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2017 ; 13
(6
): e1006848
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Identification of Padi2 as a novel angiogenesis-regulating gene by genome
association studies in mice
#MMPMID28617813
Khajavi M
; Zhou Y
; Birsner AE
; Bazinet L
; Rosa Di Sant A
; Schiffer AJ
; Rogers MS
; Krishnaji ST
; Hu B
; Nguyen V
; Zon L
; D'Amato RJ
PLoS Genet
2017[Jun]; 13
(6
): e1006848
PMID28617813
show ga
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.