Cadmium (Cd) is a metallic pollutant which has been classified as a possible pancreatic carcinogen. Cd uses similar ion channels than divalent cations to accumulate into the cells. These include the Transient Receptor Potential Cation Channel Subfamily M Member 7 (TRPM7) which has been also shown as a biomarker of pancreatic cancer. Pancreatic carcinogenesis is associated with the establishment of a fibrous stroma induced by pancreatic stellate cell (PSC) activation. Although several stress factors have been identified as activators of PSCs, the impact of pollutants, particularly Cd, is still unknown. Here, we chronically exposed human PSCs to Cd and we observed that Cd-exposed cells acquired a myofibroblast-like phenotype. Moreover, TRPM7 expression and activity were upregulated following Cd exposure. Both TRPM7 inhibition by silencing or NS8593 treatment prevented the Cd-induced PSC cell migration indicating that TRPM7 regulated PSC activation. We used a model of indirect co-culture to study the impact of PSC on MIA PaCa-2 cancer cell migration. Interestingly, we showed that Cd-exposed PSCs stimulated MIA PaCa-2 cancer cell migration to a greater extent than non-exposed PSCs. TRPM7 inhibition in PSCs abolished the migration of cancer cells. Finally, in a mouse model with the KRASG12D mutation inducing spontaneous pancreatic intraepithelial neoplasia, Cd exposure aggravates collagen deposition in fibrotic areas showing high α-SMA and TRPM7 expressions. In summary, our study showed that Cd exposure upregulates TRPM7 leading to PSC activation and aggravation of precancerous pancreatic fibrosis in vivo.

Timing-Dependent Effects of ω-3 Fatty Acids on Adipose Tissue and Tumorigenesis in Obesity-Driven Pancreatic Cancer in Mice.

Pancreatic cancer (PC) is a lethal malignancy with limited early detection strategies and poor therapeutic response. Emerging evidence implicates the gut microbiota in carcinogenesis, yet whether microbial alterations are causal or secondary remains uncertain. In this study, we integrated cross-sectional 16S rDNA sequencing, two-sample Mendelian randomization (MR), and mediation analysis to investigate the causal role of gut microbiota in PC risk. We profiled fecal microbiota in a Beijing-based cohort of 26 newly diagnosed PC patients and 9 healthy controls, revealing significant dysbiosis characterized by reduced microbial diversity, depletion of butyrate-producing genera (e.g., Faecalibacterium), and enrichment of pro-inflammatory taxa such as Olsenella. Using European GWAS summary data, MR analysis identified 17 gut microbial taxa causally associated with PC risk, including Olsenella and Pauljensenia sp000411415. Notably, higher abundance of Pauljensenia sp000411415 was associated with increased PC risk, an effect partially mediated by reduced circulating levels of octanoylcarnitine (C8) and glutarylcarnitine (C5-DC)-metabolites independently linked to lower PC risk. Population-matched MR in East Asian cohorts validated several causal associations, enhancing ancestral relevance. Our findings support a causal role for specific gut microbes in pancreatic carcinogenesis and highlight a Pauljensenia-acylcarnitine axis whereby microbial suppression of protective metabolites may contribute to disease development. This integrative approach bridges microbial dysbiosis with functional mechanisms, offering novel insights for microbiome-informed strategies in PC prevention and early detection.

Accumulating evidence has demonstrated that distinct tumour-promoting and tumour-restraining cancer-associated fibroblast (CAF) subtypes coexist in pancreatic ductal adenocarcinoma. To develop targeted CAF therapeutic strategies by reprogramming tumour-promoting CAF subtypes. We leveraged multiomics technologies to systematically identify and characterise CAF subtypes transcriptionally, epigenetically and spatially and correlate them with clinicopathological features. We found that complement-secreting CAFs (csCAFs), initially identified by our group and inflammatory CAFs (iCAFs) share significant overlap in their transcriptional profiles and chromatin accessibility. iCAFs specifically express transcription factors from the heme and oxidative homeostasis pathway and the activator protein 1 family, which are both involved in cellular response to oxidative stress. Notably, the composition of csCAFs among all CAFs declined during pancreatic carcinogenesis, while trajectory analysis showed that csCAFs could potentially differentiate into iCAFs. Spatially resolved analysis indicated that tumour regions with a higher csCAF composition were associated with lower levels of TGF-β ligands, fewer M2 tumour-associated macrophages and increased levels of lipid mediators. Additionally, we identified a spatially defined CXCL12-CXCR4 ligand-receptor interaction between csCAFs and T cells, but in distinct patterns between different metastatic organs. Patients with a higher composition of csCAFs have significantly longer overall survival and recurrence-free survival through multiplex immunohistochemistry and bulk RNA-seq deconvolution. Our study demonstrates that csCAFs may represent an early-stage iCAF subtype and suggests a promising strategy for reprogramming iCAFs into csCAFs.

712 Background: Pancreatic cancer is one of the most lethal gastrointestinal malignancies, with obesity and smoking as key risk factors. The widespread use of glucagon-like peptide-1 receptor agonists (GLP-1RAs) for diabetes and obesity has raised concern about pancreatic carcinogenesis. While early data were inconclusive, recent large cohorts suggest reduced risk. We conducted a meta-analysis to provide a pooled hazard ratio quantifying this association. Methods: Randomized trials and large observational cohorts reporting pancreatic cancer outcomes in GLP-1RA users were reviewed through February 2025. Hazard ratios (HRs) were extracted, log-transformed, and pooled using fixed-effects models. New-user, active-comparator, and landmark designs were prioritized. Studies without HRs were summarized qualitatively but excluded from pooling. Results: Dankner et al. reported HR 0.52 (95% CI 0.19–1.41) in a landmark new-user design versus basal insulin, with HR 0.75 (0.37–1.53) in a prevalent new-user analysis. Alchirazi et al. found HR 0.56 (0.44–0.72) versus insulin, HR 0.80 (0.73–0.89) versus DPP-4 inhibitors, HR 0.78 (0.69–0.89) versus SGLT2 inhibitors, and HR 0.84 (0.74–0.95) versus sulfonylureas. Wang et al. reported HRs from 0.42–0.82 across six comparator groups, with stronger effects in obese and smoking populations. Ayoub showed a seven-year risk of 0.1% in GLP-1RA users versus 0.2% in non-users, but no HR was reported. Kristensen’s randomized trial meta-analysis found HR 1.12 (0.77–1.63), consistent with no excess risk but limited by shorter follow-up. When seven HRs from four real-world studies and one trial meta-analysis were combined, GLP-1RA use was associated with a pooled HR of 0.79 (95% CI 0.72–0.86), representing a 21% risk reduction. Absolute incidence remained very low, not exceeding 0.2% at seven years. Conclusions: To our knowledge, this is one of the first pooled synthesis of hazard ratios showing that GLP-1RA therapy is not linked to increased pancreatic cancer risk and instead correlates with a significant reduction, particularly in obese and smoking populations. By integrating large real-world datasets with trial evidence, this analysis reframes GLP-1RAs from a potential liability to a therapy with possible risk-modifying effects on pancreatic carcinogenesis. Further research in GI cancer prevention and survivorship is warranted.

Digital polymerase chain reaction (dPCR) is widely recognized for its high sensitivity in detecting low-frequency variants; however, conventional 2-color systems have limited multiplex capacity. Expanding this capability is essential for simultaneous detection of multiple driver mutations in cancer-related genes. KRAS and GNAS are key driver genes in the early development of pancreatic cancer and its precursor lesions, and mutations in these genes are often present at low abundance in clinical samples. Two 6-color dPCR assays were developed using a droplet-based platform. PlexScreen-dPCR is a multicolored drop-off assay designed to screen for mutations in KRAS codons 12/13 and 61 and GNAS codon 201, without specifying individual variants. PlexID-dPCR employs variant-specific probes to distinguish among 14 predefined KRAS and GNAS mutations in a single reaction. The assays were validated using synthetic DNA, cell lines, 23 tissue samples, and 12 duodenal fluid samples. Customized primer/probe sets with 6 fluorophores were employed in a 6-color droplet dPCR system, and the limits of detection (LOD) were determined. PlexScreen-dPCR, applied in contrived samples, demonstrated LODs as low as 0.03% to 0.06%, enabling high-sensitivity detection of low-abundance mutations. PlexID-dPCR accurately identified all 14 variants in a single well. Both assays showed complete concordance with conventional methods, exhibiting a strong correlation for variant allele frequency quantification. These 6-color dPCR assays offer scalable solutions for improved throughput detection of KRAS and GNAS mutations. Their compatibility with commercially available platforms and streamlined workflow support their integration into clinical practice. Further optimization can enhance cluster interpretation in high-plex settings and facilitate expansion toward broader genomic targets.

ENPP1-Regulated Extracellular Purine Metabolism Drives Pancreatitis-Mediated Pancreatic Cancer.

Chronic pancreatitis (CP) is a risk factor for pancreatic cancer, with inherited cases conferring a markedly increased risk. The underlying mechanisms driving malignant transformation by CP remain poorly understood. Combining a recently developed mouse model of CP carrying the human carboxypeptidase A1 (CPA1) p.N256K mutation with the established KrasG12D pancreatic cancer model, we characterised mechanisms linking chronic inflammation to early pancreatic carcinogenesis. We crossed Cpa1 N256K mice (Cpa1) with Ptf1aCre;KrasLSL-G12D (KC). In Cre, Cpa1, KC and KC-Cpa1 mice, we performed phenotypical characterisation at five early time points and in an ageing cohort. Assessment of histology combined with both RNA-sequencing and single-cell RNA-sequencing was performed to analyse metaplasia, preneoplastic lesions and cellular heterogeneity. KC-Cpa1 pancreata displayed a stark increase in remodelling, fibrosis and formation of metaplastic lesions as compared with KC. Cpa1N256K induced extensive plasticity in both the acinar and ductal compartment, including an early acinar-to-ductal metaplasia state in acinar cells characterised by an upregulation of endoplasmic reticulum stress markers and an inflammatory ductal phenotype (iDucts). We characterised the complex cell-cell communication networks underlying both pancreatic inflammation and early carcinogenesis, revealing disease-specific signalling between ductal cells, granulocytes and fibroblasts. The humanised KC-Cpa1 mouse model reveals the interplay of inflammation in hereditary CP and carcinogenesis. Cpa1N256K -induced plasticity in acinar and ductal cells, inflammation and cell-cell interaction networks cooperate with KrasG12D in early pancreatic carcinogenesis.

Although single-cell analyses have advanced our understanding of pancreatic ductal adenocarcinoma (PDAC), most studies to date have focused on primary and metastatic tumors. Here, we map cell composition, phenotypic plasticity, and microenvironmental remodeling from human normal pancreas through preneoplastic lesions to PDAC, with the preneoplastic phase recognized as a critical window for carcinogenesis. We pinpoint genes that are persistently dysregulated throughout malignant transformation and are associated with a poor prognosis. Focusing on ductal and acinar cells as the principal origins of PDAC, we delineate malignant preneoplastic cell clusters that exhibit strong carcinogenic potential. Immune profiling reveals marked expansion and functional reprogramming of macrophages during disease progression. Integrative analysis with human PDAC bulk transcriptomic cohorts identifies candidate compounds, such as Brefeldin A, with potential for intervention in preneoplastic disease. Together, our study elucidates dynamic molecular and cellular mechanisms underlying PDAC carcinogenesis and provides actionable insights for early intervention and targeted therapy.

Abstract Introduction: The increasing incidence of early-onset pancreatic ductal adenocarcinoma (PDAC), defined as diagnosis before age 50, coincides with rising global exposures to carcinogens such as alcohol and tobacco during critical developmental windows. Emerging evidence suggests these exposures reprogram epithelial and stromal compartments early in life, priming the pancreas for oncogenic transformation. However, the molecular events linking these exposures to early tumorigenesis remain underexplored. Methods: To investigate alcohol- and tobacco-driven mechanisms of early-onset pancreatic carcinogenesis, we utilized both syngeneic orthotopic models and genetically engineered mouse models (GEMMs), including Ptf1aCreERTM/+, Ptf1aCreERTM/+;LSL-KrasG12D/+ (KC), KPC (KC with mutant p53) and Ptf1aCre/+;LSL-KrasG12D/+;Tgfbr2flox/flox (PKT) lines (both tumor-bearing and non-tumor). Chronic cigarette smoke exposure and alcoholic chronic pancreatitis (ACP) were initiated in early adulthood. single-cell RNA sequencing (scRNA-seq), bulk RNA-seq, immunophenotyping, and histopathology were performed to delineate epithelial, stromal, and immune cell changes. Acinar lineage tracing using Rosa26tdTomato/+ was integrated to track acinar-to-ductal metaplasia (ADM). Human PDAC (smokers and non-smokers) and chronic pancreatitis tissues were used for cross-species validation. Results: Both alcohol and tobacco exposure accelerated neoplastic progression and induced pronounced fibroinflammatory transformation of the tumor microenvironment (TME). scRNA-seq revealed the early emergence of ductal-like epithelial clusters with elevated Sox9, Pdx1, and Hnf1b-hallmarks of ADM. Ethanol- and smoke-exposed mice showed impaired acinar regeneration, persistent epithelial reprogramming, and an augmented PanIN burden. Immune profiling showed expanded MDSCs, M2 macrophages, and regulatory T cells in exposed models, mirroring immune-suppressive signatures seen in early-onset human PDAC samples. We show the molecular mechanism involved and the differences in cellular signaling between tobacco-induced and alcohol-induced carcinogenesis. Lineage tracing confirmed that acinar cells serve as a source for ductal lesions under repeated inflammatory insults, especially in the presence of oncogenic Kras. Conclusions: Early exposure to carcinogens from external sources, like tobacco and alcohol, triggers molecular changes in pancreatic epithelium and immune stroma, which accelerate neoplastic transformation. Such exposures result in persistent fibroinflammatory damage, encourage ADM, and establish a pro-tumorigenic microenvironment. Our research provides a mechanistic understanding of the increase in early-onset PDAC and advocates for using integrated technologies-GEMMs, scRNA-seq, and lineage tracing-to model early-life exposures as crucial for intervention targets. These findings emphasize the importance of prevention strategies aimed at modifiable exogenous risk factors during the critical stages of PDAC development. Citation Format: Nagaraj Nagathihalli. Exogenous carcinogenic exposures in early-onset pancreatic carcinogenesis: Insights from alcohol and tobacco-driven models [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: The Rise in Early-Onset Cancers—Knowledge Gaps and Research Opportunities; 2025 Dec 10-13; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2025;31(23_Suppl):Abstract nr C017.

Pancreatic cancer (PC) presents unique challenges to traditional immunotherapy due to its immunosuppressive microenvironment and low mutation burden. Research into antibody-mediated immune responses, which has been extensively applied in various cancer types, is also applicable to the study of pancreatic cancer. Using public Genome-wide association study (GWAS) data and genetic instruments, we conducted a two-sample Mendelian randomization (MR) analysis to explore causal relationships between 46 antibody-mediated immune responses and PC. We employed five MR methods. Cochran's Q statistic and corresponding p-values from the MR Egger and Inverse Variance Weighted methods assessed instrument heterogeneity. Egger intercept and MR pleiotropy residual sum and outlier (MR-PRESSO) methods detected potential horizontal pleiotropy, ensuring the robustness of our findings. Our study identified causal links between five antibody-mediated immune responses and pancreatic cancer risk. Chlamydia trachomatis momp A antibody levels were inversely correlated, while Anti-helicobacter pylori IgG seropositivity, Polyomavirus 2 JC VP1 antibody levels, and Merkel cell polyomavirus VP1 antibody levels showed positive correlations. Notably, Anti- Merkel cell polyomavirus IgG seropositivity exhibited strong positive associations across Inverse variance weighting (IVW), weighted median, and MR-Egger analyses (p < 0.05). These results suggest that specific Antibody-Mediated Immune Responses may modulate pancreatic carcinogenesis through immune activation or chronic inflammation. The protective association for Chlamydia trachomatis momp A contrasts with the risk-enhancing effects observed for anti-helicobacter pylori IgG and polyomaviruses, highlighting heterogeneous immunobiological pathways that warrant mechanistic investigation. Our study demonstrated a randomized causal effect between antibody-mediated immune responses and pancreatic cancer, offering a new perspective for clinical diagnosis and treatment of the disease.

The function of cytosolic aldolase A (ALDOA) in glycolysis is well recognized. However, the cytosol-to-nucleus redistribution of ALDOA and its nuclear function is poorly understood. Here, we uncover inflammatory factor-stimulated nuclear function of ALDOA in augmenting pancreatic carcinogenesis by activating NF-κB signaling in a ubiquitination-dependent manner. TNF-α-triggered K11- and K29-linked ubiquitination of ALDOA at Lys200 promotes its interaction with RelA/p65 and facilitates importin-β-dependent nuclear translocation, establishing a positive feedback regulation in the tumor microenvironment by elevating the TNF-α expression in pancreatic ductal adenocarcinoma (PDAC). USP4 is identified as a negative regulator that deubiquitinates ALDOA. Instead of broadly targeting ALDOA, which causes glycolysis impairment, the specific elimination of ALDOA ubiquitination enhances chemosensitivity and the synergistic effect of chemotherapy combined with p65-specific anti-inflammatory therapy by selectively suppressing inflammation-induced proliferation in cancer cells. Collectively, we unveil the multifaceted mechanisms by which ALDOA promotes PDAC carcinogenesis, from metabolic to gene regulatory perspectives, providing potential therapies combatting cancer.

Digital PCR-based detection of Cryptosporidium in Pancreatic Tissue and Saliva Samples of Cancer patients; Pancreatic cryptosporidiosis.

Abstract Pancreatic ductal adenocarcinoma (PDAC) remains a highly lethal cancer with hereditary chronic pancreatitis (CP) conferring a significantly increased risk. Hereditary CP is caused by mutations in genes, such as PRSS1, SPINK1, CTRC, or CPA1 and presents with variable onset. However, the underlying mechanisms through which CP contributes to oncogenic transformation remain poorly understood. To elucidate this, we generated a genetically engineered mouse model carrying CP-associated human p.N256K carboxypeptidase A1 (CPA1) mutation and oncogenic Kras mutation resulting in Ptf1a Cre ;Kras LSL-G12D ;Cpa1 N256K (KC-Cpa1) mouse strain. Histological analyses of the KC-Cpa1 pancreas revealed accelerated development of early steps of PDAC including acinar-to-ductal metaplasia (ADM), pancreatic intraepithelial neoplasia (PanIN) lesions, and extensive fibrosis. Ex vivo 3D acinar cultures from 8-week-old KC-Cpa1 mice pancreata demonstrated enhanced ADM formation compared to Ptf1a Cre (Cre), Cpa1 N256K/N256K (Cpa1), and Ptf1a Cre ;Kras LSL-G12D (KC). These findings suggest that inflammation induced by the Cpa1 N256K mutation synergizes with oncogenic Kras mutation to promote the early initiation of PDAC. To explore the cellular heterogeneity and transcriptional program of metaplastic cells, we performed single cell RNA sequencing of pancreata of KC-Cpa1, KC, Cpa1, and Cre which revealed Cpa1 N256K -induced exocrine plasticity marked by an early ADM state and inflammatory phenotype of ductal cells (iDucts). Furthermore, single cell RNA sequencing also suggested disease-specific signaling between ductal cells, granulocytes, and fibroblasts. These results support the utility of KC-Cpa1 mouse model for studying early stages of CP-induced PDAC. Citation Format: Tanvi V. Inamdar, Ferdinand Krannich, Nico Hesselbarth, Atul Verma, Teresa Vauti, Ghanem El Kassem, Jasmine Hillmer, Michael Boettcher, Ivonne Regel, Heidi Griesmann, Irene Esposito, Markus Glaß, Monika Hämmerle, Patrick Michl, Helmut Laumen, Jonas Rosendahl. Hereditary chronic pancreatitis induced plasticity cooperates with mutant Kras in early pancreatic carcinogenesis [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research—Emerging Science Driving Transformative Solutions; Boston, MA; 2025 Sep 28-Oct 1; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2025;85(18_Suppl_3):Abstract nr A026.

Protocol for adeno-associated virus-mediated gene delivery to accelerate pancreatic carcinogenesis in mice.

Pancreatic ductal adenocarcinoma (PDAC), one of the most aggressive forms of pancreatic cancer, is associated with poor survival outcomes and currently ranks as the third leading cause of cancer-related death in the United States. Despite its clinical significance, the mechanisms of PDAC development and progression remain, in part, poorly understood. In this study, we provide evidence of a novel role of sorting nexin 10 (SNX10), a member of the sorting nexin family, in the regulation of KRAS-induced pancreatic carcinogenesis. We demonstrate that SNX10 is downregulated in PDAC, especially in advanced cases. Furthermore, mutational analysis revealed SNX10 genetic alterations in PDAC cases. Functional studies demonstrated that SNX10 overexpression in human PDAC cells inhibited cell proliferation and colony formation. Moreover, SNX10 overexpression induced G1-phase cell-cycle arrest and decreased KRAS signaling activity. Using a novel Snx10 knockout mouse crossed with a Kras-driven PDAC model, we observed reduced survival, increased tumor cell proliferation, enhanced aggression, and heightened inflammation. Collectively, these findings highlight SNX10 as a tumor suppressor candidate in PDAC and underscore its promise as a foundation for new therapeutic approaches.Significance:SNX10 plays a crucial role in reducing pancreatic tumorigenesis. This discovery offers valuable insights into PDAC’s biology and the development of new effective treatments.

Normalizing Body Weight with a Dietary Strategy Mitigates Obesity-Accelerated Pancreatic Carcinogenesis in Mice.

Genomic instability, DNA damage response and telomere homeostasis in pancreatic cancer.

Pancreatic cancer is the third leading cause of cancer-related death in Northern America and fourth in Europe. Emerging evidence suggests that the pancreatic microbiome may play a significant role in the development and progression of this disease. Although the human microbiota contributes to health by supporting nutritional and hormonal homeostasis, modulating inflammation, detoxifying harmful compounds, and producing beneficial metabolites, several studies have implicated its crucial modulatory role in numerous diseases, including cancer. The main objective of this review is to investigate the specific relationship between the microbiome and pancreatic carcinogenesis. A comprehensive literature search will identify studies examining the microbiome in human samples of saliva, pancreatic fluid, bile, pancreatic tissue, and feces of patients with chronic pancreatitis, precancerous pancreatic lesions, and pancreatic cancer. Studies differentiating bacteria to at least the genus level will be prioritized. Eligible studies include randomized controlled trials and observational studies analyzing the human microbiome in patients with chronic pancreatitis, pancreatic precursor lesions, or pancreatic cancer compared to healthy controls. Studies analyzing nonhuman samples, single bacterial strains, or lacking comparator groups will be excluded. The following databases will be searched without any restrictions to the publication date up until December 2024: the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE (via PubMed), Embase, and Web of Science. Animal studies, case reports, and studies not reporting analyses of human samples are excluded. Details regarding blinding, risk of bias, and funding sources will be extracted and assessed. The main outcomes include the bacterial diversity in each sample type (stool, saliva, bile, intratumoral, and tissue) itemized for each diagnosis, identifying differentially abundant or depleted taxa, and evaluating the correlation of specific bacteria with disease prevention or progression and clinical outcomes. Data extraction will be performed independently by two reviewers. Risk-of-bias assessment will be performed using Cochrane tools appropriate for each study design. Comparisons will be analyzed by descriptive statistics, and meta-analyses will be performed when applicable. The review will be conducted according to the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. In summary, this systematic review aims to synthesize studies analyzing microbiome profiles in patients with chronic pancreatitis, precursor lesions, and pancreatic cancer, focusing on identifying bacterial diversity and specific taxa related to disease progression and development of cancer in comparison to healthy controls and will include a thorough critical appraisal of the available literature. Anticipated limitations include heterogeneity in microbiome sampling methods and potential variability in taxonomic resolution across studies. PROSPERO CRD42023487995.

Warthin tumours (WT), the second most common benign salivary gland tumour, are histopathologically composed of bilayered oncocytic epithelial cells with occasional metaplastic epithelium. Tuft cells, which are chemosensory epithelial cells, are present in WT. Tuft cells serve various roles by producing physiologically active substances, such as prostaglandins (PGs). PGD2 released from tuft cells is crucial for tissue repair and inhibiting pancreatic carcinogenesis. However, whether or not tuft cells in WT produce PGD2 has not yet been elucidated. The present study aimed to investigate the production of PGD2 in POU class 2 homeobox 3 (POU2F3; a specific tuft cell marker)‑positive cells of WT and normal salivary glands. Consecutive patients with WT who underwent surgical resection were selected. Dual immunohistochemical staining for POU2F3 and haematopoietic PGD synthase (H‑PGDS) was performed. The present study included 28 patients with WT of the parotid gland (all male patients; median age, 68 years). The conventional bilayered oncocytic epithelium was present in all tumours; squamous metaplastic epithelium and conventional bilayered oncocytic epithelium were observed in 16 patients. Dual immunohistochemical analysis revealed that POU2F3+/H‑PGDS‑ cells were exclusively present in the striated duct of the normal salivary gland, and abundant POU2F3‑positive tuft cells were observed in both the conventional bilayered oncocytic and metaplastic squamous epithelia of WT. The median ratio of POU2F3‑positive cells expressing H‑PGDS was significantly higher in the conventional oncocytic epithelium (89.9%) than in the metaplastic squamous epithelium (10.6%) of WT (P=0.00044). The present results suggest a link between tissue injury to the striated duct and the pathogenesis of WT, and that PGD2 released from POU2F3‑positive cells in the conventional bilayered oncocytic epithelium is associated with ongoing tissue injury. Further studies are warranted to clarify the function of tuft cells in WT and gain deeper insights into the pathogenesis of WT.