Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies, and approximately 15% of cases arise from intraductal papillary mucinous neoplasms (IPMNs). These precursor lesions can progress to invasive cancer through the accumulation of molecular alterations. To identify pathways associated with this transition, we performed comparative transcriptomic analyses of patient-derived IPMN and PDAC samples, followed by protein-level validation in tissue specimens and pancreatic cancer cell lines. Functional studies were conducted using the selective p38α inhibitor ralimetinib in two-dimensional cultures, three-dimensional tumorspheres, and patient-derived tumor organoids (PDTOs). Transcriptomic profiling revealed significant downregulation of the p38 pathway in IPMNs compared with PDAC, suggesting progressive pathway activation during pancreatic tumorigenesis. Immunohistochemical analyses confirmed increased phospho-activated p38α in PDAC tissues, particularly in lesions with advanced malignant features, while PDAC cell lines displayed higher levels of activated p38α than non-tumorigenic pancreatic epithelial cells. Pharmacological inhibition of p38α suppressed downstream MAPKAPK2 phosphorylation, induced apoptotic cell death, and reduced tumorsphere growth and viability. Consistently, PDTOs exhibiting elevated p38α activation showed increased sensitivity to ralimetinib treatment. Collectively, these findings identify p38α signaling as a pathway activated during PDAC progression and support further investigation of p38α-targeted therapies as a potential precision medicine strategy for pancreatic cancer.

Leucine-rich-repeat kinase 2 (LRRK2) is a multidomain protein highly expressed in immune cells and implicated in the regulation of immune functions including immune signaling, cytokine release and autophagy. LRRK2 is one of the therapeutic targets in Parkinson’s Disease (PD). Aberrant activation of LRRK2 can also contribute to intestinal inflammation, mainly in inflammatory bowel disease (IBD). Hence the modulation of LRRK2 may influence gut inflammation providing an improvement in disease outcomes. Over the years, microRNAs (miRNAs) have acquired much attention thanks to their potential as therapeutic targets in several pathological conditions, including inflammatory disorders. In this study, we aimed to examine the ability of miR-369-3p in the modulation of autophagy targeting LRRK2 expression. Bioinformatics analysis revealed that Lrrk2 is a target gene of miR-369-3p, and LRRK2 expression was increased in ulcerative colitis patients compared with that in a healthy control. In in vitro analysis, we validated that mimic transfection with miR-369-3p in Raw264.7 significantly reduced the expression of LRRK2 both in basal and inflammatory conditions. Moreover, the inhibition of LRRK2 limited the nuclear translocation of Nuclear factor kappa B (NF-κB) induced by lipopolysaccharide (LPS) stimulation. In turn, we found that, in inflammatory conditions, the intracellular increase in miR-369-3p precluded the inhibition of autophagy by LRRK2 by restoring autophagy marker light chain 3 (LC3)II/I ratio, BECLIN-1 and decreasing p62 expression. Furthermore, we detected that the upregulation of miR-369-3p decreased the release of pro-inflammatory mediators Tumor necrosis factor (TNF), C-C motif ligand 2/Monocyte chemoattractant protein-1 (CCL2/MCP-1), C-C motif ligand 3/Macrophage inflammatory protein-1 alpha (CCL3/MIP-1α) and C-C motif ligand 5/Regulated on activation, normal T-cell expressed and secreted (CCL5/RANTES) and increased the anti-inflammatory cytokine interleukin 10 (IL-10) in response to LPS. This study supports the anti-inflammatory potential of miR-369-3p in immune cells and suggests the potential of miR-369-3p as a therapeutic agent in the treatment of acute intestinal inflammatory conditions such as ulcerative colitis.

Angiogenesis is a key process in colorectal cancer (CRC) progression, with vascular endothelial growth factor A (VEGFA) being a major pro-angiogenic mediator. miR-195-5p has previously been identified by our group as a tumor-suppressive miRNA downregulated in CRC. Here, we aimed to investigate whether miR-195-5p contributes to CRC-associated angiogenenic processes through VEGFA regulation. Potential miR-195-5p targets were investigated through bioinformatic predictions and miRNA Target RNA immunoprecipitation. VEGFA expression was evaluated in public CRC datasets and paired tumor/adjacent tissues of a cohort of CRC patients by quantitative real-time PCR and immunohistochemistry. miR-195-5p gain- and loss-of-function experiments were performed in CRC cells to assess VEGFA and VEGFR2 expression and extracellular VEGF-A levels by ELISA. Endothelial tube formation assays using HUVECs were conducted to evaluate paracrine angiogenic effects. VEGFA expression was also analyzed in an AOM/DSS-induced murine CRC model treated with a miR-195-5p mimic. VEGFA was predicted as a conserved miR-195-5p target and was enriched in miR-195-5p-associated RISCs. VEGFA was overexpressed in CRC tissues compared to normal or adjacent non-tumor tissues. miR-195-5p restoration reduced VEGFA mRNA and protein expression, extracellular VEGF-A levels, and VEGFR2 protein levels in the HCT116 CRC cell line, whereas miR-195-5p inhibition produced the opposite molecular profile. Conditioned media from miR-195-5p-overexpressing CRC cells significantly impaired endothelial tube formation. Consistently, miR-195-5p in vivo administration reduced colonic Vegfa expression in AOM/DSS-treated mice. Overall, these findings indicate that miR-195-5p negatively modulates VEGFA expression and reduces the ability of CRC cell-derived conditioned media to support endothelial tube formation. This study extends the tumor-suppressive functions previously attributed to miR-195-5p by supporting its involvement in the regulation of angiogenesis-related processes in CRC.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease with marked molecular heterogeneity, resulting in highly variable responses to biologic therapies. Despite the efficacy of Infliximab (IFX), many patients fail to achieve sustained remission. Identifying biomarkers predictive of treatment response is essential for advancing precision medicine in UC. Gene expression profiling of colonic mucosal biopsies from IFX responders, IFX non-responders, and healthy controls was performed using a multiplex array, followed by technical confirmation by RT-qPCR and immunohistochemical assessment to further characterize tissue expression and biological relevance. IFX non-responders displayed a stromal–immune molecular signature marked by extracellular matrix remodeling, stromal remodeling, integrin-mediated signaling, and persistent inflammatory pathways. SPARC and SPP1 emerged as the most prominent biomarkers, with significantly increased gene and protein expression in non-responder patients, highlighting their association with chronic tissue remodeling and therapeutic resistance. Mucosal resistance to IFX is associated with a specific stromal–immune profile characterized by SPARC and SPP1 upregulation. These findings suggest that both molecules may serve as promising candidate markers for patient stratification, highlighting the need for future prospective studies to explore their role in personalized management strategies for ulcerative colitis.

Pancreatic ductal adenocarcinoma frequently metastasises to the liver, although the mechanisms underlying hepatic pre-metastatic niche formation remain unclear. Small extracellular vesicles mediate tumour–host communication and may drive hepatic microenvironment reprogramming. This study investigated the effects of pancreatic ductal adenocarcinoma-derived small extracellular vesicles on extracellular matrix remodelling and epithelial–mesenchymal transition-related plasticity in hepatic cells. Small extracellular vesicles were isolated from pancreatic ductal adenocarcinoma cell lines (MIAPaCa-2, PANC-1) and from the serum of 25 patients, characterized, and administered to hepatic stellate (LX-2) and hepatocyte-like (HEPA-RG) cells. Cell viability and migration were evaluated by functional assays, morphology by scanning electron microscopy, and molecular changes by RT-PCR, Western blotting, and immunofluorescence. In LX-2 cells, small extracellular vesicles exposure increased metabolic activity, adhesion, and migration, while inducing morphological and molecular changes associated with extracellular matrix remodelling, including reduced collagen type I alpha 2 chain, vimentin, and E-cadherin expression. In HEPA-RG cells, viability was minimally affected, whereas migration and EMT-related plasticity were enhanced. Patient-derived small extracellular vesicles induced similar but less pronounced effects. Overall, pancreatic ductal adenocarcinoma-derived small extracellular vesicles induced early hepatic microenvironmental remodelling, supporting a potential role for tumour–liver crosstalk in pre-metastatic niche-associated processes, highlighting tumour–liver crosstalk as a potential therapeutic target.