title: Next-generation sequencing analysis of gene regulation in the rat model of retinopathy of prematurity.
abstract: OBJECTIVE
  The purpose of this study was to identify the genes, biochemical signaling pathways, and biological themes involved in the pathogenesis of retinopathy of prematurity (ROP).
  
  
  METHODS
  Next-generation sequencing (NGS) was performed on the RNA transcriptome of rats with the Penn et al. (Pediatr Res 36:724-731, 1994) oxygen-induced retinopathy model of ROP at the height of vascular abnormality, postnatal day (P) 19, and normalized to age-matched, room-air-reared littermate controls. Eight custom-developed pathways with potential relevance to known ROP sequelae were evaluated for significant regulation in ROP: The three major Wnt signaling pathways, canonical, planar cell polarity (PCP), and Wnt/Ca(2+); two signaling pathways mediated by the Rho GTPases RhoA and Cdc42, which are, respectively, thought to intersect with canonical and non-canonical Wnt signaling; nitric oxide signaling pathways mediated by two nitric oxide synthase (NOS) enzymes, neuronal (nNOS) and endothelial (eNOS); and the retinoic acid (RA) signaling pathway. Regulation of other biological pathways and themes was detected by gene ontology using the Kyoto Encyclopedia of Genes and Genomes and the NIH's Database for Annotation, Visualization, and Integrated Discovery's GO terms databases.
  
  
  RESULTS
  Canonical Wnt signaling was found to be regulated, but the non-canonical PCP and Wnt/Ca(2+) pathways were not. Nitric oxide signaling, as measured by the activation of nNOS and eNOS, was also regulated, as was RA signaling. Biological themes related to protein translation (ribosomes), neural signaling, inflammation and immunity, cell cycle, and cell death were (among others) highly regulated in ROP rats.
  
  
  CONCLUSIONS
  These several genes and pathways identified by NGS might provide novel targets for intervention in ROP.
authors.affiliation: Department of Ophthalmology, Boston Children's Hospital, 300 Longwood Avenue, Fegan 4, Boston, MA 02115, USA.
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Griffith
authors.forename: Hu
authors.initials: H
authors.lastname: Li
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
authors.forename: Tara L
authors.initials: TL
authors.lastname: Favazza
authors.forename: Anne B
authors.initials: AB
authors.lastname: Fulton
authors.forename: Ronald M
authors.initials: RM
authors.lastname: Hansen
authors.forename: James D
authors.initials: JD
authors.lastname: Akula
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007231:Infant, Newborn; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012171:Retinal Vessels / Q000378:metabolism / Q000473:pathology; D012178:Retinopathy of Prematurity / Q000235:genetics*; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D059467:Transcriptome / Q000235:genetics*
chemical_list: D012333:RNA, Messenger