PMID- 31413262
OWN - NLM
STAT- MEDLINE
DCOM- 20201022
LR  - 20210110
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 9
IP  - 1
DP  - 2019 Aug 14
TI  - Intrabody against prolyl hydroxylase 2 promotes angiogenesis by stabilizing 
      hypoxia-inducible factor-1alpha.
PG  - 11861
LID - 10.1038/s41598-019-47891-1 [doi]
LID - 11861
AB  - Hypoxia-inducible factor (HIF)-1alpha is a crucial transcription factor that 
      regulates the expression of target genes involved in angiogenesis. Prolyl 
      hydroxylase 2 (PHD2) dominantly hydroxylates two highly conserved proline 
      residues of HIF-1alpha to promote its degradation. This study was designed to 
      construct an intrabody against PHD2 that can inhibit PHD2 activity and promote 
      angiogenesis. Single-chain variable fragment (scFv) against PHD2, INP, was 
      isolated by phage display technique and was modified with an endoplasmic 
      reticulum (ER) sequence to obtain ER-retained intrabody against PHD2 (ER-INP). 
      ER-INP was efficiently expressed and bound to PHD2 in cells, significantly 
      increased the levels of HIF-1alpha, and decreased hydroxylated HIF-1alpha in human 
      embryonic kidney cell line (HEK293) cells and mouse mononuclear macrophage 
      leukaemia cell line (RAW264.7) cells. ER-INP has shown distinct angiogenic 
      activity both in vitro and in vivo, as ER-INP expression significantly promoted 
      the migration and tube formation of human umbilical vein endothelial cells 
      (HUVECs) and enhanced angiogenesis of chick chorioallantoic membranes (CAMs). 
      Furthermore, ER-INP promoted distinct expression and secretion of a range of 
      angiogenic factors. To the best of our knowledge, this is the first study to 
      report an ER-INP intrabody enhancing angiogenesis by blocking PHD2 activity to 
      increase HIF-1alpha abundance and activity. These results indicate that ER-INP may 
      play a role in the clinical treatment of tissue injury and ischemic diseases in 
      the future.
FAU - Zhao, Liangzhong
AU  - Zhao L
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
AD  - Department of Immunoassay Technology, Jilin Medical University, Jilin, 132013, 
      China.
FAU - Liu, Ziyu
AU  - Liu Z
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
FAU - Yang, Fang
AU  - Yang F
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
FAU - Zhang, Ying
AU  - Zhang Y
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
AD  - Department of Pediatrics, The First Hospital of Jilin University, Changchun, 
      Jilin, 130021, P.R. China.
FAU - Xue, Ying
AU  - Xue Y
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
FAU - Miao, Haipeng
AU  - Miao H
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
FAU - Liao, Xiangzhi
AU  - Liao X
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
FAU - Huang, Hongli
AU  - Huang H
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China.
FAU - Li, Guiying
AU  - Li G
AD  - Key Laboratory for Molecular Enzymology and Engineering of the Ministry of 
      Education, School of Life Sciences, Jilin University, Changchun, 130012, China. 
      ligy@jlu.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190814
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - EC 1.14.11.- (Prolyl Hydroxylases)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cell Movement
MH  - Chickens
MH  - Chorioallantoic Membrane/metabolism
MH  - Human Umbilical Vein Endothelial Cells/metabolism
MH  - Humans
MH  - Hypoxia-Inducible Factor 1, alpha Subunit/*metabolism
MH  - Mice
MH  - *Neovascularization, Physiologic
MH  - Prolyl Hydroxylases/*metabolism
MH  - Protein Stability
MH  - Up-Regulation
PMC - PMC6694103
COIS- The authors declare no competing interests.
EDAT- 2019/08/16 06:00
MHDA- 2020/10/23 06:00
PMCR- 2019/08/14
CRDT- 2019/08/16 06:00
PHST- 2019/01/04 00:00 [received]
PHST- 2019/07/25 00:00 [accepted]
PHST- 2019/08/16 06:00 [entrez]
PHST- 2019/08/16 06:00 [pubmed]
PHST- 2020/10/23 06:00 [medline]
PHST- 2019/08/14 00:00 [pmc-release]
AID - 10.1038/s41598-019-47891-1 [pii]
AID - 47891 [pii]
AID - 10.1038/s41598-019-47891-1 [doi]
PST - epublish
SO  - Sci Rep. 2019 Aug 14;9(1):11861. doi: 10.1038/s41598-019-47891-1.