Neuropilin-1 inhibition suppresses nerve growth factor signaling and nociception in pain models
Chloe J. Peach, … , Rajesh Khanna, Nigel W. Bunnett
Chloe J. Peach, … , Rajesh Khanna, Nigel W. Bunnett
Published November 26, 2024
Citation Information: J Clin Invest. 2025;135(4):e183873. https://doi.org/10.1172/JCI183873.
View: Text | PDF
Research Article Cell biology Neuroscience

Neuropilin-1 inhibition suppresses nerve growth factor signaling and nociception in pain models

Abstract

Nerve growth factor (NGF) monoclonal antibodies inhibit chronic pain, yet failed to gain approval due to worsened joint damage in osteoarthritis patients. We report that neuropilin-1 (NRP1) is a coreceptor for NGF and tropomyosin-related kinase A (TrkA) pain signaling. NRP1 was coexpressed with TrkA in human and mouse nociceptors. NRP1 inhibitors suppressed NGF-stimulated excitation of human and mouse nociceptors and NGF-evoked nociception in mice. NRP1 knockdown inhibited NGF/TrkA signaling, whereas NRP1 overexpression enhanced signaling. NGF bound NRP1 with high affinity and interacted with and chaperoned TrkA from the biosynthetic pathway to the plasma membrane and endosomes, enhancing TrkA signaling. Molecular modeling suggested that the C-terminal R/KXXR/K NGF motif interacts with the extracellular “b” NRP1 domain within a plasma membrane NGF/TrkA/NRP1 of 2:2:2 stoichiometry. G α interacting protein C-terminus 1 (GIPC1), which scaffolds NRP1 and TrkA to myosin VI, colocalized in nociceptors with NRP1/TrkA. GIPC1 knockdown abrogated NGF-evoked excitation of nociceptors and pain-like behavior. Thus, NRP1 is a nociceptor-enriched coreceptor that facilitates NGF/TrkA pain signaling. NRP binds NGF and chaperones TrkA to the plasma membrane and signaling endosomes via the GIPC1 adaptor. NRP1 and GIPC1 antagonism in nociceptors offers a long-awaited nonopioid alternative to systemic antibody NGF sequestration for the treatment of chronic pain.

Authors

Chloe J. Peach, Raquel Tonello, Elisa Damo, Kimberly Gomez, Aida Calderon-Rivera, Renato Bruni, Harsh Bansia, Laura Maile, Ana-Maria Manu, Hyunggu Hahn, Alex R.B. Thomsen, Brian L. Schmidt, Steve Davidson, Amedee des Georges, Rajesh Khanna, Nigel W. Bunnett

×

Figure 6

NRP1 modulates TrkA-mediated kinase signaling.

Options: View larger image (or click on image) Download as PowerPoint
NRP1 modulates TrkA-mediated kinase signaling. (A and B) Effect of mouse...
(A and B) Effect of mouse NGF (100 nM, 15 minutes) and NRP1 inhibitor EG00229 (30 μM, 30 minutes preincubation) on phosphorylated TrkA Y785 staining in mouse DRG neurons. n = 34–44 neurons from 3 independent experiments. Scale bars: 20 μm. (C–F) Effect of mouse NGF (100 nM) and NRP1 inhibitors EG00229 (30 μM, 30 minutes preincubation) (C and D) and CendR (1 μM, 30 minutes preincubation) (E and F) on phosphorylated ERK Thr202/Tyr204 staining in mouse DRG neurons. n = 15–432 neurons from 4 independent experiments. Scale bars: 20 μm. (G–N) NGF-induced ERK signaling measured using FRET-based EKAR biosensors (H–M) or a downstream luciferase reporter (N). (H–M) NGF-induced modulation of ERK activity using biosensors localized to the cytosol (H and I) or nucleus (H and J) in neuron-like CAD cells expressing human TrkA. Kinetics of NGF-induced ERK monitored in CAD cells (H), comparing increasing NGF concentrations after preincubation with EG00229 (30 μM, 30 minutes) (I–J). (K–M) ERK signaling in HEK293T cells expressing TrkA alone or expressing both TrkA and NRP1 (L and M). (N) Effect of increasing NGF concentrations on ERK transcription in cells expressing TrkA, NRP1, or both (% positive control, 10 μM PDBu). RFU, relative fluorescence units. Data from 4–8 independent experiments with triplicate wells. Data are represented as mean ± SEM. *P < 0.05. (B, M, and N) One-way ANOVA, Sídák’s multiple comparisons. (D and F) Two-way ANOVA, Tukey’s multiple comparison.