title: Antinociceptive effect of antisense oligonucleotides against the vanilloid receptor VR1/TRPV1.
abstract: To examine the role of the vanilloid receptor TRPV1 in neuropathic pain, we assessed the effects of the receptor antagonist thioxo-BCTC and antisense oligonucleotides against the TRPV1 mRNA in a rat model of spinal nerve ligation. In order to identify accessible sites on the mRNA of TRPV1, the RNase H assay was used, leading to the successful identification of binding sites for antisense oligonucleotides. Cotransfection studies using Cos-7 cells were employed to identify the most effective antisense oligonucleotide efficiently inhibiting the expression of a fusion protein consisting of TRPV1 and the green fluorescent protein in a specific and concentration-dependent manner. In an in vivo rat model of spinal nerve ligation, intravenous application of the TRPV1 antagonist thioxo-BCTC reduced mechanical hypersensitivity yielding an ED(50) value of 10.6mg/kg. Intrathecal administration of the antisense oligonucleotide against TRPV1, but not the mismatch oligonucleotide or a vehicle control, reduced mechanical hypersensitivity in rats with spinal nerve ligation in a similar manner. Immunohistochemical analysis revealed neuropathy- and antisense-associated regulation of TRPV1 protein expression in spinal cord and dorsal root ganglia. Our data demonstrate comparative analgesic effects of a TRPV1 anatagonist and a rationally designed TRPV1 antisense oligonucleotide in a spinal nerve ligation model of neuropathic pain and thus, lend support to the validation of TRPV1 as a promising target for the treatment of neuropathic pain.
authors.affiliation: Research & Development, Grünenthal GmbH, Zieglerstr. 6, 52078 Aachen, Germany. thomas.christoph@grunenthal.de
authors.forename: Thomas
authors.initials: T
authors.lastname: Christoph
authors.forename: Clemens
authors.initials: C
authors.lastname: Gillen
authors.forename: Joanna
authors.initials: J
authors.lastname: Mika
authors.forename: Arnold
authors.initials: A
authors.lastname: Grünweller
authors.forename: Martin K-H
authors.initials: MK
authors.lastname: Schäfer
authors.forename: Klaus
authors.initials: K
authors.lastname: Schiene
authors.forename: Robert
authors.initials: R
authors.lastname: Frank
authors.forename: Ruth
authors.initials: R
authors.lastname: Jostock
authors.forename: Gregor
authors.initials: G
authors.lastname: Bahrenberg
authors.forename: Eberhard
authors.initials: E
authors.lastname: Weihe
authors.forename: Volker A
authors.initials: VA
authors.lastname: Erdmann
authors.forename: Jens
authors.initials: J
authors.lastname: Kurreck
mesh_terms: D000700:Analgesics / Q000494:pharmacology*; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers; D007150:Immunohistochemistry; D008297:Male; D008856:Microscopy, Fluorescence; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism; D050916:TRPV Cation Channels / Q000187:drug effects*
chemical_list: D000700:Analgesics; D017931:DNA Primers; D016376:Oligonucleotides, Antisense; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat