Treatment with methyl-β-cyclodextrin prevents mechanical allodynia in resiniferatoxin neuropathy in a mouse model

Abstract

Specialized microdomains which have cholesterol-rich membrane regions contain transient receptor potential vanilloid subtype 1 (TRPV1) are involved in pain development. Our previous studies have demonstrated that the depletion of prostatic acid phosphatase (PAP) – a membrane-bound ectonucleotidase ­– and disordered adenosine signaling reduce the antinociceptive effect. The role of membrane integrity in the PAP-mediated antinociceptive effect in small-fiber neuropathy remains unclear, especially with respect to whether TRPV1 and PAP are colocalized in the same microdomain which is responsible for PAP-mediated antinociception. Immunohistochemistry was conducted on the dorsal root ganglion to identify the membrane compositions, and pharmacological interventions were conducted using methyl-β-cyclodextrin (MβC) – a membrane integrity disruptor that works by depleting cholesterol – in pure small-fiber neuropathy with resiniferatoxin (RTX). Immunohistochemical evidence indicated that TRPV1 and PAP were highly colocalized with flotillin 1 (66.7%±9.7%) and flotillin 2 (73.7%±6.0%), which reside in part in the microdomain. MβC mildly depleted PAP, which maintained the ability to hydrolyze phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] and delayed the development of mechanical allodynia. MβC treatment had no role in thermal transduction and neuronal injury following RTX neuropathy. In summary, this study demonstrated the following: (1) membrane cholesterol depletion preserves PAP-mediated antinociception through PI(4,5)P2 hydrolysis and (2) pain hypersensitivity that develops after TRPV1(+) neuron depletion-mediated neurodegeneration following RTX neuropathy is attributable to the downregulation of PAP analgesic signaling.

Antinociceptive Effects of Lipid Raft Disruptors, a Novel Carboxamido-Steroid and Methyl β-Cyclodextrin, in Mice by Inhibiting Transient Receptor Potential Vanilloid 1 and Ankyrin 1 Channel Activation

“…Although there are several in vitro evidence that lipid raft disruption affected TRP channel activation (Szőke et al, 2010;Sághy et al, 2015), there are only sporadic, recent in vivo reports. MCD-related cholesterol depletion induced antinociception in RTX-induced mononeuropathy through PI(4,5)P2 hydrolysis in mice (Lin et al, 2019). Intraplantar injection of MCD attenuated the PGE2-, but not cyclopentyladenosine-evoked mechanical hyperalgesia.…”

“…TRPA1 is also considered to be a promising analgesic target based on experimental and human studies which seem to be free of severe side effects ( Romanovsky et al, 2009 ; Botz et al, 2017 ). These data clearly suggest the drug developmental potential of TRPV1 and TRPA1 blockade, therefore alternative mechanisms in addition to the direct antagonism have been proposed as promising inhibitors options ( Ferrari and Levine, 2015 ; Sághy et al, 2015 , 2018 ; Lin et al, 2019 ).…”

“…TRPA1 is also considered to Abbreviations: C1, carboxamido-steroid compound; CAPS, capsaicin; CRAC, Cholesterol Recognition/interaction Amino acid Consensus; DHEA, dehydroepiandrosterone; DRG, dorsal root ganglion; E2, 17-β estradiol; MCD, methyl β-cyclodextrin; PGE2, prostaglandin E2; PI(4,5)P2, phosphatidylinositol 4,5-bisphosphate; PS, pregnenolone sulfate; RAMEB, random methylated β-cyclodextrins; RTX, resiniferatoxin; TrkA, tropomyosin-related kinase A; TRP, be a promising analgesic target based on experimental and human studies which seem to be free of severe side effects (Romanovsky et al, 2009;Botz et al, 2017). These data clearly suggest the drug developmental potential of TRPV1 and TRPA1 blockade, therefore alternative mechanisms in addition to the direct antagonism have been proposed as promising inhibitors options (Ferrari and Levine, 2015;Sághy et al, 2015Sághy et al, , 2018Lin et al, 2019).…”

“…Although several in vitro data show that lipid raft decomposition inhibits TRP channel opening, there are only two recent in vivo evidence. Methyl β-cyclodextrin (MCD)-induced membrane cholesterol depletion led to antinociception in the RTX-evoked mononeuropathy model via phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) hydrolysis (Lin et al, 2019) and significantly attenuated the prostaglandin E2 (PGE2)-evoked mechanical hyperalgesia in rats (Ferrari and Levine, 2015).…”

Analgesic Effects of Lipid Raft Disruption by Sphingomyelinase and Myriocin via Transient Receptor Potential Vanilloid 1 and Transient Receptor Potential Ankyrin 1 Ion Channel Modulation

“…Despite all these in vitro data showing that lipid raft disruption inhibits TRP channel activation, there are only few very recent reports investigating this phenomenon in vivo . Hyperalgesia responses in the RTX-evoked mouse neuropathy model ( Lin et al, 2019 ) and prostaglandin E2 (PGE2) ( Ferrari and Levine, 2015 ) administration were significantly attenuated by MCD. We recently reported the antihyperalgesic actions of MCD and a novel carboxamido-steroid compound in TRPV1 and TRPA1 activation-related mouse pain models ( Horváth et al, 2020 ).…”