Transcription factor network downstream of protease activated receptors (PARs) modulating mouse bladder inflammation.
BACKGROUND
All four PARs are present in the urinary bladder, and their expression is altered during inflammation. In order to search for therapeutic targets other than the receptors themselves, we set forth to determine TFs downstream of PAR activation in the C57BL/6 urinary bladders.


METHODS
For this purpose, we used a protein/DNA combo array containing 345 different TF consensus sequences. Next, the TF selected was validated by EMSA and IHC. As mast cells seem to play a fundamental role in bladder inflammation, we determined whether c-kit receptor deficient (Kit w/Kit w-v) mice have an abrogated response to PAR stimulation. Finally, TFEB antibody was used for CHIP/Q-PCR assay and revealed up-regulation of genes known to be downstream of TFEB.


RESULTS
TFEB, a member of the MiTF family of basic helix-loop-helix leucine zipper, was the only TF commonly up-regulated by all PAR-APs. IHC results confirm a correlation between inflammation and TFEB expression in C57BL/6 mice. In contrast, Kit w/Kit w-v mice did not exhibit inflammation in response to PAR activation. EMSA results confirmed the increased TFEB binding activity in C57BL/6 but not in Kit w/Kit w-v mice.


CONCLUSIONS
This is the first report describing the increased expression of TFEB in bladder inflammation in response to PAR activation. As TFEB belongs to a family of TFs essential for mast cell survival, our findings suggest that this molecule may influence the participation of mast cells in PAR-mediated inflammation and that targeting TFEB/MiTF activity may be a novel approach for the treatment of bladder inflammatory disorders.
Saban
Ricardo
R
Department of Physiology, The University Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA. ricardo-saban@ouhsc.edu
Simpson
Cindy
C
Davis
Carole A
CA
Dozmorov
Igor
I
Maier
Julie
J
Fowler
Ben
B
Ihnat
Michael A
MA
Hurst
Robert E
RE
Wershil
Barry K
BK
Saban
Marcia R
MR
D000818:Animals; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000378:metabolism*; D003556:Cystitis / Q000378:metabolism*; D005260:Female; D053263:Gene Regulatory Networks; D007249:Inflammation / Q000378:metabolism*; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL / Q000378:metabolism; D008817:Mice, Mutant Strains; D009092:Mucous Membrane / Q000166:cytology / Q000378:metabolism; D044462:Receptors, Proteinase-Activated / Q000302:isolation & purification / Q000378:metabolism*; D001743:Urinary Bladder / Q000378:metabolism*
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D044462:Receptors, Proteinase-Activated; C484068:Tcfeb protein, mouse
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