title: Regulation of TIMP-1 in Human Placenta and Fetal Membranes by lipopolysaccharide and demethylating agent 5-aza-2'-deoxycytidine.
abstract: BACKGROUND
  An appropriate transcriptional profile in the placenta and fetal membranes is required for successful pregnancy; any variations may lead to inappropriate timing of birth. Epigenetic regulation through reversible modification of chromatin has emerged as a fundamental mechanism for the control of gene expression in a range of biological systems and can be modified by pharmacological intervention, thus providing novel therapeutic avenues. TIMP-1 is an endogenous inhibitor of MMPs, and hence is intimately involved in maintaining the integrity of the fetal membranes until labor.
  
  
  OBJECTIVE
  To determine if TIMP-1 is regulated by DNA methylation in gestational tissues we employed an in vitro model in which gestational tissue explants were treated with demethylating agent 5-aza-2'-deoxycytidine (AZA) and lipopolysaccharide (LPS).
  
  
  RESULTS
  Quantitative Real-Time PCR (qRT-PCR) revealed that TIMP-1 transcription was significantly increased by combined treatment of AZA and LPS, but not LPS alone, in villous, amnion and choriodecidua explants after 24 and 48 hrs, whilst western blotting showed protein production was stimulated after 24 hrs only. Upon interrogation of the TIMP-1 promoter using Sequenom EpiTyper MassARRAY, we discovered sex-specific differential methylation, in part explained by x-linked methylation in females. Increased TIMP-1 in the presence of LPS was potentiated by AZA treatment, signifying that a change in chromatin structure, but not in DNA methylation at the promoter region, is required for transcriptional activators to access the promoter region of TIMP-1.
  
  
  CONCLUSIONS
  Collectively, these observations support a potential role for pharmacological agents that modify chromatin structure to be utilized in the therapeutic targeting of TIMP-1 to prevent premature rupture of the fetal membranes in an infectious setting.
authors.affiliation: Liggins Institute, University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand.|Gravida: National Centre for Growth and Development, Palmerston North, New Zealand.
authors.forename: Zoë L
authors.initials: ZL
authors.lastname: Vincent
authors.affiliation: Liggins Institute, University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand.|University of Queensland Centre for Clinical Research, Brisbane, Australia.
authors.forename: Murray D
authors.initials: MD
authors.lastname: Mitchell
authors.affiliation: Liggins Institute, University of Auckland, Private Bag 92019, Auckland, 1142, New Zealand. a.ponnampalam@auckland.ac.nz.|Gravida: National Centre for Growth and Development, Palmerston North, New Zealand. a.ponnampalam@auckland.ac.nz.|The Heart Foundation, Auckland, New Zealand. a.ponnampalam@auckland.ac.nz.
authors.forename: Anna P
authors.initials: AP
authors.lastname: Ponnampalam
mesh_terms: D000650:Amnion / Q000378:metabolism; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002823:Chorion / Q000378:metabolism; D002824:Chorionic Villi / Q000378:metabolism; D002843:Chromatin / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects; D003656:Decidua / Q000378:metabolism; D000077209:Decitabine; D044127:Epigenesis, Genetic; D005321:Extraembryonic Membranes / Q000378:metabolism*; D005260:Female; D005322:Premature Rupture of Fetal Membranes / Q000517:prevention & control; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology*; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D060888:Real-Time Polymerase Chain Reaction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D002843:Chromatin; D008070:Lipopolysaccharides; C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D000077209:Decitabine; D001374:Azacitidine