Journal Article Ahead of print 2 citations
PubMed: 40856314 DOI: 10.1177/17448069251376200
EXPRESS: Humanized anti-P2X4 scFv Reduces ATP-Induced P2X4 Currents and Modulates Excitability in Human DRG Neurons.
Mol Pain
Authors: Nesia Zurek, Mark W Shilling, Jenna B Demeter, Reza Ehsanian, Ian M Adams, Aleyah E Goins, Sachin Goyal, Philippe Séguéla, June Bryan de la Pena, Adinarayana Kunamneni, Karin N Westlund, Sascha Ra Alles
Affiliations
- Department of Anesthesiology & Critical Care Medicine, University of New Mexico Health Sciences Center, Albuquerque, NM.
- Department of Anesthesia, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.
- Department of Anesthesiology & Critical Care Medicine, University of New Mexico Health Sciences Center, Albuquerque, NM.
- Department of Anesthesiology & Critical Care Medicine, University of New Mexico Health Sciences Center, Albuquerque, NM.
- Department of Neurology and Neurosurgery, Montreal Neurological Institute, Alan Edwards Centre for Research on Pain, McGill University, Montreal, QC, H3A 2B4, Canada.
- Department of Internal Medicine, Mayo Clinic, Jacksonville, FL 32224, USA.
- Biomedical Laboratory Research & Development (121F), New Mexico VA Health Care System, Albuquerque, NM 87108, USA.
Abstract
Chronic pain affects nearly 100 million adults in the U.S., yet few novel therapeutics have emerged in recent decades. P2X4 receptor (P2X4R), implicated in pain signaling, represents a promising target. We evaluated a humanized single-chain variable fragment (hscFv) targeting P2X4R for its ability to reduce ATP-induced currents and modulate excitability in human dorsal root ganglion (hDRG) neurons. Voltage-clamp recordings confirmed that human P2X4R (hP2X4R) hscFv significantly reduced ATP-evoked currents in HEK-293T cells expressing human P2X4, likely by relocalization of the receptor to the perinuclear region after hscFv treatment. Immunohistochemistry and transcriptomic analyses demonstrated widespread P2X4R (P2RX4) expression across hDRG neuronal subtypes in both male and female donors. Current-clamp recordings revealed that hP2X4R hscFv selectively increased action potential (AP) threshold in multi-firing hDRG neurons, without affecting single-firing neurons. Spontaneous activity at rest and depolarizing spontaneous fluctuation (DSF) amplitude were also reduced. Analysis confirmed consistent effects of hP2X4R hscFv on excitability parameters. These findings suggest that hP2X4 hscFv exerts modest but targeted effects on human sensory neurons, supporting its potential as a novel therapeutic for chronic pain.
MeSH
Neurons, Pain, Ion Channels, Diagnosis-Related Groups, Therapeutics, Sensory Receptor Cells