IPMN are pancreatic cysts with malignant potential, deriving from epithelium of the main or collateral branch ducts. The progression of this lesion from nonmalignant Low-grade IPMN to High-grade IPMN and finally PDAC is marked by the acquisition of distinct molecular and histological features [
4]. Moreover, Bernard et al. [
6] showed by single-cell RNA sequencing that Low-grade IPMN possess a pro-inflammatory stroma that is gradually lost during the degeneration to PDAC. The prevalence of these cysts is currently in constant increase reaching around 8% of the global population [
25]. Thus, the identification of earliest molecular events responsible for PDAC onset remains critical not only for early detection and prevention, but also to provide an opportunity for the targeting of novel potential vulnerabilities.
Several papers reported the role of mucins in forming the physical barrier that protects normal epithelia from injury [
26]. It has been reported an aberrant expression of mucins in many types of disease including solid tumors [
27] and that mucins could modulate directly immune response [
11,
28‐
32]. It has also been demonstrated that mucin-specific
O-Glycosylation influences a broad range of signalling pathways to promote disease onset and progression sustaining proliferative and pro-tumorigenic signalling, contributing to pancreatic cancer phenotypes [
33,
34] and facilitating tumor immune escape [
11,
35]. In particular, aberrant mucin glycosylation on cancer cells leads to expression of atypical epitope resulting in a specific recognition and binding of cancer cell membrane glycosylation patterns, leading to apoptosis of cancer specific effector T cells [
36]. Aberrant expression of Glycosyltransferase enzyme GCNT3, a master regulator of mucin-specific
O-Glycosylation, was associated with increased production of mucins, reduced patient survival and chemoresistance in colon and ovarian cancer [
37]. Specific inhibition of GCNT3 by talniflumate, an orally available, small-molecule inhibitor and a muco-regulator, blocks mucous overproduction in patients with chronic respiratory diseases, and reduces cystic fibrosis [
14]. Moreover, it has been reported its activity in blocking pancreatic cancer cell proliferation and pancreatic intraepithelial neoplasia (PanIN) in vivo formation, mainly by downregulation of mucins expression [
13]. Gupta and colleagues [
10] also showed that talniflumate inhibits PDAC cell lines proliferation by downregulation of mucins and inhibition of their cell-intrinsic functions. On the basis of these promising results, we wanted to evaluate the effect on cancer immunity of talniflumate-mediated mucin suppression using organoids as models for both in vitro
, and in vivo experiments. To test the immune suppressive role of mucin barrier in a realistic model and to address the role of GCNT3 and mucins in immune escape, we established an in vitro recognition platform to track the cellular interactions between cancer and T cells. The organoid platform experiments, with both mouse and human 3D PDAC models, demonstrated that talniflumate abrogates mucins secretion facilitating T cells functions and highlighting the strong effect of this small molecule on immune modulation (Fig.
4 and Additional file
4: Figure S4). We confirmed these results in an orthotopic syngeneic mouse model with low immunogenic potential, in which talniflumate enhanced Gem/Txl efficacy provoking a significant reduction in tumor growth (Fig.
5a and b) and increas in overall survival (Fig.
5c). IF analyses suggested that the main effect of talniflumate was an increase of T cell infiltration into the tumor (Fig.
6b and c). However, these are proof of concept findings and further studies are needed for clinical applicability. Randomized clinical trials with the administration of talniflumate in addiction or not to standard chemotherapy could definitively demonstrate the efficacy of
O-glycosylation inhibition on chemotherapy response and finally on patient outcome. This translational path can guide future efforts to successfully translate our findings into healthcare.