Analysis of the role of the HIF hydroxylase family members in erythropoiesis.
HIF (hypoxia-inducible factor) hydroxylases have been implicated in EPO (erythropoietin) production and erythropoiesis. Here we examined the role of each of the three EGLN family members and the HIF asparaginyl hydroxylase FIH (factor inhibiting HIF) in EPO production. We examined the effect of inhibiting individual as well as combinations of HIF hydroxylases with RNAi. We found that inhibition of EGLN1 (egl nine homolog 1) as well as other members of the EGLN family (EGLN2 and EGLN3) led to accumulative EPO production in vitro. We then knocked down EGNL1 in vivo by injecting one-cell murine zygotes with lentivirus-containing RNAi. Progeny with demonstrated EGLN1 inhibition had elevated EPO production and erythropoiesis in vivo. Among all the in vitro and in vivo studies, no or minimal VEGF (vascular endothelial growth factor) mRNA or protein stimulation resulted from inhibition of EGLN1.
Fisher
Timothy S
TS
Department of Antibacterials, Immunology and Cancer, Pfizer Global Research and Development, Groton, CT 06340, USA.
Lira
Paul D
PD
Stock
Jeffrey L
JL
Perregaux
David G
DG
Brissette
William H
WH
Ozolins
Terence R S
TR
Li
Baiyong
B
D000818:Animals; D002460:Cell Line; D049308:Dioxygenases / Q000235:genetics / Q000502:physiology*; D004920:Erythropoiesis / Q000235:genetics; D004921:Erythropoietin / Q000096:biosynthesis*; D006801:Humans; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases; D051379:Mice; D008822:Mice, Transgenic; D006899:Mixed Function Oxygenases; D009687:Nuclear Proteins / Q000235:genetics / Q000502:physiology*; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000502:physiology*
D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D004921:Erythropoietin; D006899:Mixed Function Oxygenases; D049308:Dioxygenases; C439790:HIF1AN protein, human; C490190:EGLN1 protein, human; D011393:Procollagen-Proline Dioxygenase; C447780:EGLN2 protein, human; C488894:EGLN3 protein, human; D064799:Hypoxia-Inducible Factor-Proline Dioxygenases