Work

Raf-MEK-ERK signaling mediates FLT3-induced mechanical allodynia and reduced morphine analgesic efficacy in male rodents

Year: 2026

Type: preprint

Abstract: The cytokine FL and its receptor, Fms-like tyrosine kinase 3 (FLT3), contribute to chronic pain and to the maladaptive effects of morphine but the intracellular mechanisms underlying these effects remain incompletely understood. Here, we investigated the contribution of the Raf-MEK-ERK pathway to FLT3dependent nociceptive signaling and to the modulation of morphine analgesia. Using HEK293T cells expressing human or mouse FLT3, primary dorsal root ganglion (DRG) neurons, and rodent models, we examined ERK activation, TRPV1 sensitization, and behavioral responses following FLT3 activation by its ligand, FL. FL induced concentration-dependent ERK phosphorylation through FLT3, while pharmacological inhibition of FLT3, Raf, or MEK reduced this response. In primary DRG neurons, MEK inhibition attenuated FLinduced potentiation of capsaicin-evoked calcium responses, supporting a role for MEK-ERK signaling in TRPV1 sensitization. In vivo, intrathecal FL increased ERK phosphorylation in DRGs and induced mechanical hypersensitivity, both of which were reduced by Raf or MEK inhibition. In FLT3/MOR-expressing cells, morphine potentiated FL-induced ERK activation, an effect reduced by naloxone and abolished by FLT3 inhibition. In rats, MEK inhibition prevented the FL-induced reduction of morphine analgesia and subsequent mechanical hypersensitivity. Together, these findings identify Raf-MEK-ERK signaling as an important downstream pathway linking neuronal FLT3 activation to nociceptor sensitization and impaired morphine analgesic efficacy. Highlights - Mouse or human FLT3 stimulation activates ERK which is reversed by FLT3, Raf or MEK inhibition - FL increases capsaicin-induced Ca2+ influx via MEK-ERK signaling in DRG neurons - FL injection activates ERK in DRG for at least 1h - FL-induced pain hypersensitivity is partially prevented by Raf or MEK inhibition - FL reduces morphine analgesia which is totally prevented by MEK inhibition

Source: bioRxiv (Cold Spring Harbor Laboratory)

DOI: 10.64898/2026.10.04.756546