title: Bladder inflammatory transcriptome in response to tachykinins: neurokinin 1 receptor-dependent genes and transcription regulatory elements.
abstract: BACKGROUND
  Tachykinins (TK), such as substance P, and their neurokinin receptors which are ubiquitously expressed in the human urinary tract, represent an endogenous system regulating bladder inflammatory, immune responses, and visceral hypersensitivity. Increasing evidence correlates alterations in the TK system with urinary tract diseases such as neurogenic bladders, outflow obstruction, idiopathic detrusor instability, and interstitial cystitis. However, despite promising effects in animal models, there seems to be no published clinical study showing that NK-receptor antagonists are an effective treatment of pain in general or urinary tract disorders, such as detrusor overactivity. In order to search for therapeutic targets that could block the tachykinin system, we set forth to determine the regulatory network downstream of NK1 receptor activation. First, NK1R-dependent transcripts were determined and used to query known databases for their respective transcription regulatory elements (TREs).
  
  
  METHODS
  An expression analysis was performed using urinary bladders isolated from sensitized wild type (WT) and NK1R-/- mice that were stimulated with saline, LPS, or antigen to provoke inflammation. Based on cDNA array results, NK1R-dependent genes were selected. PAINT software was used to query TRANSFAC database and to retrieve upstream TREs that were confirmed by electrophoretic mobility shift assays.
  
  
  RESULTS
  The regulatory network of TREs driving NK1R-dependent genes presented cRel in a central position driving 22% of all genes, followed by AP-1, NF-kappaB, v-Myb, CRE-BP1/c-Jun, USF, Pax-6, Efr-1, Egr-3, and AREB6. A comparison between NK1R-dependent and NK1R-independent genes revealed Nkx-2.5 as a unique discriminator. In the presence of NK1R, Nkx2-5 _01 was significantly correlated with 36 transcripts which included several candidates for mediating bladder development (FGF) and inflammation (PAR-3, IL-1R, IL-6, alpha-NGF, TSP2). In the absence of NK1R, the matrix Nkx2-5_02 had a predominant participation driving 8 transcripts, which includes those involved in cancer (EYA1, Trail, HSF1, and ELK-1), smooth-to-skeletal muscle trans-differentiation, and Z01, a tight-junction protein, expression. Electrophoretic mobility shift assays confirmed that, in the mouse urinary bladder, activation of NK1R by substance P (SP) induces both NKx-2.5 and NF-kappaB translocations.
  
  
  CONCLUSIONS
  This is the first report describing a role for Nkx2.5 in the urinary tract. As Nkx2.5 is the unique discriminator of NK1R-modulated inflammation, it can be imagined that in the near future, new based therapies selective for controlling Nkx2.5 activity in the urinary tract may be used in the treatment in a number of bladder disorders.
authors.affiliation: Department of Physiology, The University Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. ricardo-saban@ouhsc.edu
authors.forename: Ricardo
authors.initials: R
authors.lastname: Saban
authors.forename: Cindy
authors.initials: C
authors.lastname: Simpson
authors.forename: Rajanikanth
authors.initials: R
authors.lastname: Vadigepalli
authors.forename: Sylvie
authors.initials: S
authors.lastname: Memet
authors.forename: Igor
authors.initials: I
authors.lastname: Dozmorov
authors.forename: Marcia R
authors.initials: MR
authors.lastname: Saban
mesh_terms: D000818:Animals; D003556:Cystitis / Q000235:genetics* / Q000276:immunology*; D005260:Female; D007155:Immunologic Factors / Q000235:genetics / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D020543:Proteome / Q000235:genetics / Q000276:immunology; D018040:Receptors, Neurokinin-1 / Q000235:genetics* / Q000378:metabolism*; D050436:Regulatory Elements, Transcriptional / Q000235:genetics*; D015320:Tachykinins / Q000235:genetics / Q000276:immunology*; D014157:Transcription Factors / Q000235:genetics / Q000276:immunology; D001743:Urinary Bladder / Q000276:immunology*
chemical_list: D007155:Immunologic Factors; D020543:Proteome; D018040:Receptors, Neurokinin-1; D015320:Tachykinins; D014157:Transcription Factors