P2X7 receptor antagonism by AZ10606120 significantly depletes glioblastoma cancer stem cells in vitro

Liyen K. Kan; Matthew Drill; Padmakrishnan C. Jayakrishnan; Richard P. Sequeira; Paul G. Sanfilippo; Catriona McLean; Martin Hunn; David A. Williams; Terence J. O’Brien; Katharine J. Drummond; Mastura Monif

Brain Research Bulletin (Sep 2024)

P2X7 receptor antagonism by AZ10606120 significantly depletes glioblastoma cancer stem cells in vitro

Liyen K. Kan,
Matthew Drill,
Padmakrishnan C. Jayakrishnan,
Richard P. Sequeira,
Paul G. Sanfilippo,
Catriona McLean,
Martin Hunn,
David A. Williams,
Terence J. O’Brien,
Katharine J. Drummond,
Mastura Monif

Affiliations

Liyen K. Kan: Department of Neuroscience, Central Clinical School, Monash University, Melbourne, Victoria, Australia
Matthew Drill: Department of Neuroscience, Central Clinical School, Monash University, Melbourne, Victoria, Australia
Padmakrishnan C. Jayakrishnan: Department of Neuroscience, Central Clinical School, Monash University, Melbourne, Victoria, Australia
Richard P. Sequeira: Department of Neuroscience, Central Clinical School, Monash University, Melbourne, Victoria, Australia
Paul G. Sanfilippo: Department of Neuroscience, Central Clinical School, Monash University, Melbourne, Victoria, Australia
Catriona McLean: Department of Pathology, The Alfred, Melbourne, Victoria, Australia
Martin Hunn: Department of Neurosurgery, The Alfred, Melbourne, Victoria, Australia
David A. Williams: Department of Physiology, The University of Melbourne, Melbourne, Victoria, Australia
Terence J. O’Brien: Department of Neuroscience, Central Clinical School, Monash University, Melbourne, Victoria, Australia; Department of Neurology, The Royal Melbourne Hospital, Melbourne, Victoria, Australia; Department of Neurology, The Alfred, Melbourne, Victoria, Australia
Katharine J. Drummond: Department of Neurosurgery, The Royal Melbourne Hospital, Melbourne, Victoria, Australia
Mastura Monif: Department of Neuroscience, Central Clinical School, Monash University, Melbourne, Victoria, Australia; Department of Physiology, The University of Melbourne, Melbourne, Victoria, Australia; Department of Neurology, The Royal Melbourne Hospital, Melbourne, Victoria, Australia; Department of Neurology, The Alfred, Melbourne, Victoria, Australia; Correspondence to: Department of Neuroscience, Level 6, The Alfred Centre, 99 Commercial Rd, Melbourne, VIC 3004, Australia.

Journal volume & issue: Vol. 215
p. 110996

Abstract

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Glioblastoma is the most aggressive and lethal primary brain malignancy with limited treatment options and poor prognosis. Self-renewing glioblastoma cancer stem cells (GSCs) facilitate tumour progression, resistance to conventional treatment and tumour recurrence. GSCs are resistant to standard treatments. There is a need for novel treatment alternatives that effectively target GSCs. The purinergic P2X receptor 7 (P2X7R) is expressed in glioblastomas and has been implicated in disease pathogenesis. However, the roles of P2X7R have not been comprehensively elucidated in conventional treatment-resistant GSCs. This study characterised P2X7R channel and pore function and investigated the effect of pharmacological P2X7R inhibition in GSCs. Immunofluorescence and live cell fluorescent dye uptake experiments revealed P2X7R expression, and channel and pore function in GSCs. Treatment of GSCs with the P2X7R antagonist, AZ10606120 (AZ), for 72 hours significantly reduced GSC numbers, compared to untreated cells. When compared with the effect of the first-line conventional chemotherapy, temozolomide (TMZ), GSCs treated with AZ had significantly lower cell numbers than TMZ-treated cultures, while TMZ treatment alone did not significantly deplete GSC numbers compared to the control. AZ treatment also induced significant lactate dehydrogenase release by GSCs, indicative of treatment-induced cytotoxic cell death. There were no significant differences in the expression of apoptotic markers, Annexin V and cleaved caspase-3, between AZ-treated cells and the control. Collectively, this study reveals for the first time functional P2X7R channel and pore in GSCs and significant GSC depletion following P2X7R inhibition by AZ. These results indicate that P2X7R inhibition may be a novel therapeutic alternative for glioblastoma, with effectiveness against GSCs resistant to conventional chemotherapy.

Published in Brain Research Bulletin

ISSN: 1873-2747 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Medicine: Internal medicine: Neurosciences. Biological psychiatry. Neuropsychiatry
Website: https://www.sciencedirect.com/journal/brain-research-bulletin

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