Functional expression of the ATP-gated P2X7 receptor in human iPSC-derived astrocytes

• Published in: Purinergic signalling 2024-06, Vol.20 (3), p.303-309
• Main Authors: Kesavan, Jaideep, Watters, Orla, de Diego-Garcia, Laura, Méndez, Aida Menéndez, Alves, Mariana, Dinkel, Klaus, Hamacher, Michael, Prehn, Jochen H. M., Henshall, David C., Engel, Tobias
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - analogs & derivatives; Adenosine Triphosphate - metabolism; Adenosine Triphosphate - pharmacology; Agonists; Antagonists; Astrocytes; Astrocytes - drug effects; Astrocytes - metabolism; Biomedical and Life Sciences; Biomedicine; Brief Report; Cancer Research; Cell death; Cell differentiation; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell lines; Cell proliferation; Cells, Cultured; Central nervous system; Drug screening; Glial fibrillary acidic protein; Human Physiology; Humans; Induced Pluripotent Stem Cells - metabolism; Inflammation; Neuromodulation; Neurosciences; Neurotransmission; Pharmacology/Toxicology; Pluripotency; Purinergic P2X Receptor Antagonists - pharmacology; Receptors, Purinergic P2X7 - metabolism
• ISSN: 1573-9538; 1573-9546; 1573-9546
• DOI: 10.1007/s11302-023-09957-8

Activation of the ATP-gated P2X7 receptor (P2X7R), implicated in numerous diseases of the brain, can trigger diverse responses such as the release of pro-inflammatory cytokines, modulation of neurotransmission, cell proliferation or cell death. However, despite the known species-specific differences in its pharmacological properties, to date, most functional studies on P2X7R responses have been analyzed in cells from rodents or immortalised cell lines. To assess the endogenous and functional expression of P2X7Rs in human astrocytes, we differentiated human-induced pluripotent stem cells (hiPSCs) into GFAP and S100 β-expressing astrocytes. Immunostaining revealed prominent punctate P2X7R staining. P2X7R protein expression was also confirmed by Western blot. Importantly, stimulation with the potent non-selective P2X7R agonist 2′,3′-O-(benzoyl-4-benzoyl)-adenosine 5′- triphosphate (BzATP) or endogenous agonist ATP induced robust calcium rises in hiPSC-derived astrocytes which were blocked by the selective P2X7R antagonists AFC-5128 or JNJ-47965567. Our findings provide evidence for the functional expression of P2X7Rs in hiPSC-derived astrocytes and support their in vitro utility in investigating the role of the P2X7R and drug screening in disorders of the central nervous system (CNS).