PUBLICATION

Identification of Padi2 as a novel angiogenesis-regulating gene by genome association studies in mice

Authors
Khajavi, M., Zhou, Y., Birsner, A.E., Bazinet, L., Di Sant, A.R., Schiffer, A.J., Rogers, M.S., Krishnaji, S.T., Hu, B., Nguyen, V., Zon, L., D'Amato, R.J.
ID
ZDB-PUB-170616-5
Date
2017
Source
PLoS Genetics   13: e1006848 (Journal)
Registered Authors
Zhou, Yi, Zon, Leonard I.
Keywords
Cornea, Inbred strains, Zebrafish, Mammalian genomics, Angiogenesis, Endothelial cells, Embryos, Small interfering RNAs
MeSH Terms
  • Animals
  • Endothelial Cells/metabolism
  • Endothelial Cells/pathology
  • Fibroblast Growth Factor 2/genetics
  • Genetic Variation
  • Genome-Wide Association Study*
  • Haplotypes
  • Humans
  • Hydrolases/biosynthesis
  • Hydrolases/genetics*
  • Mice
  • Mice, Inbred Strains
  • Neovascularization, Pathologic/genetics*
  • Phenotype
  • Zebrafish
PubMed
28617813 Full text @ PLoS Genet.
Abstract
Recent findings indicate that growth factor-driven angiogenesis is markedly influenced by genetic variation. This variation in angiogenic responsiveness may alter the susceptibility to a number of angiogenesis-dependent diseases. Here, we utilized the genetic diversity available in common inbred mouse strains to identify the loci and candidate genes responsible for differences in angiogenic response. The corneal micropocket neovascularization assay was performed on 42 different inbred mouse strains using basic fibroblast growth factor (bFGF) pellets. We performed a genome-wide association study utilizing efficient mixed-model association (EMMA) mapping using the induced vessel area from all strains. Our analysis yielded five loci with genome-wide significance on chromosomes 4, 8, 11, 15 and 16. We further refined the mapping on chromosome 4 within a haplotype block containing multiple candidate genes. These genes were evaluated by expression analysis in corneas of various inbred strains and in vitro functional assays in human microvascular endothelial cells (HMVECs). Of these, we found the expression of peptidyl arginine deiminase type II (Padi2), known to be involved in metabolic pathways, to have a strong correlation with a haplotype shared by multiple high angiogenic strains. In addition, inhibition of Padi2 demonstrated a dosage-dependent effect in HMVECs. To investigate its role in vivo, we knocked down Padi2 in transgenic kdrl:zsGreen zebrafish embryos using morpholinos. These embryos had disrupted vessel formation compared to control siblings. The impaired vascular pattern was partially rescued by human PADI2 mRNA, providing evidence for the specificity of the morphant phenotype. Taken together, our study is the first to indicate the potential role of Padi2 as an angiogenesis-regulating gene. The characterization of Padi2 and other genes in associated pathways may provide new understanding of angiogenesis regulation and novel targets for diagnosis and treatment of a wide variety of angiogenesis-dependent diseases.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping