2018 - A Novel Gain-Of-Function Mutation Of Piezo1 Is Functionally Affirmed In Red Blood Cells By High-Throughput Patch Clamp
SyncroPatch 384PE publication in Haematologica (2018)
Rotordam G.M., Fermo E., Becker N., Barcellini W., Brüggemann A., Fertig N., Egée S., Rapedius M., Bianchi P., Kaestner L.
Journal of Pharmacological and Toxicological Methods (2018) doi: 10.3324/haematol.2018.201160
Piezo1 is a mechanosensitive ion channel that is believed to be expressed in red blood cells (RBCs), mainly supported by the findings that mutations of PIEZO1 gene are associated with the RBC disease Hereditary Xerocytosis. So far several mutations have been reported, e.g. R2456H, T2127M and E2496ELE, to exhibit a partial gain-of-function phenotype with generation of mechanically activated currents that inactivate more slowly than wild type. However, characterisation of the mutated ion channel has almost exclusively been performed based on heterologous expression in cell lines and recordings in RBCs were rather of episodic character.
Here we present a patient with a novel PIEZO1 mutation (R2110W) and a patch clamp based high-throughput screening assay for Piezo1 activity. It is the first electrophysiologic single-cell based screening ever performed on RBCs, demonstrating the Piezo1 gain-of-function mutation directly on RBCs. Thus we provide a putative routine approach for detecting functional (Piezo1) channel mutations as the molecular cause of rare anaemia that can become a standard method in specialised haematological centres.