Human P2X7 receptor variants Gly150Arg and Arg276His polymorphisms have differential effects on risk association and cellular functions in pancreatic cancer

Lara Magni, Haoran Yu, Nynne M. Christensen, Mette H. Poulsen, Alexander Frueh, Ganga Deshar, Astrid Z. Johansen, Julia S. Johansen, Stephan A. Pless, Niklas R. Jørgensen, Ivana Novak*

*Corresponding author af dette arbejde

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Abstract

Background
The purinergic P2X7 receptor (P2X7R) plays an important role in the crosstalk between pancreatic stellate cells (PSCs) and cancer cells, thus promoting progression of pancreatic ductal adenocarcinoma (PDAC). Single nucleotide polymorphisms (SNPs) in the P2X7R have been reported for several cancers, but have not been explored in PDAC.

Materials and methods
Blood samples from PDAC patients and controls were genotyped for 11 non-synonymous SNPs in P2X7R and a risk analysis was performed. Relevant P2X7R-SNP GFP variants were expressed in PSCs and cancer cells and their function was assayed in the following tests. Responses in Ca2+ were studied with Fura-2 and dye uptake with YO-PRO-1. Cell migration was monitored by fluorescence microscopy. Released cytokines were measured with MSD assay.

Results
Risk analysis showed that two SNPs 474G>A and 853G>A (rs28360447, rs7958316), that lead to the Gly150Arg and Arg276His variants, had a significant but opposite risk association with PDAC development, protecting against and predisposing to the disease, respectively. In vitro experiments performed on cancer cells and PSCs expressing the Gly150Arg variant showed reduced intracellular Ca2+ response, fluorescent dye uptake, and cell migration, while the Arg276His variant reduced dye uptake but displayed WT-like Ca2+ responses. As predicted, P2X7R was involved in cytokine release (IL-6, IL-1β, IL-8, TNF-α), but the P2X7R inhibitors displayed varied effects.

Conclusion
In conclusion, we provide evidence for the P2X7R SNPs association with PDAC and propose that they could be considered as potential biomarkers.
OriginalsprogEngelsk
Artikelnummer148
TidsskriftCancer Cell International
Vol/bind24
Udgave nummer1
Antal sider16
ISSN1475-2867
DOI
StatusUdgivet - 2024

Bibliografisk note

Funding Information:
Expert technical assistance of Dorthe Nielsen and Majken Gudmundsson is greatly appreciated. Imaging data were collected at the Center for Advanced Bioimaging (CAB) Denmark, University of Copenhagen. IN and NRJ are members of PRESTO COST Action (CA21130).

Funding Information:
Open access funding provided by Copenhagen University. This research was funded by the Danish Council for Independent Research Medical Sciences, Grant Number 8020-00254B, Marie Sk\u0142odowska-Curie COFUND Doctoral Programme TALENT, Grant Number H2020-MSCA-COFUND-2017-801199.

Publisher Copyright:
© The Author(s) 2024.

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