title: Parallel changes in bladder suburothelial vanilloid receptor TRPV1 and pan-neuronal marker PGP9.5 immunoreactivity in patients with neurogenic detrusor overactivity after intravesical resiniferatoxin treatment.
abstract: OBJECTIVE
  To compare PGP9.5 and transient receptor potential vanilloid receptor (TRPV1) suburothelial immunoreactivity between controls and patients with spinal neurogenic detrusor overactivity (NDO) before and after treatment with intravesical resiniferatoxin, as suburothelial PGP9.5-staining nerve fibres decrease in patients with spinal NDO who respond to intravesical capsaicin, and TRPV1 is present on these suburothelial nerve fibres in normal and overactive human urinary bladder.
  
  
  METHODS
  Patients with refractory NDO were enrolled in a prospective, randomized, parallel-group, double-blind, placebo-controlled trial using escalating doses of resiniferatoxin to a maximum of 1 micro mol/L. Flexible cystoscopic bladder biopsies obtained at baseline, 4 weeks after each instillation and at the time of maximum clinical response were compared with biopsies taken from control subjects. Frozen sections were incubated with rabbit antibodies to TRPV1 and PGP9.5, and assessed using standard immunohistochemical methods. PGP9.5 nerve density was analysed using a nerve-counting graticule by an observer unaware of sample origin. Another two independent observers unaware of each other's results used a random grading scale to evaluate TRPV1 nerve fibre density and intensity. The immunohistochemistry results were compared with histology findings (haematoxylin-eosin), and the Mann-Whitney test used to assess any differences (P < 0.05 significant) and the Pearson test for correlation.
  
  
  RESULTS
  There were eight controls and 20 patients with spinal NDO, 14 (five clinical responders and nine not) who received the maximum dose of resiniferatoxin. There were more PGP9.5 and TRPV1 nerve fibres in patients with NDO than in controls (P = 0.007 and 0.002, respectively). Immunoreactivity before resiniferatoxin was similar in both groups for both PGP9.5 and TRPV1. In responders there were fewer PGP9.5 and TRPV1-positive fibres after treatment (P = 0.008 for each) but no change in those not responding. Changes after treatment for TRPV1 correlated well with those for PGP9.5 (r = 0.88, P < 0.001).
  
  
  CONCLUSIONS
  The decrease of PGP9.5 and TRPV1 immunoreactive nerve fibres in responders to resiniferatoxin (to levels in control tissues) suggests that the increased numbers of nerve fibres in patients with NDO are mainly of sensory origin and express TRPV1. As baseline nerve fibre values were similar in responders and nonresponders, an additional factor may account for the difference in treatment outcome.
authors.affiliation: Department of Uro-Neurology, Institute of Neurology, Queen Square, London, UK.
authors.forename: C M
authors.initials: CM
authors.lastname: Brady
authors.forename: A N
authors.initials: AN
authors.lastname: Apostolidis
authors.forename: M
authors.initials: M
authors.lastname: Harper
authors.forename: Y
authors.initials: Y
authors.lastname: Yiangou
authors.forename: A
authors.initials: A
authors.lastname: Beckett
authors.forename: T S
authors.initials: TS
authors.lastname: Jacques
authors.forename: A
authors.initials: A
authors.lastname: Freeman
authors.forename: F
authors.initials: F
authors.lastname: Scaravilli
authors.forename: C J
authors.initials: CJ
authors.lastname: Fowler
authors.forename: P
authors.initials: P
authors.lastname: Anand
mesh_terms: D000283:Administration, Intravesical; D015415:Biomarkers; D001706:Biopsy / Q000379:methods; D004224:Diterpenes / Q000627:therapeutic use; D004311:Double-Blind Method; D006801:Humans; D007150:Immunohistochemistry; D007473:Ion Channels; D008875:Middle Aged; D009498:Neurotoxins / Q000627:therapeutic use; D011446:Prospective Studies; D011955:Receptors, Drug / Q000378:metabolism*; D050916:TRPV Cation Channels; D043222:Ubiquitin Thiolesterase / Q000378:metabolism*; D001743:Urinary Bladder / Q000378:metabolism* / Q000473:pathology; D001750:Urinary Bladder, Neurogenic / Q000188:drug therapy / Q000378:metabolism*; D014549:Urinary Incontinence / Q000188:drug therapy / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D004224:Diterpenes; D007473:Ion Channels; D009498:Neurotoxins; D011955:Receptors, Drug; D050916:TRPV Cation Channels; C476722:Trpv2 protein, rat; C482860:UCHL1 protein, human; C024353:resiniferatoxin; D043222:Ubiquitin Thiolesterase