Peripherally targeted analgesia via AAV-mediated sensory neuron–specific inhibition of multiple pronociceptive sodium channels
Seung Min Shin, … , Quinn H. Hogan, Hongwei Yu
Seung Min Shin, … , Quinn H. Hogan, Hongwei Yu
Published May 9, 2024
Citation Information: J Clin Invest. 2024;134(13):e170813. https://doi.org/10.1172/JCI170813.
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Research Article Neuroscience

Peripherally targeted analgesia via AAV-mediated sensory neuron–specific inhibition of multiple pronociceptive sodium channels

Abstract

This study reports that targeting intrinsically disordered regions of the voltage-gated sodium channel 1.7 (NaV1.7) protein facilitates discovery of sodium channel inhibitory peptide aptamers (NaViPA) for adeno-associated virus–mediated (AAV-mediated), sensory neuron–specific analgesia. A multipronged inhibition of INa1.7, INa1.6, INa1.3, and INa1.1 — but not INa1.5 and INa1.8 — was found for a prototype and named NaViPA1, which was derived from the NaV1.7 intracellular loop 1, and is conserved among the TTXs NaV subtypes. NaViPA1 expression in primary sensory neurons (PSNs) of dorsal root ganglia (DRG) produced significant inhibition of TTXs INa but not TTXr INa. DRG injection of AAV6-encoded NaViPA1 significantly attenuated evoked and spontaneous pain behaviors in both male and female rats with neuropathic pain induced by tibial nerve injury (TNI). Whole-cell current clamp of the PSNs showed that NaViPA1 expression normalized PSN excitability in TNI rats, suggesting that NaViPA1 attenuated pain by reversal of injury-induced neuronal hypersensitivity. IHC revealed efficient NaViPA1 expression restricted in PSNs and their central and peripheral terminals, indicating PSN-restricted AAV biodistribution. Inhibition of sodium channels by NaViPA1 was replicated in the human iPSC-derived sensory neurons. These results summate that NaViPA1 is a promising analgesic lead that, combined with AAV-mediated PSN-specific block of multiple TTXs NaVs, has potential as a peripheral nerve–restricted analgesic therapeutic.

Authors

Seung Min Shin, Brandon Itson-Zoske, Fan Fan, Yucheng Xiao, Chensheng Qiu, Theodore R. Cummins, Quinn H. Hogan, Hongwei Yu

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Figure 5

NaViPA1 binds to full-length NaV1.7 protein and phosphoinositides.

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NaViPA1 binds to full-length NaV1.7 protein and phosphoinositides. (A) I...
(A) IBs show selectivity of NaV1.7 antibody using cell lysates from naive HEK cells, HEK1.5, HEK1.7, HEK1.6, HEK1.1, HEK1.3, HEK1.8 cells, and 50B11 cells. (B–E) Representative IHC images show NaV1.7 detection (red) in SDH (red), sciatic nerve (green), DRG neurons (red), and cutaneous nerve fibers (red). Scale bars: 100 μm. (F) IBs of NaV1.7, GFP, NKA1α, and Gapdh in the cytosol and membrane samples extracted from HEK1.7 cells transfected with sham (transfection without plasmid), GFP, GFPNaViPA1, and GFP1.7iPA2. A vertical white line in GFP panel denotes that the lanes were run on the same gel but were noncontiguous. (G) NaV1.7 IB (left) and silver stain (right) of inputs (cell lysates, 20 μg for each lane) and pulldown beads (10 μL for each lane) prepared by a nondenaturing lysis buffer from HEK1.7 cells transfected with GFP, GFPNaViPA1, and GFP1.7iPA2. (G, right) Stained gel pieces ranging 100–300 kDa (G, red asterisk denotes NaV1.7 site) from GFPNP and GFPNaViPA1 excised for mass spectrometry. (H) Silver stain on 1D SDS-PAGE gel of GFP-affinity pulldown beads in the NG108-15 cells transfected with GFPNaViPA1 and GFP and cell lysates prepared using denaturing RIPA buffer (I) and the results of PIP strip analysis.