Purinergic GPCR-integrin interactions drive pancreatic cancer cell invasion

Elena Tomas Bort; Megan D Joseph; Qiaoying Wang; Edward P Carter; Nicolas J Roth; Jessica Gibson; Ariana Samadi; Hemant M Kocher; Sabrina Simoncelli; Peter J McCormick; Richard P Grose

doi:10.7554/elife.86971

Purinergic GPCR-integrin interactions drive pancreatic cancer cell invasion

Elena Tomas Bort, Megan D Joseph, Qiaoying Wang, Edward P Carter, Nicolas J Roth, Jessica Gibson, Ariana Samadi, Hemant M Kocher, Sabrina Simoncelli, Peter J McCormick, Richard P Grose

Department of Life Sciences

Research output: Contribution to journal › Article › peer-review

12 Citations (SciVal)

Abstract

Pancreatic ductal adenocarcinoma (PDAC) continues to show no improvement in survival rates. One aspect of PDAC is elevated ATP levels, pointing to the purinergic axis as a potential attractive therapeutic target. Mediated in part by highly druggable extracellular proteins, this axis plays essential roles in fibrosis, inflammation response, and immune function. Analyzing the main members of the PDAC extracellular purinome using publicly available databases discerned which members may impact patient survival. P2RY2 presents as the purinergic gene with the strongest association with hypoxia, the highest cancer cell-specific expression, and the strongest impact on overall survival. Invasion assays using a 3D spheroid model revealed P2Y2 to be critical in facilitating invasion driven by extracellular ATP. Using genetic modification and pharmacological strategies, we demonstrate mechanistically that this ATP-driven invasion requires direct protein-protein interactions between P2Y2 and αV integrins. DNA-PAINT super-resolution fluorescence microscopy reveals that P2Y2 regulates the amount and distribution of integrin αV in the plasma membrane. Moreover, receptor-integrin interactions were required for effective downstream signaling, leading to cancer cell invasion. This work elucidates a novel GPCR-integrin interaction in cancer invasion, highlighting its potential for therapeutic targeting.

Original language	English
Article number	e86971
Journal	eLife
Volume	12
DOIs	https://doi.org/10.7554/elife.86971
Publication status	Published - 21 Mar 2023

Bibliographical note

Funding Information:
We thank the Barts Pancreatic Tissue Bank (BPTB) for providing pancreatic tissue slides presented in this work. BPTB is supported by Pancreatic Cancer Research Fund and we thank all its members, in particular, Prof. Claude Chelala, Christine Hughes, Ahmet Imrali, and Amina Hughes for help, as well as Consultant Pathologist Dr. Joanne Chin-Aleong and members of Tissue Access Committee and Operations Group. We thank Dr. Ann-Marie Baker for her expertise in RNAscope experiments. This work was supported by a Medical Research Council (MRC) iCase award to PJM and RPG from Barts Charity and the MRC Doctoral Training Programme for ETB at Queen Mary University of London (Project MRC0227). NJR acknowledges the QMUL MRC Doctoral Training Program (MR/N014308/1). MDJ acknowledges support from the BBSRC (BB/T008709/1) via the London Interdisciplinary Doctoral Programme and SS acknowledges financial support from the Royal Society through a Dorothy Hodgkin fellowship (DHF\R1\191019) and a Research Grant (RGS\R2\202038). This work was supported by Cancer Research UK (CRUK) awarded to EPC and RPG (A27781) and a CRUK Centre grant to Barts Cancer Institute (A25137). Diagrams were generated using BioRender.

Data Availability
All data generated or analysed during this study are included in the manuscript and supporting file, or online resources are fully referenced. Human PDAC tumour data were generated by TCGA Research Network (https://www.cancer.gov/tcga) and by the Clinical Proteomic Tumour Analysis Consortium (https://www.proteomics.cancer.gov). The Genotype-Tissue Expression (GTEx) Project was used for the analysis of normal pancreatic tissue samples (https://gtexportal.org).

Keywords

Humans
Cell Line, Tumor
Pancreatic Neoplasms/pathology
Carcinoma, Pancreatic Ductal/pathology
Neoplasm Invasiveness/genetics
Adenosine Triphosphate/metabolism
Integrins/metabolism
Cell Proliferation/genetics
Cell Movement
Gene Expression Regulation, Neoplastic
Receptors, Purinergic P2Y2/genetics

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.7554/elife.86971Licence: CC BY

Cite this

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title = "Purinergic GPCR-integrin interactions drive pancreatic cancer cell invasion",

abstract = "Pancreatic ductal adenocarcinoma (PDAC) continues to show no improvement in survival rates. One aspect of PDAC is elevated ATP levels, pointing to the purinergic axis as a potential attractive therapeutic target. Mediated in part by highly druggable extracellular proteins, this axis plays essential roles in fibrosis, inflammation response, and immune function. Analyzing the main members of the PDAC extracellular purinome using publicly available databases discerned which members may impact patient survival. P2RY2 presents as the purinergic gene with the strongest association with hypoxia, the highest cancer cell-specific expression, and the strongest impact on overall survival. Invasion assays using a 3D spheroid model revealed P2Y2 to be critical in facilitating invasion driven by extracellular ATP. Using genetic modification and pharmacological strategies, we demonstrate mechanistically that this ATP-driven invasion requires direct protein-protein interactions between P2Y2 and αV integrins. DNA-PAINT super-resolution fluorescence microscopy reveals that P2Y2 regulates the amount and distribution of integrin αV in the plasma membrane. Moreover, receptor-integrin interactions were required for effective downstream signaling, leading to cancer cell invasion. This work elucidates a novel GPCR-integrin interaction in cancer invasion, highlighting its potential for therapeutic targeting.",

keywords = "Humans, Cell Line, Tumor, Pancreatic Neoplasms/pathology, Carcinoma, Pancreatic Ductal/pathology, Neoplasm Invasiveness/genetics, Adenosine Triphosphate/metabolism, Integrins/metabolism, Cell Proliferation/genetics, Cell Movement, Gene Expression Regulation, Neoplastic, Receptors, Purinergic P2Y2/genetics",

author = "\{Tomas Bort\}, Elena and Joseph, \{Megan D\} and Qiaoying Wang and Carter, \{Edward P\} and Roth, \{Nicolas J\} and Jessica Gibson and Ariana Samadi and Kocher, \{Hemant M\} and Sabrina Simoncelli and McCormick, \{Peter J\} and Grose, \{Richard P\}",

note = "Funding Information: We thank the Barts Pancreatic Tissue Bank (BPTB) for providing pancreatic tissue slides presented in this work. BPTB is supported by Pancreatic Cancer Research Fund and we thank all its members, in particular, Prof. Claude Chelala, Christine Hughes, Ahmet Imrali, and Amina Hughes for help, as well as Consultant Pathologist Dr. Joanne Chin-Aleong and members of Tissue Access Committee and Operations Group. We thank Dr. Ann-Marie Baker for her expertise in RNAscope experiments. This work was supported by a Medical Research Council (MRC) iCase award to PJM and RPG from Barts Charity and the MRC Doctoral Training Programme for ETB at Queen Mary University of London (Project MRC0227). NJR acknowledges the QMUL MRC Doctoral Training Program (MR/N014308/1). MDJ acknowledges support from the BBSRC (BB/T008709/1) via the London Interdisciplinary Doctoral Programme and SS acknowledges financial support from the Royal Society through a Dorothy Hodgkin fellowship (DHF\textbackslash{}R1\textbackslash{}191019) and a Research Grant (RGS\textbackslash{}R2\textbackslash{}202038). This work was supported by Cancer Research UK (CRUK) awarded to EPC and RPG (A27781) and a CRUK Centre grant to Barts Cancer Institute (A25137). Diagrams were generated using BioRender. Data Availability All data generated or analysed during this study are included in the manuscript and supporting file, or online resources are fully referenced. Human PDAC tumour data were generated by TCGA Research Network (https://www.cancer.gov/tcga) and by the Clinical Proteomic Tumour Analysis Consortium (https://www.proteomics.cancer.gov). The Genotype-Tissue Expression (GTEx) Project was used for the analysis of normal pancreatic tissue samples (https://gtexportal.org).",

year = "2023",

month = mar,

day = "21",

doi = "10.7554/elife.86971",

language = "English",

volume = "12",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications Ltd",

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TY - JOUR

T1 - Purinergic GPCR-integrin interactions drive pancreatic cancer cell invasion

AU - Tomas Bort, Elena

AU - Joseph, Megan D

AU - Wang, Qiaoying

AU - Carter, Edward P

AU - Roth, Nicolas J

AU - Gibson, Jessica

AU - Samadi, Ariana

AU - Kocher, Hemant M

AU - Simoncelli, Sabrina

AU - McCormick, Peter J

AU - Grose, Richard P

N1 - Funding Information: We thank the Barts Pancreatic Tissue Bank (BPTB) for providing pancreatic tissue slides presented in this work. BPTB is supported by Pancreatic Cancer Research Fund and we thank all its members, in particular, Prof. Claude Chelala, Christine Hughes, Ahmet Imrali, and Amina Hughes for help, as well as Consultant Pathologist Dr. Joanne Chin-Aleong and members of Tissue Access Committee and Operations Group. We thank Dr. Ann-Marie Baker for her expertise in RNAscope experiments. This work was supported by a Medical Research Council (MRC) iCase award to PJM and RPG from Barts Charity and the MRC Doctoral Training Programme for ETB at Queen Mary University of London (Project MRC0227). NJR acknowledges the QMUL MRC Doctoral Training Program (MR/N014308/1). MDJ acknowledges support from the BBSRC (BB/T008709/1) via the London Interdisciplinary Doctoral Programme and SS acknowledges financial support from the Royal Society through a Dorothy Hodgkin fellowship (DHF\R1\191019) and a Research Grant (RGS\R2\202038). This work was supported by Cancer Research UK (CRUK) awarded to EPC and RPG (A27781) and a CRUK Centre grant to Barts Cancer Institute (A25137). Diagrams were generated using BioRender. Data Availability All data generated or analysed during this study are included in the manuscript and supporting file, or online resources are fully referenced. Human PDAC tumour data were generated by TCGA Research Network (https://www.cancer.gov/tcga) and by the Clinical Proteomic Tumour Analysis Consortium (https://www.proteomics.cancer.gov). The Genotype-Tissue Expression (GTEx) Project was used for the analysis of normal pancreatic tissue samples (https://gtexportal.org).

PY - 2023/3/21

Y1 - 2023/3/21

N2 - Pancreatic ductal adenocarcinoma (PDAC) continues to show no improvement in survival rates. One aspect of PDAC is elevated ATP levels, pointing to the purinergic axis as a potential attractive therapeutic target. Mediated in part by highly druggable extracellular proteins, this axis plays essential roles in fibrosis, inflammation response, and immune function. Analyzing the main members of the PDAC extracellular purinome using publicly available databases discerned which members may impact patient survival. P2RY2 presents as the purinergic gene with the strongest association with hypoxia, the highest cancer cell-specific expression, and the strongest impact on overall survival. Invasion assays using a 3D spheroid model revealed P2Y2 to be critical in facilitating invasion driven by extracellular ATP. Using genetic modification and pharmacological strategies, we demonstrate mechanistically that this ATP-driven invasion requires direct protein-protein interactions between P2Y2 and αV integrins. DNA-PAINT super-resolution fluorescence microscopy reveals that P2Y2 regulates the amount and distribution of integrin αV in the plasma membrane. Moreover, receptor-integrin interactions were required for effective downstream signaling, leading to cancer cell invasion. This work elucidates a novel GPCR-integrin interaction in cancer invasion, highlighting its potential for therapeutic targeting.

AB - Pancreatic ductal adenocarcinoma (PDAC) continues to show no improvement in survival rates. One aspect of PDAC is elevated ATP levels, pointing to the purinergic axis as a potential attractive therapeutic target. Mediated in part by highly druggable extracellular proteins, this axis plays essential roles in fibrosis, inflammation response, and immune function. Analyzing the main members of the PDAC extracellular purinome using publicly available databases discerned which members may impact patient survival. P2RY2 presents as the purinergic gene with the strongest association with hypoxia, the highest cancer cell-specific expression, and the strongest impact on overall survival. Invasion assays using a 3D spheroid model revealed P2Y2 to be critical in facilitating invasion driven by extracellular ATP. Using genetic modification and pharmacological strategies, we demonstrate mechanistically that this ATP-driven invasion requires direct protein-protein interactions between P2Y2 and αV integrins. DNA-PAINT super-resolution fluorescence microscopy reveals that P2Y2 regulates the amount and distribution of integrin αV in the plasma membrane. Moreover, receptor-integrin interactions were required for effective downstream signaling, leading to cancer cell invasion. This work elucidates a novel GPCR-integrin interaction in cancer invasion, highlighting its potential for therapeutic targeting.

KW - Humans

KW - Cell Line, Tumor

KW - Pancreatic Neoplasms/pathology

KW - Carcinoma, Pancreatic Ductal/pathology

KW - Neoplasm Invasiveness/genetics

KW - Adenosine Triphosphate/metabolism

KW - Integrins/metabolism

KW - Cell Proliferation/genetics

KW - Cell Movement

KW - Gene Expression Regulation, Neoplastic

KW - Receptors, Purinergic P2Y2/genetics

UR - https://www.scopus.com/pages/publications/85151541434

U2 - 10.7554/elife.86971

DO - 10.7554/elife.86971

M3 - Article

C2 - 36942939

SN - 2050-084X

VL - 12

JO - eLife

JF - eLife

M1 - e86971

ER -

Purinergic GPCR-integrin interactions drive pancreatic cancer cell invasion

Abstract

Bibliographical note

Keywords

UN SDGs

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