Antinociceptive effect of antisense oligonucleotides against the vanilloid receptor VR1/TRPV1

• Published in: Neurochemistry international 2007, Vol.50 (1), p.281-290
• Main Authors: Christoph, Thomas, Gillen, Clemens, Mika, Joanna, Grünweller, Arnold, Schäfer, Martin K.-H., Schiene, Klaus, Frank, Robert, Jostock, Ruth, Bahrenberg, Gregor, Weihe, Eberhard, Erdmann, Volker A., Kurreck, Jens
• Format: Article
• Language: English
• Subjects: Analgesics - pharmacology; Animals; Antisense oligonucleotides; Base Sequence; Biological and medical sciences; Capsaicin receptor; Cercopithecus aethiops; CHO Cells; COS Cells; Cranial nerves. Spinal roots. Peripheral nerves. Autonomic nervous system. Gustation. Olfaction; Cricetinae; Cricetulus; DNA Primers; Fundamental and applied biological sciences. Psychology; Immunohistochemistry; Male; Medical sciences; Microscopy, Fluorescence; Nervous system (semeiology, syndromes); Neurology; Neuropathic pain; Oligonucleotides, Antisense - pharmacology; Rats; Rats, Sprague-Dawley; Spinal Cord - drug effects; Spinal Cord - metabolism; Thioxo-BCTC; TRPV Cation Channels - drug effects; Vanilloid receptor; Vertebrates: nervous system and sense organs
• ISSN: 0197-0186; 1872-9754
• DOI: 10.1016/j.neuint.2006.08.017

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 10.6 mg/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.