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Piro, G.

Publications and source records attributed to Piro, G..

3 recordsLinked to original sources

The axon guidance cue SEMA3A promotes the aggressive phenotype of basal-like PDAC

Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with few available therapeutic options. Two transcriptional cancer cell states have been consistently reported in PDAC, with the basal-like/squamous phenotype displaying a more aggressive biological behavior. Genetic and epigenetic dysregulation of the axon guidance pathway are common in PDAC, yet our understanding of its biological relevance is limited. Here, we investigated the functional role of the axon guidance cue SEMA3A in sustaining the progression of PDAC. We integrated available transcriptomic datasets of human PDAC with in situ hybridization analyses of patients tissues to find that SEMA3A is expressed by stromal cells and selectively enriched in epithelial cells of the basal-like/squamous subtype. We found that both cell-intrinsic and cell extrinsic factors instructing the basal-like/squamous subtype induce expression of SEMA3A in PDAC cells. In vitro, SEMA3A promoted cell migration as well as anoikis resistance. At molecular level, these phenotypes were associated with increased FAK signaling and enrichment of gene programs related to cytoskeleton remodeling. Accordingly, SEMA3A provided mouse PDAC cells with greater metastatic competence. In mouse orthotopic allografts, SEMA3A remodeled the TME by favoring infiltration of tumor-associated macrophages and exclusion of T cells. Mechanistically, SEMA3A functioned as chemoattractant for macrophages and favored their polarization towards an M2-like phenotype. In SEMA3Ahigh tumors, depletion of macrophages resulted in greater intratumor infiltration by CD8+ T cells and increased sensitivity of these tumors to chemotherapy. Overall, we show that SEMA3A contributes to the malignant phenotype of basal-like PDAC.

cancer biology↗

Transcriptomic dissection of Intraepithelial Papillary Mucinous Neoplasms progression by spatial technologies identified novel markers of pancreatic carcinogenesis.

Intraductal papillary mucinous neoplasms (IPMN) are one of the main precursor lesions of Pancreatic Ductal Adenocarcinoma (PDAC). The number of patients diagnosed with IPMN is constantly increasing. While in most of the cases IPMN present as indolent and nonmalignant entities, some degenerate into PDAC. The main mechanisms behind the IPMN progression to malignancy is still not fully understood. This is mainly due to the technological limit of the analyzes and to cysts heterogeneity whose malignant transformation potential is estimated based on size and degree of dysplasia without take in consideration the transformation time and therefore the real malignancy potential. Moreover, there is a general lack of consensus diagnostic markers to discern the Low-grade nonmalignant from High-grade malignant IPMN. In this study, we used two different Spatial Transcriptomic technologies (Visium, and GeoMx) to investigate the transcriptome of Low-grade dysplasia nonmalignant IPMN, Borderline IPMN, and High-grade dysplasia malignant IPMN to dissect the main mechanism that drives carcingenesis and to find specific markers associated to risk of tumor progression. We performed Visium spatial transcriptomics on two TMAs containing three Low-grade dysplasia nonmalignant IPMN, one Borderline IPMN, two High-grade dysplasia malignant IPMN, and two PDAC. We identified three specific epithelial cell clusters that characterize Low-grade dysplasia IPMN, Borderline IPMN, and High-grade dysplasia malignant IPMN and three transcription factors whose expression is associated with each grade. High-grade malignant IPMN were characterized by high expression levels of NKX6-2 and other markers of gastric isthmus cell lineage such as MUC5AC, PSCA, FERIL6. The SPDEF high IPMN cluster was found in Borderline IPMN and spotted in some regions of High-grade malignant IPMN. This cluster was characterized by high expression levels of SPDEF and other goblet cell lineage markers such as TFF2, AQP5, and MUC6. Low-grade nonmalignant IPMN were characterized by high expression levels of HOXB3, HOXB5, ZNF117. The association of these markers with the different grades was validated by GeoMx spatial transcriptomics on 43 additional IPMN samples divided according to their grade of dysplasia and malignancy. To better understand the transcriptomic changes along IPMN progression we performed spatial trajectory inference and we found that SPDEF high IPMN cluster cells are likely to evolve into NKX6-2 high malignant IPMN, and we found that this switch is characterized by the expression of NKX6-2 and other gastric markers. Taken together, the results presented here not only shed more light in to IPMN and PDAC oncogenesis, but also provided a plethora of novel malignancy-associated markers to be tested in diagnostic routine, to better delineate IPMN progression in patients and improve clinical management.

cancer biology↗

Subtilases turn on Pectin Methylesterase activity for a robust apoplastic immunity against pathogens

Plants involve a fine modulation of pectin methylesterase (PME) activity against microbes. PME activity can promote the cell wall stiffening and the production of damage signals able to induce defense responses. However, to date, the knowledge about the molecular mechanisms triggering PME activity during disease remains largely unknown. In this study, we explored the role of subtilases (SBTs), serine proteases consisting of 56 isoforms in Arabidopsis thaliana, as activators of PME activity in plant immunity. By using biochemical and reverse genetic approaches, we found that SBT3.3 and SBT3.5 are required to control PME activity and resistance to the fungus Botrytis cinerea. Arabidopsis sbt3.3 and sbt3.5 knockout mutants showed a reduced induction of PME activity and an increased susceptibility to B. cinerea. SBT3.3 expression is controlled by the damage-associated molecular patterns Oligogalacturonides. The SBT3.3 overexpression overactivates PME activity, but only during fungal infection, resulting in an increased expression of the defense-related genes and in an enhanced resistance to B. cinerea. We revealed that SBT3.3 and the Pro-PME17 isoforms are both secreted in the cell wall exploiting distinct protein secretion pathways and a different kinetic. Our findings point to SBTs as a mechanism to switch on PME activity and the related pectin integrity signaling to strengthen plant immunity against pests, in a timely manner to avoid the growth-defense trade-off. One sentence SummarySubtilases arm pectin methylesterase activity against pathogens to switch on pectin integrity signalling, reinforcing plant immunity and avoiding the growth-defense trade-offs

plant biology↗