Deliv Rev. 2022;188(114442):114442.

Introduction

Intraductal papillary mucinous neoplasms (IPMNs) are a common type of mucinous pancreatic cystic lesions and represent a precursor of pancreatic cancer. The WHO classification distinguishes three grades of IPMN — low-grade/high-grade intraepithelial neoplasia and IPMN-associated invasive carcinoma—with distinct clinical management. The correct estimation of IPMN biology in clinical practice is often challenging and relies particularly on the assessment of lesion morphology, as histological confirmation is seldom possible. Therefore, improved biomarkers are needed to guide personalized management of IPMN patients.
In our preliminary analysis of public RNA-seq datasets (healthy controls n=7; LGD IPMN n=6; HGD IPMN n=6; invasive IPMN n=3), DLGAP5 expression increased significantly with IPMN grade (ANOVA p=5.48×10⁻⁶). DLGAP5 is a microtubule-associated protein essential for mitosis and vesicular trafficking—processes implicated in pancreatic cancer progression.

Aims & Methods

We aimed to assess the utility of DLGAP5 as a marker of IPMN progression as well as identify potential surrogate cystic-fluid biomarkers. Because of the intracellular localization of DLGAP5, we did not expect it to be present in the cyst fluid. However, given the role of microtubules in protein secretion, we hypothesized that DLGAP5 regulates the secretion of distinct set of proteins, which are highly correlated with its expression.
Protein expression of DLGAP5 was evaluated by immunohistochemistry (Santa Cruz Biotechnology, sc-377044) on retrospective cohort of patients who underwent pancreatic resection for IPMN at our institution (n=45). Functional studies were conducted in vitro by modulating DLGAP5 expression in pancreatic cancer cell lines (PaTu 8902, MiaPaCa2) and IPMN precursor cell line (ASAN-PaCa). Mass spectrometry-based secretome analysis of IPMN precursor cells was performed to identify DLGAP5-associated secreted proteins with potential for clinical testing in cyst fluid.

Results

DLGAP5 staining density and intensity differed significantly among LGD, HGD, and carcinoma (Fisher–Freeman–Halton p<0.0001 for both). Post-hoc Fisher’s exact tests showed LGD vs HGD (density p<0.001; intensity p<0.001) and LGD vs carcinoma (density p<0.001; intensity p<0.001) to be significant; HGD vs carcinoma was not (density p=0.673; intensity p=0.414). Functional assays demonstrated that DLGAP5 knockdown in pancreatic cancer cells decreased proliferation, while its overexpression in IPMN precursor cells increased proliferation. Secretome profiling revealed eight proteins whose secretion was significantly increased in IPMN precursor cells overexpressing DLGAP5 compared to empty vector expressing cells. Among them, PRDX1, PRDX2, FABP5, and SCGB1D2 (p=0.03, 0.038, 0.035 and 0.009 respectively) have previously been identified in cyst fluid or serum of IPMN patients, indicating translational relevance.

Conclusion

In conclusion, we validated DLGAP5 as a marker for IPMN progression and identified several DLGAP5-dependent secreted proteins that may serve as surrogate biomarkers for stratification of the patients with IPMN. Prospective validation in cystic fluid samples is ongoing.

Introduction

Lumen-apposing metal stents (LAMSs) have significantly advanced endoscopic ultrasound (EUS)-guided treatment of pancreatic fluid collections (PFCs). However, their impact on clinical outcomes in real-world settings remains incompletely understood.

Aims & Methods

Using the Diagnosis Procedure Combination database—a nationwide inpatient database in Japan where LAMSs were approved in 2018—we identified 5,885 patients who underwent EUS-guided treatment for PFCs at 550 hospitals between 2010 and 2023. We examined the association between treatment period (before vs. after LAMS implementation at each hospital) and clinical outcomes. Multivariable logistic regression analyses were conducted to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for in-hospital mortality, bleeding, length of stay, and total healthcare costs, adjusting for potential confounders.

Results

Compared with patients treated in the pre-LAMS period, those treated in the post-LAMS period had a significantly higher risk of bleeding (adjusted OR, 1.56; 95% CI, 1.35–1.81). Additionally, the post-LAMS period was associated with an increase of approximately $3,300 (USD) in total healthcare costs, with an adjusted OR for being in the highest quartile of total costs of 1.55 (95% CI, 1.34–1.79). There was no significant difference in the risk of in-hospital mortality (adjusted OR, 1.21; 95% CI, 0.95–1.55) or prolonged hospitalisation (adjusted OR for top quartile, 0.98; 95% CI, 0.86–1.12).

Conclusion

The introduction of LAMSs was associated with an increased risk of bleeding and higher healthcare costs, without significant improvements in mortality or hospital stay duration. These findings suggest that routine use of LAMSs for EUS-guided PFC treatment should be reconsidered.

Disclosure

HI declares research funding from Boston Scientific Japan, Fujifilm, and Piolax Medical Devices; honoraria from Boston Scientific Japan, Century Medical, Create Medic, Fujifilm, Gadelius Medical, Hitachi Medical, Japan Lifeline, Kaneka, Kawasumi Laboratories, Olympus Medical, Piolax Medical Devices, Sumitomo Bakelite, UMIDAS, and Zeon Medical; and contributions from Boston Scientific Japan, Gadelius Medical, Japan Lifeline, and Zeon Medical. YN declares research funding from Boston Scientific Japan, Century Medical, Fujifilm, Gadelius Medical, HOYA Pentax Medical, Hitachi Medical, Kaneka, and Medico’s Hirata; and honoraria from Boston Scientific Japan, Fujifilm, Gadelius Medical, Hitachi Medical, J-MIT, Medico’s Hirata, and Olympus Medical. This work was not supported by any of those companies. No other conflicts of interest exist. The other authors declare that they have no conflicts of interest.

Introduction

Pancreatic fibrosis (PF) is a prominent histological feature of chronic pancreatitis and influences on vast majority of pancreatic disorders. So, PF early diagnosis gives a great opportunity to be a part of the strategy for preventing pancreatic diseases progression. It makes non-invasive PF assessment especially via biomarkers level measurement research perspective.

Aims & Methods

Aim was to evaluate the possibility of biomarkers levels measurement in PF grade assessment.
Methods. We measured interleukin-10 (IL-10) and interleukin-36 (IL-36), matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-9 (MMP-9), fibronectin (FN) and hyaluronic acid (HA) plasma levels from 165 patient who underwent surgery from April 2022 to February 2024 due to pancreatic tumors and advanced chronic pancreatitis. We included 69 (41.8%) male and 96 (58.2%) female patients, mean age was 57.91±13.5 (25-89). Serum levels of specified biomarkers were determined by enzyme immunoassay using commercial kits; data registration performed was carried out on a semi-automatic analyzer. PF morphological grade was assessed according to Klöppel & Maillet’s scoring system [1] and scaled into three grades: mild, moderate and severe PF.
This study was conducted within the healthcare research project “Assessment of pancreatic fibrosis as a prognostic factor for its diseases course” funded by Autonomous non-profit organization "Moscow Center for Innovative Technologies in Healthcare" administered by the Moscow Healthcare Department.

Results

Plasma FN (pFN) level was significantly (p=0.029) higher in patients with mild PF (pFN=110.7±72.7 μg/mL), than in patients with severe PF (pFN=74.9±32.8 μg/mL). The median pFN levels were significantly varied in groups with different grades of intralobular PF (p<0.05), but not in groups divided by perilobular PF grade (p>0.05). Moreover, pFN level significantly negatively correlated with PF integrative grade (Spearmen’s rho = -0.21, p=0.007). Regarding IL-10, IL36, MMP-1, MMP-9 and HA levels, there were no significant differences either in patients with different PF integrative grade, or in groups divided by perilobular and intralobular PF (p>0.05). We revealed no significant correlations between specified biomarkers levels and PF grade (p>0.05).

Conclusion

The FN serum level significantly decreased in patients with severe PF in contrast to patients with mild PF who had comparatively higher serum FN level. Oppositely, IL-10, IL36, MMP-1, MMP-9 and HA had shown no effect in PF grade assessment. It could be useful for noninvasive pancreatic fibrosis diagnosis and even for patients’ selection for advanced diagnostics.

References

1 Esposito I, Hruban RH, Verbeke C, et al. Guidelines on the histopathology of chronic pancreatitis. Recommendations from the working group for the international consensus guidelines for chronic pancreatitis in collaboration with the International Association of Pancreatology, the American Pancreatic Association, the Japan Pancreas Society, and the European Pancreatic Club. Pancreatology. 2020;20(4):586-593. doi:10.1016/j.pan.2020.04.009

Introduction

​Pancreatic fibrosis (PF), a pathological hallmark of chronic pancreatitis, serves as a critical mediator in the progression of numerous pancreatic disorders. Early identification of fibrotic changes presents a strategic opportunity to intervene in disease pathways, potentially curbing the advancement of pancreatic pathologies. It makes research of non-invasive radiological PF assessment perspective.

Aims & Methods

Aim of our study was to evaluate the possibility of multidetector computed tomography (MDCT) post-processing parameters in PF grade assessment.
Methods. Results of contrast-enhanced 128-row MDCT from 165 patients (mean age 57.9±13.5) who underwent surgery from April 2022 to February 2024 due to pancreatic tumors and advanced chronic pancreatitis were analyzed. We calculated contrast enhancement ratio (CR) between venous phase (VP) and non-contrast MDCT, and normalized CR (NCR) during the pancreatic phase (PP) and VP, also we measured pancreas attenuation during MDCT examination. PF morphological grade was assessed according to Klöppel & Maillet’s scoring system [1] and scaled into three grades: mild, moderate and severe PF.
This study was conducted within the healthcare research project “Assessment of pancreatic fibrosis as a prognostic factor for its diseases course” funded by Autonomous non-profit organization "Moscow Center for Innovative Technologies in Healthcare" administered by the Moscow Healthcare Department.

Results

The values of post-processing CT results were compared in groups with different degrees of PF. Mean CR were significantly higher (p=0.001) in patients with severe PF (CR 1.16±0.65 HU) than in patients with mild PF (CR 0.78±0.31 HU). CR significant increase (p=0.03) was found in patients with signs of inflammatory changes in the pancreas tissue (CR 1.14±0.6 HU) than in those without them (CR 0.81±0.3 HU). There were no significant differences between the NCRPP and NCRVP, and the degree of PF, which we had shown in publications of our preliminary results. We suppose the main reason of this fact is study groups enlargement as well as measurement technique improvement, led to results clarification.

Conclusion

The CR value increased in patients with severe degree of PF and decreased in patients with mild PF. Also, there was a significant relation between CR increase and presence of early signs of pancreatic inflammatory changes. Thus, there was a relationship between MDCT postprocessing results and morphological signs of PF, which can be used for pancreatic fibrosis non-invasive diagnosis and identification of additional signs of early chronic pancreatitis.

References

1. Esposito I, Hruban RH, Verbeke C, et al. Guidelines on the histopathology of chronic pancreatitis. Recommendations from the working group for the international consensus guidelines for chronic pancreatitis in collaboration with the International Association of Pancreatology, the American Pancreatic Association, the Japan Pancreas Society, and the European Pancreatic Club. Pancreatology. 2020;20(4):586-593. doi:10.1016/j.pan.2020.04.009