title: Comprehensive RNA profiling of villous trophoblast and decidua basalis in pregnancies complicated by preterm birth following intra-amniotic infection.
abstract: We performed RNA sequencing with the primary goal of discovering key placental villous trophoblast (VT) and decidua basalis (DB) transcripts differentially expressed in intra-amniotic infection (IAI)-induced preterm birth (PTB).
  
  
  
  RNA was extracted from 15 paired VT and DB specimens delivered of women with: 1) spontaneous PTB in the setting of amniocentesis-proven IAI and histological chorioamnionitis (n = 5); 2) spontaneous idiopathic PTB (iPTB, n = 5); and 3) physiologic term pregnancy (n = 5). RNA sequencing was performed using the Illumina HiSeq 2500 platform, and a spectrum of computational tools was used for gene prioritization and pathway analyses.
  
  
  
  In the VT specimens, 128 unique long transcripts and 7 mature microRNAs differed significantly between pregnancies complicated by IAI relative to iPTB (FDR<0.1). The up-regulated transcripts included many characteristic of myeloblast-derived cells, and bioinformatic analyses revealed enrichment for multiple pathways associated with acute inflammation. In an expanded cohort including additional IAI and iPTB specimens, the expression of three proteins (cathepsin S, lysozyme, and hexokinase 3) and two microRNAs (miR-133a and miR-223) was validated using immunohistochemistry and quantitative PCR, respectively. In the DB specimens, only 11 long transcripts and no microRNAs differed significantly between IAI cases and iPTB controls (FDR<0.1). Comparison of the VT and DB specimens in each clinical scenario revealed signatures distinguishing these placental regions.
  
  
  
  IAI is associated with a transcriptional signature consistent with acute inflammation in the villous trophoblast. The present findings illuminate novel signaling pathways involved in IAI, and suggest putative therapeutic targets and potential biomarkers associated with this condition.
authors.affiliation: Department of Obstetrics and Gynecology, The Ohio State College of Medicine, Columbus, OH, USA. Electronic address: william.ackerman@osumc.edu.
authors.forename: William E
authors.initials: WE
authors.lastname: Ackerman
authors.affiliation: Center for Perinatal Research, Nationwide Children's Hospital, Columbus, OH, USA. Electronic address: irina.buhimschi@nationwidechildrens.org.
authors.forename: Irina A
authors.initials: IA
authors.lastname: Buhimschi
authors.affiliation: Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA. Electronic address: haley.eidem@vanderbilt.edu.
authors.forename: Haley R
authors.initials: HR
authors.lastname: Eidem
authors.affiliation: Program in Human Genetics, Vanderbilt University Medical Center, Nashville, TN, USA. Electronic address: david.rinker@vanderbilt.edu.
authors.forename: David C
authors.initials: DC
authors.lastname: Rinker
authors.affiliation: Department of Biological Sciences, Vanderbilt University, Nashville, TN, USA; Program in Human Genetics, Vanderbilt University Medical Center, Nashville, TN, USA. Electronic address: antonis.rokas@vanderbilt.edu.
authors.forename: Antonis
authors.initials: A
authors.lastname: Rokas
authors.affiliation: Department of Obstetrics and Gynecology, The Ohio State College of Medicine, Columbus, OH, USA. Electronic address: kara.rood@osumc.edu.
authors.forename: Kara
authors.initials: K
authors.lastname: Rood
authors.affiliation: Center for Perinatal Research, Nationwide Children's Hospital, Columbus, OH, USA. Electronic address: lisa.zhao@nationwidechildrens.org.
authors.forename: Guomao
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Department of Obstetrics and Gynecology, The Ohio State College of Medicine, Columbus, OH, USA. Electronic address: taryn.summerfield@osumc.edu.
authors.forename: Taryn L
authors.initials: TL
authors.lastname: Summerfield
authors.affiliation: Department of Obstetrics and Gynecology, The Ohio State College of Medicine, Columbus, OH, USA. Electronic address: mark.landon@osumc.edu.
authors.forename: Mark B
authors.initials: MB
authors.lastname: Landon
authors.affiliation: Department of Obstetrics and Gynecology, The Ohio State College of Medicine, Columbus, OH, USA. Electronic address: catalin.buhimschi@osumc.edu.
authors.forename: Catalin S
authors.initials: CS
authors.lastname: Buhimschi
mesh_terms: D000293:Adolescent; D000328:Adult; D000653:Amniotic Fluid / Q000378:metabolism; D002821:Chorioamnionitis / Q000235:genetics / Q000378:metabolism*; D003656:Decidua / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007231:Infant, Newborn; D011247:Pregnancy; D047928:Premature Birth / Q000235:genetics / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D014327:Trophoblasts / Q000378:metabolism*; D055815:Young Adult
chemical_list: D012313:RNA
keywords: Infection; Placenta; Preterm birth; RNA-sequencing; Transcriptomics; microRNA