A novel basal DeltaNP63 (P40)-expressing pancreatic cell type undergoes expansion and reorganization in chronic pancreatitis

Sandrina Martens, Mathias Van Bulck, Katarina Coolens, Farzad Esni, Gunter Leuckx, Henry Heimberg, Luc Bouwens, Patrick Jacquemin, Francesca Spagnoli, Francisco X Real, Pieter in 't Veld, Pierre Lesfevre, Ilse Rooman

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Introduction: The basal-like/squamous molecular subtype of pancreatic ductal adenocarcinoma (PDAC) is characterised by expression of basal cell markers such as deltaNp63 (p40), an isoform of the p53 homologue p63. In other organs, tumours that display basal features arise from normal epi-thelia in which a subset of cells with a basal phenotype can be identified. These deltaNp63-expressing basal cells have stem cell capacity that can be activated in tissue regeneration and can be the cell of origin of cancer. Hitherto, in the normal pancreatic epithelium, such basal cells have not been reported.Aims & Methods: We used multiplex immunofluorescence for protein detection and BaseScope for RNA detection and quantified the stainings with HALO software. We assessed (deltaN)p63 expression in histologically normal human pancreas tissue of subjects without a history of pancreatic disease (n=120, obtained for an islet transplant program) or with chronic pancreatitis (n=11). In addition, we analysed the pancreas of normal mice (n=14) and mice with caerulein-induced acute (n=5) or chronic (n=5) pancreatitis. In addi-tion to deltaNp63, other basal cell markers were assessed in a subset of these human and mouse samples. We developed in house a method (see results) for sample clearing and 3D protein detection. Samples were imaged with light sheet fluorescence mi-croscopy and analysed with Arivis software.Results: We detected rare (approx. 1/8000) deltaNp63+ cells in the healthy adult human pancreas, but not in mouse pancreas. In the human samples, their frequency increased approximately 30-fold in chronic pancreatitis. The deltaNp63+ cells were located in or near ducts and were positive for cytokeratin (CK) 19 but lacked markers of differentiated pancreatic duct cells such as CA19.9 and lacked MUC6, a marker of pancreatic ductal glands. Moreover, they had low or undetectable levels of SOX9 and exhibited cyto-plasmic expression of HNF1b, in contrast to the strong nuclear expression of these transcription factors in the neighbouring duct cells. In addition, the deltaNp63+ cells co-expressed other typical markers of basal cells such as CK5, CK14 and S100A2. To image this rare basal cell type in 3D at single cell resolution, we developed a method, based on FLASH, CUBIC and ECi principles, that could be completed in two weeks. This method used formalin-fixed paraffin embedded archival human pan-creatic specimens and preserved the native 3D pancreatic morphology and volume. We revealed that the deltaNp63+/ CK5+ cells were located as singular cells in the duct lining, between the basal lamina and the lumi-nal duct cells, or as clusters branching from these ducts. Unlike in normal pancreas, in chronic pancreatitis samples these basal cells were organised in more abundant gland-like structures containing several layers around a larger lumen.Conclusion: For the first time, we identified and imaged in 3D a novel cell population in the adult human pancreas, i.e. basal cells that increase upon chronic inflammation. The identification of a basal cell population in the pancreas raises important questions about their developmental origin, fate, and role in regeneration and disease, including the basal-type PDAC. Disclosure: Nothing to disclose
Originele taal-2English
Pagina's (van-tot)123-124
Aantal pagina's2
TijdschriftUnited European Gastroenterology Journal
Volume8
DOI's
StatusPublished - okt 2020
EvenementUEG Week Virtual 2020 -
Duur: 11 okt 202013 okt 2020

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