Characterizing NaV1.7 antisense analgesia through antagonist binding, mRNA expression, and neuraxial distribution
Kaue Malange, Julia Borges Paes Lemes, Briana Noble, Yuvicza Anchondo, Carlos E. Morado-Urbina, Saee Jadhav, Saurabh Yadav, Jingrui Ma, Vineet Augustine, Sara A. Dochnal, Chiraag Kambalimath, Marie Duong, Pedro Alvarez, Bethany Fitzsimmons, Curt Mazur, Holly B. Kordasiewicz, Kim Dore, Hien T. Zhao, Tony Yaksh
Kaue Malange, Julia Borges Paes Lemes, Briana Noble, Yuvicza Anchondo, Carlos E. Morado-Urbina, Saee Jadhav, Saurabh Yadav, Jingrui Ma, Vineet Augustine, Sara A. Dochnal, Chiraag Kambalimath, Marie Duong, Pedro Alvarez, Bethany Fitzsimmons, Curt Mazur, Holly B. Kordasiewicz, Kim Dore, Hien T. Zhao, Tony Yaksh
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Research Article Cell biology Neuroscience

Characterizing NaV1.7 antisense analgesia through antagonist binding, mRNA expression, and neuraxial distribution

Abstract

Genome-targeting strategies to address NaV1.7-mediated signaling in nociceptive afferents are of great clinical interest given its high selectivity and persistent analgesic outcomes. In the present work, we analyze concentration-dependent effects of suppressing primary afferent NaV1.7 using the intrathecal delivery of an antisense oligonucleotide (ASO) on pain behaviors and demonstrate that NaV1.7 ASOs are likely to modulate pain traffic by blocking afferent signaling at the dorsal root ganglion (DRG) level. Intrathecal NaV1.7 ASOs induced a concentration-dependent gradient of knockdown in DRGs (lumbar to cervical) of Scn9a mRNA and ATTO488-ProTx-II binding in small DRG neurons, and a suppression of pain behaviors initiated by mechanical compression, inflammation, and the NaV1.7 agonist OD-1. The rostro-caudal reduction in mRNA correlated with ASO distribution, with implications for dose-related analgesic profiles. Marked reduction in DRG NaV1.7 binding was seen after intrathecal delivery of an ASO optimal dose, with high selectivity for small DRG neurons. Changes in sciatic nerve NaV1.7 binding were not detected, indicating a pain traffic modulation likely happening due to a block of afferent signaling at the DRG level.

Authors

Kaue Malange, Julia Borges Paes Lemes, Briana Noble, Yuvicza Anchondo, Carlos E. Morado-Urbina, Saee Jadhav, Saurabh Yadav, Jingrui Ma, Vineet Augustine, Sara A. Dochnal, Chiraag Kambalimath, Marie Duong, Pedro Alvarez, Bethany Fitzsimmons, Curt Mazur, Holly B. Kordasiewicz, Kim Dore, Hien T. Zhao, Tony Yaksh

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Figure 4

NaV1.7 ASO affects formalin behavior in a dose-dependent way with a different profile for 488PTx-II binding.

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NaV1.7 ASO affects formalin behavior in a dose-dependent way with a diff...
The lumbar intrathecal delivery of NaV1.7 ASO (300 or 1000 μg in 10 μL) using spinal catheters led to a different response in formalin nociceptive behavior. (A, C, and D) The injection of formalin (2.5%, 50 μL, intradermal) at the hind paw led to flinching responses (A), with an increase in 488PTx-II binding in ipsilateral DRGs from mismatch animals (C and D). A similar profile was followed by the NaV1.7 ASO smaller dose (A–D). The optimal dose of NaV1.7 ASO (1000 μg/10 μL) was able to reduce the formalin-induced flinching at phase 2 (A and B) and decrease 488PTx-II binding in ipsilateral DRGs in comparison with mismatch and the smaller antisense dose (C and D). At lumbar DRGs (E, contralateral and ipsilateral) and at the lumbar spinal cord (F) a significant reduction in Scn9a mRNA is seen for both NaV1.7 ASO doses in comparison with mismatch controls. Scale bars: 20 μm. Results shown as mean ± SD. In D, mismatch (Contra = 147 cells/Ipsi = 161 cells), NaV1.7 ASO 300 μg (Contra = 125 cells/Ipsi =166 cells), Nav 1.7 ASO 1000 μg (Contra = 147 cells/Ipsi = 101 cells). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 by 2-way ANOVA with Tukey’s post hoc test (B), Kruskal-Wallis with Dunn’s post hoc test (D), or 1-way ANOVA with Dunnett’s post hoc test (E and F).