Calcium-sensing receptor regulates Kv7 channels via Gi/o protein signalling and modulates excitability of human induced pluripotent stem cell-derived nociceptive-like neurons

Br J Pharmacol. 2024 Aug;181(15):2676-2696. doi: 10.1111/bph.16349. Epub 2024 Apr 16.

Abstract

Background and purpose: Neuropathic pain, a debilitating condition with unmet medical needs, can be characterised as hyperexcitability of nociceptive neurons caused by dysfunction of ion channels. Voltage-gated potassium channels type 7 (Kv7), responsible for maintaining neuronal resting membrane potential and thus excitability, reside under tight control of G protein-coupled receptors (GPCRs). Calcium-sensing receptor (CaSR) is a GPCR that regulates the activity of numerous ion channels, but whether CaSR can control Kv7 channel function has been unexplored until now.

Experimental approach: Experiments were conducted in recombinant cell models, mouse dorsal root ganglia (DRG) neurons and human induced pluripotent stem cell (hiPSC)-derived nociceptive-like neurons using patch-clamp electrophysiology and molecular biology techniques.

Key results: Our results demonstrate that CaSR is expressed in recombinant cell models, hiPSC-derived nociceptive-like neurons and mouse DRG neurons, and its activation induced depolarisation via Kv7.2/7.3 channel inhibition. The CaSR-Kv7.2/7.3 channel crosslink was mediated via the Gi/o protein-adenylate cyclase-cyclicAMP-protein kinase A signalling cascade. Suppression of CaSR function demonstrated a potential to rescue hiPSC-derived nociceptive-like neurons from algogenic cocktail-induced hyperexcitability.

Conclusion and implications: This study demonstrates that the CaSR-Kv7.2/7.3 channel crosslink, via a Gi/o protein signalling pathway, effectively regulates neuronal excitability, providing a feasible pharmacological target for neuronal hyperexcitability management in neuropathic pain.

Keywords: Ca2+‐sensing receptors; G protein pathways; Kv7.2/7.3 channels; electrophysiology; stem cells.

MeSH terms

  • Animals
  • Cells, Cultured
  • GTP-Binding Protein alpha Subunits, Gi-Go / metabolism
  • Ganglia, Spinal* / cytology
  • Ganglia, Spinal* / metabolism
  • HEK293 Cells
  • Humans
  • Induced Pluripotent Stem Cells* / metabolism
  • Mice
  • Nociceptors / metabolism
  • Receptors, Calcium-Sensing* / metabolism
  • Signal Transduction*

Substances

  • Receptors, Calcium-Sensing
  • GTP-Binding Protein alpha Subunits, Gi-Go