FXYD2 antisense oligonucleotide provides an efficient approach for long-lasting relief of chronic peripheral pain
Alexandre Derre, … , Alexandre Pattyn, Stephanie Venteo
Alexandre Derre, … , Alexandre Pattyn, Stephanie Venteo
Published May 8, 2023
Citation Information: JCI Insight. 2023;8(9):e161246. https://doi.org/10.1172/jci.insight.161246.
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Research Article Neuroscience

FXYD2 antisense oligonucleotide provides an efficient approach for long-lasting relief of chronic peripheral pain

Abstract

Chronic pain, whether of inflammatory or neuropathic origin, affects about 18% of the population of developed countries, and most current treatments are only moderately effective and/or cause serious side effects. Therefore, the development of novel therapeutic approaches still represents a major challenge. The Na,K-ATPase modulator FXYD2 is critically required for the maintenance of neuropathic pain in rodents. Here, we set up a therapeutic protocol based on the use of chemically modified antisense oligonucleotides (ASOs) to inhibit FXYD2 expression and treat chronic pain. We identified an ASO targeting a 20-nucleotide stretch in the FXYD2 mRNA that is evolutionarily conserved between rats and humans and is a potent inhibitor of FXYD2 expression. We used this sequence to synthesize lipid-modified forms of ASO (FXYD2-LASO) to facilitate their entry into dorsal root ganglia neurons. We established that intrathecal or intravenous injections of FXYD2-LASO in rat models of neuropathic or inflammatory pain led to a virtually complete alleviation of their pain symptoms, without causing obvious side effects. Remarkably, by using 2′-O-2-methoxyethyl chemical stabilization of the ASO (FXYD2-LASO-Gapmer), we could significantly prolong the therapeutic action of a single treatment up to 10 days. This study establishes FXYD2-LASO-Gapmer administration as a promising and efficient therapeutic strategy for long-lasting relief of chronic pain conditions in human patients.

Authors

Alexandre Derre, Noelian Soler, Valentine Billoux, Sebastien Benizri, Brune Vialet, Cyril Rivat, Philippe Barthélémy, Patrick Carroll, Alexandre Pattyn, Stephanie Venteo

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Figure 5

2′-MOE–modified FXYD2-LASO-Gapmer provides long-lasting pain relief in neuropathic pain and inflammatory pain models.

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2′-MOE–modified FXYD2-LASO-Gapmer provides long-lasting pain relief in n...
(A–C) After induction of mechanical hypersensitivity by SNL, animals were intrathecally injected weekly — until full recovery — for 56 days with 30, 100, or 300 μg of FXYD2- or control-LASO-Gapmer (A); daily for 14 days with 2 μg of FXYD2- or control-LASO or FXYD2- or control-LASO-Gapmer — until full recovery — followed by an injection every 8–11 days of 2 μg of FXYD2- or control-LASO-Gapmer (B); or daily for 14 days with 2 μg followed by a 300 μg single injection of FXYD2- or control-LASO-Gapmer (C). (D) Mechanical hypersensitivity was induced by intraplantar injection of CFA, and animals were intrathecally injected daily for 15 days with 2 μg of FXYD2- or control-LASO-Gapmer and reinjected with a 2 μg single injection of FXYD2- or control-LASO-Gapmer when a return of hypersensitivity to mechanical stimuli was observed. (E–H) To test spontaneous pain, the Rat Grimace Scale (RGS) (E and G) and Static Weight Bearing (SWB) test (F and H) were used when complete attenuation of pain behavior was reached and assessed by Randall-Selitto evoked behavioral test in cohort of SNL (E and F) or CFA (G and H) rats intrathecally injected daily for 14 days with 2 μg of FXYD2-LASO-Gapmer. (I–N) After induction of mechanical hypersensitivity by SNL (I, K, and L) or CFA (J, M, and N), animals were intravenously injected daily for 16 days with 40 mg/kg of FXYD2- or control-LASO-Gapmer (I and J). After the last injection (day 28 in I, day 25 in J), RGS (K and M) and SWB test (L and N) were performed to evaluate spontaneous pain. Mechanical hypersensitivity was evaluated with the Randall-Selitto test (A–D, I, and J). Means ± SEM of data from 6 animals (A–N). Two-way (A–D, I, and J) and 1-way (E–H and K–N) ANOVA and post hoc Bonferroni’s test. **P < 0.01; ***P < 0.001; ****P < 0.0001.