The effect of epigenetic therapy on congenital neurogenic bladders--a pilot study.
OBJECTIVE
To demonstrate that human smooth muscle cells derived from neurogenic bladders produce more collagen in vitro than smooth muscle cells derived from normal bladders, and that epigenetic therapy may normalize this increased collagen production.


METHODS
Human smooth muscle cells from normal (n = 3) and neurogenic bladders (n = 3) were cultured in normal culture media and at different concentrations of the histone deacetylase inhibitors trichostatin A, valproic acid, and the DNA methylation inhibitor 5-azacytidine (5-aza). Collagen type I and III gene expression was measured using real-time quantitative reverse transcription-polymerase chain reaction after varying doses of drug exposure. Cell viability was measured using trypan blue.


RESULTS
The smooth muscle cells from neurogenic bladders released significantly more collagen than the normal bladder cells (mean 4.1 vs 1.8 microg/mL in control media) when grown in normal conditions. Treatment with trichostatin A at 50 ng/mL decreased the collagen level in cells from neurogenic bladders to almost normal levels (2.1 microg/mL). In addition, valproic acid treatment decreased collagen types I and III gene expression relative to controls, with maximal effect at 300 mg/mL. These treatments had little effect on cell viability.


CONCLUSIONS
Histone deacetylase inhibitors decreased collagen production of smooth muscle cells from neurogenic bladders in vitro. These agents may be a means of effectively preventing bladder fibrosis in patients with this condition.
Department of Urology, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27157, USA. shodges@wfubmc.edu
Steve J
SJ
Hodges
James J
JJ
Yoo
Nilamadhab
N
Mishra
Anthony
A
Atala
D001374:Azacitidine / Q000494:pharmacology; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D044127:Epigenesis, Genetic; D015316:Genetic Therapy; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010865:Pilot Projects; D001743:Urinary Bladder / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001750:Urinary Bladder, Neurogenic / Q000151:congenital* / Q000378:metabolism / Q000628:therapy*; D014635:Valproic Acid / Q000494:pharmacology
D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C012589:trichostatin A; D014635:Valproic Acid; D003094:Collagen; D001374:Azacitidine