ATP in the tumour microenvironment drives expression of nfP2X7, a key mediator of cancer cell survival

The ATP-gated receptor P2X₇ is expressed in multiple malignant tumours including neuroblastoma, melanoma, prostate, lung and breast. P2X₇ has a significant role in mediating diverse cell responses, which upon dysregulation are associated with tumour initiation and development. The rapid, ATP-mediated activation of P2X₇ induces a fast-inward cation current in cells. However, prolonged ATP-mediated activation of P2X₇ leads to formation of a pore that increases membrane permeability and eventually causes cell death. This presents a potential paradox, as the tumour microenvironment contains extracellular ATP at levels sufficient to activate the P2X₇ pore and trigger cell death. However, P2X₇ expression is associated with enhanced cancer cell survival, proliferation and metastatic potential. At least one distinct conformational form of P2X₇, termed non-pore functional P2X₇ (nfP2X₇), has been described, which is not able to form a functional pore. We demonstrate for the first time in this study that exposure to a high ATP concentration, equivalent to those measured in the tumour microenvironment, drives nfP2X₇ expression and also that nfP2X₇ is essential for tumour cell survival. We show that monoclonal antibodies raised against a P2X₇ amino acid sequence (200-216), whose conformation is distinct from that of wild-type (WT) P2X₇, bind specifically to nfP2X₇ expressed on the surface of tumour cells. We also show that nfP2X₇ is broadly expressed in patient-derived tumour sections from a wide range of cancers. Therefore, antibodies raised against E200 provide tools that can differentiate between forms of the P2X₇ receptor that have a key role in cancer.